NZ554250A - Isolated terminal flower 1 gene (TFL1) from festuca arundinacea and methods for its use - Google Patents

Abstract

Disclosed is an isolated polynucleotide encoding a TFL1 polypeptide active in the regulation of flowering in the Festuca arundinacea plant (common name tall fescue). Further disclosed are expression vectors and host cells comprising such isolated polynucleotide. Methods for the use of such polynucleotide and polypeptide are also provided.

Description

*10053447564* NEW ZEALAND PATENTS ACT, 1953 intellectual property office of n.z.

No: 3 0 MAR 2007 received Date: COMPLETE SPECIFICATION COMPOSITIONS ISOLATED FROM FORAGE GRASSES AND METHODS FOR THEIR USE We, GENESIS RESEARCH & DEVELOPMENT CORPORATION LTD, of 1 Fox Street, Parnell, Auckland, New Zealand AND WRIGHTSON SEEDS LTD., of Wrightson House, 14 Hartham Place, Porirua, New Zealand, do hereby declare the invention for which we pray that a patent may be granted to us, and the method by which it is to be performed, to be particularly described in and by the following statement: (followed by page 1a) COMPOSITIONS ISOLATED FROM FORAGE GRASSES AND METHODS FOR THEIR USE Technical Field of the Invention This invention relates to polynucleotides isolated from forage grass tissues, specifically from Lolium perenne (perennial ryegrass) and Festuca arundinacea (tall fescue), as well as oligonucleotide probes and primers, genetic constructs comprising the 10 polynucleotides, biological materials (including host cells and plants) incorporating the polynucleotides, polypeptides encoded by the polynucleotides, and methods for using the polynucleotides and polypeptides. More particularly, the invention relates to polypeptides involved in the regulation of flowering, and to polynucleotides encoding such polypeptides.

Background of the Invention Over the past 50 years, there have been substantial improvements in the genetic production potential of ruminant animals (sheep, cattle and deer). Levels of meat, milk or fiber production that equal an animal's genetic potential may be attained within controlled 20 feeding systems, where animals are fully fed with energy dense, conserved forages and grains. However, the majority of temperate farming systems worldwide rely on the in situ grazing of pastures. Nutritional constraints associated with temperate pastures can prevent the fall expression of an animal's genetic potential. This is illustrated by a comparison between milk production by North American grain-fed dairy cows and New 25 Zealand pasture-fed cattle. North American dairy cattle produce, on average, twice the milk volume of New Zealand cattle, yet the genetic base is similar within both systems (New Zealand Dairy Board and United States Department of Agriculture figures). Significant potential therefore exists to improve the efficiency of conversion of pasture nutrients to animal products through the correction of nutritional constraints associated 30 with pastures.

The ability to control flowering in C3 monocotyledonous plants, such as forage grasses (e.g. perennial ryegrass and tall fescue) and cereals (e.g. wheat and barley), has wide ranging applications. For example, controlling flowering in forage grasses offers the ability to halt the increase in syringyl lignin that is associated with the decrease in la digestibility of forage at this time. In addition, it offers the ability to control the spread of genetically modified organisms, as well as lowering the incidence of allergies associated with ryegrass pollen levels. Other advantages include the ability to induce the time of flowering to suit farming practices better. To achieve this a flowering control gene would 5 have to be placed under the control of an inducible promoter and the endogenous flowering genes would need to be silenced. A number of genes are known to control flowering in a range of species.

A simple model has been proposed for the genetic network regulating flowering time and flower development in Arabidopsis. In Arabidopsis there are three genetic 10 pathways that control flowering time (Reeves and Coupland, Curr. Opin. Plant Biol. 3:37-42, 2000). The long-day pathway represented by GIGANTEA (GI) and CONSTANS (CO) and the autonomous pathway represented by LUMINIDEPENDENS (LD), FLOWERING TIME CONTROL PROTEIN (FCA) and FLOWERING LOCUS C (FLC) are likely integrated through FLOWERING LOCUS T (FT) and AGAMOUS-15 LIKE20 (AGL20) to promote activation of meristem identity genes LEAFY (LFY), APETALA1 (API) and CAULIFLOWER (CAL). The vernalization pathway represented by FRIGIDA (FRI), feeds into the autonomous pathway upstream of FLC. The giberellin pathway (GA) is represented by gibberellic acid insensitive (GAI) that leads to the activation of LFY. The TERMINAL FLOWER 1 (TFL1) restricts the expression of the 20 meristem identity genes to the floral meristems, thereby promoting the patterned expression of floral organ identity genes such as APETALA2 (AP2), APETALA3 (AP3), PISTILATA (PI), and AGAMOUS (AG). These floral identity genes are also affected by other regulatory genes" such as AINTEGUMENTA (ANT), UNUSUAL FLORAL ORGANS (UFO) and SUPERMAN (SUP). Homologs of some of these genes have been 25 identified in other monocots such as maize and rice as well as the dicot species Antirrhinum, where they play a role that is either similar or divergent to that of the Arabidopsis gene in flowering. For example, some key regulatory flowering genes are conserved between rice and Arabidopsis, however, the regulation of FT by CO is reversed in the two species under long day conditions (Hayama et al., Nature 422, 719 - 722, 30 2003).

Both genetic and molecular studies have led to the proposal of the ABC model for floral organ identity (Ma and DePamphilis, Cell 101:5-8, 2000). The Arabidopsis B function genes, APETALA3 (AP3) and PISTILATA (PI), are required to specify petal and stamen identities. The Arabidopsis meristem identity gene, LFY, is required for 2 normal levels of AP3 and PI expression (Weigel and Meyerowitz, Science 261:1723-1726, 1993). The Arabidopsis gene UFO plays a role in controlling floral meristem development and B function, and the activation of AP3 by LFY requires UFO (Lee et al, Curr. Biol. 7:95-104, 1997). The ASK1 gene regulates B function gene expression in 5 cooperation with UFO and LFY in Arabidopsis (Zhao et al., Development 128:2735-2746,2001; Durfee etal.,Proc. Natl. Acad. Sci. USA 100:8571-8576,2003).

It has been suggested that UFO and ASK1 may be suburdts of a three-component SCF (SKP1, cullin, F-box) ubiquitin ligase. In addition, ASK1 shows high sequence identity to the yeast SKP1 protein. Ubiquitin ligase is part of the ubiquitin-dependent 10 protein degradation pathway; this suggests that UFO and ASK1 may regulate the level of other regulatory proteins that control cell division and transcription during floral development.

FCA encodes a strong promoter of the transition to flowering in Arabidopsis. Arabidopsis fca mutants flower late in both long days and short days. FCA has been 15 cloned and shown to encode a protein containing two RNA-binding domains and a WW protein interaction domain (Macknight et al., Cell 89:737-745, 1997). The regulation of FCA expression is complex. FCA pre-mRNA is alternatively processed resulting in four types of transcripts of which FCA-y is the active form. Recent studies have shown that FCA functions with FY, a WD-repeat protein, to regulate 3' end formation of mRNA and 20 control the floral transition (Simpson et al., Cell, 113:777-787,2003). Plants carrying the FCA gene fused to the strong constitutive 35S promoter flowered earlier, and the ratio and abundance of the different FCA transcripts were altered. The rice genome contains a single copy homolog of FCA (Goff et al., Science 296:92-100, 2002).

The FT/TFL gene family encodes proteins with homology to phosphatidy-25 ethanolamine binding proteins that have been shown to be involved in major aspects of whole-plant architecture. FT acts in parallel with the meristem-identity gene LFY to induce flowering of Arabidopsis (Kardailsky et al, Science 286:1962-1965, 1999), it is similar in sequence to TFL1, an inhibitor of flowering (Ohshima et al., Mol. Gen. Genet. 254:186-194, 1997). The crystal structure of the Antirrhinum FT/TFL homolog, 30 CENTRORADIALIS (CEN) suggests that it has a role as a kinase regulator (Banfield and Brady, J. Mol. Biol. 14:1159-1170, 2000). The rice genome contains 17 members of the FT/TFL gene family; one member is most similar to TFL, and nine are more similar to FT. A functional FT ortholog from rice, Hd3a, was detected as a heading date QTL and 3 has the same regulatory relationship with rice CONSTANS homolog, Hdl, that Arabidopsis FT has with CO (Kojima et al, Plant Cell Physiol. 43:1096-1105, 2002). A TFLl-like gene from Lolium perenne has been isolated and characterized (Jensen et al., Plant Physiol. 125:1517-1528, 2001). Arabidopsis plants over-expressing the LpTFLl 5 gene were significantly delayed in flowering and the LpTFLl gene was able to complement the severe tfll-14 mutant of Arabidopsis.

The Arabidopsis gai (gibberellic acid insensitive) mutant allele confers a reduction in gibberellin (GA) responsiveness, thereby playing a role in the GA regulated control of flowering. GAI contains nuclear localization signals, a region of homology to a 10 putative transcription factor, and motifs characteristic of transcriptional co-activators (Peng et al., Genes Dev. 11:3194-3205, 1997). Homologs from other plant species have been identified, for example, RHT from wheat, D8 from maize and SLR1 from rice (Ikeda et al., Plant Cell 13:999-1010, 2001). Four rice sequence homologs of the Arabidopsis GAI gene have been identified in the rice genome (Goff et al., Science 15 296:92-100,2002).

Alongside CONSTANS (CO), GIGANTEA (GI) exerts major control over the promotion of flowering under long days in Arabidopsis. Mutations in the Arabidopsis thaliana GI gene cause photoperiod-insensitive flowering and alteration of circadian rhythms. GI, originally described as a putative membrane protein (Fowler et al, EMBO 20 J. 18:4679-4688, 1999), was recently determined to be a nuclear protein involved in photochrome signaling (Huq et al, Proc. Natl Acad. Sci. USA 97:9789-9794, 2000). GI is believed to function upstream of CO, because the late-flowering phenotype of gi mutants is corrected by CO over expression (Fowler et al, EMBO J. 18:4679-4688, 1999). A single putative GI ortholog exists in rice, based on the similarity of the 25 predicted GI amino acid sequence. Overexpression of OsGI, an ortholog of the Arabidopsis GIGANTEA (GI) gene in transgenic rice, caused late flowering under both SD and LD conditions (Hayama et al., Nature 422, 719 - 722,2003).

The indeterminate 1 (idl) mutation in maize results in plants that are unable to undergo a normal transition to flower development and remain in a prolonged state of 30 vegetative growth. The ID1 gene plays an important role in controlling the transition to flowering and maintaining the florally determined state. The ID1 gene was cloned by transposon mapping in maize (Colasanti et al, Cell 93:593-603, 1998). The ID1 gene encodes a protein with zinc finger motifs, indicating that it functions by transcriptional regulation of flowering. Expression studies showed that ID1 is expressed in immature 4 leaves and not the shoot apex, and may therefore mediate the transition to flowering by regulating the transmission or synthesis of a signal for flowering. ID1 functional homologs have not been in identified in Arabidopsis but putative ID1 gene sequences have been identified from rice (Goff et al., Science 296:92-100, 2002).

LEUNIG (LUG) is a key regulator of the Arabidopsis floral homeotic gene AGAMOUS. Mutations in LEUNIG cause ectopic AGAMOUS mRNA expression in the outer two whorls of a flower, leading to homeotic transformations of floral organ identity as well as loss of floral organs. LEUNIG is a glutamine-rich protein with seven WD repeats and is similar in motif structure to a class of functionally related transcriptional co-repressors. The nuclear localization of LEUNIG is consistent with a role of LEUNIG as a transcriptional regulator (Conner and Liu, Proc. Natl. Acad. Sci. USA 97:12902-12907, 2000). Another regulatory gene, SEUSS, has recently been identified that functions together with LEUNIG to regulate AGAMOUS (Franks et al., Development 129:253-263, 2002).

Summary of the Invention In one aspect the invention provides an isolated polynucleotide comprising the sequence of SEQ ID NO: 16.

In a further aspect the invention provides an isolated polynucleotide comprising a sequence selected from the group consisting of: (a) the complement of SEQ ID NO: 16; (b) the reverse complement of SEQ ID NO: 16; (c) the reverse sequence of SEQ ID NO: 16; and (d) an operable antisense fragment of SEQ ID NO: 16.

In a further aspect the invention provides an isolated polynucleotide comprising a sequence selected from the group consisting of: (a) sequences having at least 75% identity to the sequence of SEQ ID NO: 16; (b) sequences having at least 90% identity to the sequence of SEQ ID NO: 16; (c) sequences having at least 95% identity to the sequence of SEQ ID NO: 16, wherein the polynucleotide encodes a polypeptide with Terminal Flower 1 (TFL1) activity.

In a further aspect the invention provides an isolated polypeptide comprising the amino acid sequence of SEQ ID NO: 36. and (followed by page 5a) - 8 JUL 2008 In a further aspect the invention provides an isolated polypeptide comprising an amino acid sequence selected from the group consisting of: (a) sequences having at least 75% identity to a sequence of SEQ ID NO: 36; (b) sequences having at least 90% identity to a sequence of SEQ ID NO: 36; and (c) sequences having at least 95% identity to a sequence of SEQ ID NO: 36, wherein the polypeptide has Terminal Flower 1 (TFLl) activity.

In a further aspect the invention provides a method for producing a plant with altered flowering, the method comprising transformation of a plant with 10 a polynucleotide capable of hybridising under stringent conditions to an endogenous gene encoding a polypeptide with at least 75% identity to the sequence of SEQ ID NO: 16 and with Terminal Flower 1 (TFLl) activity, such that expression of the polynucleotide results in reduced expression of the endogenous gene, reduced levels of TFLl and altered flowering.

In a further aspect the invention provides a method for producing a plant with altered flowering, the method comprising transformation of a plant with a polynucleotide capable of hybridising under stringent conditions to an endogenous nucleic acid comprising a sequence with at least 75% sequence identity to SEQ ID NO: 16 encoding a polypeptide with TFLl activity, such that expression of the polynucleotide 20 results in reduced expression of the endogenous nucleic acid, reduced levels of TFLl and altered flowering.

Disclosed herein are polypeptides involved in the flowering pathway that are encoded by polynucleotides isolated from forage grass tissues. The polynucleotides were isolated from Lolium perenne (perennial ryegrass) and Festuca arundinacea (tall fescue) 25 tissues taken at different times of the year, specifically in winter and spring, and from different parts of the plants, including: leaf blades, leaf base, pseudostems, inflorescence, roots and stems. The present invention also provides genetic constructs, expression vectors and host cells comprising the inventive polynucleotides, and methods for using the inventive polynucleotides and genetic constructs to modulate flowering. 30 The isolated polynucleotides disclosed comprise a sequence selected from the group consisting of: (a) SEQ ID NOS: 1-20; (b) complements of SEQ ID NOS: 1-20; (c) reverse complements of SEQ ID NOS: 1-20; (d) reverse sequences of SEQ ID NOS: 1-20; (e) sequences having a 99% probability of being functionally or evolutionarily related to a sequence of (a)-(d), determined as described below; and (f) sequences having at least 5a |~V~| (followed by page 5b) I - 8 JUL 2008 RFHP l\/nrrv 75%, 80%, 90%, 95% or 98% identity to a sequence of (a)-(d), the percentage identity being determined as described below. Polynucleotides comprising at least a specified number of contiguous residues ("x-mers") of any of SEQ ID NOS: 1-20, and oligonucleotide probes and primers corresponding to 5b (followed by page 6) INTEnJ§^AL ^OPERTY office of l\j.z. - 8 JUL 2008 receive SEQ ID NOS: 1-20, are also provided. All of the above polynucleotides are referred to herein as "polynucleotides disclosed." Also disclosed are isolated polypeptides comprising an amino acid sequence of SEQ ID NOS: 21-40, together with polypeptides comprising a sequence having at least 5 75%, 80%, 90%, 95% or 98% identity to a sequence of SEQ ID NOS: 21-40, wherein the polypeptide possesses the same functional activity as the polypeptide comprising a sequence of SEQ ID NOS: 21-40. Also disclosed are isolated polypeptides comprising at least a functional portion of a polypeptide comprising an amino acid sequence selected from the group consisting of: (a) SEQ ID NOS: 21-40; and (b) sequences having at least 10 75%, 80%, 90%, 95% or 98% identity to a sequence of SEQ ID NOS: 21-40.

Also disclosed are genetic constructs comprising a polynucleotide disclosed, either alone or in combination with one or more of the disclosed sequences, or in combination with one or more known polynucleotides.

Also disclosed are genetic constructs comprising, in the 5'-3' direction: a gene 15 promoter sequence; an open reading frame coding for at least a functional portion of a polypeptide disclosed; and a gene termination sequence. An open reading frame may be orientated in either a sense or anti-sense direction. Genetic constructs comprising a non-coding region of a polynucleotide of the present invention or a polynucleotide complementary to a non-coding region, together with a gene promoter sequence and a 20 gene termination sequence, are also provided. Preferably, the gene promoter and termination sequences are functional in a host cell, such as a plant cell. Most preferably, the gene promoter and termination sequences are those of the original enzyme genes but others generally used in the art, such as the Cauliflower Mosaic Virus (CMV) promoter, with or without enhancers, such as the Kozak sequence or Omega enhancer, and 25 Agrobacterium tumefaciens nopalin synthase terminator may be usefully employed in the present invention. Tissue-specific promoters may be employed in order to target expression to one or more desired tissues. The construct may further include a marker for the identification of transformed cells.

Also disclosed are transgenic cells, such as transgenic plant cells, comprising the 30 constructs provided, together with tissues and plants comprising such transgenic cells, and fruits, seeds and other products, derivatives, or progeny of such plants. office of m.z 1 - 8 JUL 2008 Methods for modulating the flowering of a target plant are also provided. Such methods include stably incorporating into the genome of the target plant a genetic construct comprising a polynucleotide disclosed. In a preferred embodiment, the target plant is a forage grass, preferably selected from the group consisting of Lolium and Festuca species, and most preferably from the group consisting of Lolium perenne and Festuca arundinacea.

A method for producing a plant having altered flowering is provided, the method comprising transforming a plant cell with a genetic construct comprising a polynucleotide disclosed to provide a transgenic cell, and cultivating the transgenic cell under conditions conducive to regeneration and mature plant growth.

Also provided are methods for modifying the activity of an enzyme in a target organism, such as a plant, comprising stably incorporating into the genome of the target organism a genetic construct disclosed. The target plant may be a forage grass, preferably selected from the group consisting of Lolium and Festuca species, and most preferably from the group consisting of Lolium perenne and Festuca arundinacea.

Brief Description of the Drawings Fig. 1 shows the amino acid sequence of SEQ ID NO: 21. The conserved dimerization domain of the SKP1 family is underlined.

Fig. 2 shows the amino acid sequence of SEQ ID NO: 22. The conserved dimerization domain of the SKP1 family is underlined.

Fig. 3 shows the amino acid sequence of SEQ ID NO: 23. The conserved dimerization domain of the SKP1 family is underlined.

Fig. 4 shows the amino acid sequence of SEQ ID NO: 24. The conserved RNA-binding region RNP-1 (RNA recognition motif) domains are underlined and the WW/Rsp5/WWP domain is in bold/italics.

Fig. 5 shows the amino acid sequence of SEQ ID NO: 25. The conserved RNA-binding region RNP-1 (RNA recognition motif) domains are underlined and the WW/Rsp5/WWP domain is in bold/italics.

Fig. 6 shows the amino acid sequence of SEQ ID NO: 26. The conserved phosphatidylethanolamine-binding protein (PBP) domain is underlined and the conserved PBP family signature is boxed. •s^tual property office of m.z. - 8 JUL 2008 D l~ A r— • % < Fig. 7 shows the amino acid sequence of SEQ ID NO: 27. The conserved phosphatidylethanolamine-binding protein (PBP) domain is underlined and the conserved PBP family signature is boxed.

Fig. 8 shows the amino acid sequence of SEQ ID NO: 28. The conserved GRAS family domain is underlined with conserved residues in the conserved C-terminus being in bold (Pysh et al., Plant J. 18:111-119,1999).

Fig. 9 shows the amino acid sequence of SEQ ID NO: 29. Predicted transmembrane domains characteristic of GIGANTEA proteins (Fowler et al., EMBO J. 18:4679-4688,1999) are underlined.

Fig. 10 shows the amino acid sequence of SEQ ID NO: 30. Predicted transmembrane domains, characteristic of GIGANTEA proteins (Fowler et ah, EMBO J. 18:4679-4688, 1999) are underlined.

Fig. 11 shows the amino acid sequence of SEQ ID NO: 31. The conserved C2H2-type zinc finger is underlined with the conserved residues being boxed (Kubo et al., Nucleic Acids Res. 26:608-615, 1998).

Fig. 12 shows the amino acid sequence of SEQ ID NO: 32. The conserved C2H2-type zinc finger is underlined with the conserved residues being boxed (Kubo et al., Nucleic Acids Res. 26:608-615,1998).

Fig. 13 shows the amino acid sequence of SEQ ID NO: 33. The conserved C2H2-type zinc finger is underlined with the conserved residues being boxed (Kubo et al., Nucleic Acids Res. 26:608-615, 1998).

Fig. 14 shows the amino acid sequence of SEQ ID NO: 34. The conserved G-protein beta WD-40 repeat domains are underlined and the conserved G-protein beta WD-40 repeat domain signature is boxed.

Fig. 15 shows the amino acid sequence of SEQ ED NO: 35. The conserved G-protein beta WD-40 repeat domains are underlined and the conserved G-protein beta WD-40 repeat domain signature is boxed.

Fig. 16 shows the amino acid sequence of SEQ ID NO: 36. The conserved phosphatidylethanolamine-binding protein (PBP) domain is underlined and the conserved PBP family signature is boxed.

Fig. 17 shows the amino acid sequence of SEQ ID NO: 37. A Gin-rich region is in bold/italics and a predicted transmembrane domain is double-underlined. 8 Fig. 18 shows the amino acid sequence of SEQ ID NO: 38. The conserved dimerization domain with similarity to the Ldb proteins (Franks et al., Development 129:253-263,2002) is underlined. A Gin-rich region is in bold/italics.

Fig. 19 shows the amino acid sequence of SEQ ID NO: 39. The conserved 5 dimerization domain with similarity to the Ldb proteins (Franks et al., Development 129:253-263, 2002) is underlined. A Gin-rich region is in bold/italics and a predicted transmembrane domain is double-underlined.

Fig. 20 shows the amino acid sequence of SEQ ID NO: 40. The conserved GRAS family domain is underlined with conserved residues in the conserved C-terminus is in 10 bold (Pysh et al., Plant J. 18:111-119, 1999).

Fig. 21 shows the time to first floral bud formation for Arabidopsis plants over-expressing the grass flowering time gene FaFT (SEQ ED NO: 6).

Fig. 22 shows the time to first open flowers for plants over-expressing the grass flowering time FLOWERING LOCUS T gene FaFT (SEQ ED NO: 6) and the grass 15 LEUNIG gene FaLUGl (SEQ ID NO: 14).

Detailed Description of the Invention The polypeptides of the present invention, and the polynucleotides encoding the 20 polypeptides, have activity in flowering pathways in plants. Using the methods and materials of the present invention, the transition to flowering in a plant may be modulated by modulating expression of polynucleotides of the present invention, or by modifying the polynucleotides or the polypeptides encoded by the polynucleotides.

The isolated polynucleotides and polypeptides of the present invention may be 25 used to reduce lignin content, control flowering, induce flowering time, control spread of seed/pollen, and reduce spread of allergenic pollen. The main decrease in forage digestibility occurs around the time of flowering in grass plants when there is a sharp increase in syringyl lignin. This appears to be a defense mechanism by the plant to avoid being grazed whilst trying to reproduce. By controlling, or preventing, flowering in 30 grasses, this decrease in forage digestibility can be avoided as there will be no increase in syringyl lignin. An added side effect of controlling or preventing flowering is that no pollen or seed will produced. This in turn will reduce the uncontrolled spread of genetically modified organisms, as well as reduce the amount of pollen produced. Ryegrass pollen is one of the most common allergens leading to hay fever in humans 9 (Bhalla et al., Proc. Nat. Acad. Sci. USA 96:11676-11680, 1999). In addition, by linking the flowering control genes of the present invention to an inducible promoter, the timing of flowering can be accurately controlled.

The flowering of a plant may be modified by incorporating additional copies of 5 flower control genes of the present invention into the genome of the target plant, or by transforming the target plant with anti-sense copies of such flower control genes. In addition, the number of copies of flowering genes can be manipulated to alter the time of transition from vegetative to floral state.

The present invention thus provides methods for modulating the polynucleotide 10 and/or polypeptide content and composition of an organism, such methods involving stably incorporating into the genome of the organism a genetic construct comprising one or more polynucleotides of the present invention. In one embodiment, the target organism is a plant species, preferably a forage plant, more preferably a grass of the Lolium or Festuca species, and most preferably Lolium perenne or Festuca arundinacea. 15 In related aspects, methods for producing a plant having an altered genotype or phenotype is provided, such methods comprising transforming a plant cell with a genetic construct of the present invention to provide a transgenic cell, and cultivating the transgenic cell under conditions conducive to regeneration and mature plant growth. Plants having an altered genotype or phenotype as a consequence of modulation of the level or content of a 20 polynucleotide or polypeptide of the present invention compared to a wild-type organism, as well as components (seeds, etc.) of such plants, and the progeny of such plants, are contemplated by and encompassed within the present invention.

The isolated polynucleotides of the present invention also have utility in genome mapping, in physical mapping, and in positional cloning of genes. Additionally, the 25 polynucleotide sequences identified as SEQ ID NOS: 1-20 and their variants, may be used to design oligonucleotide probes and primers. Oligonucleotide probes and primers have sequences that are substantially complementary to the polynucleotide of interest over a certain portion of the polynucleotide. Oligonucleotide probes designed using the polynucleotides of the present invention may be employed to detect the presence and 30 examine the expression patterns of genes in any organism having sufficiently similar DNA and RNA sequences in their cells using techniques that are well known in the art, such as slot blot DNA hybridization techniques. Oligonucleotide primers designed using the polynucleotides of the present invention may be used for PCR amplifications. Oligonucleotide probes and primers designed using the polynucleotides of the present invention may also be used in connection with various microarray technologies, including the microarray technology of Affymetrix Inc. (Santa Clara, CA).

Disclosed herein are isolated polynucleotide sequences identified in the attached Sequence Listing as SEQ ID NOS: 1-20, and polypeptide sequences identified in the 5 attached Sequence Listing as SEQ ID NOS: 21-40. The polynucleotides and polypeptides disclosed have demonstrated similarity to the following polypeptides that are known to be involved in flowering pathways: TABLE 1 SEQ ID NO: DNA SEQ ID NO polypeptide Category Description 1-3 21-23 Transcriptional regulation/ Floral (. development Homologs isolated from L. perenne of ASK1 Arabidopsis SKP-like), which regulates B function gene expression in cooperation with UFO and LFY in Arabidopsis (Zhao et al, Development 128:2735-2746, 2001). 24 Transcriptional regulation/ Floral development Homolog isolated from F. arundinacea of the Arabidopsis thaliana transcription factor FCA that is involved in control of flowering time. FCA encodes a RNA binding protein. The protein contains two RNA-binding domains and a WW protein interaction domain suggesting that FCA functions in the posttranscriptional regulation of transcripts involved in the flowering process. FCA appears to be a component of a posttranscriptional cascade involved in the control of flowering time (Koornneef, Curr. Biol 7:R651-652, 1997).

Transcriptional regulation/ Floral development Homolog isolated from L. perenne of the Arabidopsis thaliana transcription factor FCA that is involved in control of flowering time. FCA encodes a RNA binding protein. The protein contains two RNA-binding domains and a WW protein interaction domain suggesting that FCA functions in the posttranscriptional regulation of transcripts involved in the flowering process. FCA appears to be a component of a posttranscriptional cascade involved in the control of flowering time (Koornneef, Curr. Biol 7:R651-652, 1997). 26 Transcriptional regulation/ Floral development Homolog isolated from F. arundinacea of the 1 Flowering locus T (FT), which together with "Suppression of overexpression of COl" (SOC1) interacts with Arabidopsis C OFFirc^ PROPERTY UFRCF OF M.Z - 8 JUL 2008 Eic£ I vedI 11 SEQ ID NO: DNA SEQ ID NO polypeptide Category Description flowering in response to day length. Ft and Socl can act independently of CO, putatively by acting within a different flowering-time pathway (Samach et al., Science 288:1613-1616,2000). 7 27 Transcriptional regulation/ Flora development Homolog isolated from L. perenne of the Flowering locus T (FT), which together with "Suppression of overexpression of COl" (SOC1) interacts with Arabidopsis CO to promote flowering in response to day length. Ft and Socl can act independently of CO, putatively by acting within a different flowering-time pathway (Samach et al., Science 288:1613-1616,2000). 8 28 Transcriptional regulation/ Floral development Homolog isolated from L. perenne of the Arabidopsis thaliana GEBBERELLIN INSENSITIVE (GAI) gene that is involved in developmental processes including seed development and germination, flower and fruit development and flowering time. Genetic studies with A thaliana have identified two genes involved in GA perception or signal transduction. A semidominant mutation at the GAI locus results in plants resembling GA-deficient mutants but exhibiting reduced sensitivity to GA (Jacobsen et al, Proc. Natl Acad. Sci. USA 93:9292-9296,1996). 9 29 Transcriptional regulation/ DNA binding/ 'lowering control homolog isolated from L. perenne of the Arabidopsis thaliana GIGANTEA (GI) gene that is involved in control of flowering time. GI is a nucleoplasmically localized protein involved in Dhytochrome signaling (Huq et al., Proc. Natl Acad Sci. USA 97:9789-9794,2000). Flowering of Arabidopsis is promoted by long days and delayed by short days. GI expression is regulated jy the circadian clock GI plays an important role in regulating the expression of flowering time genes during the promotion of flowering by jhotoperiod (Fowler et al., EMBO J. 18:4679-4-688, 1999).

Transcriptional regulation/ DNA binding/ 'lowering control Homolog isolated fromi7. arundinacea of the Arabidopsis thaliana GIGANTEA (GI) gene that is involved in control of flowering time. GI is a nucleoplasmically localized protein involved in phytochrome signaling (Huq et al., Proc. Natl Acad. Sci. USA 97:9789-9794,2000). Flowering if Arabidopsis is promoted by long days and ielayed by short days. GI expression is regulated jy the circadian clock GI plays an important role 12 SEQ ID NO: DNA SEQ ID NC polypeptide ► Category .

Description in regulating the expression of flowering time genes during the promotion of flowering by photoperiod (Fowler et al, EMBO J. 18:4679-4688, 1999). 11 31 Transcriptional regulation/ DNA binding/ Flowering development Homolog isolated from F. arundinacea of the maize Indeterminate 1 gene (ID1) that controls the transition to flowering. ID1 encodes a protein with zinc finger motifs and functions as a transcriptional regulator of the floral transition (Colasanti et ah, Cell 93:593-603,1998). 12 32 Transcriptional regulation/ DNA binding/ Flowering development Homolog isolated from L. perenne of the maize Indeterminate 1 gene (TDl) that controls the transition to flowering. ID1 encodes a protein with zinc finger motifs and functions as a transcriptional regulator of the floral transition (Colasanti et al., Cell 93:593-603, 1998). 13 33 Transcriptional regulation/ DNA binding/ Flowering development Homolog isolated from F. arundinacea of the maize Indeterminatel gene (ID1) that controls the transition to flowering. ID1 encodes a protein with zinc finger motifs and functions as a transcriptional regulator of the floral transition Colasanti et al., Cell 93:593-603, 1998). 14,15 34,35 Transcriptional regulation/ Floral development .

