BRPI0714552A2 - identification of a type of sucrose synthase and its use in fiber modification - Google Patents

Abstract

Identification of a type of sucrose synthase and use of same in fiber modification. The present invention relates to methods and means for modifying fiber characteristics in a fiber producing plant such as cotton based on the use of a new type of sucrose synthase protein or related nucleic acids.

Description

Report of the Invention Patent for "IDENTIFICATION OF A TYPE OF SACAROSIS SYNTASE AND USE OF THE SAME IN FIBER MODIFICATION".

Field of the Invention

The invention relates to the field of agriculture, more specifically, to the use of molecular biology techniques to alter fiber producing plants, particularly cotton plants, and / or to accelerate the generation of such fiber containing plants. Methods and means are provided for modifying fiber quality, for example by increasing or decreasing cellulose contents, fiber strength, fiber uniformity or micronaire (fiber diameter index). Methods are also provided to identify molecular markers associated with such characteristics in a population of cotton varieties and related parent plants. Prior art

Much of the high quality fiber for the textile industry is supplied by cotton. About 90% of cotton grown worldwide is Gossypium hirsutum L., while Gossypium barbadense accounts for about 8%. Therefore, modifying cotton fiber characteristics to better meet industry needs is a major generation effort through either classical methods or genetically altering the genome of cotton plants. Objectives to be achieved include longer cotton fiber length, strength, dyeing capacity, lower down fiber production, fiber maturity ratio, immature fiber content, fiber uniformity, and micronaire.

US 6472588 and WO 0117333 provide methods for enhancing the quality of cotton fiber produced from a cotton plant by transformation with a DNA encoding sucrose phosphate synthase. Fiber qualities include strength, length, fiber maturity ratio, immature fiber content, fiber uniformity and micro-grain.

W09508914 describes a fiber producing plant comprising in its genome a heterologous genetic construct. The genetic construct comprises a fiber specific promoter and a coding sequence encoding a plant peroxidase, such as a cotton peroxidase.

W09626639 provides methods whereby preference coding sequence directing gene expression in ovarian tissue, particularly very early in fruit development, are used to express plant growth modifying hormones in cotton egg tissue. The methods allow modification of the cotton plant capsule characteristics and provide a mechanism for altering fiber quality characteristics such as fiber size and strength.

US5981834, US5597718, US5620882, US5521708 and US 5495070 all describe a method for genetically engineering a fiber producing plant and identifying cDNA clones useful for the identification of cotton fiber genes. CDNA clones are useful in developing corresponding genomic clones of fiber producing plants to allow genetic engineering of cotton and other plants using these genes. Coding sequences of these isolated genes are used in sense and antisense orientation to alter the fiber characteristics of transgenic fiber producing plants.

Published US patent applications US2002049999 and US2003074697 both disclose Gossypium cotton plants with improved cotton fiber characteristics. The cotton plant has an expression cassette containing a gene encoding an enzyme selected from the group consisting of endoxyloglucan transferase, catalase and peroxidase so that the gene is expressed in cotton fiber cells to improve the characteristics of the cotton fiber.

US5880110 produces cotton fibers with improved fiber characteristics through treatment with brassinoids.

WO 01/40250 provides methods for improving the quality of cotton fiber by modulating transcription factor gene expression. WO 96/40924 provides new DNA constructs that can be used as molecular probes or inserted into a plant host to provide transcription modification of a DNA sequence of interest during various stages of cotton fiber development. DNA constructs comprise a cotton fiber transcriptional initiation regulatory region associated with a gene, which is expressed in cotton fiber. It is also new cotton having a cotton fiber that has a natural color introduced by expression in pigment fiber cells using such a construct of pigment genes.

EP0834566 provides a gene that controls the fiber forming mechanism in a cotton plant and which can be used for industrially useful improvement.

Ruan et al 2003) (The Plant Celll vol. 15, 952-964) report that suppression of sucrose synthase gene expression suppresses cell initiation, elongation and seed development of cotton fiber. The results described here provide direct evidence that sucrose synthase expression in the epidermis of the egg is crucial for initiation and elongation of single cell fibers. The results described also allow us to conclude that suppression of Sus in the seed coat only reduces fiber growth without affecting embryo development and seed size. The paper concludes that new insights into the role of Sus in controlling plant and seed cell development offer opportunities for modifying fiber and seed development through genetic expression of Sus.

W00245485 describes methods and means for modulating fiber quality in fiber producing plants such as cotton by modulating and / or expressing sucrose synthase activity in such plants.

Baud et al 2004 (Journal of Experimental Botany, Vol. 55, N-396, pp. 397-409) describe the structure and expression profile of the multigene family of sucrose synthase having six members in Arabidopsis.

No prior art document describes the recognition of the currently recognized type of sucrose synthase, nor its involvement in developing secondary plant cell wall, such as type C in plants. This new type of Sus protein opens the possibility for new methods and means for altering fiber characteristics of fiber producing plants such as cotton, which may be further combined with any of the methods for altering fiber characteristics. Such methods and means are described in the embodiments and claims described below. Summary of the Invention

In one embodiment of the invention, a new type of protein saccharose synthase is provided having an amino acid sequence comprising an amino acid sequence selected from an amino acid sequence having at least 50% sequence homology to the amino acid sequence. amino acid frequency of any SEQ ID NO .: 2, 3, 5, 7, 9, 11 or 13; or an amino acid sequence comprising the amino acid sequence of any SEQ ID NO .: 2, 3, 5, 7, 9, 11 or 13; an amino acid sequence located at the amino terminal part of said protein, said amino acid sequence comprising at least about 60% sequence identity with the amino acid sequence of SEQ ID NO .: 16; or an amino acid sequence located at the carboxy terminal portion of said protein, said amino acid sequence comprising at least about 60% sequence identity with the amino acid sequence of SEQ ID NO. The protein sucrose synthase may comprise a hydrophobic N-terminal sequence. It may also comprise any of the following amino acid sequences: the amino acid sequence of SEQ ID NO: 3 from amino acid 383 to amino acid 394; the amino acid sequence of SEQ ID NO: 3 from amino acid 270 to amino acid 329; the amino acid sequence of SEQ ID NO: 3 from amino acid 549 to amino acid 737; the amino acid sequence of SEQ ID NO: 3 from amino acid 1 to amino acid 545; or the amino acid sequence of SEQ ID NO: 3 from amino acid 18 to amino acid 794.

In another embodiment of the invention, an antibody recognizing isolated new sucrose synthase protein is provided.

In yet another embodiment, the invention provides an isolated DNA or nucleic acid molecule encoding the new sucrose synthase protein, such as nucleic acid comprising the nucleotide sequence of any SEQ ID N9: 1, 4, 6, 8, 10, 12 or 14

The invention further provides an expression cassette comprising the operably linked DNA molecules that follow: a plant expressible promoter such as a plant expressible promoter preferably controlling transcription in fiber cells of a producing plant fiber; a DNA encoding the new sucrose synthase proteins produced; and optionally a transcription termination region and polyadenylation.

The invention also provides an expression cassette comprising operably linked DNA molecules: a plant expressible promoter a DNA which when transcribed results in an RNA molecule said RNA molecule comprising or a nucleotide sequence of at least 19 consecutive nucleotides having at least about 94% sequence identity with the nucleotide sequence of a gene encoding endogenous sucrose synthase C isoform or complementing said nucleotide sequence of a gene encoding sucrose C isoform endogenous synthase or nucleic acid sequence of at least 19 consecutive nucleotides having at least about 94% sequence identity with a new type C isoform sucrose synthase nucleotide sequence; and optionally a transcription and polyadenylation termination region.

Another embodiment of the invention is an expression cassette comprising the operably linked DNA molecules that follow: a plant expressible promoter, preferably a plant expressible promoter that preferably controls transcription in fiber cells; a DNA, which when transcribed gives a double stranded RNA molecule capable of reducing the expression of an endogenous SusC gene to the fiber producing plant, and the RNA molecule comprising a first and a second region of RNA where the first region of RNA comprises a nucleotide sequence of at least 19 consecutive nucleotides having at least about 94% sequence identity with the nucleotide sequence of an endogenous SusC gene or nucleotide sequence of the provided nucleotide sequences encoding proteins. SusC; the second RNA region comprising a nucleotide sequence complementary to at least 19 consecutive nucleotides from the first RNA region; the first and second RNA regions are capable of base pairing to form a double stranded RNA molecule between at least 19 consecutive nucleotides of the first and second regions; and optionally a 3 'end region comprising transcription termination and polyadenylation signals functioning in plant cells.

The invention further provides a plant cell comprising a heterologous plant expressible promoter operably linked to a DNA molecule selected from the following DNA molecules: a DNA encoding a new isoform C sucrose synthase or encoding an inhibitor RNA for a gene encoding SusC. The cell may be a cell of a fiber producing plant such as cotton. Also encompassed by the invention are plants, seeds or plant parts or tissues which comprise or consist essentially of such plant cells as well as fibers produced by such plants.