Homolog isolated from F. arundinacea of _,EUNIG, a key regulator of the Arabidopsis floral homeotic gene AGAMOUS. LEUNIG encodes a glutamine-rich protein with seven WD repeats and is similar in motif structure to a class of functionally related transcriptional co-repressors. The nuclear localization of LEUNIG is consistent with a role of LEUNIG as a transcriptional regulator (Conner and Liu, Proc. Natl. Acad. Sci. USA 97:12902-12907,2000). Another regulatory gene, SEUSS, has been identified that functions together with LEUNIG to regulate AGAMOUS (Franks et al, Development 129:253-263,2002). 16 36 Transcriptional -egulation/ Floral Ievelopment lomolog isolated from F. arundinacea of the Arabidopsis TERMINAL FLOWER1 (TFLl) *ene involved in initiation of flowering. TFKL1 5 controlled by the MADS box proteins CAULIFLOWER, LEAFY and APETALA1 'Liljegren et al., Plant Cell 11:1007-1018, 1999). 17-19 37-39 transcriptional •egulation/ Floral Ievelopment e iomologs isolated from F. arundinacea of the SEUSS transcription factor that plays a role in •egulation of plant development. The SEUSS jrotein contains two glutamine-rich domains and i conserved domain with similarity to 13 SEQ ID NO: DNA SEQ ID NO polypeptide Category Description dimerization domain of the LIM-domain-binding transcription co-regulators in animals. SEUSS encodes a regulator of AGAMOUS and functions together with LEUNIG (Franks et al., Development. 129:253-263,2002). 40 Transcriptional regulation/ Floral development Homolog isolated from F. arundinacea of the Arabidopsis thaliana GD3BERELLIN INSENSITIVE (GAI) gene that is involved in developmental processes including seed development and germination, flower and fruit development and flowering time. Genetic studies with A. thaliana have identified two genes involved in GA perception or signal transduction. A semidominant mutation at the GAI locus results in plants resembling GA-deficient mutants but exhibiting reduced sensitivity to GA (Jacobsen et al., Proc. Natl. Acad. Sci. USA 93:9292-9296,1996).

All the polynucleotides and polypeptides provided by the present invention are isolated and purified, as those terms are commonly used in the art. Preferably, the 5 polypeptides and polynucleotides are at least about 80% pure, more preferably at least about 90% pure, and most preferably at least about 99% pure.

The word "polynucleotide(s)," as used herein, means a polymeric collection of nucleotides, and includes DNA and corresponding RNA molecules and both single and double stranded molecules, including HnRNA and mRNA molecules, sense and anti-10 sense strands of DNA and RNA molecules, and comprehends cDNA, genomic DNA, and wholly or partially synthesized polynucleotides.

In analyzing the phloem-mobile RNA populations of cucurbits, the presence of microRNA-like molecules (miRNAs) in phloem sap and vascular strands of cucurbits has been detected. miRNAs have been reported in other organisms including C. elegans, 15 Droscphila and humans, and are proposed to act as regulators of processes involved in early development and synaptic plasticity of neurons (for a review see Ruvkun, Science 294:191 (1999)). These small RNAs are derived from double-stranded RNA precursors by cellular machinery that produces small RNAs associated with PTGS/RNAi (Hutvagner et al., Science 293:834-838 (2001); Grishok et al., Cell 106: 23-34 (2001)). The presence 20 of this small RNA population in phloem sap of plants suggests that miRNA may play a 14 regulatory role in flowering and other processes that act systemically using long distance signaling mechanisms.

While not wishing to be held to theory, the inventors believe that the small RNA population of the phloem is produced by components of cellular processes involved in the 5 maturation of siRNA (Hamilton and Baulcombe, Science 286:950-2 (1999)). These components may include homologs of the plant genes Argonaute (Bohmert et al., EMBO J. 17: 170-180 (1998), Carpel Factory (Jacobsen et al., Development 126: 5231-5243 (1999); SDE1/SGS2 (Mourrain, Cell 101: 533-542 (2000); Dalmay et al., Cell 101: 543-553 (2000)) and SDE3 (Dalmay et al., EMBO J. 20: 2069-2078 (2001)). miRNAs 10 corresponding to the inventive polynucleotide sequences are contemplated by the present invention and encompassed within the term "polynucleotide".

A polynucleotide of the present invention may be an entire gene or any portion thereof. As used herein, a "gene" is a DNA sequence that codes for a functional protein or RNA molecule. Operable anti-sense polynucleotides may comprise a fragment of the 15 corresponding polynucleotide, and the definition of "polynucleotide" therefore includes all operable anti-sense fragments. Anti-sense polynucleotides and techniques involving anti-sense polynucleotides are well known in the art and are described, for example, in Robinson-Benion et al., Methods in Enzymol. 254(23): 363-375, 1995 and Kawasaki et al.,Artific. Organs 20(8): 836-848, 1996.

Disclosed are isolated polynucleotides comprising a sequence of SEQ ID NO: 1- ; polynucleotides comprising variants of SEQ ID NO: 1-20; polynucleotides comprising extended sequences of SEQ ID NO: 1-20 and their variants, oligonucleotide primers and probes corresponding to the sequences set out in SEQ ID NO: 1-20 and their variants, polynucleotides comprising at least a specified number of contiguous residues of 25 any of SEQ ID NO: 1-20 (x-mers), and polynucleotides comprising extended sequences which include portions of the sequences set out in SEQ ID NO: 1-20, all of which are referred to herein, collectively, as "polynucleotides disclosed." Polynucleotides that comprise complements of such polynucleotide sequences, reverse complements of such polynucleotide sequences, or reverse sequences of such polynucleotide sequences, 30 together with variants of such sequences, are also provided.

The definition of the terms "complement(s)," "reverse complement(s)," and "reverse sequence(s)," as used herein, is best illustrated by the following example. For intellectual property OFFICE of N.Z - 8 JUL 2008 r> .r~../r\ r» the sequence 5' AGGACC 3', the complement, reverse complement, and reverse sequence are as follows: Preferably, sequences that are complements of a specifically recited polynucleotide sequence are complementary over the entire length of the specific polynucleotide sequence.

As used herein, the term "x-mer," with reference to a specific value of "x," refers to a polynucleotide comprising at least a specified number ("x") of contiguous residues of: any of the polynucleotides provided in SEQ ID NOS: 1-20. The value of x may be from about 20 to about 600, depending upon the specific sequence.

Polynucleotides disclosed comprehend polynucleotides comprising at least a specified number of contiguous residues (x-mers) of any of the polynucleotides identified as SEQ ID NOS: 1-20, or their variants. Similarly, polypeptides disclosed comprehend polypeptides comprising at least a specified number of contiguous residues (x-mers) of any of the polypeptides identified as SEQ ID NOS: 21-40. The value of x may be at least 20, more preferably at least 40, more preferably yet at least 60, and most preferably at least 80. Thus, polynucleotides disclosed include polynucleotides comprising a 20-mer, a 40-mer, a 60-mer, an 80-mer, a 100-mer, a 120-mer, a 150-mer, a 180-mer, a 220-mer, a 250-mer; or a 300-mer, 400-mer, 500-mer or 600-mer of a polynucleotide provided in SEQ ID NOS: 1-20, or a variant of one of the polynucleotides corresponding to the polynucleotides provided in SEQ ID NOS: 1-20. Polypeptides disclosed include polypeptides comprising a 20-mer, a 40-mer, a 60-mer, an 80-mer, a 100-mer, a 120-mer, a 150-mer, a 180-mer, a 220-mer, a 250-mer; or a 300-mer, 400-mer, 500-mer or 600-mer of a polypeptide provided in SEQ ID NOS: 21-40, or a variant thereof.

Polynucleotides of the present invention were isolated by high throughput sequencing of cDNA libraries comprising forage grass tissue collected from Lolium perenne and Festuca arundinacea. Some of the polynucleotides of the present invention may be "partial" sequences, in that they do not represent a full-length gene encoding a full-length polypeptide. Such partial sequences may be extended by analyzing and sequencing various DNA libraries using primers and/or probes and well known hybridization and/or PCR techniques. Partial sequences may be extended until an open complement reverse complement reverse sequence 3' TCCTGG 5' 3' GGTCCT 5' ' CCAGGA 3'. 16 reading frame encoding a polypeptide, a full-length polynucleotide and/or gene capable of expressing a polypeptide, or another useful portion of the genome is identified. Such extended sequences, including full-length polynucleotides and genes, are described as "corresponding to" a sequence identified as one of the sequences of SEQ ID NOS: 1-20 5 or a variant thereof, or a portion of one of the sequences of SEQ ID NOS: 1-20 or a variant thereof, when the extended polynucleotide comprises an identified sequence or its variant, or an identified contiguous portion (x-mer) of one of the sequences of SEQ ID NOS: 1-20 or a variant thereof. Similarly, RNA sequences, reverse sequences, complementary sequences, anti-sense sequences and the like, corresponding to the 10 polynucleotides of the present invention, may be routinely ascertained and obtained using the cDNA sequences identified as SEQ ID NOS: 1-20.

The polynucleotides identified as SEQ ID NOS: 1-20 may contain open reading frames ("ORFs") or partial open reading frames encoding polypeptides and functional portions of polypeptides. Partial open reading frames are encoded by SEQ ID NOS: 3-5, 15 7, 8, 17 and 19, while SEQ ID NOS: 1, 2, 6, 9-16, 18 and 20 represent full-length sequences. Additionally, open reading frames encoding polypeptides may be identified in extended or full-length sequences corresponding to the sequences disclosed herein. Open reading frames may be identified using techniques that are well known in the art. These techniques include, for example, analysis for the location of known start and stop codons, 20 most likely reading frame identification based on codon frequencies, etc. These techniques include, for example, analysis for the location of known start and stop codons, most likely reading frame identification based on codon frequencies, etc. Suitable tools and software for ORF analysis are well known in the art and include, for example, GeneWise, available from The Sanger Center, Wellcome Trust Genome Campus, 25 Hinxton, Cambridge, CB10 ISA, United Kingdom; Diogenes, available from Computational Biology Centers, University of Minnesota, Academic Health Center, UMHG Box 43 Minneapolis MN 55455; and GRAIL, available from the Informatics Group, Oak Ridge National Laboratories, Oak Ridge, Tennessee TN. Once a partial open reading frame is identified, the polynucleotide may be extended in the area of the partial 30 open reading frame using techniques that are well known in the art until the polynucleotide for the full open reading frame is identified.

Once open reading frames are identified in the polynucleotides of the present invention, the open reading frames may be isolated and/or synthesized. Expressible genetic constructs comprising the open reading frames and suitable promoters, initiators, 17 terminators, etc., which are well known in the art, may then be constructed. Such genetic constructs may be introduced into a host cell to express the polypeptide encoded by the open reading frame. Suitable host cells may include various prokaryotic and eukaryotic cells, including plant cells, mammalian cells, bacterial cells, algae and the like.

The polynucleotides of the present invention may be isolated by high throughput sequencing of cDNA libraries prepared from forage grass tissue, as described below in Example 1. Alternatively, oligonucleotide probes and primers based on the sequences provided in SEQ ID NOS: 1-36 can be synthesized as detailed below, and used to identify positive clones in either cDNA or genomic DNA libraries from forage grass tissue cells 10 by means of hybridization or polymerase chain reaction (PCR) techniques. Hybridization and PCR techniques suitable for use with such oligonucleotide probes are well known in the art (see, for example, Mullis et al, Cold Spring Harbor Symp. Quant. Biol., 51:263, 1987; Erlich, ed., PCR technology, Stockton Press: NY, 1989; and Sambrook et al., eds., Molecular cloning: a laboratory manual, 2nd ed., CSHL Press: Cold Spring Harbor, NY, 15 1989). In addition to DNA-DNA hybridization, DNA-RNA or RNA-RNA hybridization assays are also possible. In the first case, the mRNA from expressed genes would then be detected instead of genomic DNA or cDNA derived from mRNA of the sample. In the second case, RNA probes could be used. Artificial analogs of DNA hybridizing specifically to target sequences could also be employed. Positive clones can be analyzed 20 by using restriction enzyme digestion, DNA sequencing or the like.

The polynucleotides of the present invention may also, or alternatively, be synthesized using techniques that are well known in the art. The polynucleotides may be synthesized, for example, using automated oligonucleotide synthesizers (e.g., Beckman Oligo 1000M DNA Synthesizer; Beckman Coulter Ltd., Fullerton, CA) to obtain 25 polynucleotide segments of up to 50 or more nucleic acids. A plurality of such polynucleotide segments may then be ligated using standard DNA manipulation techniques that are well known in the art of molecular biology. One conventional and exemplary polynucleotide synthesis technique involves synthesis of a single stranded polynucleotide segment having, for example, 80 nucleic acids, and hybridizing that 30 segment to a synthesized complementary 85 nucleic acid segment to produce a 5 nucleotide overhang. The next segment may then be synthesized in a similar fashion, with a 5 nucleotide overhang on the opposite strand. The "sticky" ends ensure proper ligation when the two portions are hybridized. In this way, a complete polynucleotide of the present invention may be synthesized entirely in vitro. 18 Oligonucleotide probes and primers complementary to and/or corresponding to SEQ ID NOS: 1-20 and variants of those sequences, are also comprehended. Such oligonucleotide probes and primers are substantially complementary to the polynucleotide of interest over a certain portion of the polynucleotide. An oligonucleotide probe or 5 primer is described as "corresponding to" a polynucleotide disclosed, including one of the sequences set out as SEQ ID NOS: 1-20 or a variant thereof, if the oligonucleotide probe or primer, or its complement, is contained within one of the sequences set out as SEQ ID NOS: 1-20 or a variant of one of the specified sequences.

Two single stranded sequences are said to be substantially complementary when 10 the nucleotides of one strand, optimally aligned and compared, with the appropriate nucleotide insertions and/or deletions, pair with at least 80%, preferably at least 90% to 95%, and more preferably at least 98% to 100%, of the nucleotides of the other strand. Alternatively, substantial complementarity exists when a first DNA strand will selectively hybridize to a second DNA strand under stringent hybridization conditions. 15 The oligonucleotide probes and/or primers comprise at least about 6 contiguous residues, more preferably at least about 10 contiguous residues, and most preferably at least about 20 contiguous residues complementary to a polynucleotide sequence of the present invention. Probes and primers of the present invention may be from about 8 to 100 base pairs in length, preferably from about 10 to 50 base pairs in length, and more 20 preferably from about 15 to 40 base pairs in length. The probes can be easily selected using procedures well known in the art, taking into account DNA-DNA hybridization stringencies, annealing and melting temperatures, potential for formation of loops, and other factors that are well known in the art. Preferred techniques for designing PCR primers are disclosed in Dieffenbach and Dyksler, PCR Primer: a laboratory manual, 25 CSHL Press: Cold Spring Harbor, NY, 1995. A software program suitable for designing probes, and especially for designing PCR primers, is available from Premier Biosoft International, 3786 Corina Way, Palo Alto, CA 94303-4504.

The isolated polynucleotides of the present invention also have utility in genome mapping, in physical mapping, and in positional cloning of genes. 30 The polynucleotides identified as SEQ ID NOS: 1-20 were isolated from cDNA clones and represent sequences that are expressed in the tissue from which the cDNA was prepared. RNA sequences, reverse sequences, complementary sequences, anti-sense sequences, and the like, corresponding to the polynucleotides of the present invention, 19 / I - 8 JUL 2008 | iRFrcn/r-^ may be routinely ascertained and obtained using the cDNA sequences identified as SEQ ID NOS: 1-20.

Identification of genomic DNA and heterologous species DNA can be accomplished by standard DNA/DNA hybridization techniques, under appropriately 5 stringent conditions, using all or part of a polynucleotide sequence as a probe to screen an appropriate library. Alternatively, PCR techniques using oligonucleotide primers that are designed based on known genomic DNA, cDNA and protein sequences can be used to amplify and identify genomic and cDNA sequences.

In another aspect, the present invention provides isolated polypeptides encoded by 10 the polynucleotides of the invention. As used herein, the term "polypeptide" encompasses amino acid chains of any length, including full-length proteins, wherein the amino acid residues are linked by covalent peptide bonds. The term "polypeptide encoded by a polynucleotide" as used herein, includes polypeptides encoded by a polynucleotide that comprises a partial isolated polynucleotide sequence provided herein. Also disclosed are 15 polypeptides comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 21-40, as well as variants of such sequences.

As noted above, polypeptides of the present invention may be produced recombinantly by inserting a polynucleotide sequence of the present invention encoding the polypeptide into an expression vector and expressing the polypeptide in an 20 appropriate host. Any of a variety of expression vectors known to those of ordinary skill in the art may be employed. Expression may be achieved in any appropriate host cell that has been transformed or transfected with an expression vector containing a polynucleotide molecule that encodes a recombinant polypeptide. Suitable host cells include prokaryotes, yeast, and higher eukaryotic cells. Preferably, the host cells 25 employed are plant, E. coli, insect, yeast, or a mammalian cell line such as COS or CHO. The polynucleotide sequences expressed in this manner may encode naturally occurring polypeptides, portions of naturally occurring polypeptides, or other variants thereof. The expressed polypeptides may be used in various assays known in the art to determine their biological activity. Such polypeptides may also be used to raise antibodies, to isolate 30 corresponding interacting proteins or other compounds, and to quantitatively determine levels of interacting proteins or other compounds.

Polypeptides are provided that comprise at least a functional portion of a polypeptide having an amino acid sequence selected from the group consisting of sequences provided in SEQ ID NO: 21-40 and variants thereof. As used inteHal property] office of m.z. - 8 JUL 2008 D nrr^t- i w _ herein, the "functional portion" of a polypeptide is that portion which contains an active site essential for affecting the function of the polypeptide, for example, a portion of the molecule that is capable of binding one or more reactants. The active site may be made up of separate portions present on one or more polypeptide chains and will generally 5 exhibit high binding affinity. Functional portions of a polypeptide may be identified by first preparing fragments of the polypeptide by either chemical or enzymatic digestion of the polypeptide, or by mutation analysis of the polynucleotide that encodes the polypeptide and subsequent expression of the resulting mutant polypeptides. The polypeptide fragments or mutant polypeptides are then tested to determine which portions 10 retain biological activity, using methods well known to those of skill in the art, including the representative assays described below.

Portions and other variants of the inventive polypeptides may be generated by synthetic or recombinant means. Synthetic polypeptides having fewer than about 100 amino acids, and generally fewer than about 50 amino acids, may be generated using 15 techniques well known to those of ordinary skill in the art. For example, such polypeptides may be synthesized using any of the commercially available solid-phase techniques, such as the Merrifield solid-phase synthesis method, where amino acids are sequentially added to a growing amino acid chain. See Merrifield, J. Am. Chem. Soc. 85: 2149-2146, 1963. Equipment for automated synthesis of polypeptides is commercially 20 available from suppliers such as Perkin Elmer/Applied Biosystems, Inc. (Foster City, California), and may be operated according to the manufacturer's instructions. Variants of a native polypeptide may be prepared using standard mutagenesis techniques, such as oligonucleotide-directed site-specific mutagenesis (Kunkel, Proc. Natl. Acad. Sci. USA 82:488-492, 1985). Sections of DNA sequences may also be removed using standard 25 techniques to permit preparation of truncated polypeptides.

As used herein, the term "variant" comprehends nucleotide or amino acid sequences different from the specifically identified sequences, wherein one or more nucleotides or amino acid residues is deleted, substituted, or added. Variants may be naturally occurring allelic variants, or non-naturally occurring variants. Variant 30 sequences (polynucleotide or polypeptide) preferably exhibit at least 75%, more preferably at least 80%, more preferably at least 90%, more preferably yet at least 95% and most preferably, at least 98% identity to a sequence of the present invention. The percentage identity is determined by aligning the two sequences to be compared as described below, determining the number of identical residues in the aligned portion, 21 dividing that number by the total number of residues in the inventive (queried) sequence, and multiplying the result by 100.

Polynucleotides and polypeptides having a specified percentage identity to a polynucleotide or polypeptide identified in one of SEQ ID NO: 1-40 thus share a high 5 degree of similarity in their primary structure. In addition to a specified percentage identity to a polynucleotide of the present invention, variant polynucleotides and polypeptides preferably have additional structural and/or functional features in common with a polynucleotide of the present invention. Polynucleotides having a specified degree of identity to, or capable of hybridizing to, a polynucleotide of the present invention 10 preferably additionally have at least one of the following features: (1) they contain an open reading frame, or partial open reading frame, encoding a polypeptide, or a functional portion of a polypeptide, having substantially the same functional properties as the polypeptide, or functional portion thereof, encoded by a polynucleotide in a recited SEQ ID NO.; or (2) they contain identifiable domains in common.

Polynucleotide or polypeptide sequences may be aligned, and percentages of identical nucleotides or amino acids in a specified region may be determined against another polynucleotide or polypeptide, using computer algorithms that are publicly available. The BLASTN and FASTA algorithms, set to the default parameters described in the documentation and distributed with the algorithm, may be used for aligning and 20 identifying the similarity of polynucleotide sequences. The alignment and similarity of polypeptide sequences may be examined using the BLASTP algorithm. BLASTX and FASTX algorithms compare nucleotide query sequences translated in all reading frames against polypeptide sequences. The FASTA and FASTX algorithms are described in Pearson and Lipman, Proc. Natl. Acad. Sci. USA 85:2444-2448, 1988; and in Pearson, 25 Methods in Enzymol. 183:63-98, 1990. The FASTA software package is available from the University of Virginia by contacting the Assistant Provost for Research, University of . Virginia, PO Box 9025, Charlottesville, VA 22906-9025. The BLASTN software is available from the National Center for Biotechnology Information (NCBI), National Library of Medicine, Building 38A, Room 8N805, Bethesda, MD 20894. The BLASTN 30 algorithm Version 2.0.11 [Jan-20-2000] set to the default parameters described in the documentation and distributed with the algorithm, is preferred for use in the determination of polynucleotide variants according to the present invention. The use of the BLAST family of algorithms, including BLASTN, BLASTP and BLASTX, is 22 described in the publication of Altschul et al., "Gapped BLAST and PSI-BLAST: a new generation of protein database search programs," Nucleic Acids Res. 25:3389-3402,1997.

The following running parameters are preferred for determination of alignments and similarities using BLASTN that contribute to the E values and percentage identity for 5 polynucleotides: Unix running command with the following default parameters: blastall -p blastn -d embldb -e 10 -G 0 -E 0 -FF -r 1 -v 30 -b 30 -i queryseq -o results; and parameters are: -p Program Name [String]; -d Database [String]; -e Expectation value (E) [Real]; -G Cost to open a gap (zero invokes default behavior) [Integer]; -E Cost to extend a gap (zero invokes default behavior) [Integer]; -FF low complexity filter; -r 10 Reward for a nucleotide match (BLASTN only) [Integer]; -v Number of one-line descriptions (V) [Integer]; -b Number of alignments to show (B) [Integer]; -i Query File [File In]; -o BLAST report Output File [File Out] Optional.

The following running parameters are preferred for determination of alignments and similarities using BLASTP that contribute to the E values and percentage identity of 15 polypeptide sequences: blastall -p blastp -d swissprottrembledb -e 10 -G 0 -E 0 -FF -v 30 -b 30 -i queryseq -o results; the parameters are: -p Program Name [String]; -d Database [String]; -e Expectation value (E) [Real]; -G Cost to open a gap (zero invokes default behavior) [Integer]; -E Cost to extend a gap (zero invokes default behavior) [Integer]; -FF low complexity filter; -v Number of one-line descriptions (v) [Integer]; -b 20 Number of alignments to show (b) [Integer]; -I Query File [File In]; -o BLAST report Output File [File Out] Optional.

The "hits" to one or more database sequences by a queried sequence produced by BLASTN, BLASTP, FASTA, or a similar algorithm, align and identify similar portions of sequences. The hits are arranged in order of the degree of similarity and the length of 25 sequence overlap. Hits to a database sequence generally represent an overlap over only a fraction of the sequence length of the queried sequence.

As noted above, the percentage identity of a polynucleotide or polypeptide sequence is determined by aligning polynucleotide and polypeptide sequences using appropriate algorithms, such as BLASTN or BLASTP, respectively, set to default 30 parameters; identifying the number of identical nucleic or amino acids over the aligned portions; dividing the number of identical nucleic or amino acids by the total number of nucleic, or amino acids of the polynucleotide or polypeptide of the present invention; and then multiplying by 100 to determine the percentage identity. By way of example, a queried polynucleotide having 220 nucleic acids has a hit to a polynucleotide sequence in 23 the EMBL database having 520 nucleic acids over a stretch of 23 nucleotides in the alignment produced by the BLASTN algorithm using the default parameters. The 23-nucleotide hit includes 21 identical nucleotides, one gap and one different nucleotide. The percentage identity of the queried polynucleotide to the hit in the EMBL database is 5 thus 21/220 times 100, or 9.5%. The percentage identity of polypeptide sequences may be determined in a similar fashion.

The BLASTN and BLASTX algorithms also produce "Expect" values for polynucleotide and polypeptide alignments. The Expect value (E) indicates the number of hits one can "expect" to see over a certain number of contiguous sequences by chance 10 when searching a database of a certain size. The Expect value is used as a significance threshold for determining whether the hit to a database indicates true similarity. For example, an E value of 0.1 assigned to a polynucleotide hit is interpreted as meaning that in a database of the size of the EMBL database, one might expect to see 0.1 matches over the aligned portion of the sequence with a similar score simply by chance. By this 15 criterion, the aligned and matched portions of the sequences then have a probability of 90% of being related. For sequences having an E value of 0.01 or less over aligned and matched portions, the probability of finding a match by chance in the EMBL database is 1% or less using the BLASTN algorithm. E values for polypeptide sequences may be determined in a similar fashion using various polypeptide databases, such as the 20 SwissProt-TrEMBLE database.

According to one embodiment, "variant" polynucleotides and polypeptides, with reference to each of the polynucleotides and polypeptides of the present invention, preferably comprise sequences having the same number or fewer nucleotides or amino acids than each of the polynucleotides or polypeptides of the present invention and 25 producing an E value of 0.01 or less when compared to the polynucleotide or polypeptide of the present invention. That is, a variant polynucleotide or polypeptide is any sequence that has at least a 99% probability of being related to the polynucleotide or polypeptide of the present invention, measured as having an E value of 0.01 or less using the BLASTN or BLASTX algorithms set at the default parameters. According to a preferred 30 embodiment, a variant polynucleotide is a sequence having the same number or fewer nucleic acids than a polynucleotide of the present invention that has at least a 99% probability of being related to the polynucleotide of the present invention, measured as having an E value of 0.01 or less using the BLASTN algorithm set at the default parameters. Similarly, according to a preferred embodiment, a variant polypeptide is a 24 sequence having the same number or fewer amino acids than a polypeptide of the present invention that has at least a 99% probability of being related as the polypeptide of the present invention, measured as having an E value of 0.01 or less using the BLASTP algorithm set at the default parameters.

In an alternative embodiment, variant polynucleotides are sequences that hybridize to a polynucleotide of the present invention under stringent conditions. Stringent hybridization conditions for determining complementarity include salt conditions of less than about 1 M, more usually less than about 500 mM, and preferably less than about 200 mM. Hybridization temperatures can be as low as 5°C, but are 10 generally greater than about 22°C, more preferably greater than about 30°C, and most preferably greater than about 37°C. Longer DNA fragments may require higher hybridization temperatures for specific hybridization. Since the stringency of hybridization may be affected by other factors such as probe composition, presence of organic solvents, and extent of base mismatching, the combination of parameters is more 15 important than the absolute measure of any one alone. An example of "stringent conditions" is prewashing in a solution of 6X SSC, 0.2% SDS; hybridizing at 65°C, 6X SSC, 0.2% SDS overnight; followed by two washes of 30 minutes each in IX SSC, 0.1% SDS at 65°C and two washes of 30 minutes each in 0.2X SSC, 0.1% SDS at 65°C.

The present invention also encompasses polynucleotides that differ from the 20 disclosed sequences but that, as a consequence of the discrepancy of the genetic code, encode a polypeptide having similar enzymatic activity to a polypeptide encoded by a polynucleotide of the present invention. Thus, polynucleotides comprising sequences that differ from the polynucleotide sequences recited in SEQ ID NO: 1-20, or complements, reverse sequences, or reverse complements of those sequences, as a result of conservative 25 substitutions are contemplated. Additionally, polynucleotides comprising sequences that differ from the polynucleotide sequences recited in SEQ ID NO: 1-20, or complements, reverse complements or reverse sequences thereof, as a result of deletions and/or insertions totaling less than 10% of the total sequence length are also contemplated. Similarly, polypeptides comprising sequences that differ from the polypeptide sequences 30 recited in SEQ ID NO: 21-40 as a result of amino acid substitutions, insertions, and/or deletions totaling less than 10% of the total sequence length are intellectualprBperty] office of n.z - 8 JUL 2008 RECElVFn contemplated, provided the variant polypeptide has activity in a flowering pathway.

Also disclosed are genetic constructs comprising, in the 5'-3' direction, a gene promoter sequence; an open reading frame coding for at least a functional portion of a polypeptide disclosed; and a gene termination sequence. The open reading frame may be 5 orientated in either a sense or anti-sense direction. For applications where amplification of enzyme activity is desired, the open reading frame may be inserted in the construct in a sense orientation, such that transformation of a target organism with the construct will lead to an increase in the number of copies of the gene and therefore an increase in the amount of enzyme. When down-regulation of enzyme activity is desired, the open 10 reading frame may be inserted in the construct in an anti-sense orientation, such that the RNA produced by transcription of the polynucleotide is complementary to the endogenous mRNA sequence. This, in turn, will result in a decrease in the number of copies of the gene and therefore a decrease in the amount of enzyme. Alternatively, regulation may be achieved by inserting appropriate sequences or subsequences (e.g., 15 DNA or RNA) in ribozyme constructs.

Genetic constructs comprising a non-coding region of a gene coding for a polypeptide of the present invention, or a nucleotide sequence complementary to a non-coding region, together with a gene promoter sequence and a gene termination sequence, are also provided. As used herein the term "non-coding region" includes both transcribed 20 sequences that are not translated, and non-transcribed sequences within about 2000 base pairs 5' or 3' of the translated sequences or open reading frames. Examples of non-coding regions that may be usefully employed in the inventive constructs include introns and 5'- non-coding leader sequences. Transformation of a target plant with such a genetic construct may lead to a reduction in the amount of enzyme synthesized by the 25 plant by the process of cosuppression, in a manner similar to that discussed, for example, by Napoli et al., Plant Cell 2:279-290, 1990; and de Carvalho Niebel et al., Plant Cell 7:347-358, 1995.

The genetic constructs of the present invention further comprise a gene promoter sequence and a gene termination sequence, operably linked to the polynucleotide to be 30 transcribed, which control expression of the gene. The gene promoter sequence is generally positioned at the 5' end of the polynucleotide to be transcribed, and is employed to initiate transcription of the polynucleotide. Gene promoter sequences are generally found in the 5' non-coding region of a gene but they may exist in introns (Luehrsen, Mol. 26 I "8 JUL 2008 Gen. Genet. 225:81-93, 1991). When the construct includes an open reading frame in a sense orientation, the gene promoter sequence also initiates translation of the open reading frame. For genetic constructs comprising either an open reading frame in an anti-sense orientation or a non-coding region, the gene promoter sequence consists only of a 5 transcription initiation site having a RNA polymerase binding site.

A variety of gene promoter sequences that may be usefully employed in the genetic constructs of the present invention are well known in the art. The promoter gene sequence, and also the gene termination sequence, may be endogenous to the target plant host or may be exogenous, provided the promoter is functional in the target host. For 10 example, the promoter and termination sequences may be from other plant species, plant viruses, bacterial plasmids and the like. Preferably, gene promoter and termination sequences are from the inventive sequences themselves.

Factors influencing the choice of promoter include the desired tissue specificity of the construct, and the timing of transcription and translation. For example, constitutive 15 promoters, such as the 35S Cauliflower Mosaic Virus (CaMV 35S) promoter, will affect the activity of the enzyme in all parts of the plant. Use of a tissue specific promoter will result in production of the desired sense or anti-sense RNA only in the tissue of interest. With genetic constructs employing inducible gene promoter sequences, the rate of RNA polymerase binding and initiation can be modulated by external physical or chemical 20 stimuli, such as light, heat, anaerobic stress, alteration in nutrient conditions and the like. Temporally regulated promoters can be employed to effect modulation of the rate of RNA polymerase binding and initiation at a specific time during development of a transformed cell. Preferably, the original promoters from the enzyme gene in question, or promoters from a specific tissue-targeted gene in the organism to be transformed, such as Lolium or 25 Festuca, are used. Grass promoters different from the original gene may also be usefully employed in the inventive genetic constructs in order to prevent feedback inhibition. Other examples of gene promoters which may be usefully employed in the present invention include, mannopine synthase (mas), octopine synthase (ocs) and those reviewed by Chua et al., Science 244:174-181,1989.

The gene termination sequence, which is located 3' to the polynucleotide to be transcribed, may come from the same gene as the gene promoter sequence or may be from a different gene. Many gene termination sequences known in the art may be usefully employed in the present invention, such as the 3' end of the Agrobacterium 27 tumefaciens nopaline synthase gene. However, preferred gene terminator sequences are those from the original enzyme gene or from the target species to be transformed.

The genetic constructs of the present invention may also contain a selection marker that is effective in plant cells, to allow for the detection of transformed cells 5 containing the inventive construct. Such markers, which are well known in the art, typically confer resistance to one or more toxins. One example of such a marker is the NPTII gene whose expression results in resistance to kanamycin or hygromycin, antibiotics which are usually toxic to plant cells at a moderate concentration (Rogers et alin Weissbach A and H, eds., Methods for Plant Molecular Biology, Academic Press 10 Inc.: San Diego, CA, 1988). Alternatively, the presence of the desired construct in transformed cells can be determined by means of other techniques well known in the art, such as Southern and Western blots.

Techniques for operatively linking the components of the inventive genetic constructs are well known in the art and include the use of synthetic linkers containing 15 one or more restriction endonuclease sites as described, for example, by Sambrook et al., Molecular cloning: a laboratory manual, CSHL Press: Cold Spring Harbor, NY, 1989. The genetic construct of the present invention may be linked to a vector having at least one replication system, for example, E. coli, whereby after each manipulation, the resulting construct can be cloned and sequenced and the correctness of the manipulation 20 determined.

The genetic constructs of the present invention may be used to transform a variety of plants, both monocotyledonous (e.g., grasses, maize/corn, grains, oats, rice, sorghum, millet, rye, sugar cane, wheat and barley), dicotyledonous (e.g., Arabidopsis, tobacco, legumes, alfalfa, oaks, eucalyptus,. maple), and gymnosperms. In a preferred 25 embodiment, the inventive genetic constructs are employed to transform grasses. Preferably the target plant is selected from the group consisting of Lolium and Festuca species, most preferably from the group consisting of Lolium perenne and Festuca arundinacea. Other plants that may be usefully transformed with the inventive genetic constructs include other species of ryegrass and fescue, including, but not limited to, 30 Lolium multiflorum (Italian ryegrass), Lolium hybridum (hybrid ryegrass), Lolium rigidum (Wimerra grass), Lolium temulentum (darnel), Festuca rubra (red fescue) and Festuca pratensis (meadow fescue). As discussed above, transformation of a plant with a genetic construct of the present invention will produce a modification in the flowering of the plant. 28 The production of RNA in target cells may be controlled by choice of the promoter sequence, or by. selecting the number of functional copies or the site of integration of the polynucleotides incorporated into the genome of the target organism. A target plant may be transformed with more than one construct of the present invention, 5 thereby modulating the flowering by affecting the activity of more than one enzyme, affecting enzyme activity in more than one tissue or affecting enzyme activity at more than one expression time. Similarly, a construct may be assembled containing more than one open reading frame coding for an enzyme encoded by a polynucleotide of the present invention or more than one non-coding region of a gene coding for such an enzyme. The 10 polynucleotides of the present invention may also be employed in combination with other known sequences encoding enzymes involved in the flowering and/or other pathways. In this manner, more than one pathway may be modulated to produce a plant having an altered phenotype.

Techniques for stably incorporating DNA constructs into the genome of target 15 plants are well known in the art and include Agrobacterium tumefaciens mediated introduction, electroporation, protoplast fusion, injection into reproductive organs, injection into immature embryos, high velocity projectile introduction and the like. The choice of technique will depend upon the target plant to be transformed. For example, dicotyledonous plants and certain monocots and gymnosperms may be transformed by 20 Agrobacterium Ti plasmid technology, as described, for example by Bevan,- Nucleic Acid Res. 12:8711-8721, 1984. ~ Targets for the introduction of the DNA constructs of the present invention include tissues, such as leaf tissue, disseminated cells, protoplasts, seeds, embryos, meristematic regions; cotyledons, hypocotyls, and the like. Transformation techniques which may be usefully employed .in the inventive methods 25 include those taught by Ellis et al, Plant Cell Reports, 8:16-20,1989, Wilson et al., Plant Cell Reports 7:704-707, 1989; Tautorus etal., Theor. Appl. Genet. 78:531-536, 1989; and Ishida et al., Nat. Biotechnol. 14:745-750,1996.

Once the cells are transformed, cells having the inventive genetic construct incorporated in their genome may be selected by means of a marker, such as the 30 kanamycin resistance marker discussed above. Transgenic cells may then be cultured in an appropriate medium to regenerate whole plants, using techniques well known in the art. In the case of protoplasts, the cell wall is allowed to reform under appropriate osmotic conditions. In the case of seeds or embryos, an appropriate germination or callus initiation medium is employed. For explants, an appropriate regeneration medium is 29 used. Regeneration of plants is well established for many species. The resulting transformed plants may be reproduced sexually or asexually, using methods well known in the art, to give successive generations of transgenic plants.

Polynucleotides of the present invention may also be used to specifically suppress 5 gene expression by methods that operate post-transcriptionally to block the synthesis of products of targeted genes, such as RNA interference (RNAi), and quelling. For a review of techniques of gene suppression see Science, 288:1370-1372, 2000. Exemplary gene silencing methods are also provided in WO 99/49029 and WO 99/53050. Posttranscriptional gene silencing is brought about by a sequence-specific RNA 10 degradation process that results in the rapid degradation of transcripts of sequence-related genes. Studies have provided evidence that double-stranded RNA may act as a mediator of sequence-specific gene silencing (see, e.g., review by Montgomery and Fire, Trends in Genetics, 14: 255-258, 1998). Gene constructs that produce transcripts with self-complementary regions are particularly efficient at gene silencing. A unique feature of 15 this posttranscriptional gene silencing pathway is that silencing is not limited to the cells where it is initiated. The gene-silencing effects may be disseminated to other parts of an organism and even transmitted through the germ line to several generations.

The polynucleotides of the present invention may be employed to generate gene silencing constructs and or gene-specific self-complementary RNA sequences that can be 20 delivered by conventional art-known methods to plant tissues, such as forage grass tissues. Within genetic constructs, sense and antisense sequences can be placed in regions flanking an intron sequence in proper splicing orientation with donor and acceptor splicing sites, such that intron sequences are removed during processing of the transcript and sense and antisense sequences, as well as splice junction sequences, bind together to 25 form double-stranded RNA. Alternatively, spacer sequences of various lengths may be employed to separate self-complementary regions of sequence in the construct. During processing of the gene construct transcript, intron sequences are spliced-out, allowing sense and anti-sense sequences, as well as splice junction sequences, to bind forming double-stranded RNA. Select ribonucleases bind to and cleave the double-stranded RNA, 30 thereby initiating the cascade of events leading to degradation of specific mRNA gene sequences, and silencing specific genes. Alternatively, rather than using a gene construct to express the self-complementary RNA sequences, the gene-specific double-stranded RNA segments are delivered to one or more targeted areas to be internalized into the cell cytoplasm to exert a gene silencing effect. Gene silencing RNA sequences comprising the polynucleotides of the present invention are useful for creating genetically modified plants with desired phenotypes as well as for characterizing genes (e.g., in high-throughput screening of sequences), and studying their functions in intact organisms.

The following examples are offered by way of illustration and not by way of limitation.

Example 1 Isolation of cDNA Sequences from L. perenne and F. arundinacea CDNA LIBRARIES L. perenne and F. arundinacea cDNA expression libraries were constructed and screened as follows. Tissue was collected from L. perenne and F. arundinacea during winter and spring, and snap-frozen in liquid nitrogen. The tissues collected include those 15 obtained from leaf blades, leaf base, pseudostem, roots and stem. Total RNA was isolated from each tissue type using TRIzol Reagent (BRL Life Technologies, Gaithersburg, MD). mRNA from each tissue type was obtained using a Poly(A) Quik mRNA isolation kit (Stratagene, La Jolla, CA), according to the manufacturer's specifications. cDNA expression libraries were constructed from the purified mRNA by 20 reverse transcriptase synthesis followed by insertion of the resulting cDNA in Lambda ZAP using a ZAP Express cDNA Synthesis Kit (Stratagene), according to the manufacturer's protocol. The resulting cDNA clones were packaged using a Gigapack II Packaging Extract (Stratagene) employing 1 jj.1 of sample DNA from the 5 |il ligation mix. Mass excision of the libraries was done using XL 1-Blue MRF' cells and XLOLR 25 cells (Stratagene) with ExAssist helper phage (Stratagene). The excised phagemids were diluted with NZY broth (Gibco BRL, Gaithersburg, MD) and plated out onto LB-kanamycin agar plates containing 5-bromo-4-chloro-3-indolyl-beta-D-galactosidase (X-gal) and isopropylthio-beta-galactoside (IPTG).

Of the colonies plated and picked for DNA preparations, the large majority 30 contained an insert suitable for sequencing. Positive colonies were cultured in NZY broth with kanamycin and DNA was purified following standard protocols. Agarose gel at 1% was used to screen sequencing templates for chromosomal contamination. Dye terminator sequences were prepared using a Biomek 2000 robot (Beckman Coulter Inc., Fullerton, CA) for liquid handling and DNA amplification using a 9700 PCR machine 31 (Perkin Elmer/Applied Biosystems, Foster City, CA) according to the manufacturer's protocol.

The DNA sequences for positive clones were obtained using a Perkin Elmer/Applied Biosystems Division Prism 377 sequencer. cDNA clones were sequenced 5 from the 5' end. The polynucleotide sequence identified as SEQ ID NO: 8 was identified from a L. perenne leaf cDNA expression library; the polynucleotide sequences identified as SEQ ID NOS: 1, 3 and 7 were identified from L. perenne leaf and pseudostem cDNA expression libraries; the polynucleotide sequences identified as SEQ ID NOS: 2 and 12 were identified from L. perenne floral stem cDNA expression libraries; the ' 10 polynucleotide sequence identified as SEQ ID NO: 9 was identified from a L.perenne stem cDNA expression library; the polynucleotide sequence identified as SEQ ID NO: 5 was identified from a L. perenne root cDNA expression library; the polynucleotide sequences identified as SEQ ID NOS: 4, 11 and 14 were identified from a F. arundinacea inflorescence (day 2) cDNA expression library; the polynucleotide sequences identified 15 as SEQ ID NOS: 13 and 16 were identified from a F. arundinacea cDNA expression library constructed from stem bases from day 7 inflorescences; the polynucleotide sequences identified as SEQ ED NOS: 10 and 20 were identified from F. arundinacea pseudostem cDNA expression libraries; the polynucleotide sequences identified as SEQ ID NOS: 15, 18 and 19 were identified from F. arundinacea leaf cDNA expression 20 libraries; the polynucleotide sequence identified as SEQ ID NO: 6 was identified from F. arundinacea inflorescence cDNA expression libraries; and the polynucleotide sequence identified as SEQ ID NO: 17 was identified from a F. arundinacea rhizome cDNA expression library. SEQ ED NOS: 1, 2, 6, 9-16, 18 and 20 represent full-length sequences, while SEQ ID NOS: 3-5, 7, 8,17 and 19 encode partial open reading frames.

BLASTN Polynucleotide Analysis The isolated cDNA sequences were compared to sequences in the EMBL DNA database using the computer algorithm BLASTN. Comparisons of DNA sequences provided in SEQ ID NOS: 1-17, 19 and 20, to sequences in the EMBL database (using 30 BLASTN) were made as of August 20, 2003, using BLASTN algorithm Version 2.2.1 [Apr-13-2001] and comparisons of the DNA sequence provided in SEQ ID NO: 18 to sequences in the EMBL database (using BLASTN) were made as of August 26, 2003, using BLASTN algorithm Version 2.0.11 [Jan-20-2000], and the following Unix running 32 command: blastall -p blastn -d embldb -e 10 -GO -E0 -FF -r 1 -v 30 -b 30 -i queryseq -o.

The sequences of SEQ ID NOS: 1-5, 7-15 and 17-19 were determined to have less than 50% identity to sequences in the EMBL database using the computer algorithm 5 BLASTN, as described above. The sequences of SEQ ID NOS: 6, 16 and 20 were determined to have less than 75% identity to sequences in the EMBL database using the computer algorithm BLASTN, as described above.

BLASTP Polypeptide Analysis 10 The isolated cDNA sequences were compared to sequences in the SwissProt- TrEMBLE database using the computer algorithm BLASTP. Comparisons of protein sequences provided in SEQ ID NOS: 21-37, 39 and 40 to sequences in the SwissProt-TrEMBLE protein database were made as of August 15, 2003, using BLASTP algorithm Version 2.2.1 [Apr-13-2001] and comparisons of the protein sequence provided in SEQ 15 ID NO: 38, to sequences in the SwissProt-TrEMBLE protein database were made as of August 26, 2003, using BLASTP algorithm Version 2.0.11 [Jan-20-2000], and the following Unix running command: blastall -p blastp -d swissprottrembledb -e 10 -GO -E0 -FF -v 30 -b 30 -i queryseq -o.

The amino acid sequences of SEQ ID NOS: 31-33 were determined to have less 20 than 50% identity to sequences in the SWISSPROT-TREMBLE database using the BLASTP computer algorithm as described above. The amino acid sequences of SEQ ID NOS: 24, 55, 34, 35 and 38 were determined to have less than 75% identity to sequences in the SWISSPROT-TrEMBLE database using the computer algorithm BLASTP, as described above. The amino acid sequences of SEQ ID NOS: 23, 26, 27, 29, 30, 36, 37 25 and 39 were determined to have less than 90% identity to sequences in the SWISSPROT-TrEMBLE database using the computer algorithm BLASTP, as described above. The amino acid sequences of SEQ ID NOS: 21, 22, 28 and 40 were determined to have less than 98% identity to sequences in the SWISSPROT-TrEMBLE database using the computer algorithm BLASTP, as described above. 33 BLASTX Polynucleotide Analysis The isolated cDNA sequences were compared to sequences in the SwissProt-TrEMBLE protein database using the computer algorithm BLASTX. Comparisons of DNA sequences provided in SEQ ID NOS: 1-17, 19 and 20, to sequences in the SwissProt-TrEMBLE database (using BLASTX) were made as of August 20, 2003 using BLAST algorithm Version 2.2.1 [Apr-13-2001] and comparisons of the DNA sequence provided in SEQ ID NO: 18 to sequences in the SwissProt-TrEMBLE protein database were made as of August 26, 2003, using BLASTP algorithm Version 2.0.11 [Jan-20-2000], and the following Unix running command: blastall -p blastx -d swissprottrembledb -e 10 -GO -E0 -FF -v 30 -b 30 -i queryseq -o.

The cDNA sequences of SEQ ID NOS: 1-5, 7 and 11-16 were determined to have less than 50% identity to sequences in the SWISSPROT-TrEMBLE database using the computer algorithm BLASTX, as described above. The cDNA sequences of SEQ ID NOS: 6, 8, 9, 10 and 17-20 were determined to have less than 75% identity to sequences in the SWISSPROT-TrEMBLE database using BLASTX, as described above.

The location of open reading frames (ORFs), by nucleotide position, contained within the sequences of SEQ ID NO: 1-20 and the corresponding amino acid sequences are provided in Table 2 below.

TABLE 2 Polynucleotide SEQ ID NO: ORF Polypeptide SEQ ID NO: 1 100*591 21 2 95-604 22 3 0-454 23 4 0-1967 24 0-1858 6 97-630 26 7 395-664 27 8 0-898 28 9 154-3600 29 189-3635 11 163-1653 31 12 336-1928 32 13 271-1671 33 14 109-2394 34 ' 140-2413 16 81-605 36 34 Polynucleotide SEQ ID NO: ORF Polypeptide SEQ ID NO: 17 0-1975 37 18 107-2533 38 19 0-2398 39 133-1962 40 Example 2 Use of Grass Flowering Genes to Control Flowering Transformation of Arabidopsis and N. benthamiana plants with grass flowering control genes Sense constructs containing a polynucleotide including the coding region of flowering control genes isolated from Lolium perenne or Festuca arundinacea (SEQ ID 10 NOS: 1, 2, 6, 11, 12, 13, 14, 15, 16) were inserted into a binary vector and used to transform Agrobacterium tumefaciens LBA4404 using published methods (see, Ah G, Ebert PR, Mitra A, Ha SB, "Binary Vectors," in Gelvin SB, Schilperoort RA, eds., Plant Molecular Biology Manual, Kluwer Academic Publishers: Dordrecht, 1988). The presence and integrity of the binary vector in A. tumefaciens was verified by polymerase 15 chain reaction (PCR) utilizing vector primers.

The A. tumefaciens containing the sense gene constructs were used to transform Arabidopsis by floral dipping (Clough and Bent, Plant J. 16:735-743, 1998). Several independent transformed plant lines were established for the sense construct for each flowering gene. Transformed plants containing the appropriate flowering gene construct 20 were verified using PCR experiments.

Effects of grass FaFT flowering control genes on flowering time in transformed Arabidopsis plants The Arabidopsis plant lines transformed with the F. arundinacea FT gene 25 FLOWERING LOCUS T (FaFT) given in SEQ ID NO: 6 were grown for 70 days with 16 hours light and 8 hour night breaks. The plants were visually scored for first floral bud formation and flower opening every 3 days.

Figure 21 shows the time to first floral bud formation for plants over-expressing the grass flowering time gene FaFT (SEQ ID NO: 6) and plants containing the empty control vector. Figure 22 shows the time to first open flowers in plants over-expressing grass flowering time gene FaFT (SEQ NO: 6) and plants containing the empty control vector. These results show that over-expression significantly reduced the time to floral bud formation and first open flowers under long day conditions.

Effects of grass FaLUG flowering control genes on flowering time in transformed Arabidopsis plants The Arabidopsis plant lines transformed with the F. arundinacea LEUNIG gene (FaLUG) given in SEQ ID NO: 14 were grown for 70 days with 16 hours light and 8 hour night breaks. The plants were visually scored for first floral bud formation and flower opening every 3 days.

Figure 22 shows the time to first open flowers in plants over-expressing grass flowering time LEUNIG gene FaLUG (SEQ NO: 14) and plants containing the empty control vector. These results show that over-expression reduced the time to first open flowers under long day conditions.

SEQ ED NOS: 1-40 are set out in the attached Sequence Listing. The codes for nucleotide sequences used in the attached Sequence Listing, including the symbol "n," conform to WIPO Standard ST.25 (1998), Appendix 2, Table 1.

All references cited herein, including patent references and non-patent publications, are hereby incorporated by reference in their entireties.

While in the foregoing specification this invention has been described in relation to certain preferred embodiments, and many details have been set forth for purposes of illustration, it will be apparent to those skilled in the art that the invention is susceptible to additional embodiments and that certain of the details described herein may be varied considerably without departing from the basic principles of the invention. 36

Claims (33)

Claims

1. An isolated polynucleotide comprising the sequence of SEQ ID NO: 16.

2. An isolated polynucleotide comprising a sequence selected from the group consisting of: (a) the complement of SEQ ID NO: 16; (b) the reverse complement of SEQ ID NO: 16; (c) the reverse sequence of SEQ ID NO: 16; and (d) an operable antisense fragment of SEQ ID NO: 16.

3. An isolated polynucleotide comprising a sequence selected from the group consisting of: (a) sequences having at least 75% identity to the sequence of SEQ ID NO: 16; (b) sequences having at least 90% identity to the sequence of SEQ ID NO: 16; and (c) sequences having at least 95% identity to the sequence of SEQ ID NO: 16, wherein the polynucleotide encodes a polypeptide with Terminal Flower 1 (TFLl) activity.

4. An isolated polypeptide comprising the amino acid sequence of SEQ ID NO: 36.

5. An isolated polypeptide comprising an amino acid sequence selected from the group consisting of: (a) sequences having at least 75% identity to a sequence of SEQ ID NO: 36; (b) sequences having at least 90% identity to a sequence of SEQ ID NO: 36; and (c) sequences having at least 95% identity to a sequence of SEQ ID NO: 36, wherein the polypeptide has Terminal Flower 1 (TFLl) activity.

6. An isolated polynucleotide that encodes a polypeptide of any one of claims 4 and 5.

7. An isolated polypeptide encoded by a polynucleotide of any one of claims 1-3.

8. A genetic construct comprising a polynucleotide of any one of claims 1-3 and 6.

9. A transgenic cell comprising a construct according to claim 8, with the proviso that the cell does not form part of a living human being. 37 -8 JUL 2008 I IVEDl ■1 "W J|

10. A genetic construct comprising, in the 5'-3' direction: (a) a gene promoter sequence; (b) a polynucleotide sequence comprising at least one of the following: (1) a polynucleotide coding for at least a functional portion of a polypeptide of any one of claims 4 and 5; and (2) a polynucleotide comprising a non-coding region of a polynucleotide of any one of claims 1-3; and (c) a gene termination sequence.

11. The genetic construct of claim 10, wherein the polynucleotide is in a sense orientation.

12. The genetic construct of claim 10, wherein the polynucleotide is in an anti-sense orientation.

13. A transgenic plant cell comprising a genetic construct of claim 10.

14. A plant comprising a transgenic plant cell according to claim 13, or fruit or seeds or progeny thereof.

15. A method for modulating flowering in a plant, comprising stably incorporating into the genome of the plant at least one polynucleotide of any one of claims 1-3 and 6.

16. The method of claim 15, wherein the plant is selected from the group consisting of: grasses.

17. The method of claim 15, comprising stably incorporating into the genome of the plant a genetic construct of any one of claims 8 and 10.

18. The method of claim 17, wherein the promoter is an inducible promoter.

19. A method for producing a plant having altered flowering, comprising: (a) transforming a plant cell with a genetic construct of any one of claims 8 and 10 to provide a transgenic cell; and (b) cultivating the transgenic cell under conditions conducive to regeneration and mature plant growth.

20. The method of claim 19, wherein the promoter is an inducible promoter and the plant cell is exposed to an inducing agent selected from the group consisting of: chemical and physical stimuli. 38

21. A method for modifying the activity of a polypeptide involved in a flowering pathway in a plant comprising stably incorporating into the genome of the plant a construct of any one of claims 8 and 10.

22. A method for producing a plant with altered flowering, the method comprising transformation of a plant with a polynucleotide capable of hybridising under stringent conditions to an endogenous gene encoding a polypeptide with at least 75% identity to the sequence of SEQ ID NO: 16 and with Terminal Flower 1 (TFLl) activity, such that expression of the polynucleotide results in reduced expression of the endogenous gene, reduced levels of TFLl and altered flowering.

23. The method of claim 22 wherein the polypeptide with TFLl activity has at least 75% identity to the sequence of SEQ ID NO: 36.

24. The method of claim 22 wherein the polypetide with TFLl activity has the sequence of SEQ ID NO: 36.

25. A method for producing a plant with altered flowering, the method comprising transformation of a plant with a polynucleotide capable of hybridising under stringent conditions to an endogenous nucleic acid comprising a sequence with at least 75% sequence identity to SEQ ID NO: 16 encoding a polypeptide with TFLl activity, such that expression of the polynucleotide results in reduced expression of the endogenous nucleic acid, reduced levels of TFLl and altered flowering.

26. The method of claim 25 wherein the endogenous nucleic acid has at least 75% identity to the sequence of SEQ ID NO: 16.

27. The method of claim 25 wherein the endogenous nucleic acid has the sequence of SEQ ID NO: 16.

28. The polynucleotide of any one of claims 1-3 and 6 substantially as herein described with reference to any example thereof.

29. The polypeptide of any one of claims 4, 5 and 7 substantially as herein described with reference to any example thereof.

30. The construct of any one of claims 8 and 10-12 substantially as herein described with reference to any example thereof.

31. The cell of any one of claims 9 and 13 substantially as herein described with reference to any example thereof. intellectual property office of n.z - 8 JUL 2008 RECEIVED