In still another embodiment of the invention, a method for modifying the fiber characteristics of a fiber producing plant is provided, said method comprising modification, such as increasing or decreasing the functional level of a sucrose C isoform. synthase or a sucrose synthase having similar characteristics in cells or cell walls or apoplastic fluid of said plant. This can conveniently be accomplished by providing the plants with the expression cassettes described herein.

The invention also comprises use of a novel SusC-like protein or coding nucleic acid to modify the characteristics of a fiber in a fiber producing plant. Brief Description of the Figures and Drawings Figure 1. Hydrophobicity graphs of SusC proteins and similar sucrose proteins. A. Chart for SusC of a variety Gossypium hirsutum; B: Chart for SusC of a variety Gossypium barbadense \ C: Chart for SusC of Gossypium arboretum; D: Gossypium raymondir SusC Chart; E: Gossypium Iongicalyx SusC Chart Subtype 1; F: Graph for SusC of Gossypium Iongicalyx subtype 2; G: Graph for sucrose synthase of Vigna radiata; H: Graph for Eucalyptus grandis sucrose synthase; I: Graph for sucrose synthase 6 isoforms from Ara-bidopsis thaliana-, K: Graph for sucrose synthase from Populus tremuloides. All hydrophobicity charts depict a hydrophobic N-terminus for different sucrose synthases.

Figure 2. Analysis of SusC gene expression in cotton fiber tissue by RT-PCR. Dpa: days after anesthesia; gDNA: control genomic DNA. Tracks 1 and 8: 1 kb marker. Figure 3: Introns and exons in genes encoding sucrose synthase

cotton The intron exon distribution is schematically represented for sucrose synthase type C and compared to sucrose synthase types BeA cotton. Black boxes represent exons.

Figure 4. 2D structure analysis of the N-terminal part of cotton sucrose synthase genes. 2D analytical structure analysis (HCA + JPRED) of the N-terminal part of the susyC gene compared to the other sucrose synthases of Gossypium hirsutum. The arrow shows the influence of the specific susyC region on the surrounding 2d structure. (P) = putative phosphorylation sites.

Figure 5: 2D structure analysis of the C-terminal part of genes.

sucrose synthase cotton. The beta sheet structure appears to be missing from the susyC 2D structure.

Figure 6: Phenotypic analysis of transgenic Arabidopsis plants. Fasciated, bifurcated, thick trunks are shown in 2 independent transgenic lineages carrying CaMV35S-SusC chimeric genes (panels A and B). CeD panels show a typical trichome phenotype in the T2 generation of CaMV 35S-SusC and S2A-SusC chimeric genes. Panel C is a three-dimensional photomontage.

Figure 7: Western blot of CaMV 35S-SusC transgene seedlings of 12 using SUS C-specific antibody. Lane A: light phenotype plants; Lane B: plant with severe phenotype; Lane C: plant with mild phenotype; Lane D: plant with severe phenotype; WT: wild type plant. Detailed Description

The present invention is based on the identification of a novel sucrose synthase isoform (called SusC or SuSyC) of fibers whose RNA level expression profile is closely related to the onset of secondary cell wall development phase in producing plants. of fiber such as cotton. The protein is abundant during secondary cell wall formation and is predominantly located in the apoplast. Its localization, abundance and expression profile indicate that this new type of sucrose synthase isoform is the main isoform present in the cell wall at the secondary cell wall synthesis stage. Without wishing to limit the invention to a particular mode of action, it is thought that this sucrose synthase isoform is involved in sequestering sugars from the apoplastic fluid surrounding the fiber and incorporating them as UDPGlycosis into cellulose and / or callose. Thus, in a first embodiment, the invention provides a

A method for modifying cell wall characteristics in a plant cell characterized by the fact that the functional level of this new sucrose synthase (SusC) or sucrose synthase isoform with similar characteristics is modified. The functional level of sucrose synthase type C or isoform

Carose synthases with similar characteristics can be modified by increased expression of such isoform in the plant cell. This can conveniently be accomplished by introducing an expression construct comprising operably linked nucleic acids that follow, for example, DNA molecules:

a) an expressible plant promoter

(b) a nucleic acid encoding a sucrose synthase isoform C or a sucrose synthase having similar characteristics; and optionally

c) a transcription termination and polyadenylation region.