32. The plant of claim 14 substantially as herein described with reference to any example thereof.

33. The method of any one of claims 15-27 substantially as herein described with reference to any example thereof. 40 SEQUENCE LISTING <110> Demmer, Jeroen Hall, Claire c C t Norriss, Michael Geoffrey 5 3 4 2 5 0 Saulsbury, Keith Martin <120> Compositions Isolated from Forage Grasses and methods.for their use. <130> JC220037/142 <150> 60/408,782 <151> 2002-09-05 <160> 40 <170> FastSEQ for Windows Version 4.0 <210> 1 <211> 808 <212> DNA <213> Lolium perenne <400> 1 gcagaagtct ctgtcgtccg cagccgcctc gctaggattt cgtttgtccc caaatcgccc 60 ccaaatccgc cgccgatccc caacctcaac ccaccaccca tggcggccga ggacaagaag 120 atcacgctca agtcctcgga cggcgagcag ttcgaggtgg acgaggcggt ggcgatggag 180 tcgcagacga tccgccacat gatcgaggat gactgcgccg acaacgggat cccgctcccc 24 0 aacgtcaacg ccaagatcct ctccaaggtc gtcgagtact gcagcaagca cgtccaggcg 300 gccgacggcg ccgcggcggc ggacggagct cccgccccgc cccccgccga ggacctcaag 360 aactgggacg ccgagttcgt caaggtcgac caggccacgc tcttcgacct catcctcgcc 420 gccaactacc tcaacatcaa gggcctgctc gacctcacct gccagaccgt cgccgacatg 4 80 atcaagggca agacacccga ggagatccgc aagacgttca acatcaagaa cgacttcacc 540 gccgaggagg aggaggagat ccgcagggag aaccagtggg ccttcgagta aatccacatc 600 gccccggtga agctgtaaat ttacatatct aattcactag ttagtcggat cgaaagagtt 660 gtaaagtgac atcaattttg atcgttggtg gttgatagtg taatctttct gtcagcactt 720 cttcattcgt tttggtttgt tttaatttta tgttgcctat tcttgattct tttagaatgg 7 80 cacaccttaa tatcatttaa aaaaaaaa 808 <210> 2 <211> 831 <212> DNA <213> Lolium perenne <400> 2 gtcctcttcc gccgcccccc tcgtttagca gctagggttt cctcccatcc caaatccccc 60 gatttcccga tctccaaccc cacctc'gccc acccatggcg gccgccgacg actccaagaa 120 gatgatcacc ctcaagtcgt ccgacgggga ggtgttcgag gtggaggagg cggtggcgat 180 ggagtcgcag accatccgcc acatgatcga ggacgactgc gccgacaacg ggatcccgct 240 ccccaacgtc aactccaaga tcctctccaa ggtcatcgag t'actgcaaca agcacgtcca 300 ggccgccaag cccgccgccg acgccgccgc cgccgacagc tcctccgccg ccgccccgcc 360 cgaggacctc aagaactggg acgccgagtt cgtcaaggtc gaccaggcca ccctcttcga 420 cctcatcctc gccgccaact acctcaacat caagggcctg ctcgacctca cctgccagac 480 cgtcgccgac atgatcaagg gcaagacacc cgaggagatc cgcaagacct tcaacatcaa 540 1 gaacgacttc accgccgagg aggaggagga gatccgcagg gagaaccagt gggcgttcga 600 gtagagcctc acaaccctgc cgcgccgcgt tgatgatgcc tagctaaaac tcgcaattta 660 cgcatctcga cgctgctact accttttatg taataattat cttcttgagt cgaggtccgg 720 tttatgaaca tctatctatc tatcttcggt ggtctgaaca aaaactatat atccttgttc 780 agtgggtttt atctatgaac atctatcgtc agtggttgtt taaaaaaaaa a 831 <210> 3 <211> 773 <212> DNA <213> Lolium perenne <400> 3 ctccgacggc gaggagtttg aggtggagga ggtgctggtg ctggagtcgc agaccatcaa 60 gcacatgatc gaggacgagt gcgacggcgt catcccgctc cccaacgtca gcgccaagat 120 cctctccaag gtcatcgagt actgcaggaa gcacgtccag acgcgcgccg ccctcgcccc 18 0 cgacggcgac atgagcacca acgccgccgg caccgagctc aagaccttcg acgaggactt 24 0 cgtcaaggtc gaccaggcca ccctcttcga cctcatcctg gctgcaaact acctggacat 300 caaggggctg ctggacctga cctgccagac ggtggctgac atgatcaagg gtaagacccc 360 agaggagatc cgcgcgacct tcaacatcaa gaacgacttc accccagagg aagaggagga 420 agtgcgcaag gagaacgcgt gggccttcga gtgaaggtcg ccgccctgac aagtaacgcg 480 aataaccagc aagaagaggt aacgatggcg ctggtagtgc ctgggagcag ctgttaaccg 540 tctgtggttc gaaaaactat gctagggtga agttgtaagt aggttctggt tccggtggat 60,0 cgggaggcct taccatttgc tgagctgact ccgttctttt ttggtggtga tatttggtgc 660 actcttgaac ctggttatga ggtgatgctg gttgctggtt attctgcact aatgctagtt 720 ggatcttatg catgactctc ttgtgctgag cttcatttgt tttaaaaaaa aaa 773 <210> 4 <211> 2404 <212> DNA <213> Festuca arundinacea <400> 4 gcggaggccg tggtgactac tcagatcatg acaacaaaag tggccatgtt aaactttttg 60 ttggatcagt tccgagaaca gcaagtgaag acgatgttcg acctttattt gagaatcatg 120 gagatgttct tgaagttgct atgatcaggg acaggaaaac tggtgaacaa caaggctgtt 180 gctttgttaa atatgcgact tccgaagagg ctgagagagc cataagagct cttcataacc 240 agtggactat acctggggcg atgggccctg ttcaggttag atacgccgac ggtgaaaagg 300 agcgtcatgg gtccattgag cacaaattat ttgtcgcatc actgaataag caggcaactg 360 caaaggagat tgaagagatt tttgctcctt ttggtcacgt ggaagatgtt tacattatga 420 aagatggcat gaagcaga'gc cgaggttgtg gctttgtcaa attctcatca aaagaacctg 480 cacttgcggc catgaattct cttagtggga cttacataat gagggggtgt gaacaaccat 540 taatagttcg atttgctgat cctaagcggc ctagacctgg agaatcaagg tggttaagaa 600 tgcatatttg ttttgcttat attccaactc tgcactattt cccgttgctg ctgtctgaat 660 tatcttgttt ggttaggggt ggccctgcat ttggaggtcc tggtgtcagt cctcgatctg 720 atgcagcact tgttatcagg ccgactgcca atcttgatga gcctagaggt cgacatatgc 780 ctcgtgacgc ttggcgccct tcaagcccaa gttcagtggc acctcatcag tttaataact 84 0 atgggtcgga caatcctatg ggcctaatgg gtggcactgg tacatcagca acagataatg 900 gtgcttttcg gcctcagatg tttcctggga atggtcagac agctgtgccg acgtcatctc 960 atatgggcat aaacacttct tcggtacaag gccatcatct aggggggcag cagatcccgc 1020 ccttgcaaaa gccacctgga ccaccacata atttctcttt acaattgcag aatcagcagg 1080 ggcagcattc cttggggcct ggtttgtttg gccagaatgt accatctatg caattacctg 114 0 gccagcttcc cacatcacag ccattgacgc agcagaatgc ttctgcaggc gctctacagg 1200 tgcctccagc catacagtcc aatcccatgc aatcggttcc cggacaacag caacttccgt 1260 ccaatgtggc agcacaaatg atgcaacaac caatccagca gataccatca caagcgccac 1320 agttgctact ccaacagcag gcagctatgc agtccagtta tcaatcttcg'cagcaggcaa 1380 tttttcagct tcagcaacag ctgcagctaa tgcaacagca gcagcaacag cagcagcaac 1440 ctaacctcaa tcagcagcca catacacaga tttctaagca acagggacag ccaaatc.aat 1500 ccagtacccc tggtgctcca gctgccatga tgccgtcaaa cattaatgca attccacagc 1560 aggtcaattc acctgtagtt tctttaactt gcaattggac ggaacatacc tcccccgaag 1620 gttttaaata ctactacaat agtattactc gagagagtaa gtgggagaag cctgaagagt 1680 2 atgtactgta cgagcaacag caacagcagc agcatcagaa acttatttta cttcaacagc 1740 accaacaaaa gcttgttgcg cagcaacttc agtcacctcc tcaggctcaa acaattcaat 1800 ctatgcaatc tatccaacaa catcctcagt cacatcaagg acataaccag atgcagatga 18 60 aacatcagga attaaactat aatcagttgc aggcaactgg caatattgat cccaatagga 1920 tccagcaggg aattcaagct gctcaagagc gttcatggaa aagttgagac tgctggtgaa 1980 tacatgttga ggtgtcagtc aaggctcaga aatgagctcc agccaagcct gccgattcca 2040 tgcgtgagag tgatggctct tgcggtcatt gtaactggat ttggcttaga tcgcagccta 2100 gatcgtagat cccatctgtg taaaatattt gcagtctagg ccttgtatca ctgtaacatt 2160 gttgattaga atatcgctct ttgtatctgt ttcctcgctt ttctttatgg caggatgtgc 2220 tgtctcattt acatcaattt ttcctccacc tgttatgttg gagctgcgct cctgaattgc 2280 tggctcgttc tttttttctt cggaacactt gagttctttg aacagccaaa tagtgcttgg 2340 agaagggaac cttttgagct ccaacggctg gttaatctca gaatcagttt catgaaaaaa 2400 aaaa 2404 <210> 5 <211> 2135 <212> DNA <213> Lolium perenne <400> 5 cggaggccgt ggtgactact cagatcatga caacaaaagt ggccatgtta aactttttgt 60 tggatcagtt ccgagaacag caagtgaaga cgatgttcga cctttatttg agaatcatgg 120 agatgttctt gaagttgcta tgatcaggga caggaaaact ggtgaacaac aaggctgttg 180 ctttgttaaa tatgcgactt ccgaagaggc tgagagagcc ataagagctc ttcataacca 240 gtggactata cctggggcga tgggccctgt tcaggttaga tacgccgacg gtgaaaagga 300 gcgtcatggg tccattgagc acaaattatt tgtcgcatca ctgaataagc aggcaactgc 360 aaaggagatt gaagagattt ttgctccttt tggtcacgtg gaagatgttt acattatgaa 420 agatggcatg aagcagagcc gaggttgtgg ctttgtcaaa ttctcatcaa aagaacctgc 480 acttgcggcc atgaattctc ttagtgggac ttacataatg aggcggccta gacctggaga 54 0 atcaaggggt ggccctgcat ttggaggtcc cggtgtcagt cctcgatctg atgcagcact 600 tgttatcagg ccgactgcca atcttgatga gcctagaggt cgacatatgc ctcgtgacgc 660 ttggcgccct tcaagcccaa gctcagtggc atctcatcag tttaataact atgggtcgga 720 caatcctatg ggcataatgg gtggcactgg tacatcagca gcagataatg gtgcttttcg 7 80 gcctcagatg tttcctggga atggtcagac agctgtgccg acgtcatctc atatgggcat 840 aaacacttca ttacaagggc atcatctagg ggggcagcag atcccgccct tgcaaaagcc 900 acctggacca ccacacaatt tctctttaca attgcagaat cagcaggggc agcattcctt" 960 ggtgcctggt ttgtttggcc agaatgtacc atctatgcaa ttacctggcc agcttcccac 1020 atcacagcca ttgacgcagc agaatgcttc tgcaggcgct ctacaagcgc ctccagccat 1080 acagtccaat cccatgcaat cagttcctgg acaacagcaa cttccgtcca atgtggcacc 1140 acaaatgatg caacaaccaa tccagcagat accatcacaa gcaccacagt tgctactcca 1200 acagcaggca gctatgcagt ccagttatca atcttcgcag caggcgattt ttcagcttca 12 60 gcaacagctg cagctaatgc aacagcagca gcaacagcag cagcaaccta acctcaatca 1320 gcagcaacct aacctcaatc agcagcaaca tacacagatt tctaagcaac agggacagcc 1380 aaatcaatcc agtacacctg gtgctccagc tgccatgatg ccgtcaaaca ttaatgcaat 1440 tccacagcag gtcaattcac ctgcagtttc tttaacttgc aattggacgg aacatacctc 1500 ccccgaaggt tttaaatact actacaatag tattactcga gagagtaagt gggagaagcc 1560 tgaagagtat gtactgtacg agcaacagca acagcagcag cagcagcaga aacttatttt 1620 acttcaacag caccaacaaa agcttgttgc gcagcaactt cagtcacctc ctcaggctca 1680 aacaattcaa tctatgcaat ctatccaaca acatcctcag tcacatcaag gacataacca 1740 gatgcagatg aaacatcagg aattaaacta taatcagttg caggcaactg gcaatattga 1800 tcccaatagg atccagcagg gaattcaagc tgctcaagag cgttcatgga aaagttgaga 1860 ctgctggtga atacatgttg aggtgtcagt caaggctcag aaatgagctc cagccaagcc 1920 tgccgattcc atgggtgaga gtgatggctc ttgcggtcat tgtaactgga tttagcttag 1980 atcgcagcct agatcgtaga tcccatctgt gtaaaatatt tgcagtttag gccttgtatc 2040 actgtaacat tgctgattag aatatcattc cggtatctgt ttcctcgctt ttctttatgg 2100 caggatgtgc tgtttcattt cccttaaaaa aaaaa 2135 <210> 6 <211> 842 <212> DNA <213> Festuca arundinacea 3 <400> 6 cattcatcca ggtagctcct gctccagatc aatatactct agctaactag ctcaactgtg 60 cctggccatc gtcaacctct agcttcaaca tacgagatgg ctgggaggga tagggacccg 120 ttggtggttg gaagggttgt gggggacgtg ctggacccct tcgtccgcac cactaacctc 180 agggtgacat tcggaaaccg ggccgtgtcc aacggctgcg agctcaagcc ctacatggtc 240 acccaccagc ccagggtcga ggtcggcggc aatgagatga ggaccttcta cacactcgtg 300 atggtagacc ccgacgcgcc aagtccaagc gatcccaacc tcagagaata cctccattgg 360 ttggtgacag atattcctgg aactactggt gcttccttcg ggcaggaggt gatgtgctac 420 gagagccctc gccccaacat gggaatccac cgcttcgtgc tcgtactctt ccagcagctg 480 ggccggcaga cggtgtacgc gcccgggtgg cgccagaact tcaataccag ggacttcgcc 54 0 gagctctaca acctcggccc ggccgtcgcc gccgtctact tcaactgcca gcgcgaggcc 600 ggctctggcg gcaggaggat gtataattga caccgccacg ccaagactca gacctacaca 660 agatcgatga tccattcaca gcgtgcctag ctaagcttaa ctaataatta ctatatacta 720 catatggtgt gtcataagaa gctagctagc cacgcaattg atcaagcatt attatacgca 78 0 tatagatatt gtgtacaacc tatatcataa caattattag ctacatatac ataaaaaaaa 840 aa 842 <210> 7 <211> 825 <212> DNA <213> Lolium perenne <400> 7 gcaccaccag cacgcgcgcg cgcgcgagta gtagtagtag ccctccagag agtccaccag 60 acagagagta aaatggacgg cgtcttggcc ggccggccga cggata'gatt ccccccactc 120 ggagcagcca tcggatcaga ccggtcagga cagccaggct gacgcactca gtacacctcg 180 gcagccagag ctgctcgtga tccagcagct agctagctag ctagcttggt cgagactcga 24 0 tcgagagaga tctcctctcc tataagtacg ccggctcgtc gtggtgcaac agcgacggga 300 gacagaaaga gcttcagctt cagcttgcaa ctgcaaccac acgcgctcag ctaagctcac 360 acacatcgat ctagccggcc ggcgatcgga gacgatggtg ggcgtgcagc gcgccgaccc 420 gctggtggtg gggcgcgtga tcggcgacgt ggtggacccg ttcgtgcgcc gggtgccgct 480 gcgggtcggc tacgcgtcca gggacgtggc caacggctgc gagctccgcc cgtccgccat 540 cgccgaccag ccgcgcgtcg aggtcggcgg cccggacatg cgcaccttct acacgctggt 600 gatggtggat ccggatgctc ccagcccgag cgatcccagc ctcagggagt acttgcactg 660 gtgagagccg agcaccaaca ccaacatcga aagatcaatc tctctctcct acctggcctg 7 20 gaaatatccc cctcccatgc ccctaccaat ccaaattcag atatttgtgt acagttagct 7 80 ggggaacagg gccaaatagc atctttccgc aaagcaaaaa aaaaa 825 <210> 8 <211> 1473 <212> DNA <213> Lolium perenne <400> 8 cgaggccttc gccggctgcc gccgcgtcca cgtcgtcgac ttcgggatca agcagggcat - 60 gcagtggccc gccctcctcc aggccctggc cctccgcccc ggcggcccgc cctcgttccg 120 cctcacgggc gtcggccccc cgcagcccga cgagaccgac gcgctgcagc aggtcgggtg 180 gaagctggcc cagttcgcgc acaccatcgg cgtcgacttc cagtaccgcg gcctcgtcgc 24 0 cgccacgctc gccgacctcg agcccttcat gctgcagccg gaggccgacg acgggcccaa 300 cgaggagccc gaggtcatcg ccgtcaactc cgtcttcgag atgcaccgcc tcctcgcgca 360 gcccggcgcc ctggagaaag tcctgggcac cgtgcgggcc gtgcggccga ggatcgtcac 420 cgtcgtggag caggaggcca accacaacac cggctccttc ctggaccgct tcaccgagtc 480 cctgcactac tactccacca tgttcgactc cctggagggc gccggctccg ccccgtccga 54 0 aatctcatct gggccttccg .ccgccgccgc caacgccgcc gctcctggca cggaccaggt 600 catgtccgag gtgtacctcg gccggcagat ctgcaatgtc gtggcctgcg agggcgccga 660 gcgcacggag cgccacgaga cgctgggcca gtggcgcggc cgcctcggcc acgccggctt 720 cgagaccgtc cacctcggct ccaacgccta caagcaggcc agcacgctgc tcgcgctctt 780 cgccggcggc gacggctaca aggtggacga gaaggaaggc tgcctcacgc tcggctggca 840 cacgcgcccg ctcatcgcca cctccgcgtg gcgcatggcc gccgccgccg cgccatgatc 900 gcaagttttg aacgctgtaa gtacaccaca cccccgagca cggaggagca caaccccccg 960 4 ccccttggct caccggcgca cttgaatgaa agctaaaacg tcgacgaacg ctggattgca 1020 gcgaccaacg atcggagtta cggatctcgc tggcgtgaag agatggacac cggacggact 1080 cccggcgacc accaccacca ccatagcctg taattcgttc ttgttctcga ttccccactt 1140 gatccgtgaa ctcctagcaa gctctattat taagttttaa aatgtctatt attgttctgt 1200 gtaattcctc caatcgctca tatttaaata aggacgggac ggatttcggt actagctctg 1260 atgatgagaa ttttgtatgc aaagcaatct aaaactgagc tttgttctgg tctttgatca 1320 ccagttatga accttagagc aatgcgttct attctcactg ctcttagtat gaacatgagg 1380 ttcttctact cttgatcagt tgtaagcaat taagtgctga gctcttgact gttcttaatt 1440 atgaacatga tgttcttctc ctcaaaaaaa aaa 1473 <210> 9 <211> 3913 <212> DNA <213> Lolium perenne <400> 9 gctcgctcca agtttctctc tcctcgcctc cggctccgtc tacccgctcg ccgccgcgcg 60 aatcccgtcg ccgccgccgc tgattcgccg ccggagcccc ggagtagagc gcgccctgtc 120 tagtttcttg agcaggatct taaactacta agtatgtctg tctcaaatgg gaagtggatc 180 gacgggctcc agttctcttc actattctgg cccccgccac acgatgcaca gcagaaacag 24 0 gcacaaactt tggcctacgt tgagtacttt ggtcagttta catctgacag tgagcaattc 300 ccggaggatg ttgctcagct catccaaagt tactatccat cgaaagaaaa acgcttggta 360 gatgaagtat tagcaacctt tgttctccat caccccgagc atggtcatgc agttgtacat 420 ccaattctt't cacgcatcat agatgggtcc ctgagttatg atagacatgg ttccccattc 480 aattctttca tctctttatt tacccaaact gctgagaaag agtattcaga gcagtgggct 540 ttggcgtgtg gagaaattct tagagttctt actcactaca ataggccaat cttcaaagtt 600 gcagaatgta acgacacctc cgacc^ggcc acaacaagtt attccttaca tgacaaagct 660 aatagctctc cagaaaatga acctgaacgg aagccattga ggccattatc tccttggatc 720 acagacattt tgttaaatgc acctttgggc attagaagtg actattttag atggtgtggt 780 ggagtcatgg gaaaatatgc agctggtgga gaactgaagc ctccaacaac tgcttacagc 840 cggggagctg gtaagcatcc acaacttatg ccatccaccc ctagatgggc tgttgccaat 900 ggagctggag tcatcttaag tgtctgtgac gaggaagtag ctcgttacga gacagcaaac 960 ttaaccgcag cagctgttcc tgcgcttctg ctacctccac cgacaacgcc cttggatgag 1020 catttggtgg cagggttgcc ccctcttgaa ccatacgctc gcttgtttca cagatactac 1080 gcaattgcta ctccaagtgc tacacaaagg ttgctctttg gtcttcttga agcaccgcct 1140 tcatgggctc cagatgcact tgatgcagca gtacagcttg ttgaactcct tcgagcagcc 1200 gaagattatg ctactggcat gcggcttccg aaaaattggc tgcatcttca tttcttgcgt 12 60 gcaatcggaa ctgcaatgtc aatgagagct ggtatggctg ctgatacggc cgctgccttg 1320 ctatttcgta tactatccca accaacgttg ctttttcctc cactaagaea tgccgaagga 1380 gttgtgcagc atgaaccact aggtggctat gtatcatcat acaaaagaca gctggagatt 14 40 cctgcatctg aaaccactat tgatgctact gcacaaggca ttgcttcctt gctgtgcgct 1500 catggtcctg atgttgagtg gagaatatgt accatctggg aagctgccta tggtttgtta 1560 cctctgaatt catcagcagt cgatttgcct gaaattgttg tagctgctcc gcttcagcca 1620 cctactttat catggagcct atatttgcca ctgttgaaag tatttgagta tctacctcgt 1680 ggaagtccat ctgaagcatg ccttatgaga atatttgtgg caactgttga agctatactc 1740 aggagaactt tcccttcgga aaccgaacca tccaaaaaac caagaagtcc atctaagagc 1800 cttgctgttg ctgaactccg tacgatgata cattcactct ttgttgaatc atgtgcctca 18 60 atgaaccttg cttcgcggtt attgtttgta gtattgactg tctcagtcag tcatcaagct 1920 ctgccggggg gcagcaaaag acctacaggc agtgagaacc attcttctga ggagtccact 1980 gaggactcaa aattaaccaa tggaagaaac agatgcaaga agaaacaagg gcctgttggt 2040 acctttgact cgtatgtgct ggctgctgtt tgtgctttat cttgtgagct tcagctgttc 2100 cctatacttt gcaagaatgt tacgaagaca aacataaaag actctataaa gattaccatg 2160 cctggaaaaa ccaatgggat cagtaatgag ctacacaata gcgttaactc agcgattctc 2220 catactcgta gaattcttgg catcctggaa gctcttttct ccttgaagcc atcatcagtt 2280 ggtacctcct ggagctatag ttcaaatgag atagttgcag cagcaatggt tgctgctcat 2340 gtttctgagt tattccgtag gtcgaggcca tgcctaaatg cactatctgc actgaagcga 2400 tgtaagtggg atgctgagat ttctaccagg gcatcatcgc tttaccatct gatcgacttg 24 60 catggtaaaa ctgtgtcatc catcgtgaac aaagctgagc ctttggaagc tcacctgaac 2520 cttacagcag taaagaaaga tgatcaacac cacattgagg aaagcaatac cagctcatcg 258 0 gattatggga acttggagaa gaagagtaag aaaaatggtt tttcaagacc actcatgaaa 264 0 5 tgtgcagaac aggctaggag aaatggtaac gttgcaagta catcggggaa agctactgca 2700 actttacagg cggaagcatc tgatttggca aacttcctta ccatggacag gaacgggggt 2760 tatggaggtt ctcaaactct cctaagaact gtaatgtcag aaaagcagga actatgcttt 2820 tctgttgtct cgttgctgtg gcataagctt attgcatctc ccgaaacaca gatgtctgca 2880 gagagtacat cagctcatca gggttggaga aaggttgcag atgcgctttg tgatgttgtt 2940 tcagcttcac cggccaaggc ttcaactgct attgtcctgc aggctgagaa ggacttgcag 3000 ccctggattg ctcgagatga tgagcaaggt cagaagatgt ggagagtcaa ccagcgaata 3060 gtgaaactga tagctgagct tatgaggaac catgatagcc cagaagcact gataattctt 3120 gcgagcgctt cagaccttct gctccgtgcc acggatggga tgcttgttga tggtgaagct 3180 ■tgtaccttgc ctcaattgga gcttctggaa gtaaccgcca gagccattca tctcatcgtt 3240 gaatggggag atccaggtgt agcagttgct gatggcctct cgaatctgct gaagtgccgg 3300 ctatcaccta ccatccgatg cctttcccac cctagtgcac atgtacgggc gctcagcatg 3360 tccgtccttc gcgacatctt gaacagtgga ccaataagtt ccaccaagat aattcaagga 3420 gagcagcgga acggcatcca aagcccaagt taccggtgcg cggcagcaag tatgaccaac 34 80 tggcaagcgg acgtcgagag atgcatagag tgggaagccc acaaccgtca ggccaccggg 354 0 atgacgcttg cctttctcac tgcagcggct aacgaactcg gatgccccct tccttgctga 3600 cacagccata tttgaagctg acatcggcga cacttgacag ttagcgcgag cagttgctgc 3660 atggtcagcg agcaggatgg ctaatccctt gctcaaggat gacttcccag tctgccccca 3720 ttatgtgatt taaaactgat gtatattagt tgacccagtc atacggagct tgctcccact 3780 gtgtgattta acttttaatc tgacattaga tgttcaagca tattgaactg cttgtgctgt 384 0 aacttgtatt tctgtagccg aaagatgtac actatggtaa atgaagacat atcatttttc 3900 gtcaaaaaaa aaa 3913 <210> 10 <211> 3980 <212> DNA <213> Festuca arundinacea <400> 10 ggaaatcttt ttctcgcctc tcctcgcccc tcgcagtttc tctctcctca ccttcgcctc 60 cgcctccgcc tccgtctacc cctcgccgcc gcgcaattcc catcaccgcc gccgctgatt 120 cgccgccgga gctccggatt agagcgcgcc ccgtctagtt tcttgagcag gatcctaaac 18 0 tactaagtat gtctgcgtca aatgggaagt ggattgatgg gctccagttc tcttcactat 240 tctggccccc gccacacgat gcacagcaga aacaggcaca aactttggcc tacgttgagt 300 actttggtca gttcacatct gacagtgagc aattcccgga ggatgtagct cagctcatcc 360 aaagttgcta tccatcgaaa gaaaaacgct tggtagatga agtattagca acctttgttc 420 tccatcaccc cgagcatggt catgcagttg tacatccaat tctttcacgc atcatagatg 4 80 ggtcactgag ttatgataga catggttccc cattcaattc tttcatctct ttatttaccc 540 aaactgctga gaaagagtat tcagagcagt gggctttggc ctgtggagaa attcttagag 600 ttcttactca ctacaatagg ccaatcttca aagttgcaga atgtaacgac acctctgacc 660 aggccacaac aagttattcc ttacaggaga aagctaatag ctctccagaa aatgaacctg 720 aacggaagcc attgaggcca ttatctcctt ggatcacaga cattttgtta aatgcacctt 780 tgggcattag aagtgactat tttagatggt gtggtggagt catgggaaaa tacgcagctg 840 gtggagaact gaagcctcca acaactgctt acagccgggg agctggtaag catccacaac 900 ttatgccatc cacccctaga tgggctgttg ccaatggagc tggagtcatc ttaagtgtgt 960 gtgacgagga agtcgctcgt tacgagacag caaacttaac cgcagcagct gttcctgcgc 1020 ttctgctacc tccaccgaca acgcccttgg atgagcattt ggtggcaggg ctgccccctc 1080 ttgaaccata cgctcgcttg tttca-cagat actacgcaat tgctactcca agtgctacac 1140 aaaggttgct ttttggtctt cttgaagcac caccttcatg ggctccagat gcacttgatg 1200 cagcagtaca gcttgttgaa ctccttcgag cagccgaaga ttatgctact ggcatgcggc 12 60 ttccgaaaaa ttggctgcat cttcatttct tgcgtgcaat tggaactgca atgtctatga 1320 gagctggtat ggctgctgat acggccgctg ccttgctatt tcgtatacta tcccaaccaa 1380 cgttgctttt tcctccacta agacacgccg aaggagttgt gcagcatgaa ccactgggtg 1440 gctatgtatc atcatacaaa agacagctgg agattcctgc atctgaaacc actattgacg 1500 ctactgcaca aggcattgct tccttgctgt gcgctcatgg tcctgatgtt gagtggagaa 1560 tatgtaccat ctgggaagct gcctatggtt tgttacctct gaattcatca gcagtcgatt 1620 tgcctgaaat tgttgtagct gctccgcttc agccacctac tttatcatgg agcctatact 1680 tgccactgtt gaaagtattt gagtatctac ctcgtggaag tccatctgaa gcatgcctta 1740 tgagaatatt tgtggcaacg gttgaagcta tactcagaag aactttccct tcggaaacct 1800 ctgaaccatc caaaaaacca agaagtccat ctaagagcct tgctgttgct gaactccgta 18 60 cgatgataca ttcactcttt gttgaatcat gtgcgtcaat gaaccttgct tcccggttgt 1920 6 tgtttgtagt attaactgtc tcagtcagtc atcaagctct gccgggaggc agcaaaagac 1980 ctacaggcag tgataaccat tcttctgagg agtccactga ggactcaaaa ttaaccaatg 2040 gaagaaacag atgcaagaag aaacaagggc ctgtcggtac ctttgactcg tatgtgctgg 2100 ctgctgtttg tgctttatct tgtgagcttc agctgttccc tatactttgc aagaatgtta 2160 caaagtcaaa cataaaagac tctataaaga ttaccatgcc tggaaaaacc aatgggatca 2220 gtaatgagct acacaatagt gttaactcag cggttctcca tacccgtaga attcttggca 2280 tcctggaagc tcttttctcc ttgaagccat catcagttgg tacctcctgg agctatagtt 2340 caaatgagat agttgcagca gcaatggttg ctgctcatgt ttctgagtta tttcgtcggt 2400 cgaggccatg cctaaatgca ctatctgcac tgaagcgatg taagtgggat gctgagattt 24 60 ccaccagggc atcatcgctt taccatctga tcgacttgca tggtaaaact gtgtcatcca 2520 tcgtgaacaa agctgagcct ttggaagctc acctgaacct tacagcagta aagaaagatg 2580 atcaacacca cattgaggaa agcaatacca gctcatcgga ttatggcaac ttggaaaaga 2 640 agagtaagaa aaatggtttt tcaagaccac tcatgaaatg tgcagaacag gctaggcgaa 2700 atggtaacgt tgcaagtaca tcggggaaag ctactgcaac tttacaggcg gaagcatctg 2760 atttggcaaa cttccttacc atggacagga atgggggtta tggaggttct caaactcttc 2820 taagaactgt aatgtcagaa aagcaggaac tatgcttctc tgttgtctcg ttgctgtggc 2880 ataagcttat tgcatctccc gaaacacaga tgtctgcaga gagtacatca gctcatcagg 2940 gttggagaaa ggttgcagat gcgctttgtg atgttgtttc agcttcaccg gccaaggctt 3000 caactgctat tgtcctgcag gctgagaagg acttgcagcc ctggattgct cgagatgacg 3060 agcaaggtca gaagatgtgg agagtcaacc agcgaatagt gaaactgata gctgagctta 3120 tgaggaacca tgatagccca gaagcactga taattcttgc gagcgcttca gatcttctgc 318 0 tccgtgccac ggatgggatg cttgttgatg gtga'agcttg taccttgcCt caattggagc 324 0 ttctggaagt aaccgccaga gccattcatc tcatcgttga atggggagat ccaggtgtag 3300 cagttgccga tggcctctcg aatctgctga agtgccgtct atcacctacc atccgatgcc 3360 tttcccaccc tagcgcacat gtacgggcgc tcagcatgtc cgtccttcgc gacatcttga 3420 acagtggacc aataagttcc accaagataa atcaaggaga gcagcggaac ggcatccaaa 348 0 gcccaagtta ccggtgcatg gcagcaagca tgaccaactg gcaggcggac gttgagagat 354 0 gcatagagtg ggaagcgcac aaccgtcagg ccaccggcat gacgcttgcc tttctcactg 3600 cagcggctaa tgaactcgga tgcccccttc cttgctgaca tggccatatt taagctgaca 3660 tcggcgacac ttgacagttg gcgcatgcag ttggtgcatg gtcagcgagc aggatggcta 3720 atcccttgct caaggatgac ttcccagtct gcccccatta ttatgtcatt taaaactgat 3780 gtatattagt tgtcccagtc atacggagct ttaatctgtg acgttagatg ttcaagcata 3840 tt'gaactact tgtgctgtaa cttgtcttcc tgtagccgaa cgatgtacac tatggtaaat 3900 gaagacatgt catttttcgt catgtaagat acatgcttat ctgcagagct tcaacctgaa 3960 cctgcctgtt aaaaaaaaaa 3980 <210> 11 <211> 1852 <212> DNA <213> Festuca arundinacea <400> 11 atagaaacct ccttcccgct agcttatata gagaccagtc gattcccgtg atccattccc 60 atggcttaga gtggtgatcg agcacgaaca agaacgtaga caagcaaact caccagagac 120 cgaggcttaa tttcctgcct tctgttcgat taggttgcca ccatgttgag tacgtcttac 180 gcgctgacgg ccgcgccgat tccggagggg gccgctgggc cacctgatcc ttttcggccg 240 atgcagatcg ccaacgacaa cgcctccgcg aagaggaagc ggcggccagc cggcactcct 300 gacccggatg cggaggtggt gtcgctgtcg ccgcggacgc tgctggagtc tgaccggtac 360 gtgtgcgaga tctgcaacca ggggttccag cgggaccaga acctgcagat gcaccggcgg 420 cggcacaagg tgccgtggaa gctgctgaag cgggaggccg gcgaggcagc gcggaagcgg 480 gtgttcgtgt gccccgagcc gacgtgcctc caccacgacc ctgcgcacgc cctcggcgac 540 ctcgtcggca tcaagaagca cttccgacgg aagcacagcg gccaccgcca gtgggcctgc 600 tcccgctgct ccaaggcgta cgccgtccac tccgactaca aggcgcacct caagacctgc 660 ggcacccgcg gccacacctg cgactgcggc cgcgtcttct cccgggtgga gagctttatc 720 gagcaccagg acatgtgcga cgccagccgg ccccggggcg gcacgacgtc gtcgtcgcca 780 ggccatggag gcggcagggt ggtaggcgct tccaacccgc agcacctgct acatgcggcg 840 tctctgtcac ggacggcgtc aagtgcaagc ccctccagcg ggggcgaact cgtggggagc 900 ccggtggcct ggccttgcgg cccggcgaca gcaagcccca cggctgccaa cgtagcagca 960 ttccaacggc tgctcgatcc cactcagtca tcgtcacctc caacgccgtc cgaccgccgc 1020 ggcgccggca cccaaaacct ggagctgcag ctcatgccgc cgcgcggggg cggagcggct 1080 cctcctggta cggctcttac gtatcgtgcg tcgccgtgtt caccttccgt tcttcacgct 1140 7 ccccgacagc tgggcgcgga cgcggtgcgg ctacagctct ccatcggctg cggcggcgcg 1200 cctgacgaca gcagcgtgga gtcggcgccg gcgccggctg caacgctgaa ggaggaggcc 12 60 cgggagcagc taaggctggc gacggccgag atggcctcgg cggaggagac gcgggcgcag 1320 gcgaggcgtc aggtggagct ggccgagcag gagctggcgg gcgcaagacg cgtgcggcag 1380 caggcgcagc tggagctcgg ccgcgcccac gcgctccgcg accacgctgt gcgccagatc 1440 gacgcaacgc tgatggagat cacctgctac ggctgccgcc acaacttccg ggcgagggcg 1500 gccgccatga actgcgaggt agccagctac gtgtcgtccg tgctgaccga gggcggcgac 1560 gccgaggtcg acaacgacgg ccaccaccag ctcctccatg ccggggacct gccaagaagc 1620 caccgtgcca tgatgaagat ggacctcaac taggtccatc tagctgccta gctgactcgt 1680 ctcacggatg tttattaacc ttcagcgttt tttaggtttc ctttaacatt cagcttgctc 174 0 tcctgtcttt tgtttcacca acgagatagg agatcgatgt gctgcgtgat ggtgtaattt 18 00 gacgagatga ttgccataat atgccctcta ggtacagact ctaaaaaaaa aa 1852 <210> 12 <211> 2219 <212> DNA <213> Lolium perenne <400> 12 gtttggattt tgtcctgtac atggttgcta cctcaatacc acagctagca ggcttctagc 60 tagatcctgg tcatatttat gtctttcctt ctcacgtaca tacgcgcgea gctgttctca 1.20 tcgatcctct cctgcttgtc tttgtcttgt agatccacaa gacgccgccg gaagcaagca 180 gtagctgcaa ttaatcgaat cccatgtcgt cgccttgtgt tcttctctag actcactgac 24 0 agactaggac tggacgactg ctcggtggcg gcgctcacct gaagccaaca acaagcaatt 300 ggaaggagta gctagctgat tgttctattc gaccgatggc cgccgcctcg tccgctccct 3 60 tcttcggcct ctccgacgcg cagatgcagc cgatggtgcc cgcgcagcct cccgctcccg 420 ttgccgccgc gccggcgccc aagaagaagc gcaaccagcc aggcaaccca aatccggacg 480 cggaggtgat cgcgctgtcg ccgcgctccc tgatggcgac gaaccggttc gtgtgcgagg 54 0 tgtgcggcaa ggggttccag cgggagcaga acctgcagct gcaccgccgc ggccacaacc 600 tgccctggaa gctgaagcag aagaacccca aggacgccct gcggcggcgc gtgtacctgt 660 gcccggagcc gacctgcgtg caccacgacc cggccagggc cctcggcgac ctcaccggga 720 tcaagaagca ctactgccgc aagcacggcg agaagaagtg gaagtgcgac aagtgcgcca 780 agcgctacgc cgtgcagtcc gactggaagg cgcactccaa gacctgcggc acaagggagt 8 40 accgctgcga ctgcggcacc ctcttctcca ggagggacag cttcatcacc caccgcgcct 900 tctgcgacgc gctggcccag gagagcgcgc gcttgcccgc gatcggcgcc agcctatacg 960 gtggcgtcgg aaacatgggc gccctcaaca ctctctccgg catgccccaa caactgccgg 1020 gcggcagctt tcctgaccag tccggccacc actcctcggc gtcggctatg gacatccaca 1080 accttggcgg tggcagcaat gccggccagt tcgaccagca cctcatgcca cagtccgcgg 1140 gatcctccat gttccgctcc caggccgcct cgtcttcccc gtactacctc ggcgccgccg 1200 ccgcccagga cttcgccgag gatgacgtcc accgctccca tggcaaccag agctctcttc 1260 tccagggcaa gtcgacggcg gccttccacg gcctgatgca acttccagac cagcaccagg 1320 gaagcgcaag caacggtaac aacaacctcc tgaaccttgg cttctattcg ggcaacggcg 1380 gcggccagga cgggcgtgtc atgttccaga accagttcaa cagcagcgcc ggaaacggca 1440 acgtcaatgc tgagaacaat ggaagcctcc tcggcggcgg tggtgggggt ttcccttcgc 1500 tgttcggttc gtctgagtca ggcggcggac tcccgcagat gtcggcgacg gcgctgctgc 15 60 agaaagcggc gcagatgggc gcgacgacga gcagccacaa cgcgagcgcc gggctgatgc 1620 gtggccctgg gatgaggggt ggcgccggag aaggcgggtc ttcgtcgtct gcgagcgaga 168 0 ggcagtcgtt ccatgacctc attatgaact ccctggcgaa cgggagcggc gctcctgcta 1740 ctacgggtgg tggcacagtg gcgttcggcg gcggcggctt ccccatcgac gacggcaagc 18 00 tgagcacgag ggacttcctg ggtgtcggtc ccggtggcgt ggtgcacgct ggcatgggcc 18 60 cgccccggcg gcacggtggc gctgccgggc tccacatcgg ctcgctggac ccggccgagc 1920 tgaagtagtc cgcaagaatc gacaaaaaac aaaacaagaa aacatgcatg catgcaaaaa 1980 aaaaatcttg aagattttca tggaacatca catcaggacg tcaaggacta gtcaggagtg 2040 aggacaaggt taatttct'tg gataatctat cagcatgtat tagttgatgc atgtgttcat 2100 gttggcatag ctagctgcgt taggtagccg gttcaataac cctgtgaggc cagaacttca 2160 gtttaatttt. gctgttcgta caaactgtca attagctgtt ttttctgtca aaaaaaaaa 2219 <210> 13 <211> 2257 <212> DNA 8 <213> Festuca arundinacea <400> 13 gcaccccaca ccgcagcagc gagcgcctca acccgatgcc gctgccgctc tgactcccat 60 ccatctcccc cagcccagcc cccagtcgaa agcaacccag ccagccagca gcgagcgaga 120 gaacaagcac ggaaaggagg ggaaaattct tccgtccgcc accgccgact cgcgccccgt 180 tcgccgacgc ggattgggag ggtggatacg gggcggctgg agggcggcgg gctgggtcga 240 gcggcggccg tggcgccaga tcgagcgggg atgccgccca atccgacgga cccggagcag 300 ccggaggcgg ccgcggcgcc ggccccgccg cccaagaaga agaggaacct gcccgggacc 360 ccagatccgg acgcggaggt gatcgcgctg tcgccgggga cgctcatggc caccaaccgc 420 ttcgtgtgcg aggtctgcgg caagggcttc cagcgggacc agaacctgca gctgcaccgc 4 80 cggggccaca acctgccctg gcgcctccgc cagcgcgggc ccggcgccgc cccgccgcgc 54 0 cgg.agggtct acgtctgccc ggagccaggg tgcgtgcacc acgccccggc ccgcgcgctc 600 ggggacctca cgggcatcaa gaagcacttc tgccgcaagc acggcgagaa gcgatgggcg 660 tgcccgcgct gcggcaagcg ctacgccgtc caggccgacc tcaaggcgca cgccaagacc 720 tgcggcaccc gcgagtaccg atgcgactgc ggcacgctct tcaccaggcg agacagcttc 78 0 gtgacacatc gagccttctg tggtgccctc gtcgaggaga ctggcagagt gctcgccgtt 840 ccggccccgc ctgctcccgg gccgcctgat ttggacgatg ttgacgagaa ttttgacaag 900 gacagtgaga agggagagga gaatgtggaa gatgaggagg agaaaggtga agtaaatgag 960 aattctgctg tggctgacgt gaatgagcct cagcgcgtcg aggcagcgtc tgaggcgccg 1020 cagcgcattc cttcgccgca gcagcagcgc attccgtcgc cgcggcgcat tccctcacca 1080 cagcgcattc ggtcgccacc atctccagta ccacaggagc agcagcagca gccgatggtg 1140 gcagtggtgc caaatttgga ggggccaaag gtggctgcgg agccaattgt ggttgtcaag 1200 caggaggagg atgacaagcg agatgaagat gtttgcttcc aggaagccga taaatacgac 1260 gacgctgaat tggaaggctc cagcctgcca gatactgata ccccgatgct tccttgtttc 1320 ctcccgtcgc cctcggatgc cattggtaca gatggcagca gcaccagctg tggcacggtc 1380 agcagtgctt ccattccatt gcgccagcaa cgacgactag cacatttgct gggctgtttg 1440 catcggccac gacaagcacc actccccaga gtagatcgct gcgtgatctt atcggtgttg 1500 atcccacctt cctttgcctt gcgattggta cgccctcctc tctgttcccg cagacaaacg 1560 cgagcaaccc tggcagcttt gctccacctc cagcaccaca catgtccgcg actgcactcc 1620 tgcagaaggc tgctgaggct ggagcttcgc aagcaggcac gtctttcttg aaggagtttg 1680 gtctggcaag ttcctcatca tcaaccccat ccaggccacc tcaagggagg tctatggata 1740 gctcaacaca atctcagcag cctcaaggaa ggtttatcga cagctcaaga cagtcgcagc 1800 tacctcaaga gaggttcatc aataactcga tgccatccag gctgtctcaa gggagattca 18 60 tggatacctc actaccatct cagcagctac ctcaaaggag attcatggat accgcactac 1920 catcccagca gctacctcaa gggagattca tggataacgc actaccgtcc cagcagcaac 1980 aggggtaagt agttgccttt attactggtt gcttgtggtg ccaaattgcc agcgcaggat 204 0 ttgcttcgta aaaggaaagg atgagactgg gacagccgca tgtgaaaggt gttttttcag 2100 ttttcgcctg ttgatgtcgg tcactatatc tgcccaactc tctccccttt gcgagtctcg 2160 cctctgcact tttgagagta ttcattgtta cattgttttg tccctgttga ccatacgaga 2220 agatattagc aagtcatttg ctttgcaaaa aaaaaaa 2257 <210> 14 <211> 2581 <212> DNA <213> Festuca arundinacea <400> 14 ggtggagccg cgtcggaggg aactgggagg ctccacaact atcgatgcgc gccaggacca- tcaagggaac agagatccaa ctggtttcaa tcacagcaac actgggccgc aatcaacaag gccggaccgg cttctcctgc ccgacaagat ccgccaaggc cggggggatt gggacaagcc aacagatgag atcatccggc aaatgatgga tcttagaggc ttcagggtag ttgacatcaa aagaggagga cggaatcccc gctggacgtg cttcatgaac cctctttgag gcaccaaggg aatccaacta cgttaacggt agaaagaaca gaataagatg ctcggtcaat aggtgatggt agacggccag ccctcccttg tacatctacg gagggcaagg tggtggtcca gcaaccgcgg ttacaacagc gctatgaaca aggaatcatg gctcttctca atgtcagctc gctatgcCac gccaggccaa ccattgccat actacctggt tcgccaccga tcttctggga cttctataga agaacgctca actctgatgt gtcccatgga agtcagcagc tgcagcagat aacgaacaat gtgaaggcgg ggcgcggagc caagcgcaac tcccgtcgcc catcttcgac tcttatgaag cctgcagaga atcggcattt ctcagaggcg agctaatcag gcaggcgaga gcctacagac 60 120 180 240 300 360 420 480 540 600 660 720 9 ccttctgcat tatacgcagc agggatgatg caaccaaaat ctggattagt tgcttctgga 780 ctaaatcaag gagttgggag tgtaccactg aaaggctggc cgctaacagt cccaggtatc 84 0 gatcaactgc ggtcaaattt aggcgcacag aagcagttga tgccatcccc aaaccaattt 900 caacttttat caccacaaca gcaattaatt gctcaagcac aaacacagaa tgaccttgct 960 agaatgggtt cgccagctcc atctggttcc ccaaagattc ggccaaatga acaggaatat 1020 ttgattaaga tgaaaatggc ccagatgcag cagtcaggtc aacggatgat ggaattgcaa 108 0 cagcagcagc atcatctgca acaacaacaa caacagcagc aacatcaaca gcagcagcag 114 0 cagcagcagc agcagcagca gatgcaacag aatactagaa aacggaagcc aacttcttct 1200 ggggctgcta atagtacagg cacaggaaat accgttggac cttctccgcc ctcaactcca 1260 tcaacacata ctcctggtgg tggaatacca gtagctagca acgcgaacat tgcgcaaaag 1320 aattcaatgg tttgcggcac ggatgggacc agtggatttg cttcatcctc aaatcagatg 138 0 gacaacttgg atagtttcgt tgattttgat gacaatgttg attcattttt gtcaaatgat 1440 gatggggatg ggcgagacat atttgctgca atgaagaaag gcccctcaga gcaggagtct 1500 ctaaagagtc tttctttgac tgaggttggt aataatcgca caagcaacaa caaggttgtt 1560 tgctgtcatt tctctacaga cgggaagtta cttgccagtg ctggtcatga aaaaaagctc 1620 ttcctctgga atatggataa ttttagcatg gacactaaag cagaagaaca tacaaatttt 1680 ataacggaca taagattcag gccaaattca actcagttgg ctacatcatc ttctgatgga 1740 actgttcgat tatggaacgc tgttgaacga accggcgctt tacagacttt ccacgggcac 1800 acctcccacg tgacttcggt agacttccac ccaaaactaa cggaggtcct ttgctcatgc 18 60 gatgacaaca gagagctccg gttctggacg gtcggtcaga acgcaccttc acgtgtcacc 1920 agggtcaaac agggcggtac tggtagggtg aggttccagc ctcggatggg gcagctcctt 1980 gcggtggctg ctgggaacac ggtgaacatc atcgatatcg agaaggacac gagtctgcat 2040 tcacagccaa aggtccactc gggcgaggtg aactgcatct gctgggatga gagcggcgag 2100 tacctggcgt cagcgagcca ggacagcgtg aaggtgtggt cagcggcgtc aggcgcgtgc 2160 gttcacgagc tgcggtccca tgggaaccag taccagtcgt gtatattcca ccctcgatac 2220 ccgaaggtct tgattgtggg cggttatcag acgatggagc tgtggagtct gtcggacaac 2280 cagaggaacg tggtggcagc gcacgagggg cttatcgcgg cgctggcgca ctccccgtcc 2340 acggggtcgg tggcctccgc cagccacgac aaatccgtga agctgtggaa gtagatggaa 2400 aggccgggaa cctgggcaaa atggtgccac gacgacgagc gtgtgtgttc tgggggtgat 24 60 gagaggttag acgcatgtac gtacgttacg ttacatagag gaggagttaa gaatgtgtaa 2520 ttaaactgag gcgactggat caatcaattt taatggaaga aactgtgcta taaaaaaaaa 2580 a 2581 <210> 15 <211> 2582 <212> DNA <213> Festuca arundinacea <400> 15 gagacagcga ggtggtgcgg gtggaggccg gaccggaagg aagaggagga agacggccag 60 gccaagtgaa ggcggcgtcg gagggcttct cctgccggaa tcccctcccc ctaccctccc 120 ctcctccctt gccattgcca tggcgcggag caactgggag gccgacaaga tgctggacgt 180 gtacatctac gactacctgg tcaagcgcaa cctccacaac tccgccaagg ccttcatgaa 24 0 cgagggcaag gtcgccaccg atcccgtcgc catcgatgcg ccggggggat tcctctttga 300 gtggtggtcc atcttctggg acatcttcga cgccaggacc agggacaagc cgCcccaagg 360 ggccaccgcg gcttctatag atcttatgaa gtcaagggaa caacagatga gaatccaact 420 gttacaacag cagaacgccc acctgcagag aagagatcca aatcatccgg ccgttaacgg 480 tgctatgaac aactctgatg tatcggcatt tctggtttca aaaatgatgg aagaaagaac 54 0 aaggaatcat ggtcccatgg actcagaggc gtcacagcaa ctcttagagg cgaataagat 600 ggctcttctc aagtcagcag cagctaatca gactgggccg cttcagggta gctcggtcaa 660 tatgtcagct ctgcagcaga tgcaggcgag aaatcagcaa gttgacatca aaggtgatgg 720 tgctatgcca caacgaacaa tgcctacaga cccttctgca ttatacgcag cagggatgat 780 gcaaccaaaa tctggattag ttgcttctgg actaaatcaa ggaattggga gtgtaccact 84 0 gaaaggctgg ccgctaacag tcccaggtat cgatcaactg cggtcaaatt taggcgcaca 900 gaagcagttg atgccatccc caaaccaatt tcaactttta tcaccacaac agcaattaat 960 tgctcaagca caaacacaga atgaccttgc t^gaatgggt tcgccagctc catctggttc 1020 cccaaagatt cggccaaatg aacaggaata tttgattaag atgaaaatgg cccagatgca 1080 gcagtcaggt caacggatga tggaattgca acagcagcag catcatctgc aacaacaaca 114 0 acaacagcag caacatcaac agcagcagca gcagcagcag atgcaacaga atactagaaa 1200 acggaagcca acttcttctg gggctgctaa tagtacaggc acaggaaata ccgttgggcc 12 60 ttctccgccc tcaactccat caacacatac tcctggtggt ggaataccag tagctagcaa' 1320 10 cgcgaacatt gcgcaaaaga attcaatggt ttgcggcacg gatgggacca gtggatttgc 1380 ttcatcctca aatcagatgg acaacttgga tagtttcgtt gattttgatg acaacgttga 14 40 ttcatttttg tcaaatgatg atggggatgg gcgagacata tttgctgcaa tgaagaaagg 1500 cccctcagag caggagtctc taaagagtct ttctttgact gaggttggta ataatcgcac 1560 aagcaacaac aaggttgttt gctgtcattt ctctacagac gggaagttac ttgccagtgc 1620 tggtcatgaa aaaaagctct tcctctggaa tatggataat tttagcatgg acactaaagc 1680 agaagaacat acaaacttta taacggacat aagattcagg ccaaattcaa ctcagttggc 1740 tacatcatct tctgatggaa ctgttcgatt atggaacgct gttgaacgaa ccggcgcttt 1800 acagactttc cacgggcaca cctcccacgt gacttcggta gacttccacc caaaactaac 18 60 ggaggtcctt tgctcatgcg atgacaacgg agagctccgg ttctggacgg tcggtcagaa 1920 cgcaccttca cgtgtcacca gggtcaaaca gggcggtact ggtagggtga ggttccagcc 1980 tcggatgggg cagctccttg cggtggctgc tgggaacacg gtgaacatca tcgatatcga 2040 gaaggacacg ggtctgcatt cacagccaaa ggtccacccg ggcgaggtga actgcatctg 2100 ctgggatgag agcggcgagt acctggcgtc agcgagccag gacagcgtga aggtgtggtc 2160 agcggcgtca ggcgcgtgcg ttcacgagct gcggtcccat gggaaccagt accagtcgtg 2220 tatattccac cctcgatacc ccaaggtctt gattgtgggc ggttatcaga cgatggagct 2280 gtggagtctg tcggacaacc agaggaacgt ggtggcagcg cacgaggggc ttatcgcggc 2340 gctggcgcac tccctgtcca cggggtcggt ggcctccgcc agccacgaca gttccgtgaa 2400 gctgtggaag tagatggaaa ggccgggaac ctgggctggt gccacgacga cgagcatgtg 24 60 tgttgtgggg gtacgtgatg agaggttaga cgcatgtacg tacgttacgt tacatagagg 2520 agttaagaat gtgtaattaa actgaggcga ctggatcaat caattttaat ggaaaaaaaa 258 0 aa 2582 <210> 16 <211> 1053 <212> DNA <213> Festuca arundinacea <400> 16 gaattacctg agcttccatt cagcaaagag gcacacacgc acactgatca tccctccggt 60 tccgatttca aggcatcaac atgtcaaggg cgttggagcc tctcgttgtg gggaaggtga 120 tcggtgaggt gctggacagc ttcaacccca ccgtgaagat ggcggcaacc tacaactcca 180 acaagcaggt gttcaacggc catgagttct tcccctcggc catcgccgcg aagccgcgtg 240 tcgaggttca ggggggcgac cttagatcct tcttcacatt ggtgatgact gaccctgatg 300 tgccaggacc cagtgatccg.tacctgaggg agcatcttca ctggattgtt actgatattc 360 ctgggactac tgatgcttct tttgggaagg aggtggtgaa ctacgagagc ccaaagccaa 420 acatcggcat ccacaggttc atcctcgtgc tgttccagca gacgcaccgg ggctcggtaa 480 agaacacacc gtcgtcgagg gaccgcttca ggacccgcga gttcgccaag gataacgagc 540 tcggcctccc tgtcgccgct gtctacttca acgcgcagcg ggagaccgcc gcccgccggc 600 gatagctcaa cggcaaccga accaaccaac aagcaacacc cccctactat gtacctgatc 660 tagctacatg ataaaacgaa ctgcgtacga tcacctatta gctagcttcg atggcctttc 720 ctgctacatc caagcatgca caatgtctga ataaaacaca ccggtaaatt agctgtttgc 780 acgagaaagc tgctccctac tagtacgtag ccgttgccca tttagttaat ttttgtgaag 840 gtgacaagat cgatgattgg gaagagattg cagtgttgac tgagaaaaaa gtgc.aagatt 900 tgaagcaata atagtcgtca gggagtataa gttacgtgtc gagtgcccaa gggaggggaa 960 gaagtggaca tggctctagt attcccctac ccactagtat tctgttatgt ggtttttctt 1020 cattggatcg aagtttgcag cgtaaaaaaa aaa 1053 <210> 17 <211> 2421 <212> DNA <213> Festuca arundinacea <400> 17 gcaacaccac catttgatgc agctcacaaa gaagaatcct caagctgctg cggctgccca 60 acttaacctc ttgcaacagc agcggatcat gcatatgcag cagcagcaac aacaacagat 120 tctgaaaaac ctgcctttac agagaaacca attacagcag cagcagcagg tgcagcagca 180 gcagcagcaa caactacaac agcagcagca gctacttcgt c'aacagagtc taaacatgag 240 aactccagga aagtcgcctc cctatgagcc aggtacctgt gcaaagagat tgacccatta 300 catgtatcac caacaaaaca ggccgcagga taacaatatc gagtactgga gaaactttgt 360 caatgagtat tttgctccaa ctgctaaaaa gaggtggtgt gtctctctct atggaagtgg 420 11 tcgtcaaact actggagttt tccctcagga tgtctggcac tgcgaaatat gcaatcggaa 4 80 gcctggccgg ggcttcgaga caacagttga ggtcttgccg cgattatgcc aaatcaaata 540 tgcgagtggt acattggaag aactactgta tatcgatatg ccacgtgagt ccaagaatgt 600 atctggtcag attgttctgg actatacaaa agcaattcaa gaaagtgtct ttgatcaatt 660 gcgtgtcgta cgtgaggggc atctgaggat aatttttaat ccagacctca agatcgcatc 720 ttgggagttc tgtgctaggc gtcatgagga acttattcca cggaggtcaa taataccgca 780 ggttagtcag cttggcgcag ttgtacagaa ataccaggct gctgctcaaa acccaaccag 840 tttatcaact caggacatgc agaataattg caactcgttt gtggcatgtg cccgtcaatt 900 ggctaaagct ctggaggtgc ctctggtaaa tgatttagga tatacaaaac gatatgtccg 960 ctgtcttcag attgcggagg tggtgaactg tatgaaagat ttgattgacc acagcaggca 1020 gactggatct ggaccaatcg atagcctgca caagtttcct cgcaggactc catcagggat 1080 caaccctctt caatcacagc agcaacagcc tgaagagcac caatctgttc cccagagttc 1140 aaaccagagt ggtcaaaatt ctgctcctat ggctggtgtg caggtttctg cctctgctaa 1200 tgcggatgcc acatcaaata attcgatcaa ctgtgcaccc tctacatctg caccctcacc 1260 aactgttgtt gggctcctcc aaggttcaat ggattctaga cacaatcatc caatgtgcag 1320 cgcaaatggc cagtataaca gtgggaataa tggcgcaatt cccagggtga actccgcaag 1380 ctcattacag tcaaatccat ctagtccttt cccttcgcag gtgcctacat cacccaataa 1440 caacatgatg ccgacccttc agaacgcaaa ccaactcagt tctcccccag cagtatcatc 1500 aaacttacct ccaattcagc ctccttcaac tcggcctcag gagtctgagc caagtgatgc 1560 ccaaagctcg gttcagagaa tcttgcaaga gatgatgtca tcacaaatga atggtgttgg 1620 ccatggaggg aatgacatga agaggccaaa tgggcttacc cctggtatta atggggttaa 1680 ctgcttagtt ggtaacgccg tcacaaatca ctccggaatg ggaggaatgg gatttggggc 1740 catgggcggg tttggttcga ctcctgcagc aagtggactc agaatggcaa tgacgaataa 1800 tgcaatggca atgaatggta ggatgggaat gcatcacagt gcacaagacc tatcacagtt 1860 gggccagcag caccagcacc agcaccagca tgacatagga aatcagctgt tgggtggact 1920 tggagcagca aacagcttca ataatattca gtatgattgg aaaccctctc aatagagtgg 198 0 ccggaaacat tagaaagtat gatgacgatg atatgcagct gtcctggctg ggctaattga 204 0 ttatggagca tcaagggcag caccataaca acgccccttg ggtcaaagcg tttgggcttt 2100 tgctccaatg gtgccatggc aaggaatcat aagcgacggc aaacacctga gctggtcact 2160 gtatgtcgca acggttagtt tagctggttc gttgtgtatt atgcaactat ggcactgagc 2220 tacctgcctc agttatctta ccaaaagatg agttaaagga ttataacctg ccagcaccgg 228 0 gcaccgttgg tgtctgtgta tggccttatt tctcacccag aaaagaagtt ttccctctct 234 0 tttttcgttg acggatgaca tccaatctgt atttatcacc accccttgct gtagtaatca 2400 tatgtgctga taaaaaaaaa a 2421 <210> 18 <211> 2833 <212> DNA <213> Festuca arundinacea <4 00> 18 agcgattgag cccgccgaag ctcgccgccc gccagccaag ctaaaagata tgaagactct 60 tagccaataa gcaagattct gtaaggctgc aacattggta acctccatgt ctggggcccc 120 acgctccaat cttggatttg ttgccaggga catgaatggt agcattccag ttagttctgc 180 aaattcctct gggccaagta tcggtgttag ctctttggtg accgatggca attcatcact 240 ctccggaggt gcccagtttc agcatagtac gagcatgaat gctgattcat tcatgcgcct 300 tcctgcctcc ccgatgtcat tttcatccaa taacatatct ggctcatcag tca'tcgatgg 360 gtccatcatg cagcaaagtc caccccaaga tcagatgcag aagcgcagat catctactgc 420 aacgtcccaa cctgggattg aggctggcgc tgcattccat gctcagaaga agccaagggt 480 cgatattagg caagacgata tcctgcaaca acacttgatt cagcaggtgc tccaaggtca 540 aagttctctc catctcccgg gccaacataa cccacagctt caagctttga tccgtcagca 600 gaaactggca catattcagc atctacagca gcagcagttg tcacaacaat ttcctcaaat 660 ccagcaatca caagttggca tacctcgtca gccgcagttg aggctgccac tagcacagcc 720 tggcatgcag ctagctggac ctgttaggac tcctgtcgag agtgggcttt gttctcgaag 780 gttaatgcag tatttgtttc ataagcggca ccggccagag gataatccca taacttactg 840 gaggaagctt attgatgaat'attttgcacc acgagcaaga gaaagatggt gtgtgtcatc 900 atatgaaaaa agagggaatt ctccagttgc tattccacag acatctcagg atacatggcg 960 ttgtgatatt tgcaatacac atgcagggaa aggacatgag gctacctatg aaatacttcc 1020 tagactatgt cagattcgat ttgaccaagg tgttatagat gaatatctat tcctggacat 1080 gcccaatgaa ttccggttgc ccaatggatt acttctcctg gagcatacta aagttgttca 1140 gaagagcatc tatgatcatc tacatgttac acacgagggg caactgagaa taatattcac 1200 12 tccagaacta aagattatgt cttgggagtt ctgttcacga cgacatgacg agtatatcac 12 60 tcgcaggttt ctaacaccac aggttaatca tatgctgcaa gttgcccaga agtatcaagc 1320 tgctgccaat gaaagtgggc ctgctggggt atcgaacaat gatgcacaag ccatttgcag 1380 catgtttgtg tctgcatcac ggcaattagc gaaaaatcta gaccaccaca gcttaaatga 14 40 gcatggtctc tctaaaagat atgttcgctg cttgcagata tcagaggtgg tgaatcacat 1500 gaaggactta attgagttca gccacaagaa caagcttggt cctatagagg gtctgaagaa 15 60 ctatcccaga caaaccggac caaagcttac aacgcagaac atgcatgatg caaagggggt 1620 ggtcaaaacg gaagaaagta cacatgtgaa taacgagggt ccagatgctg gacccgctgg 1680 tagcagtcct cagaatgctg gagcacaaaa caactaccag aatatgctga gaagcccaag 17 40 cccaaatcag ggactgactc accaggaggc atcccagaat gccgcggcac tgaacaacta 18 00 ccagaatatg cttagaagct caagcgcaaa ccagggtttg cttcagcagg aggcttcaca 18 60 gaatgtgtcg gggttaaata attaccagaa tatgcttaga agctcgagtg cgaaccagag 1920 tatccttcag caggaggcat cgagcatctt taaaggccct acaggagtgc acagtagcat 1980 tcagctggaa gcggctagat ccttccgcgc ggctcagctt gggcccatgt cgtttcagca 204 0 agctgtgccc'ctgtatcagc agaacaggtt tggggctggt gtgagtccgc agtaccagca 2100 gcatgtcatg cagcagctgc tgcaagaagc caacaggagt accaacaacc gggttctggc 2160 gcagcagcag cctcttagca ctcccaatgc aaacggaggt ctcacgatca ccaacagcgg 2220 tgctagtgga gatcaggcac aacacatgaa taataacgga gccgcaaagg gcgtggcagc 2280 tccaatgggt atggcgggaa ccagcaatct gatcaacagc ggatcagctg gggtcgtcca 234 0 gcgatgcagc agcttcaagt cggtgactag caaccccgct gctgccgcgg ctggcaacct 2400 gctgaccccc aaggccgagt ccatgcacga gatggacgag cttgaccatc tcatcactag 24 60 cgagctcgcg gagagcgggc tgttcatggg ggagcagcag ggaggtggtg gcggctactc 2520 atggcacatg tgagagagac tgctaaatta acctatatag ttcatctgtt ctgcgagttg 258 0 tgtttgatgt gtaaccgccg tagattattc ggagtctttc ttcctttttt tcgagcttcc 2640 igtgtagctga ctggaacgga tggaaccttg agttatgtga gtgtgagctg gcttgggaat 2700 ttgtgagcag tgcagcccag tgttattatc tatggaatga catggtgtgg ttgtcgtttg 2760 tgctgcaaca ttgctgattt cccgtgtccc tagaaaattg ctgatttttt cctgtgggct 2820 tttaaaaaaa aaa 2833 <210> 19 <211> 2780 <212> DNA <213> Festuca arundinacea <400> 19 gtctgatcct ctgtcattcc catcatcctc ccatgttagt ttgggcaatc acataagttc 60 agataatttg cagcagcagc agcagatgga tatgccggat ttgcagcagc agcagcaaca 12 0 acaacaacgt caactaccaa tgtcttacaa ccaacagcac ttgccaatgc aacggccgca 18 0 gccacaggct acagtgaagt tggagaatgg tggcagtatg ggtggagtta aaatggagca 24 0 gcagacaggg catcctgatc agaatggccc agcccagatg atgcacaatt ctggcaatgt 300 aaaatttgag ccacagcagt tgcaggcgtt gaggggtttg ggcacggtga agatggagca 360 accgaattca gacccgtcag cattcttgca gcaacagcag caacaacagc agcaacacca 420 ccatttgatg cagctcacaa agcagaatcc tcaagctgct gcggctgccc aacttaacct 480 cttgcaacag cagcggatca tgcatatgca gcagcagcaa caacaacata ttctgaaaaa 540 catgccttta cagagaaacc aattacaaca gcagcagcag caacaacaac aactacaaca 600 acagcagcat cagcagctac ttcgtcaaca gagtctaaac atgagaactc caggaaagtc 660 gcctccctat gagccaggta cctgtgcaaa gagattgacc cattacatgt atcaccagca 720 aaacaggcca caggataaca atgtcgagta ctggagaaac tttgtcaatg agtattttgc 7 80 tccaactgct aaaaagaggt ggtgtgtctc tctctatgga agtggtcgtc aaactactgg 840 agttttccct caggatgtct ggcactgcga aatatgcaat cggaagcctg gccggggctt 900 cgagacaaca gttgaggtct taccgcgatt atgccaaatc aaatatgcga gtggtacatt 960 ggaagaacta ctgtatatcg atatgccacg tgagtccaag aacgtatctg gtcagattgt 1020 tctggactat acaaaagcaa ttcaagaaag tgtctttgat caattgcgtg tcgtacgtga 1080 ggggcatctg aggataattt ttaatccaga cctcaagatt gcatcttggg agttctgtgc 1140 taggcgtcat gaggaactta ttccacggag gtcaataata ccgcaggtta gtcagcttgg 1200 cgcggttgta cagaaatacc aggctgctgc tcaaaaccca accagtttat caactcagga 1260 cctgcagaat aattgcaact cgtttgtggc atgtgcccgt caattggcta aagctctgga 1320 ggtgcctctg gtaaatgatt taggatatac caaacgatac gtccgctgtc ttcagattgc 1380 ggaggtggtg aactgtatga aagatttgat tgaccacagc aggcagactg gatctggacc 1440 aattgatagc ctgcacaagt ttcctcgcag gactccatca gggatcaacc ctcttcaatc 1500 acagcagcaa ccgcctgaag agcaacaatc tgttccccag agttcaaacc agagtggtca 1560 13 aaattctgct aaataattcg cctccaaggt taacagtggg tccatctagt ccttcagaac gcagcctcca gagaatcttg catgaagagg cgccgtcaca ttcgaatcct tggtaggatg gcaccagcac acttagagca tggccggaaa taattcatta gggcttttgc ggccactgta actgagctac gcacccggca cctccttttt cctatggctg gtgtgcaggt ttctgcctct gctaatgcgg atgccacatc ctcaactgtg caccctctac atctgcaccc tcaccaacag ttgttgggct tcaatggatt ctagacaaga tcatccaatg tgcagcgcaa atggccagta aataatggtg caattcccag ggtgaactcc gcaagctcgt tacagtcaaa cctttccctt tgcaggtgcc tacgtcaccc aataacaaca tgatgccgac gcaaaccaac tcagttctcc cccagcagta tcaccaaact tacctccaat tcaactcggc ctcaggagtc tgagccaagt gatgcccaaa gctcagttca caagagatga tgtcatcaca aatgaatggt gttggccatg cagggaatga ccaaatgggc ttacccctgg tattaatggg gttaactgct tagttggtaa aatcactccg gaatgggagg aatgggattt ggggccatgg gcgggttcgg gcagccagtg gactcagaat ggcaatgacg aataatacaa tggcaatgaa ggaatgcacc acagtgcaca tgacctatca cagttgggcc agcagcacca cagcaccagc accagcacca gcatgacata ggaaatcagc tgttgggtgg acaaatagct tcaataatat tcagtatgat tggaaaccct ctcaatagag cattagaaag tcagtatgat gaagatgata tgcagctgtc ctggctgggc tggagaatca agggcagcgc cataacaacg ccccttgggt caaagcgttt tccaatggtg ccatggcaag gaatcataag cgacggcaaa cacctgagct tgtctcaacg gttagtttag ctggttcgtt gtgtattatg caactatggc cggcctcagt tatcttaccc aaagatgagt tgaaggatta taacctgcct ccgttggtgt ctgtgtatgg ccttatttct cacccagaaa agaagttttc aaaaaaaaaa 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2780 <210> 20 <211> 2302 <212> DNA <213> Festuca arundinacea <400> 20 cctcgtgccg cttcctccct ttcccacgcc cgcttcccaa ccctggatcc aaatcccaac 60 ctatcccaaa accgaaaccg aggcaaggaa aagcatcgcg cagttattag ctagctagct 12 0 caaggcgaga tcatgaagcg tgagtaccaa gacgccggcg ggagcagcgc cggcggtgac 180 atgggcatgt ccaaggacaa gatgatgtcg gcgccgccgg cgcaggagga cgaggacgtc 240 gacgagctcc tcgcggcgct cgggtacaag gtgcgctcct ccgacatggc ggacgtcgcg 300 cagaagctgg agcagctgga gatggccatg gggatgggcg gcgtgcctgc gccggacgac 360 ggcttcacca cgcacctggc caccgagacc gtgcactaca accccaccga cctctcctcc 420 tgggtcgaga gcatgctctc cgagctcaac gcgccgccgc cgctcccgcc ggccccgagg 4 80 ctcgctcccg cct.ccgccag cgtcacggcc gacggcttct tcgatatccc gccgccatcc 54 0 gtcgactcct ccagcagcac ctacgcgctg aggccgatcc cctcgccggc cgacctgtcc 600 gccgacctgt ctgccgactc cccgcgggac cccaagcgga tgcgtaccgg cggcggcagc 660 acgtcctcct cctcatcatc gtcatcctcc ctcggcggct gcgtggtgga ggccgctccg 720 ccggcggccg cggaggccaa cgccatcgcg ctgccggtcg tggtggccga cacgcaggag 7 80 gcagggatcc ggctggtgca cgcgctgctg gcgtgcgcgg aggccgtgca gcaggagaac 840 ttctcggccg ccgaggcgct ggtgaagcag ataccctfcgc tggcggcctc ccagggcggc 900 gccatgcgca aggtcgcggc ctact'tcggc gaggccctcg cccgccgcgt cttccgcttc 960 cgcccgcagc ccgacagctc ccacctcgac gccgccttcg ccgacctcct ccacgcgcac 1020 ttctacgagt cctgccccta cctcaagttc gcccacttca ccgccaacca ggccatcctc 1080 gaggccttcg ccggctgccg ccgcgtccac gtcgtcgact tcggcatcaa gcaagggatg 1140 cagtggcccg ctcttctcca ggccctcgcc ctccgccccg gcggccctcc gtcgttccgc. .1200 ctcaccggcg tgggcccacc gcagccggac gagaccgacg ccctgcagca ggtgggctgg 1260 aagctggccc agttcgcgca caccatcggc gtcgatttcc agtaccgcgg cctcgtcgcc 1320 gccacgctcg ccgacctgga gccgttcatg ctgcagccag aggccgagga cggccccaac 1380 gaagaacccg aggtaatcgc cgtgaactca atcttcgaga tgcaccggct gctcgcgcag 1440 cccggcgccc tcgagaaggt cctgggcacc gtgcgcgccg tgcggccgag gatcgtgacc 1500 gtggtagagc aggaggccaa ccacaacgcc ggctcgttcc tggaccgatt caccgagtcc 1560 ctgcactact actccaccat gttcgattcg ctggagggcg ccggctccgg cccgtccgaa 1620 atctcgtcgg ggcctgctgc tgctgccgct gctcctggca cggaccaggt catgtccgag 1680 gtgtacctcg gccggcagat ctgcaatgtc gtggcctgcg agggcgcgga gcgcacggag 1740 cgccacgaga cgctggggca ttggcgcggc cgcctcggcc acgccgggtt cgagaccgtg 1800 cacctgggct ccaacgccta caagcaggcg agcacgctgc tggcgctctt cgccggcggc 18 60 gacgggtaca aggtggacga gaaggaaggc tgcctcacgc tcggctggca cacccgcccg 1920 ctgatcgcca cctccgcatg gcgcatggcc gccgcgccct gatcgcaagt tttgaacgct 1980 14 gtaagtacac cacaccccga tgaatgaagc taaaacgtcg gcctcgctgg cgtgaagaga cgttcttgtt ctcgattccc aaaatgtcta ttattgttct acggatttcg gtaaaaaaaa gcacggaaca caacccccgc ccttggctca- ccggcgcact acgaacgctg gattgcagcg accaacgatc ggagttaagg tggacaccgg atcgctccga ccacaccaga gcctgtaatt cacttgatcc gtgaactcta gcagcctatt attaagtttt gtgtaattcc tgcaatcgct catatttaaa taaggacggg aa 2040 2100 2160 2220 2280 2302 <210> 21 <211> 163 <212> PRT <213> Lolium perenne <400> 21 Met Ala Ala Glu Asp Lys Lys lie Thr Leu Lys Ser Ser Asp Gly Glu 15 10 15 Gin Phe Glu Val Asp Glu Ala Val Ala Met Glu Ser Gin Thr lie Arg 20 25 30 His Met lie Glu Asp Asp Cys Ala Asp Asn Gly lie Pro Leu Pro Asn 35 40 45 Val Asn Ala Lys lie Leu Ser Lys Val Val Glu Tyr Cys Ser Lys His 50 55 60 Val Gin Ala Ala Asp Gly Ala Ala Ala Ala Asp Gly Ala Pro Ala Pro 65 70 75 80 Pro Pro Ala Glu Asp Leu Lys Asn Trp Asp Ala Glu Phe Val Lys Val 85 90 95 Asp Gin Ala Thr Leu Phe Asp Leu lie Leu Ala Ala Asn Tyr Leu Asn 100 105 110 lie Lys Gly Leu Leu Asp Leu Thr Cys Gin Thr Val Ala Asp Met lie 115 120 125 Lys Gly Lys Thr Pro Glu Glu lie Arg Lys Thr Phe Asn lie Lys Asn 130 135 140 Asp Phe Thr Ala Glu Glu Glu Glu Glu lie Arg Arg Glu Asn Gin Trp 145 150 155 160 Ala Phe Glu <210> 22 <211> 169 <212> PRT <213> Lolium perenne <400> 22 Met Ala Ala Ala Asp Asp Ser Lys Lys Met lie Thr Leu Lys Ser Ser 15 10 15 Asp Gly Glu Val Phe Glu Val Glu Glu Ala Val Ala Met Glu Ser Gin 20 25 30 Thr lie Arg His Met lie Glu Asp Asp Cys Ala Asp Asn Gly lie Pro 35 40 45 Leu Pro Asn Val Asn Ser Lys lie Leu Ser Lys Val lie Glu Tyr Cys 50 55 60 Asn Lys His Val Gin Ala Ala Lys Pro Ala Ala Asp Ala Ala Ala Ala 65 70 75 80 Asp Ser Ser Ser Ala Ala Ala Pro Pro Glu Asp Leu Lys Asn Trp Asp 85 ' 90 95 Ala Glu Phe Val Lys Val Asp Gin Ala Thr Leu Phe Asp Leu lie Leu 100 105 110 Ala Ala Asn Tyr Leu Asn lie Lys Gly Leu Leu Asp Leu Thr Cys Gin 115 120 125 Thr Val Ala Asp Met lie Lys Gly Lys Thr Pro Glu Glu lie Arg Lys 130 135 140 Thr Phe Asn lie Lys Asn Asp Phe Thr Ala Glu Glu Glu Glu Glu lie 15 145 150 155 160 Arg Arg Glu Asn Gin Trp Aia Phe Glu 165 <210> 23 <211> 150 <212> PRT <213> Lolium perenne <400> 23 Ser Asp Gly Glu Glu Phe Glu Val Glu Glu Val Leu Val Leu Glu Ser 1 5 10 15 Gin Thr He Lys His Met lie Glu Asp Glu Cys Asp Gly Val lie Pro 20 25 30 Leu Pro Asn Val Ser Ala Lys lie Leu Ser Lys Val lie Glu Tyr Cys 35 40 45 Arg Lys His Val Gin Thr Arg Ala Ala Leu Ala Pro Asp Gly Asp Met 50 55 60 Ser Thr Asn Ala Ala Gly Thr Glu Leu Lys Thr Phe Asp Glu Asp Phe 65 70 75 80 Val Lys Val Asp Gin Ala Thr Leu Phe Asp Leu lie Leu Ala Ala Asn 85 90 95 Tyr Leu Asp lie Lys Gly Leu Leu Asp Leu Thr Cys Gin Thr Val Ala 100 105 110 Asp Met lie Lys Gly Lys Thr Pro Glu Glu lie Arg Ala Thr Phe Asn 115 120 125 lie Lys Asn Asp Phe Thr Pro Glu Glu Glu Glu Glu Val Arg Lys Glu 130 135 140 Asn Ala Trp Ala Phe Glu 145 150 <210>.24 <211> 654 <212> PRT <213> Festuca arundinacea <400> 24 Gly Gly Arg Gly Asp Tyr Ser Asp His Asp Asn Lys Ser Gly His Val 1 5 10 15 Lys Leu Phe Val Gly Ser Val Pro Arg Thr Ala Ser Glu Asp Asp Val 20 25 30 Arg Pro Leu Phe Glu Asn His Gly Asp Val Leu Glu Val Ala Met lie 35 40 45 Arg Asp Arg Lys Thr Gly Glu Gin Gin Gly Cys Cys Phe Val Lys Tyr 50 55 60 Ala Thr Ser Glu Glu Ala Glu Arg Ala lie Arg Ala Leu His Asn Gin 65 70 75 80 Trp Thr lie Pro Gly Ala Met Gly Pro Val Gin Val Arg Tyr Ala Asp 85 90 95 Gly Glu Lys Glu Arg His Gly Ser lie Glu His Lys Leu Phe Val Ala 100 105 110 Ser Leu Asn Lys Gin Ala Thr Ala Lys Glu lie Glu Glu lie Phe Ala 115 120 125 Pro Phe Gly His Val Glu Asp Val Tyr lie Met Lys Asp Gly Met Lys 130 135 140 Gin Ser Arg Gly Cys Gly Phe Val Lys Phe Ser Ser Lys Glu Pro Ala 145 150 155 160 Leu Ala Ala Met Asn Ser Leu Ser Gly Thr Tyr lie Met Arg Gly Cys 165 170 175 Glu Gin Pro Leu lie Val Arg Phe Ala Asp Pro Lys Arg Pro Arg Pro 16 180 185 190 Gly Glu Ser Arg Trp Leu Arg Met His lie Cys Phe Ala Tyr lie Pro 195 200 205 Thr Leu His Tyr Phe Pro Leu Leu Leu Ser Glu Leu Ser Cys Leu Val 210 215 220 Arg Gly Gly Pro Ala Phe Gly Gly Pro Gly Val Ser Pro Arg Ser Asp 225 230 235 240 Ala Ala Leu Val lie Arg Pro Thr Ala Asn Leu Asp Glu Pro Arg Gly 245 250 255 Arg His Met Pro Arg Asp Ala Trp Arg Pro Ser Ser Pro Ser Ser Val 260 265 ' 270 Ala Pro His Gin Phe Asn Asn Tyr Gly Ser Asp Asn Pro Met Gly Leu 275 280 285 ' Met Gly Gly Thr Gly Thr Ser Ala Thr Asp Asn Gly Ala Phe Arg Pro 290 295 300 Gin Met Phe Pro Gly Asn Gly Gin Thr Ala Val Pro Thr Ser Ser His 305 310 315 320 Met Gly lie Asn Thr Ser Ser Val Gin Gly His His. Leu Gly Gly Gin 325 330 335 Gin lie Pro Pro Leu Gin Lys Pro Pro Gly Pro Pro His Asn Phe Ser 340 345 350 Leu Gin Leu Gin Asn Gin Gin Gly Gin His Ser Leu Gly Pro Gly Leu 355 360 365 Phe Gly Gin Asn Val Pro Ser Met Gin Leu Pro Gly Gin Leu Pro Thr 370 375 380 Ser Gin Pro Leu Thr Gin Gin Asn Ala Ser Ala Gly Ala Leu Gin Val 385 390 395 400 Pro Pro Ala lie Gin Ser Asn Pro Met Gin Ser Val Pro Gly Gin Gin 405 410 415 Gin Leu Pro Ser Asn Val Ala Ala Gin Met Met Gin Gin Pro lie Gin 420 425 430 Gin lie Pro Ser Gin Ala Pro Gin Leu Leu Leu Gin Gin Gin Ala Ala 435 440 445 Met Gin Ser Ser Tyr Gin Ser Ser Gin Gin Ala lie Phe Gin Leu Gin 450 455 460 Gin Gin Leu Gin Leu Met Gin Gin Gin Gin Gin Gin Gin Gin Gin Pro 465 470 475 480 Asn Leu Asn Gin Gin Pro His Thr Gin lie Ser Lys Gin Gin Gly Gin 485 490 495 Pro Asn Gin Ser Ser Thr Pro Gly Ala Pro Ala Ala Met Met Pro Ser 500 505 510 Asn. lie Asn Ala lie Pro Gin Gin Val Asn Ser Pro Val Val Ser Leu 515 520 525 Thr Cys Asn Trp Thr Glu His Thr Ser Pro Glu Gly Phe Lys Tyr Tyr 530 535 540 Tyr Asn Ser lie Thr Arg Glu Ser Lys Trp Glu Lys Pro Glu Glu Tyr 545 550 555 560 Val Leu Tyr Glu Gin Gin Gin Gin Gin Gin His Gin Lys Leu lie Leu 565 570 575 Leu Gin Gin His Gin Gin Lys Leu Val Ala Gin Gin Leu Gin Ser Pro 580 585 590 Pro Gin Ala Gin Thr lie Gin Ser Met Gin Ser lie Gin Gin His Pro 595 600 605 Gin Ser His Gin Gly His Asn Gin Met Gin Met Lys His Gin Glu Leu 610 615 620' Asn Tyr Asn Gin Leu Gin Ala Thr Gly Asn lie Asp Pro Asn Arg lie 625 630 635 640 Gin Gin' Gly lie Gin Ala Ala Gin Glu Arg Ser Trp Lys Ser 645 650 <210> 25 17 <211> 618 <212> PRT <213> Lolium perenne <400> 25 Gly Gly Arg Gly Asp Tyr Ser Asp His Asp Asn Lys Ser Gly His Val 15 10 15 Lys Leu Phe Val Gly Ser Val Pro Arg Thr Ala Ser Glu Asp Asp Val 20 25 30 Arg Pro Leu Phe Glu Asn His Gly Asp Val Leu Glu Val Ala Met lie 35 40 45 Arg Asp Arg Lys Thr Gly Glu Gin Gin Gly Cys Cys Phe Val Lys Tyr 50 55 60 Ala Thr Ser Glu Glu Ala Glu Arg Ala lie Arg Ala Leu His Asn Gin 65 70 75 80 Trp Thr lie Pro Gly Ala Met Gly Pro Val Gin Val Arg Tyr Ala Asp 85 90 95 Gly Glu Lys Glu Arg His Gly Ser lie Glu His Lys Leu Phe Val Ala 100 105 110 Ser Leu Asn Lys Gin Ala Thr Ala Lys Glu lie Glu Glu lie Phe Ala 115 120 125 Pro Phe Gly His Val Glu Asp Val Tyr lie Met Lys Asp Gly Met Lys 130 135 140 Gin Ser Arg Gly Cys Gly Phe Val Lys Phe Ser Ser Lys Glu Pro Ala 145 150 155 160 Leu Ala Ala Met Asn Ser Leu Ser Gly Thr Tyr lie Met Arg Arg Pro ' 165 170 175 Arg Pro Gly Glu Ser Arg Gly Gly Pro Ala Phe Gly Gly Pro Gly Val 180 185 190 Ser Pro Arg Ser Asp Ala Ala Leu Val lie Arg Pro Thr Ala Asn Leu 195 200 205 Asp Glu Pro Arg Gly Arg His Met Pro Arg Asp Ala Trp Arg Pro Ser 210 215 220 Ser Pro Ser Ser Val Ala Ser His Gin Phe Asn Asn Tyr Gly Ser Asp 225 230 235 240 Asn Pro Met Gly lie Met Gly Gly Thr Gly Thr Ser Ala Ala Asp Asn 245 250 255 Gly Ala Phe Arg Pro Gin Met Phe Pro Gly Asn Gly Gin Thr Ala Val 260 265 270 Pro Thr Ser Ser His Met Gly lie Asn Thr Ser Leu Gin Gly His His 275 280 285 Leu Gly Gly Gin Gin lie Pro Pro Leu Gin Lys Pro Pro Gly Pro Pro 290 295 300 His Asn Phe Ser Leu Gin Leu Gin Asn Gin Gin Gly Gin His Ser Leu 305 310 315 -320 Val Pro Gly Leu Phe Gly Gin Asn Val Pro Ser Met Gin Leu Pro Gly 325 330 335 Gin Leu Pro Thr Ser Gin Pro Leu Thr Gin Gin Asn Ala Ser Ala Gly 340 345 350 Ala Leu Gin Ala Pro Pro Ala lie Gin Ser Asn Pro Met Gin Ser Val 355 360 365 Pro Gly Gin Gin Gin Leu Pro Ser Asn Val Ala Pro Gin Met Met Gin 370 375 380 Gin Pro lie Gin Gin lie Pro Ser Gin Ala Pro Gin Leu Leu Leu Gin 385 390 395 400 Gin Gin Ala Ala Met Gin Ser Ser Tyr Gin Ser Ser Gin Gin Ala lie 405 410 415' Phe Gin Leu Gin Gin Gin Leu Gin Leu Met Gin Gin Gin Gin Gin Gin 420 425 ' 430 Gin Gin Gin Pro Asn Leu Asn Gin Gin Gin Pro Asn Leu Asn Gin Gin 435 440 445 18 Gin His Thr Gin lie Ser . Lys Gin Gin Gly Gin Pro Asn Gin Ser Ser 450 455 460 Thr Pro Gly Ala Pro Ala Ala Met Met Pro Ser Asn lie Asn Ala lie 465 470 475 480 Pro Gin Gin Val Asn 485 Ser Pro Ala Val Ser 490 Leu Thr Cys Asn Trp 495 Thr Glu His Thr Ser 500 Pro Glu Gly Phe Lys 505 Tyr Tyr Tyr Asn Ser 510 lie Thr Arg Glu Ser 515 Lys Trp Glu Lys Pro 520 Glu Glu Tyr Val Leu 525 Tyr Glu Gin Gin Gin 530 Gin Gin Gin Gin Gin 535 Gin Lys Leu lie Leu 540 Leu Gin Gin His Gin Gin Lys Leu Val Ala Gin Gin Leu Gin Ser Pro Pro Gin Ala Gin 545 550 555 560 Thr lie Gin Ser Met 565 Gin Ser lie Gin Gin 570 His Pro Gin Ser His 575 Gin Gly His Asn Gin 580 Met Gin Met Lys His 585 Gin Glu Leu Asn Tyr 590 Asn Gin Leu Gin Ala 595 Thr Gly Asn lie Asp 600 Pro Asn Arg lie Gin 605 Gin Gly lie Gin Ala 610 Ala Gin Glu Arg Ser 615 Trp Lys Ser <210> 26 <211> 177 <212> PRT <213> Festuca arundinacea <400> 26 Met Ala Gly Arg Asp Arg Asp Pro Leu Val Val Gly Arg Val Val Gly 1 5 10 15 Asp Val Leu Asp Pro Phe Val Arg Thr Thr Asn Leu Arg Val Thr Phe 20 25 30 Gly Asn Arg Ala Val Ser Asn Gly Cys Glu Leu Lys Pro Ser Met Val 35 40 45 Thr His Gin Pro Arg Val Glu Val Gly Gly Asn Glu Met Arg Thr Phe 50 55 60 Tyr Thr Leu Val Met Val Asp Pro Asp Ala Pro Ser Pro Ser Asp Pro 65 70 75 80 Asn Leu Arg Glu Tyr Leu His Trp Leu Val Thr Asp lie Pro Gly Thr 85 90 95 Thr Gly Ala Ser Phe Gly Gin Glu Val Met Cys Tyr Glu Ser Pro Arg 100 105 110 Pro Asn Met 115 Gly lie His Arg Phe 120 Val Leu Val Leu Phe 125 Gin Gin Leu Gly Arg Gin Thr Val Tyr Ala Pro Gly Trp Arg Gin Asn Phe Asn Thr 130 135 140 Arg Asp Phe Ala Glu Leu Tyr Asn Leu Gly Pro Ala Val Ala Ala Val 145 150 155 160 Tyr Phe Asn Cys Gin Arg Glu Ala Gly Ser Gly Gly Arg Arg Met Tyr 165 170 175 ' Asn <210> 27 <211> 89 <212> PRT <213> Lolium perenne <400> 27 Met Val Gly Val Gin Arg Ala Asp Pro Leu Val Val Gly Arg Val lie 19 1 5 10 15 Gly Asp Val Val Asp Pro Phe Val Arg Arg Val Pro Leu Arg Val Gly 20 25 30 Tyr Ala Ser Arg Asp Val Ala Asn Gly Cys Glu Leu Arg Pro Ser Ala 35 40 45 lie Ala Asp Gin Pro Arg Val Glu Val Gly Gly Pro Asp Met Arg Thr 50 55 60 Phe Tyr Thr Leu Val Met Val Asp Pro Asp Ala Pro Ser Pro Ser Asp 65 70 75 80 Pro Ser Leu Arg Glu 85 Tyr Leu His' Trp <210> 28 <211> 298 <212> PRT <213> Lolium perenne <400> 28 Glu Ala Phe Ala Gly Cys Arg Arg Val His Val Val Asp Phe Gly lie 1 5 10 15 Lys Gin Gly Met Gin Trp Pro Ala Leu Leu Gin Ala Leu Ala Leu Arg 20 25 30 Pro Gly Gly Pro Pro Ser Phe Arg Leu Thr Gly Val Gly Pro Pro Gin 35 40 45 Pro Asp Glu Thr Asp Ala Leu Gin Gin Val Gly Trp Lys Leu Ala Gin 50 55 60 Phe Ala His Thr lie Gly Val Asp Phe Gin Tyr Arg Gly Leu Val Ala 65 70 75 80 Ala Thr Leu Ala Asp Leu Glu Pro Phe Met Leu Gin Pro Glu Ala Asp 85 90 95 Asp Gly Pro Asn Glu Glu Pro Glu Val He Ala Val Asn Ser Val Phe 100 105 110 Glu Met His Arg Leu Leu Ala Gin Pro Gly Ala Leu Glu Lys Val Leu 115 120 125 Gly Thr Val Arg Ala Val Arg Pro Arg lie Val Thr Val Val Glu Gin 130 135 140 Glu Ala Asn His Asn Thr Gly Ser Phe Leu Asp Arg Phe Thr Glu Ser 145 150 155 160 Leu His Tyr Tyr Ser Thr Met Phe Asp Ser Leu Glu Gly Ala Gly Ser 165 170 175 Ala Pro Ser Glu lie Ser Ser Gly Pro Ser Ala Ala Ala Ala Asn Ala 180 185 190 Ala Ala Pro Gly Thr Asp Gin Val Met Ser Glu Val Tyr Leu Gly Arg 195 200 205 Gin lie Cys Asn Val Val Ala Cys Glu Gly Ala Glu Arg Thr Glu Arg 210 215 220 His Glu Thr Leu Gly Gin Trp Arg Gly Arg Leu Gly His Ala Gly Phe 225 230 235 ' 240 Glu Thr Val His Leu Gly Ser Asn Ala Tyr Lys Gin Ala Ser Thr Leu 245 250 '255 Leu Ala Leu Phe Ala Gly Gly Asp Gly Tyr Lys Val Asp Glu Lys Glu 260 265 . 270 Gly Cys Leu Thr Leu Gly Trp His Thr Arg Pro Leu He Ala Thr Ser 275 280 285 Ala Trp Arg Met Ala Ala Ala Ala Ala Pro • 290 295 <210> 29 <211> 1148 <212> PRT <213> Lolium perenne 20 <400> 29 Met Ser Val Ser Asn Gly Lys Trp lie Asp Gly Leu Gin Phe Ser Ser 15 10 15 Leu Phe Trp Pro Pro Pro His Asp Ala Gin Gin Lys Gin Ala Gin Thr 20 25 30 Leu Ala -Tyr Val Glu Tyr Phe Gly Gin Phe Thr Ser Asp Ser Glu Gin 35 40 45 Phe Pro Glu Asp Val Ala Gin Leu lie Gin Ser Tyr Tyr Pro Ser Lys 50 55 60 Glu Lys Arg Leu Val Asp Glu Val Leu Ala Thr Phe Val Leu His His 65 70 75 80 Pro Glu His Gly His Ala Val Val His Pro lie Leu Ser Arg lie lie 85 90 95 Asp Gly Ser Leu Ser Tyr Asp Arg His Gly Ser Pro Phe Asn Ser Phe 100 105 110 lie Ser Leu Phe Thr Gin Thr Ala Glu Lys Glu Tyr Ser Glu Gin Trp 115 120 125 Ala Leu Ala Cys Gly Glu lie Leu Arg Val Leu Thr His Tyr Asn Arg 130 135 140 Pro lie Phe Lys Val Ala Glu Cys Asn Asp Thr Ser Asp Gin Ala Thr 145 150 155 160 Thr Ser Tyr Ser Leu His Asp Lys Ala Asn Ser Ser Pro Glu Asn Glu 165 170 175 Pro Glu Arg Lys Pro Leu Arg Pro Leu Ser Pro Trp lie Thr Asp lie 180 185 190 Leu Leu Asn Ala Pro Leu Gly lie Arg Ser Asp Tyr Phe Arg Trp Cys 195 200 205 Gly Gly Val Met Gly Lys Tyr Ala Ala Gly Gly Glu Leu Lys Pro Pro 210 215 220 Thr Thr Ala Tyr Ser Arg Gly Ala Gly Lys His Pro Gin Leu Met Pro 225 230 235 240 Ser Thr Pro Arg Trp Ala Val Ala Asn Gly Ala Gly Val lie Leu Ser 245 250. 255 Val Cys Asp Glu Glu Val Ala>Arg Tyr Glu Thr Ala Asn Leu Thr Ala 260 265 270 Ala Ala Val Pro Ala Leu Leu Leu Pro Pro Pro Thr Thr Pro Leu Asp 275 280 285 Glu His Leu Val Ala Gly Leu Pro Pro Leu Glu Pro Tyr Ala Arg Leu 290 295 300 Phe His Arg Tyr Tyr Ala lie Ala Thr Pro Ser Ala Thr Gin Arg Leu 305 310 315 320 Leu Phe Gly Leu Leu Glu Ala Pro Pro Ser Trp Ala Pro Asp Ala Leu 325 330 335 Asp Ala Ala Val Gin Leu Val Glu Leu Leu Arg Ala Ala Glu Asp Tyr 340 345 350 Ala Thr Gly Met Arg Leu Pro Lys Asn Trp Leu His Leu His Phe Leu 355 360 365 Arg Ala lie Gly Thr Ala Met Ser Met Arg Ala Gly Met Ala Ala Asp 370 375 380 Thr Ala Ala Ala Leu Leu Phe Arg lie Leu Ser Gin Pro Thr Leu Leu 385 390 395 400 Phe Pro Pro Leu Arg His Ala Glu Gly Val Val Gin His Glu Pro Leu 405 410 415 Gly Gly Tyr Val Ser Ser Tyr Lys Arg Gin Leu Glu lie Pro Ala Ser 420 425 430 Glu Thr Thr lie Asp Ala Thr Ala Gin Gly lie Ala Ser Leu Leu Cys 435 440 445 Ala His Gly Pro Asp Val Glu Trp Arg lie Cys Thr lie Trp Glu Ala 450 455 460 21 Ala Tyr Gly Leu Leu Pro Leu Asn Ser Ser Ala Val Asp Leu Pro Glu 465 470 475 480 lie Val Val Ala Ala Pro Leu Gin Pro Pro Thr Leu Ser Trp Ser Leu 485 490 495 Tyr Leu Pro Leu Leu Lys Val Phe Glu Tyr Leu Pro Arg Gly Ser Pro 500 505 510 Ser Glu Ala Cys Leu Met Arg lie Phe Val Ala Thr Val Glu Ala lie 515 520 525 Leu Arg Arg Thr Phe Pro Ser Glu Thr Glu Pro Ser Lys Lys Pro Arg 530 535 540 Ser Pro Ser Lys Ser Leu Ala Val Ala Glu Leu Arg Thr Met lie His 545 550 555 560 Ser Leu Phe Val Glu Ser Cys Ala Ser Met Asn Leu Ala Ser Arg Leu 565 570 575 Leu Phe Val Val Leu Thr Val Ser Val Ser His Gin Ala Leu Pro Gly 580 585 590 Gly Ser Lys Arg Pro Thr Gly Ser Glu Asn His Ser Ser Glu Glu Ser 595 600 605 Thr Glu Asp Ser Lys Leu Thr Asn Gly Arg Asn Arg Cys Lys Lys Lys 610 615 620 Gin Gly Pro Val Gly Thr Phe Asp Ser Tyr Val Leu Ala Ala Val Cys 625 630 635 640 Ala Leu Ser Cys Glu Leu Gin Leu Phe Pro He Leu Cys Lys Asn Val 645 650 655 Thr Lys Thr Asn lie Lys Asp Ser lie Lys lie Thr Met Pro Gly Lys 660 665 670 Thr Asn Gly lie Ser Asn Glu Leu His Asn Ser Val Asn Ser Ala lie 675 680 685 Leu His Thr Arg Arg lie Leu Gly lie Leu Glu Ala Leu Phe Ser Leu 690 695 700 Lys Pro Ser Ser Val Gly Thr Ser Trp Ser Tyr Ser Ser Asn Glu lie 705 710 715 720 Val Ala Ala Ala Met Val Ala Ala His Val Ser Glu Leu Phe Arg Arg 725 730 735 Ser Arg Pro Cys Leu Asn Ala Leu Ser Ala Leu Lys Arg Cys Lys Trp 740 745 750 Asp Ala Glu lie Ser Thr Arg Ala Ser Ser Leu Tyr His Leu lie Asp 755 760 765 Leu His Gly Lys Thr Val Ser Ser lie Val Asn Lys Ala Glu Pro Leu 770 775 780 Glu Ala His Leu Asn Leu Thr Ala Val Lys Lys Asp Asp Gin His His 785 790 795 800 lie Glu Glu Ser Asn Thr Ser Ser Ser Asp Tyr Gly Asn Leu Glu Lys 805 • 810 815 Lys Ser Lys Lys Asn Gly Phe Ser Arg Pro Leu Met Lys Cys Ala Glu 820 825 830 Gin Ala Arg Arg Asn Gly Asn Val Ala Ser Thr Ser Gly Lys Ala Thr 835 840 845 Ala Thr Leu Gin Ala Glu Ala Ser Asp Leu Ala Asn Phe Leu Thr Met 850 • 855 860 Asp Arg Asn Gly Gly Tyr Gly Gly Ser Gin Thr Leu Leu Arg Thr Val 865 870 875 880 • Met Ser Glu Lys Gin Glu Leu Cys Phe Ser Val Val Ser Leu Leu Trp 885 890 895 His Lys Leu lie Ala Ser Pro Glu Thr Gin Met Ser Ala Glu Ser Thr 900 905 910 Ser Ala His Gin Gly Trp Arg Lys Val Ala Asp Ala Leu Cys Asp Val 915 920 925 Val Ser Ala Ser Pro Ala Lys Ala Ser Thr Ala lie Val Leu Gin Ala 930 935 940 Glu Lys Asp Leu Gin Pro Trp lie Ala Arg Asp Asp Glu Gin Gly Gin 22 945 950 955 960 Lys Met Trp Arg Val Asn Gin Arg lie Val Lys Leu lie Ala Glu Leu 965 970 975 Met Arg Asn His Asp Ser Pro Glu Ala Leu He lie Leu Ala Ser Ala 980 985 990 Ser Asp Leu Leu Leu Arg Ala Thr Asp Gly Met Leu Val Asp Gly Glu 995 1000 1005 Ala Cys Thr Leu Pro Gin Leu Glu Leu Leu Glu Val Thr Ala Arg Ala 1010 1015 1020 lie His Leu He Val Glu Trp Gly Asp Pro Gly Val Ala Val Ala Asp 1025 1030 1035 1040 Gly Leu Ser Asn Leu Leu Lys Cys Arg Leu Ser Pro Thr lie Arg Cys 1045 1050 1055 Leu Ser 'His Pro Ser Ala His Val Arg Ala Leu Ser Met Ser Val Leu 1060 1065 1070 Arg Asp lie Leu Asn Ser Gly Pro lie Ser Ser Thr Lys lie lie Gin 1075 1080 1085 -Gly Glu Gin Arg Asn Gly lie Gin Ser Pro Ser Tyr Arg Cys Ala Ala 1090 1095 1100 Ala Ser Met Thr Asn Trp Gin Ala Asp Val Glu Arg Cys lie Glu Trp 1105 1110 1115 1120 Glu Ala His Asn Arg Gin Ala Thr Gly Met Thr Leu Ala Phe Leu Thr 1125 1130 1135 Ala Ala Ala Asn Glu Leu Gly Cys Pro Leu Pro Cys 1140 1145 <210> 30 ' <211> 1149 <212> PRT <213> Festuca arundinacea <400> 30 Met Ser Ala Ser Asn Gly Lys Trp lie Asp Gly Leu Gin Phe Ser Ser 1 5 10 15 Leu Phe Trp Pro Pro Pro His Asp Ala Gin Gin Lys Gin Ala Gin Thr 20 25 30 Leu Ala Tyr Val Glu Tyr Phe Gly Gin Phe Thr Ser Asp Ser Glu Gin 35 40 45 Phe Pro 50 Glu Asp Val Ala Gin 55 Leu lie Gin Ser Cys 60 Tyr Pro Ser Lys Glu Lys Arg Leu Val Asp Glu Val Leu Ala Thr Phe Val Leu His His 65 70 75 80 Pro Glu His Gly His Ala Val Val His Pro lie Leu Ser Arg lie lie 85 90 95 Asp Gly Ser Leu 100 Ser Tyr Asp Arg His 105 Gly Ser Pro Phe Asn 110 Ser Phe lie Ser Leu 115 Phe Thr Gin Thr Ala 120 Glu Lys Glu Tyr Ser 125 Glu Gin Trp Ala Leu Ala Cys Gly Glu lie Leu Arg Val Leu Thr His Tyr Asn Arg 130 135 140 Pro lie Phe Lys Val Ala Glu Cys Asn Asp Thr Ser Asp Gin Ala Thr 145 150 155 160 Thr Ser Tyr Ser Leu 165 Gin Glu Lys Ala Asn 170 Ser Ser Pro Glu Asn 175 Glu Pro Glu Arg Lys 180 Pro Leu Arg Pro Leu 185 Ser Pro Trp lie Thr 190 Asp lie Leu Leu Asn Ala Pro Leu Gly lie Arg Ser Asp Tyr Phe Arg Trp Cys 195 200 205 Gly Gly 210 Val Met Gly Lys Tyr 215 Ala Ala Gly Gly Glu 220 Leu Lys Pro Pro Thr Thr Ala Tyr Ser Arg Gly Ala Gly Lys His Pro Gin Leu Met Pro 23 225 230 235 240 Ser Thr Pro Arg Trp Ala Val Ala Asn Gly Ala Gly Val lie Leu Ser 245 250 255 Val Cys Asp Glu Glu Val Ala Arg Tyr Glu Thr Ala Asn Leu Thr Ala 260 265 270 Ala Ala Val Pro Ala Leu Leu Leu Pro Pro Pro Thr Thr Pro Leu Asp 275 280 285 Glu His Leu Val Ala Gly Leu Pro Pro Leu Glu Pro Tyr Ala Arg Leu 290 295 300 Phe His Arg Tyr Tyr Ala lie Ala Thr Pro Ser Ala Thr Gin Arg Leu 305 310 315 320 Leu Phe Gly Leu Leu Glu Ala Pro Pro Ser Trp Ala Pro Asp Ala Leu 325 330 335 Asp Ala Ala Val Gin Leu Val Glu Leu Leu Arg Ala Ala Glu Asp Tyr 340 345 350 Ala Thr Gly Met Arg Leu Pro Lys Asn Trp Leu His Leu His Phe Leu 355 '360 365 Arg Ala lie Gly Thr Ala Met Ser Met Arg Ala Gly Met Ala Ala Asp 370 375 380 Thr Ala Ala Ala Leu Leu Phe Arg lie Leu Ser Gin Pro Thr Leu Leu 385 390 395 400 Phe Pro Pro Leu Arg His Ala Glu Gly Val Val Gin His Glu Pro Leu 405 410 415 Gly Gly Tyr Val Ser Ser Tyr Lys Arg Gin Leu Glu lie Pro Ala Ser 420 425 430 Glu Thr Thr lie Asp Ala Thr Ala Gin Gly lie Ala Ser Leu Leu Cys 435 440 445 Ala His Gly Pro Asp Val Glu Trp Arg lie Cys Thr lie Trp Glu Ala 450 455 460 Ala Tyr Gly Leu Leu Pro Leu Asn Ser Ser Ala Val Asp Leu Pro Glu 465 470 475 480 lie Val Val Ala Ala Pro Leu Gin Pro Pro Thr Leu Ser Trp Ser Leu 485 490 495 Tyr Leu Pro Leu Leu Lys Val Phe Glu Tyr. Leu Pro Arg Gly Ser Pro 500 505 510 Ser Glu Ala Cys Leu Met Arg He Phe Val Ala Thr Val Glu Ala lie 515 520 525 Leu Arg Arg Thr Phe Pro Ser Glu Thr Ser Glu Pro Ser Lys Lys Pro 530 535 540 Arg Ser Pro Ser Lys Ser Leu Ala Val Ala Glu Leu Arg Thr Met lie 545 550 555 560 His Ser Leu Phe Val Glu Ser Cys Ala Ser Met Asn Leu Ala Ser Arg 565 570 575 Leu Leu Phe Val Val Leu Thr Val Ser Val Ser His Gin Ala Leu Pro 580 585 590 Gly Gly Ser Lys Arg Pro Thr Gly Ser Asp Asn His Ser Ser Glu Glu 595 600 605 Ser Thr Glu Asp Ser Lys Leu Thr Asn Gly Arg Asn Arg Cys Lys Lys 610 615 620 Lys Gin Gly Pro Val Gly Thr Phe Asp Ser Tyr Val Leu Ala Ala Val 625 630 635 640 Cys Ala Leu Ser Cys Glu Leu Gin Leu Phe Pro lie Leu Cys Lys Asn 645 650 655 Val Thr Lys Ser Asn lie Lys Asp Ser lie Lys lie Thr Met Pro Gly 660 665 670 Lys Thr Asn Gly lie Ser Asn Glu Leu His Asn Ser Val Asn Ser Ala 675 680 685 Val Leu His Thr Arg Arg lie Leu Gly He Leu Glu Ala Leu Phe Ser 690 695 700 Leu Lys Pro Ser Ser Val Gly Thr Ser Trp Ser Tyr Ser Ser Asn Glu 705 710 715 720 24 lie Val Ala Ala Ala Met Val Ala Ala His Val Ser Glu Leu Phe Arg 725 730 735 Arg Ser Arg Pro Cys Leu Asn Ala Leu Ser Ala Leu Lys Arg Cys Lys 740 745 750 Trp Asp Ala Glu lie Ser Thr Arg Ala Ser Ser Leu Tyr His Leu lie 755 760 765 Asp Leu His Gly Lys Thr Val Ser Ser lie Val Asn Lys Ala Glu Pro 770 775 780 Leu Glu Ala His Leu Asn Leu Thr Ala Val Lys Lys Asp Asp Gin His 785 790 795 800 His lie Glu Glu Ser Asn Thr Ser Ser Ser Asp Tyr Gly Asn Leu Glu 805 810 815 Lys Lys Ser Lys Lys Asn Gly Phe Ser Arg Pro Leu Met Lys Cys Ala 820 825 830 Glu Gin Ala Arg Arg Asn Gly Asn Val Ala Ser Thr Ser Gly Lys Ala 835 840 845 Thr Ala Thr Leu Gin Ala Glu Ala Ser Asp Leu Ala Asn Phe Leu Thr 850 855 860 Met Asp Arg Asn Gly Gly Tyr Gly Gly Ser Gin Thr Leu Leu Arg Thr 865 870 875 880 Val Met Ser Glu Lys Gin Glu Leu Cys Phe Ser Val Val Ser Leu Leu 885 890 895 Trp His Lys Leu lie Ala Ser Pro Glu Thr Gin Met Ser Ala Glu Ser 900 905 910 Thr Ser Ala His Gin Gly Trp Arg Lys Val Ala Asp Ala Leu Cys Asp 915 920 925 Val Val Ser Ala Ser Pro Ala Lys Ala Ser Thr Ala lie Val Leu Gin 930 935 940 Ala Glu Lys Asp Leu Gin Pro Trp lie Ala Arg Asp Asp Glu Gin Gly 945 950 955 960 Gin Lys Met Trp Arg Val Asn Gin Arg lie Val Lys Leu lie Ala Glu 965 970 975 Leu Met Arg Asn His Asp Ser Pro Glu Ala Leu lie lie Leu Ala Ser 980 985 990 Ala Ser Asp Leu Leu Leu Arg Ala Thr Asp Gly Met Leu Val Asp Gly 995 1000 1005 Glu Ala Cys Thr Leu Pro Gin Leu Glu Leu Leu Glu Val Thr Ala Arg 1010 1015 1020 Ala lie His Leu lie Val Glu Trp Gly Asp Pro Gly Val Ala Val Ala 1025 1030 1035 1040 Asp Gly Leu Ser Asn Leu Leu Lys Cys Arg Leu Ser Pro Thr lie Arg 1045 1050 1055 Cys Leu Ser His Pro Ser Ala His Val Arg Ala Leu Ser Met Ser Val 1060 1065 1070 Leu Arg Asp lie Leu Asn Ser Gly Pro lie Ser Ser Thr Lys lie Asn 1075 1080 1085 Gin Gly Glu Gin Arg Asn Gly lie Gin Ser Pro Ser Tyr Arg Cys Met 1090 1095 1100 Ala Ala Ser Met Thr Asn Trp Gin Ala Asp Val Glu Arg Cys lie Glu 1105 1110 1115 1120 Trp Glu Ala His Asn Arg Gin Ala Thr Gly Met Thr Leu Ala Phe Leu 1125 1130 1135 Thr Ala Ala Ala Asn Glu Leu Gly Cys Pro Leu Pro Cys 1140 1145 <210> 31 <211> 496 <212> PRT <213> Festuca arundinacea <400> 31 25 Met Leu Ser Thr Ser Tyr Ala Leu Thr Ala Ala Pro lie Pro Glu Gly 1 5 10 15 Ala Ala Gly Pro Pro Asp Pro Phe Arg Pro Met Gin lie Ala Asn Asp 20 25 30 Asn Ala Ser Ala Lys Arg Lys Arg Arg Pro Ala Gly Thr Pro Asp Pro 35 40 45 Asp Ala Glu Val Val Ser Leu Ser Pro Arg Thr Leu Leu Glu Ser Asp 50 55 60 Arg Tyr Val Cys Glu lie Cys Asn Gin Gly Phe Gin Arg Asp Gin Asn 65 70 75 80 Leu Gin Met His Arg Arg Arg His Lys Val Pro Trp Lys Leu Leu Lys 85 90 95 Arg Glu Ala Gly Glu Ala Ala Arg Lys Arg Val Phe Val Cys Pro Glu 100 105 110 Pro Thr Cys Leu His His Asp Pro Ala His Ala Leu Gly Asp Leu Val 115 120 125 Gly lie Lys Lys His Phe Arg Arg Lys His Ser Gly His Arg Gin Trp 130 135 140 Ala Cys Ser Arg Cys Ser Lys Ala Tyr Ala Val His Ser Asp Tyr Lys 145 150 155 160 Ala His Leu Lys Thr Cys Gly Thr Arg Gly His -Thr Cys Asp Cys Gly 165 170 175 Arg Val Phe Ser Arg Val Glu Ser Phe lie Glu His Gin Asp Met Cys 180 185 190 Asp Ala Ser Arg Pro Arg Gly Gly Thr Thr Ser Ser Ser Pro Gly His 195 200 205 Gly Gly Gly Arg Val Val Gly Ala Ser Asn Pro Gin His Leu Leu His 210 215 220 Ala Ala Ser Leu Ser Arg Thr Ala Ser Ser Ala Ser Pro Ser Ser Gly 225 230 235 240 Gly Glu Leu Val Gly Ser Pro Val Ala Trp Pro Cys Gly Pro Ala Thr 245 250 255 Ala Ser Pro Thr Ala Ala Asn Val Ala Ala Phe Gin Arg Leu Leu Asp 260 " 265 270 Pro Thr Gin Ser Ser Ser Pro Pro Thr Pro Ser Asp Arg Arg Gly Ala 275 280 285 Gly Thr Gin Asn Leu Glu Leu Gin Leu Met Pro Pro Arg Gly Gly Gly 290 295 300 Ala Ala Pro Pro Gly Thr Ala Leu Thr Tyr Arg Ala Ser Pro Cys Ser 305 310 315 320 Pro Ser Val Leu His Ala Pro Arg Gin Leu Gly Ala Asp Ala Val Arg 325 330 335 Leu Gin Leu Ser lie Gly Cys Gly Gly Ala Pro Asp Asp Ser Ser Val 340 345 350 Glu Ser Ala Pro Ala Pro Ala Ala Thr Leu Lys Glu Glu Ala Arg Glu 355 360 365 Gin Leu Arg Leu Ala Thr Ala Glu Met Ala Ser Ala Glu Glu Thr Arg 370 375 380 Ala Gin Ala Arg Arg Gin Val Glu Leu Ala Glu Gin Glu Leu Ala Gly 385 390 395 400 Ala Arg Arg Val Arg Gin Gin Ala Gin Leu Glu Leu Gly Arg Ala His 405 410 415 Ala Leu Arg Asp His Ala Val Arg Gin lie Asp Ala Thr Leu Met Glu 420 425 430 lie Thr Cys Tyr Gly Cys Arg His Asn Phe Arg Ala Arg Ala Ala Ala 435 440 445 Met Asn Cys Glu Val Ala Ser Tyr Val Ser Ser Val Leu Thr Glu Gly 450 455 460 Gly Asp Ala Glu Val Asp Asn Asp Gly His His Gin Leu Leu His Ala 465 470 475 480 Gly Asp Leu Pro Arg Ser His Arg Ala Met Met Lys Met Asp Leu Asn 26 485 490 495 <210> 32 <211> 530 <212> PRT <213> Lolium perenne <400> 32 Met Ala Ala Ala Ser Ser Ala Pro Phe Phe Gly Leu Ser Asp Ala Gin 15 10 15 Met Gin Pro Met Val Pro Ala Gin Pro Pro Ala Pro Val Ala Ala Ala 20 25 30 Pro Ala Pro Lys Lys Lys Arg Asn Gin Pro Gly Asn Pro Asn Pro Asp 35 40 45 Ala Glu Val lie Ala Leu Ser Pro Arg Ser Leu Met Ala Thr Asn Arg 50 55 60 Phe Val Cys Glu Val Cys Gly Lys Gly Phe Gin Arg Glu Gin Asn Leu 65 70 75 80 Gin Leu His Arg Arg Gly His Asn Leu Pro Trp Lys Leu Lys Gin Lys 85 90 95 Asn Pro Lys Asp Ala Leu Arg Arg Arg Val Tyr Leu Cys Pro Glu Pro 100 105 110 Thr Cys Val His His Asp Pro Ala Arg Ala Leu Gly Asp Leu Thr Gly 115 120 125 lie Lys Lys His Tyr Cys Arg Lys His Gly Glu Lys Lys Trp Lys Cys 130 135 140 Asp Lys Cys Ala Lys Arg Tyr Ala Val Gin Ser Asp Trp Lys Ala His 145 150 155 160 Ser Lys Thr Cys Gly Thr Arg Glu Tyr Arg Cys Asp Cys Gly Thr Leu 165 170 175 Phe Ser Arg Arg Asp Ser Phe lie Thr His Arg Ala Phe Cys Asp Ala 180 185 190 Leu Ala Gin Glu Ser Ala Arg Leu Pro Ala lie Gly Ala Ser Leu Tyr 195 200 205 Gly Gly Val Gly Asn Met Gly Ala Leu Asn Thr Leu Ser Gly Met Pro 210 215 220 Gin Gin Leu Pro Gly Gly Ser Phe Pro Asp Gin Ser Gly His His Ser 225 230 235 240 Ser Ala Ser Ala Met Asp lie His Asn Leu Gly Gly Gly Ser Asn Ala 245 250 255 Gly Gin Phe Asp Gin His Leu Met Pro Gin Ser Ala Gly Ser Ser Met 260 265 270 Phe Arg Ser Gin Ala Ala Ser Ser Ser Pro Tyr Tyr Leu Gly Ala Ala 275 280 285 Ala Ala Gin Asp Phe Ala Glu Asp Asp Val His Arg Ser His Gly Asn 290 295 300 Gin Ser Ser Leu Leu Gin Gly Lys Ser Thr Ala Ala Phe His Gly Leu 305 310 315 320 Met Gin. Leu Pro Asp Gin His Gin Gly Ser Ala Ser Asn Gly Asn Asn 325 330 335 Asn Leu Leu Asn Leu Gly Phe Tyr Ser Gly Asn Gly Gly Gly Gin Asp 340 345 350 Gly Arg Val Met Phe Gin Asn Gin Phe Asn Ser Ser Ala Gly Asn Gly 355 360 365 Asn Val Asn Ala Glu Asn Asn Gly Ser Leu Leu Gly Gly Gly Gly Gly 370 375 380 Gly Phe Pro Ser Leu Phe Gly Ser Ser Glu Ser Gly Gly Gly Leu Pro 385 390 395 400 Gin Met Ser Ala Thr Ala Leu Leu Gin Lys Ala Ala Gin Met Gly Ala 405 410 415 Thr Thr Ser Ser His Asn Ala Ser Ala Gly Leu Met Arg Gly Pro Gly 27 420 425 430 Met Arg Gly 435 Gly Ala Gly Glu Gly Gly Ser 440 Ser Ser Ser 445 Ala Ser Glu Arg Gin 450 Ser Phe His Asp Leu lie Met Asn 455 Ser Leu Ala 460 Asn Gly Ser Gly Ala Pro Ala Thr Thr Gly Gly Gly Thr Val Ala Phe Gly Gly Gly 4 65 470 475 480 Gly Phe Pro lie Asp Asp Gly Lys Leu Ser Thr Arg Asp Phe Leu Gly 485 490 495 Val Gly Pro Gly Gly Val Val His Ala Gly Met Gly Pro Pro Arg Arg 500 505 510 His Gly Gly 515 Ala Ala Gly Leu His lie Gly 520 Ser Leu Asp 525 Pro Ala Glu Leu Lys 530 <210> 33 <211> 466 ■ <212> PRT <213> Festuca arundinacea <400> 33 Met Pro Pro Asn Pro Thr Asp ' Pro Glu Gin Pro Glu Ala Ala Ala Ala 1 5 10 15 Pro Ala Pro Pro Pro Lys Lys Lys Arg Asn Leu Pro Gly Thr Pro Asp 20 25 30 Pro Asp Ala Glu Val lie Ala Leu Ser Pro Gly Thr Leu Met Ala Thr 35 40 45 Asn Arg Phe Val Cys Glu Val Cys Gly Lys Gly Phe Gin Arg Asp Gin 50 55 60 Asn Leu Gin Leu His Arg Arg Gly His Asn Leu Pro Trp Arg Leu Arg 65 70 75 80 Gin Arg Gly Pro Gly Ala Ala Pro Pro Arg Arg Arg Val Tyr Val Cys 85 90 95 Pro Glu Pro Gly Cys Val His His Ala Pro Ala Arg Ala Leu Gly Asp 100 105 110 Leu Thr Gly lie Lys Lys His Phe Cys Arg Lys His Gly Glu Lys Arg 115 120 125 Trp Ala Cys Pro Arg Cys Gly Lys Arg Tyr Ala Val Gin Ala Asp Leu 130 135 140 Lys Ala His Ala Lys Thr Cys Gly Thr Arg Glu Tyr Arg Cys Asp Cys 145 150 155 160 Gly Thr Leu Phe Thr Arg Arg Asp Ser Phe Val Thr His Arg Ala Phe 165 170 175 Cys Gly Ala Leu Val Glu Glu Thr Gly Arg Val Leu Ala Val Pro Ala 180 185 190 Pro Pro Ala Pro Gly Pro Pro Asp Leu Asp Asp Val Asp Glu Asn Phe 195 200 205 Asp Lys Asp Ser Glu Lys Gly Glu Glu Asn Val Glu Asp Glu Glu Glu 210 215 220 Lys Gly Glu Val Asn Glu Asn Ser Ala Val Ala Asp Val Asn Glu Pro 225 230 235 240 Gin Arg Val Glu Ala Ala Ser Glu Ala Pro Gin Arg lie Pro Ser Pro 245 250 255 Gin Gin Gin Arg lie Pro Ser Pro Arg Arg lie Pro Ser Pro Gin Arg 260 265 270 lie Arg Ser Pro Pro Ser Pro Val Pro Gin Glu Gin Gin Gin Gin Pro 275 280 285 Met Val Ala Val Val Pro Asn Leu Glu Gly Pro Lys Val Ala Ala Glu 290 295 300 28 Pro lie Val Val Val Lys Gin Glu . Glu Asp Asp Lys Arg Asp > Glu . Asp 305 310 315 320 Val Cys Phe Gin Glu Ala Asp Lys Tyr Asp Asp Ala Glu Leu Glu Gly 325 330 335 Ser Ser Leu Pro Asp Thr Asp Thr Pro Met Leu Pro Cys Phe Leu Pro 340 345 350 Ser Pro Ser Asp Ala lie Gly Thr Asp Gly Ser Ser Thr Ser Cys Gly 355 360 365 Thr Val Ser Ser Ala Ser lie Pro Leu Arg Gin Gin Arg Arg Leu Ala 370 375 380 His Leu Leu Gly Cys Leu His Arg Pro Arg Gin Ala Pro Leu Pro Arg 385 390 395 400 Val Asp Arg Cys Val lie Leu Ser Val Leu lie Pro Pro Ser Phe Ala 405 410 415 Leu Arg Leu Val Arg Pro Pro Leu Cys Ser Arg Arg Gin Thr Arg Ala 420 425 430 Thr Leu Ala Ala Leu Leu His Leu Gin His His Thr Cys Pro Arg Leu 435 440 445 His Ser Cys Arg Arg Leu Leu Arg Leu Glu Leu Arg Lys Gin Ala Arg 450 455 460 Leu Ser 4 65 <210> 34 <211> 761 <212> PRT <213> Festuca arundinacea <400> 34 Met Ala Arg Ser Asn Trp Glu Ala Asp Lys Met Leu Asp Val Tyr lie 1 5 10 15 Tyr Asp Tyr Leu Val Lys Arg Asn Leu His Asn Ser Ala Lys Ala Phe 20 25 30 Met Asn Glu Gly Lys Val Ala Thr Asp Pro Val Ala He Asp Ala Pro 35 40 45 Gly Gly Phe Leu Phe Glu Trp Trp Ser lie Phe Trp Asp lie Phe Asp 50 55 60 Ala Arg Thr Arg Asp Lys Pro His Gin Gly Ala Thr Ala Ala Ser lie 65 70 75 80 Asp Leu Met Lys Ser Arg Glu Gin Gin Met Arg lie Gin Leu Leu Gin 85 90 95 Gin Gin Asn Ala His Leu Gin Arg Arg Asp Pro Asn His Pro Ala Val 100 105 110 Asn Gly Ala Met Asn Asn Ser Asp Val Ser Ala Phe Leu Val Ser Lys 1-15 120 125 Met Met Glu Glu Arg Thr Arg Asn His Gly Pro Met Asp Ser Glu Ala 130 135 140 Ser Gin Gin Leu Leu Glu Ala Asn Lys Met Ala Leu Leu Lys Ser Ala 145 150 155 160 Ala Ala Asn Gin Thr Gly Pro Leu Gin Gly Ser Ser Val Asn Met Ser 165 170 175 Ala Leu Gin Gin Met Gin Ala Arg Asn Gin Gin Val Asp lie Lys Gly 180 185 190 Asp Gly Ala Met Pro Gin Arg Thr Met Pro Thr Asp Pro Ser Ala Leu 195 200 205 Tyr Ala Ala Gly Met Met Gin Pro Lys Ser Gly Leu Val Ala Ser Gly 210 215 220 Leu Asn Gin Gly Val Gly Ser Val Pro Leu Lys Gly Trp Pro Leu Thr 225 230 235 240 Val Pro Gly lie Asp Gin Leu Arg Ser Asn Leu Gly Ala Gin Lys Gin 245 250 255 Leu Met Pro Ser Pro Asn Gin Phe Gin Leu Leu Ser Pro Gin Gin Gin 260 265 270 Leu lie Ala Gin Ala Gin Thr Gin Asn Asp Leu Ala Arg Met Gly Ser 275 280 285 Pro Ala Pro Ser Gly Ser Pro Lys lie Arg Pro Asn Glu Gin Glu Tyr 290 295 300 Leu lie Lys Met Lys Met Ala Gin Met Gin Gin Ser Gly Gin Arg Met 305 310 315 320 Met Glu Leu Gin Gin Gin Gin His His Leu Gin Gin Gin Gin Gin Gin 325 330 335 Gin Gin His Gin Gin Gin Gin Gin Gin Gin Gin Gin Gin Gin Gin Met 340 345 350 Gin Gin Asn Thr Arg Lys Arg Lys Pro Thr Ser Ser Gly Ala Ala Asn 355 360 365 Ser Thr Gly Thr Gly Asn Thr Val Gly Pro Ser Pro Pro Ser Thr Pro 370 375 380 Ser Thr His Thr Pro Gly Gly Gly lie Pro Val Ala Ser Asn Ala Asn 385 390 395 400 lie Ala Gin Lys Asn Ser Met Val Cys Gly Thr Asp Gly Thr Ser Gly 405 410 415 Phe Ala Ser Ser Ser Asn Gin Met Asp Asn Leu Asp Ser Phe Val Asp 420 425 430 Phe Asp Asp Asn Val Asp Ser Phe Leu Ser Asn Asp Asp Gly Asp Gly 435 440 445 Arg Asp lie Phe Ala Ala Met Lys Lys Gly Pro Ser Glu Gin Glu Ser 450 455 460 Leu Lys Ser Leu Ser Leu Thr Glu Val Gly Asn Asn Arg Thr Ser Asn 465 470 475 480 Asn Lys Val Val Cys Cys His Phe Ser Thr Asp Gly Lys Leu Leu Ala 485 490 495 Ser Ala Gly His Glu Lys Lys Leu Phe Leu Trp Asn Met Asp Asn Phe 500 505 510 Ser Met Asp Thr Lys Ala Glu Glu His Thr Asn Phe lie Thr Asp lie 515 520 525 Arg Phe Arg Pro Asn Ser Thr Gin Leu Ala Thr Ser Ser Ser Asp Gly 530 535 540 Thr Val Arg Leu Trp Asn Ala Val Glu Arg Thr Gly Ala Leu Gin Thr 545 550 555 • 560 Phe His Gly His Thr Ser His Val Thr Ser Val Asp Phe His Pro Lys 565 570 575 Leu Thr Glu Val Leu Cys Ser Cys Asp Asp Asn Arg Glu Leu Arg Phe 580 585 590 Trp Thr Val Gly Gin Asn Ala Pro Ser Arg Val Thr Arg Val Lys Gin 595 600 605 " Gly Gly Thr Gly Arg Val Arg Phe Gin Pro Arg Met Gly Gin Leu Leu 610 615 620 Ala Val Ala Ala Gly Asn Thr Val Asn lie lie Asp lie Glu Lys Asp 625 630 635 640 Thr Ser Leu His Ser Gin Pro Lys Val His Ser Gly Glu Val Asn Cys 645 650 655 lie Cys Trp Asp Glu Ser Gly Glu Tyr Leu Ala Ser Ala Ser Gin Asp 660 665 670 Ser Val Lys Val Trp Ser Ala Ala Ser Gly Ala Cys Val His Glu Leu 675 680 685 Arg Ser His Gly Asn Gin Tyr Gin Ser Cys lie Phe His' Pro Arg Tyr 690 695 700 Pro Lys Val Leu lie Val Gly Gly Tyr Gin Thr Met Glu Leu Trp Ser 705 710 715 720 30 Leu Ser Asp Asn Gin Arg Asn Val Val Ala Ala His Glu Gly Leu lie 725 730 735 Ala Ala Leu Ala His Ser Pro Ser Thr Gly Ser Val Ala Ser Ala Ser 740 745 750 His Asp Lys Ser Val Lys Leu Trp Lys 755 760 <210> 35 <211> 757 <212> PRT <213> Festuca arundinacea <400> 35 Met Ala Arg Ser Asn Trp Glu Ala Asp Lys Met Leu Asp Val Tyr lie 1 5 10 15 Tyr Asp Tyr Leu Val Lys Arg Asn Leu His Asn Ser Ala Lys Ala Phe 20 25 30 Met Asn Glu Gly Lys Val Ala Thr Asp Pro Val Ala lie Asp Ala Pro 35 40 45 Gly Gly Phe Leu Phe Glu Trp Trp Ser -lie Phe Trp Asp lie Phe Asp 50 55 60 Ala Arg Thr Arg Asp Lys Pro Pro Gin Gly Ala Thr Ala Ala Ser lie 65 70 75 80 Asp Leu Met Lys Ser Arg Glu Gin Gin Met Arg lie Gin Leu Leu Gin 85 90 95 Gin Gin Asn Ala His Leu Gin Arg Arg Asp Pro Asn His Pro Ala Val 100 105 110 Asn Gly Ala Met Asn Asn Ser Asp Val Ser Ala Phe Leu Val Ser Lys 115 120 125 Met Met Glu Glu Arg Thr Arg Asn His Gly Pro Met Asp Ser Glu Ala 130 135 140 Ser Gin Gin Leu Leu Glu Ala Asn Lys Met Ala Leu Leu Lys Ser Ala 145 150 155 160 Ala Ala Asn Gin Thr Gly Pro Leu Gin Gly Ser Ser Val Asn Met Ser 165 170 175 Ala Leu Gin Gin Met Gin Ala Arg Asn Gin Gin Val Asp lie Lys Gly 180 185 190 Asp Gly Ala Met Pro Gin Arg Thr Met Pro Thr Asp Pro Ser Ala Leu 195 200 205 Tyr Ala Ala Gly Met Met Gin Pro Lys Ser Gly Leu Val Ala Ser Gly 210- 215 220 Leu Asn Gin Gly lie Gly Ser Val Pro Leu Lys Gly Trp Pro Leu Thr 225 230 ' 235 240 Val Pro Gly lie Asp Gin Leu Arg Ser Asn Leu Gly Ala Gin Lys Gin 245 250 255 Leu Met Pro Ser Pro Asn Gin Phe Gin Leu Leu Ser Pro Gin Gin Gin 260 265 270 Leu lie Ala Gin Ala Gin Thr Gin Asn Asp Leu Ala Arg Met Gly Ser 275 280 285 Pro Ala Pro Ser Gly Ser Pro Lys lie Arg Pro Asn Glu Gin Glu Tyr 290 295 300 Leu lie Lys Met Lys Met Ala Gin Met Gin Gin Ser Gly Gin Arg Met 305 . 310 315 320 Met Glu Leu Gin Gin Gin Gin His His Leu Gin Gin Gin Gin. Gin Gin 325 330 335 Gin Gin His Gin Gin Gin Gin Gin Gin Gin Gin Met Gin Gin Asn Thr 340 345 350 Arg Lys Arg Lys Pro Thr Ser Ser Gly Ala Ala Asn Ser Thr Gly Thr 355 360 365 Gly Asn Thr Val Gly Pro Ser Pro Pro Ser Thr Pro Ser Thr His Thr 370 375 380 31 Pro Gly Gly Gly lie Pro Val Ala Ser Asn Ala Asn lie Ala Gin Lys 385 390 395 400 Asn Ser Met Val Cys Gly Thr Asp Gly Thr Ser Gly Phe Ala Ser Ser 405 410 415 Ser -Asn Gin Met Asp Asn Leu Asp Ser Phe Val Asp Phe Asp Asp Asn 420 425 430 Val Asp Ser Phe Leu Ser Asn Asp Asp Gly Asp Gly Arg Asp lie Phe 435 440 445 Ala Ala Met Lys Lys Gly Pro Ser Glu Gin Glu Ser Leu Lys Ser Leu 450 455 460 Ser Leu Thr Glu Val Gly Asn Asn Arg Thr Ser Asn Asn Lys Val Val 465 470 475 480 Cys Cys His Phe Ser Thr Asp Gly Lys Leu Leu Ala Ser Ala Gly His 485 490 495 Glu Lys Lys Leu Phe Leu Trp Asn Met Asp Asn Phe Ser Met Asp Thr 500 505 510 Lys Ala Glu Glu His Thr Asn Phe lie Thr Asp lie Arg Phe Arg Pro 515 520 525 Asn Ser Thr Gin Leu Ala Thr Ser Ser Ser Asp Gly Thr Val Arg Leu 530 535 540 Trp Asn Ala Val Glu Arg Thr Gly Ala Leu Gin Thr Phe His Gly His 545 550 555 560 Thr Ser His Val Thr Ser Val Asp Phe His Pro Lys Leu Thr Glu Val 565 570 575 Leu Cys Ser Cys Asp Asp Asn Gly Glu Leu Arg Phe Trp Thr Val Gly 580 585 590 Gin Asn Ala Pro Ser Arg Val Thr Arg Val Lys Gin Gly Gly Thr Gly 595 600 605 Arg Val Arg Phe Gin Pro Arg Met Gly Gin Leu Leu Ala Val Ala Ala 610 615 620 Gly Asn Thr Val Asn lie lie Asp lie Glu Lys Asp Thr Gly Leu His 625 630 . 635 640 Ser Gin Pro Lys Val His Pro Gly Glu Val Asn Cys lie Cys Trp Asp 645 650 655 Glu Ser Gly Glu Tyr Leu Ala Ser Ala Ser Gin Asp Ser Val Lys-Val 660 665 670 Trp Ser Ala Ala Ser Gly Ala Cys Val His Glu Leu Arg Ser His Gly 675 680 685 Asn Gin Tyr Gin Ser Cys lie Phe His Pro Arg Tyr Pro Lys Val Leu 690 695 700 lie Val Gly Gly Tyr Gin Thr Met Glu Leu Trp Ser Leu Ser Asp Asn 705 710 715 720 Gin Arg Asn Val Val Ala Ala His Glu Gly Leu lie Ala Ala Leu Ala 725 730 735 His Ser Leu Ser Thr Gly Ser Val Ala Ser Ala Ser His Asp Ser Ser 740 745 750 Val Lys Leu Trp Lys 755 <210> 36 <211> 174 <212> PRT <213> Festuca arundinacea <400> 36 Met Ser Arg Ala Leu Glu Pro Leu Val Val Gly Lys Val lie Gly Glu 15 10 15 Val Leu Asp Ser Phe Asn Pro Thr Val Lys Met Ala Ala Thr Tyr Asn 20 25 30 Ser Asn Lys Gin Val Phe Asn Gly His Glu Phe Phe Pro Ser Ala lie 35 40 45 32 Ala Ala Lys Pro Arg Val Glu Val Gin Gly Gly Asp Leu Arg Ser Phe 50 55 60 Phe Thr Leu Val Met Thr Asp Pro Asp Val Pro Gly Pro Ser Asp Pro 65 70 75 80 Tyr Leu Arg Glu His Leu His Trp lie Val Thr Asp lie Pro Gly Thr 85 90 95 Thr Asp Ala Ser Phe Gly Lys Glu Val Val Asn Tyr Glu Ser Pro Lys 100 105 110 Pro Asn lie Gly lie His Arg Phe lie Leu Val Leu Phe Gin Gin Thr 115 120 125 His Arg Gly Ser Val Lys Asn Thr Pro Ser Ser Arg Asp Arg Phe Arg 130 135 140 Thr Arg Glu Phe Ala Lys Asp Asn Glu Leu Gly Leu Pro Val Ala Ala 145 150 155 160 Val Tyr Phe Asn Ala Gin Arg Glu Thr Ala Ala Arg Arg Arg 165 170 <210> 37 <211> 657 <212> PRT <213> Festuca arundinacea <400> 37 Gin His His His Leu Met Gin Leu Thr Lys Lys Asn Pro Gin Ala Ala 1 5 10 15 Ala Ala Ala Gin Leu Asn Leu Leu Gin Gin Gin Arg lie Met His Met 20 25 30 Gin Gin Gin 35 Gin Gin Gin Gin lie 40 Leu Lys Asn Leu Pro 45 Leu Gin Arg Asn Gin 50 Leu Gin Gin Gin Gin 55 Gin Val Gin Gin Gin 60 Gin Gin Gin Gin Leu Gin Gin Gin Gin Gin Leu Leu Arg Gin Gin Ser Leu Asn Met Arg 65 70 75 80 Thr Pro Gly Lys Ser Pro Pro Tyr Glu Pro Gly Thr Cys Ala Lys Arg 85 90 95 Leu Thr His Tyr 100 Met Tyr His Gin Gin 105 Asn .Arg Pro Gin Asp 110 Asn Asn lie Glu Tyr 115 Trp Arg Asn Phe Val 120 Asn Glu Tyr Phe Ala 125 Pro Thr Ala Lys Lys Arg Trp Cys Val Ser Leu Tyr Gly Ser Gly Arg Gin Thr Thr 130 135 140 Gly Val Phe Pro Gin Asp Val Trp His Cys Glu lie Cys Asn Arg Lys 145 150 155 160 Pro Gly Arg Gly Phe 165 Glu Thr Thr Val Glu 170 Val Leu Pro Arg Leu 175 Cys Gin lie Lys Tyr 180 Ala Ser Gly Thr Leu 185 Glu Glu Leu Leu Tyr 190 lie Asp Met Pro Arg Glu Ser Lys Asn Val Ser Gly Gin lie Val Leu Asp Tyr 195 200 205 Thr Lys Ala lie Gin Glu Ser Val Phe Asp Gin Leu Arg Val Val Arg 210 215 220 Glu Gly His Leu Arg lie lie Phe Asn Pro Asp Leu Lys lie Ala Ser 225 230 235 240 Trp Glu Phe Cys Ala Arg Arg His Glu Glu Leu lie Pro Arg Arg Ser 245 250 255 lie lie Pro Gin Val Ser Gin Leu Gly Ala Val Val Gin Lys Tyr Gin 260 265 270 Ala Ala Ala Gin Asn Pro Thr Ser Leu Ser Thr Gin Asp Met Gin Asn 275 280 285 Asn Cys 290 Asn Ser Phe Val Ala 295 Cys Ala Arg Gin Leu 300 Ala Lys Ala Leu 33 Glu Val Pro Leu Val Asn Asp Leu Gly Tyr Thr Lys Arg Tyr Val Arg 305 310 315 320 Cys Leu Gin lie Ala Glu Val Val Asn Cys Met Lys Asp Leu lie Asp 325 330 335 His Ser Arg Gin Thr Gly Ser Gly Pro lie Asp Ser Leu His Lys Phe 340 345 350 Pro Arg Arg Thr Pro Ser Gly lie Asn Pro Leu Gin Ser Gin Gin Gin 355 360 365 Gin Pro Glu Glu His Gin Ser Val Pro Gin Ser Ser Asn Gin Ser Gly 370 375 380 Gin Asn Ser Ala Pro Met Ala Gly Val Gin Val Ser Ala Ser Ala Asn 385 390 395 400 Ala Asp Ala Thr Ser Asn Asn Ser lie Asn Cys Ala Pro Ser Thr Ser 405 410 415 Ala Pro Ser Pro Thr Val Val Gly Leu Leu Gin Gly Ser Met Asp Ser 420 425 430 Arg His Asn His Pro Met Cys Ser Ala Asn Gly Gin Tyr Asn Ser Gly 435 440 445 Asn Asn Gly Ala lie Pro Arg Val Asn Ser Ala Ser Ser Leu Gin Ser 450 455 460 Asn Pro Ser Ser Pro Phe Pro Ser Gin Val Pro Thr Ser Pro Asn Asn 465 470 475 480 Asn Met Met Pro Thr Leu Gin Asn Ala Asn Gin Leu Ser Ser Pro Pro 485 490 495 Ala Val Ser Ser Asn Leu Pro Pro lie Gin Pro Pro Ser Thr Arg Pro 500 505 510 Gin Glu Ser Glu Pro Ser Asp Ala Gin Ser Ser Val Gin Arg lie Leu 515 520 525 Gin Glu Met Met Ser Ser Gin Met Asn Gly Val Gly His Gly Gly Asn 530 535 540 Asp Met Lys Arg Pro Asn Gly Leu Thr Pro Gly lie Asn Gly Val Asn 545 550 555 560 Cys Leu Val Gly Asn Ala Val Thr Asn His Ser Gly Met Gly Gly Met 565 570 575 Gly Phe Gly Ala Met Gly Gly Phe Gly Ser Thr Pro Ala Ala Ser Gly 580 585 590 Leu Arg Met Ala Met Thr Asn Asn Ala Met Ala Met Asn Gly Arg Met 595 600 605 Gly Met His His Ser Ala Gin Asp Leu Ser Gin Leu Gly Gin Gin His 610 615 620 Gin His Gin His Gin His Asp lie Gly Asn Gin Leu Leu Gly Gly Leu 625 630 635 640 Gly Ala Ala Asn Ser Phe Asn Asn lie Gin Tyr Asp Trp Lys Pro Ser 645 650 655 Gin <210> 38 <211> 808 <212> PRT <213> Festuca arundinacea <400> 38 Met Ser Gly Ala Pro Arg Ser Asn Leu Gly Phe Val Ala Arg Asp Met 1 5 10 15 Asn Gly Ser lie ■Pro Val Ser Ser Ala Asn Ser Ser Gly Pro Ser lie 20 25 30 Gly Val Ser Ser Leu Val Thr Asp Gly Asn Ser Ser Leu Ser Gly Gly 35 40 45 Ala Gin Phe Gin His Ser Thr Ser Met Asn Ala Asp Ser Phe Met Arg 50 55 60 34 Leu Pro Ala Ser Pro Met Ser Phe Ser Ser Asn Asn lie Ser Gly Ser 65 70 75 80 Ser Val lie Asp Gly Ser lie Met Gin Gin Ser Pro Pro Gin Asp Gin 85 90 95" Met Gin Lys Arg Arg Ser Ser Thr Ala Thr Ser Gin Pro'Gly lie Glu 100 105 110 Ala Gly Ala Ala Phe His Ala Gin Lys Lys Pro Arg Val Asp I-le Arg 115 120 125 Gin Asp Asp lie Leu Gin Gin His Leu lie Gin Gin Val Leu Gin Gly 130 135 140 Gin Ser Ser Leu His Leu Pro Gly Gin His Asn Pro Gin Leu Gin Ala 145 150 155 160 Leu lie Arg Gin Gin Lys Leu Ala His lie Gin His Leu Gin Gin Gin 165 170 175 Gin Leu Ser Gin Gin Phe Pro Gin lie Gin Gin Ser Gin Val Gly lie 180 185 190 Pro Arg Gin Pro Gin Leu Arg Leu Pro Leu Ala Gin Pro Gly Met Gin 195 200 205 Leu Ala Gly Pro Val Arg Thr Pro Val Glu Ser Gly Leu Cys Ser Arg 210 215 220 Arg Leu Met Gin Tyr Leu Phe His Lys Arg His Arg Pro Glu Asp Asn 225 230 235 240 Pro He Thr Tyr Trp Arg Lys Leu lie Asp Glu Tyr Phe Ala Pro Arg 245 250 255 Ala Arg Glu Arg Trp Cys Val Ser Ser Tyr Glu' Lys Arg Gly Asn Ser 260 265 270 Pro Val Ala lie Pro Gin Thr Ser Gin Asp Thr Trp Arg Cys Asp lie 275 280 285 Cys Asn Thr His Ala Gly Lys Gly His Glu Ala Thr Tyr Glu lie Leu 290 295 300 Pro Arg Leu Cys Gin lie Arg Phe Asp Gin Gly Val lie Asp Glu Tyr 305 310 315 320 Leu Phe Leu Asp Met Pro Asn Glu Phe Arg Leu Pro Asn Gly Leu Leu 325 330 335 Leu Leu Glu His Thr Lys Val Val Gin Lys Ser lie Tyr Asp His Leu 340 345 ■ ■ 350 His Val Thr His Glu Gly Gin Leu Arg lie lie Phe Thr Pro Glu Leu 355 360 365 Lys lie Met Ser Trp Glu Phe Cys Ser Arg Arg His Asp Glu Tyr lie 370 375 380 Thr Arg Arg Phe Leu Thr Pro Gin Val Asn His Met Leu Gin Val Ala 385 390 395 400 Gin Lys Tyr Gin Ala Ala Ala Asn Glu Ser Gly Pro Ala Gly Val Ser 405 410 415 Asn Asn Asp Ala Gin Ala lie Cys Ser Met Phe Val Ser Ala Ser Arg 420 425 430 Gin Leu Ala Lys Asn Leu Asp His His Ser Leu Asn Glu His Gly Leu 435 440 445 Ser Lys Arg Tyr Val Arg Cys Leu Gin lie Ser Glu Val Val Asn His 450 455 460 Met Lys Asp Leu lie Glu Phe Ser His Lys Asn Lys Leu Gly Pro lie 465 470 475 480 Glu Gly Leu Lys Asn Tyr Pro Arg Gin Thr Gly Pro Lys Leu Thr Thr 485 490 495 Gin Asn Met His Asp Ala Lys Gly Val Val Lys Thr Glu Glu Ser Thr 500 505 510 His Val Asn Asn Glu Gly Pro Asp Ala Gly Pro Ala Gly Ser Ser Pro 515 520 525 Gin Asn Ala Gly Ala Gin Asn Asn Tyr Gin Asn Met Leu Arg Ser Pro 530 535 540 Sex Pro Asn Gin Gly Leu Thr His Gin Glu Ala Ser Gin Asn Ala Ala 35 545 550 555 560 Ala Leu Asn Asn Tyr Gin Asn Met Leu Arg Ser Ser Ser Ala Asn Gin 565 570 575 Gly Leu Leu Gin Gin Glu Ala Ser Gin Asn Val Ser Gly Leu Asn Asn 580 585 590 Tyr Gin Asn Met Leu Arg Ser Ser Ser Ala Asn Gin Ser lie Leu Gin 595 600 605 Gin Glu Ala Ser Ser lie Phe Lys Gly Pro Thr Gly Val His Ser Ser 610 615 620 lie Gin Leu Glu Ala Ala Arg Ser Phe Arg Ala Ala Gin Leu Gly Pro 625 630 635 640 Met Ser Phe Gin Gin Ala Val Pro Leu Tyr Gin Gin Asn Arg Phe Gly 645 650 655 Ala Gly Val Ser Pro Gin Tyr Gin Gin His Val Met Gin Gin Leu Leu 660 665 670 Gin Glu Ala Asn Arg Ser Thr Asn Asn Arg Val Leu Ala Gin Gin Gin 675 680 685 Pro Leu Ser Thr Pro Asn Ala Asn Gly Gly Leu Thr lie Thr Asn Ser 690 695 700 Gly Ala Ser Gly Asp Gin Ala Gin His Met Asn Asn Asn Gly Ala Ala 705 710 715 . 720 Lys Gly Val Ala Ala Pro Met Gly Met Ala Gly Thr Ser Asn Leu lie 725 730 735 Asn Ser Gly Ser Ala Gly Val Val Gin Arg Cys Ser Ser Phe Lys Ser 740 745 750 Val Thr Ser Asn Pro Ala Ala Ala Ala Ala Gly Asn Leu Leu Thr Pro 755 760 765 Lys Ala Glu Ser Met His Glu Met Asp Glu Leu Asp His Leu lie Thr 770 775 780 Ser Glu Leu Ala Glu Ser Gly Leu Phe Met Gly Glu Gin Gin Gly Gly 785 790 795 800 Gly Gly Gly Tyr Ser Trp His Met 805 <210> 39 <211> 798 <212> PRT <213> Festuca arundinacea <4 00> 39 Ser Asp Pro Leu Ser Phe Pro Ser Ser Ser His Val Ser Leu Gly Asn 1 5 10 15 His lie Ser Ser 20 Asp Asn Leu Gin Gin Gin Gin 25 Gin Met Asp 30 Met Pro Asp Leu Gin 35 Gin Gin Gin Gin Gin 40 Gin Gin Arg Gin Leu 45 Pro Met Ser Tyr Asn 50 Gin Gin His Leu Pro 55 Met Gin Arg Pro Gin 60 Pro Gin Ala Thr Val Lys Leu Glu Asn Gly Gly Ser Met Gly Gly Val Lys Met Glu Gin 65 70 75 80 Gin Thr Gly His Pro Asp Gin Asn Gly Pro Ala Gin Met Met His Asn 85 90 95 Ser Gly Asn Val Lys Phe Glu Pro Gin Gin Leu Gin Ala Leu Arg Gly 100 105 110 Leu Gly Thr Val Lys Met Glu Gin Pro Asn Ser Asp Pro Ser Ala Phe 115 120 125 Leu Gin 130 Gin Gin Gin Gin Gin 135 Gin Gin Gin His His 140 His Leu Met Gin Leu Thr Lys Gin Asn Pro Gin Ala Ala Ala Ala Ala Gin Leu Asn Leu 145 150 155 160 Leu Gin Gin Gin Arg lie Met His Met Gin Gin Gin Gin Gin Gin His 36 165 170 175 lie Leu Lys Asn Met Pro Leu Gin Arg Asn Gin Leu Gin Gin Gin Gin 180 185 190 Gin Gin Gin Gin Gin Leu Gin Gin Gin Gin His Gin Gin Leu Leu Arg 195 200 205 Gin Gin Ser Leu Asn Met Arg Thr Pro Gly Lys Ser Pro Pro Tyr Glu 210 215 220 Pro Gly Thr Cys Ala Lys Arg Leu Thr His Tyr Met Tyr His Gin Gin 225 230 235 240 Asn Arg Pro Gin Asp Asn Asn Val Glu Tyr Trp Arg Asn Phe Val Asn 245 250 255 Glu Tyr Phe Ala Pro Thr Ala Lys Lys Arg Trp Cys Val Ser Leu Tyr 260 265 270 Gly Ser Gly Arg Gin Thr Thr Gly Val Phe Pro Gin Asp Val Trp His 275 280 285 Cys Glu lie Cys Asn Arg Lys Pro Gly Arg Gly Phe Glu Thr Thr Val 290 295 300 Glu Val Leu Pro Arg Leu Cys Gin lie Lys Tyr Ala Ser Gly Thr Leu 305 310 315 320 Glu Glu Leu Leu Tyr lie Asp Met Pro Arg Glu Ser Lys Asn Val Ser 325 330 335 Gly Gin lie Val Leu Asp Tyr Thr Lys Ala lie Gin Glu Ser Val Phe 340 345 350 Asp Gin Leu Arg Val Val Arg Glu Gly His Leu Arg lie He Phe Asn 355 360 365 Pro Asp Leu Lys lie Ala Ser Trp Glu Phe Cys Ala Arg Arg His Glu 370 375 380 Glu Leu lie Pro Arg Arg Ser lie lie Pro Gin Val Ser Gin Leu Gly 385 390 395 400 Ala Val Val Gin Lys Tyr Gin Ala Ala Ala Gin Asn Pro Thr Ser Leu 405 410 415 Ser Thr Gin Asp Leu Gin Asn Asn Cys Asn Ser Phe Val Ala Cys Ala 420 425 430 Arg Gin Leu Ala Lys Ala Leu Glu Val Pro Leu Val Asn Asp Leu Gly 435 440 445 Tyr Thr Lys Arg Tyr Val Arg Cys Leu Gin lie Ala Glu Val Val Asn 450 455 460 Cys Met Lys Asp Leu lie Asp His Ser Arg Gin Thr Gly Ser Gly Pro 465 470 475 480 lie Asp Ser Leu His Lys Phe Pro Arg Arg Thr Pro Ser Gly lie Asn 485 ' 490 495 Pro Leu Gin Ser Gin Gin Gin Pro Pro Glu Glu Gin Gin Ser Val Pro 500 505 510 Gin Ser Ser Asn Gin Ser Gly Gin Asn Ser Ala Pro Met Ala Gly Val 515 520 525 Gin Val Ser Ala Ser Ala Asn Ala Asp Ala Thr Ser Asn Asn Ser Leu 530 535 540 Asn Cys Ala Pro Ser Thr Ser Ala Pro Ser Pro Thr Val Val Gly Leu 545 550 555 560 Leu Gin Gly Ser Met Asp Ser Arg Gin Asp His Pro Met Cys Ser Ala 565 570 575 Asn Gly Gin Tyr Asn Ser Gly Asn Asn Gly Ala lie Pro Arg Val Asn 580 585 590 Ser Ala Ser Ser Leu Gin Ser Asn Pro Ser Ser Pro Phe Pro Leu Gin 595 600 . 605 Val Pro Thr Ser Pro Asn Asn'Asn Met Met Pro Thr Leu Gin Asn Ala 610 615 620 Asn Gin Leu Ser Ser Pro Pro Ala Val Ser Pro Asn Leu Pro Pro Met 625 630 635 640 Gin Pro Pro Ser Thr Arg Pro Gin Glu Ser Glu Pro Ser Asp Ala Gin 645 650 655 37 Ser Ser Val Gin Arg lie Leu Gin Glu Met Met Ser Ser Gin Met Asn 660 665 670 Gly Val Gly His Ala Gly Asn Asp Met Lys Arg Pro Asn Gly Leu Thr 675 680 685 Pro Gly lie Asn Gly Val Asn Cys Leu Val Gly Asn Ala Val Thr Asn 690 695 700 His Ser Gly Met Gly Gly Met Gly Phe Gly Ala Met Gly Gly Phe Gly 705 710 715 720 Ser Asn Pro Ala Ala Ser Gly Leu Arg Met Ala Met Thr Asn Asn Thr 725 730 735 Met Ala Met Asn Gly Arg Met Gly Met His His Ser Ala His Asp Leu 740 745 750 Ser Gin Leu Gly Gin Gin His Gin His Gin His Gin His Gin His Gin 755 760 765 His Gin His Asp lie Gly Asn Gin Leu Leu Gly Gly Leu Arg Ala Thr 770 77-5 780 Asn Ser Phe Asn Asn lie Gin Tyr Asp Trp Lys Pro Ser Gin 785 790 795 <210> 40 <211> 609 <212> PRT <213> Festuca arundinacea <400> 40 Met Lys Arg Glu Tyr Gin Asp Ala Gly Gly Ser Ser Ala Gly Gly Asp 1 5 10 15 Met Gly Met Ser 20 Lys Asp Lys Met Met 25 Ser Ala Pro Pro Ala 30 Gin Glu Asp Glu Asp Val Asp Glu Leu Leu Ala Ala Leu Gly Tyr Lys Val Arg 35 40 45 Ser Ser 50 Asp Met Ala Asp Val 55 Ala Gin Lys Leu Glu 60* Gin Leu Glu Met Ala Met Gly Met Gly Gly Val Pro Ala Pro Asp Asp Gly Phe Thr Thr 65 70 75 80 His Leu Ala Thr Glu Thr Val His Tyr Asn Pro Thr Asp Leu Ser Ser 85 90 95 Trp Val Glu Ser Met Leu Ser Glu Leu Asn Ala Pro Pro Pro Leu Pro 100 105 110 Pro Ala Pro 115 Arg Leu Ala Pro Ala 120 Ser Ala Ser Val Thr 125 Ala Asp Gly Phe Phe 130 Asp lie Pro Pro Pro 135 Ser Val Asp Ser Ser 140 Ser Ser Thr Tyr Ala Leu Arg Pro lie Pro Ser Pro Ala Asp Leu Ser Ala Asp Leu Ser 145 150 155 160 Ala Asp Ser Pro Arg Asp Pro Lys Arg Met Arg Thr Gly Gly Gly Ser 165 170 175 Thr Ser Ser Ser Ser Ser Ser Ser Ser Ser Leu Gly Gly Cys Val Val 180 185 190 Glu Ala Ala 195 Pro Pro Ala Ala Ala 200 Glu Ala Asn Ala lie 205 Ala Leu Pro Val Val 210 Val Ala Asp Thr Gin 215 Glu Ala Gly lie Arg 220 Leu Val His Ala Leu Leu Ala Cys Ala Glu Ala Val Gin Gin Glu Asn Phe Ser Ala Ala 225 230 235 240 Glu Ala Leu Val Lys 245 Gin lie Pro Leu Leu 250 Ala Ala Ser Gin Gly 255 Gly Ala Met Arg Lys Val Ala Ala Tyr Phe Gly Glu Ala Leu Ala Arg Arg 260 265 270 Val Phe Arg Phe Arg Pro Gin Pro Asp Ser Ser His Leu Asp Ala Ala 275 280 285 38 Phe Ala Asp 290 Lys Phe Ala 305 Gly Cys Arg Gin Trp Pro Pro Ser Phe 355 Asp Ala Leu 370 lie Gly Val 385 Asp Leu Glu Glu Glu Pro Leu Leu Ala 435 Ala Val Arg 450 Asn Ala Gly 4 65 Ser Thr Met lie Ser Ser Val Met Ser 515 Cys Glu Gly 530 Arg Gly Arg .545 Asn Ala Tyr Asp Gly Tyr His Thr Arg 595 Pro Leu Leu His Ala 295 His Phe Thr Ala 310 Arg Val His Val 325 Ala Leu Leu Gin 340 Arg Leu Thr Gly Gin Gin Val Gly 375 Asp Phe Gin Tyr 390 Pro Phe Met Leu 405 Glu Val lie Ala 420 Gin Pro Gly Ala Pro Arg lie Val 455 Ser Phe Leu Asp 470 Phe Asp Ser Leu 485 Gly Pro Ala Ala 500 Glu Val Tyr Leu Ala Glu Arg Thr 535 Leu Gly His Ala 550 Lys Gin Ala Ser 5 65 Lys Val Asp Glu 580 Pro Leu lie Ala His Phe Tyr Glu Ser 300 Asn Gin Ala lie Leu 315 Val Asp Phe Gly lie 330 Ala Leu Ala Leu Arg 345 Val Gly Pro Pro Gin 360 ' Trp Lys Leu Ala Gin 380 Arg Gly Leu Val Ala 395 Gin Pro Glu Ala Glu 410 Val Asn Ser lie Phe 425 Leu Glu Lys Val Leu 440 Thr Val Val Glu Gin 460 Arg Phe Thr Glu Ser 475 Glu Gly Ala' Gly Ser 490 Ala Ala Ala Ala Pro 505 Gly Arg Gin lie Cys 520 Glu Arg His Glu Thr 540 Gly Phe Glu Thr Val 555 Thr Leu Leu Ala Leu 570 Lys Glu Gly Cys Leu 585 Thr Ser Ala Trp Arg 600 Cys Pro Tyr Leu Glu Ala Phe Ala 320 Lys Gin Gly Met 335 Pro Gly Gly Pro 350 Pro Asp Glu Thr 365 Phe Ala His Thr Ala Thr Leu Ala 400 Asp Gly Pro Asn 415 Glu Met His Arg 430 Gly Thr Val Arg 445 Glu Ala Asn His Leu His Tyr Tyr 480 Gly Pro Ser Glu 495 Gly Thr Asp Gin 510 Asn Val Val Ala 525 Leu Gly His Trp His Leu Gly Ser 560 Phe Ala Gly Gly 575 Thr Leu Gly Trp 590 Met Ala Ala Ala 605 39