As used herein, "sucrose synthase" refers to an enzyme that is capable of catalyzing the synthesis of sucrose from NDP-glucose (such as uridine diphosphate glucose) and D-fructose. The enzyme may also catalyze sucrose hydrolysis into glucose and fructose. The enzyme is classified as EC 2.4.1.13. Synonyms for sucrose synthase are glycosyltransferase, uridine diphosphoglycosis fructose, sucrose synthetase, sucrose-UDP glycosyltransferase, sucrose uridine diphosphate glycosyltransferase, Sus, SuSy, UDP-glucose-fructose glycosyltransferase, UDP-glucose 2-D: alpha glycosyltransferase and uridine diphosphoglycosis-fructose glycosyltransferase.

Several plant genes encoding sucrose synthase enzymes have been described. W002 / 45485 (pages 16 to 20) provides an extensive list of Genbank accession numbers for nucleotide sequences encoding plant sucrose synthase genes, parts thereof or nucleotide sequences having sequence similarity to nucleotide genes. sucrose synthase.

Enzymatic assays for sucrose synthase activity have been described (see, for example, Counce and Gravois, 2006, Crop Science, Vol. 46: pp. 1501-1507, particularly pages 1502 and 1503; Anthon and Emerich, 1990, Vol. 92: pp. 346-351, particularly page 347; both documents incorporated herein by reference).

The "sucrose synthase isoform C" or "SusC" or SuSyC "is characterized by the presence of a hydrophobic N-terminal amino acid sequence (See Figure 1). The first 44 amino acids form a hydrophobic region, particularly residues 26. at 44 da, for example, SEQ ID NO.:3 Other plant sucrose synthase enzymes characterized by such hydrophobic N-terminal amino acid sequence are Arabidopsis thaliana sucrose synthase 6 (Accession number At1g73370); mungbean bean (Accession number D10266), Sucrose synthase from Eucalyptus spp. (Accession number DD014303) and Sucrose synthase (Accession number AY341026) (all sequences incorporated herein by reference).

Alignment of different SusC amino acid sequences and comparison with other plant sucrose synthases disclose the presence of 20-25 consecutive amino acid extensions located at the N (SEQ ID NO: 16) or C terminal (SEQ ID NO: 15) respectively, which are conserved within the amino acid sequences of SusC and absent from the other plant sucrose synthases. The N-terminal sequence (HKSQKLLSVLDKEAGNQALDGMVV; SEQ ID No. 16) is generally located between amino acid position 36 and amino acid position 59, while the C-terminal sequence (AYQEQRGRKRYIEMLHAWMYNNRVKT; SEQ ID Nq 15) is generally located between. amino acid positions 765 and 790.

Sucrose synthase proteins have also been reported to have a putative actin binding region (Winter H et al, 1998, FEBS Ietters 430, 205-208; Winter, H. and Huber SC, 2000 Critical Reviews in Plant Sciences 19 (1), 31 -67) which in SusC isoforms has the following consensus amino acid sequence: KDVAAE [V / I] TKEFQ (SEQ ID NO: 53 from amino acid position 383 to amino acid position 394). The lysine residue (K) generally found at position 383 and the phenylalanine residue (F) 1 generally found at position 393 are also indicative of a SusC-like protein.

Sucrose synthase proteins have also been described to have a putative Uridine binding region which in SusC isoforms has the amino acid sequence that follows VVIMTPHGYFAQDNVLGYPDTGGQVVYLLDQVRALEEELLHRFKLQGLDIT PRILVITRL to amino acid position SEQ ID 3 at position 270

Using computer-based searches the following domains or signatures could be identified in the amino acid sequence of the SusC isoforms. These domains or signatures are shared with other plant sucrose synthases.

Glycosyl Transferase, Domain Group 1: (1PR001296; PF00534) Proteins containing this domain transfer UDP1 ADP, GDP, or CMP-linked sugars to a variety of substrates, including glycogen, fructose-6-phosphate, and lipopolysaccharides. Bacterial enzymes are involved in various biosynthetic processes that include exopolysaccharide biosynthesis, lipopolysaccharide core biosynthesis, and polysaccharide biosynthesis of cholanic acid saliva. This domain corresponds to the amino acid sequence of SEQ ID Ne: 3, for example, amino acid positions 549 to 737.

Sucrose Synthase Family Signature (IPR000368; PF00862): This signature characterizes a family including most sucrose synthase proteins. However, the terminal carboxyl region of sucrose synthase belongs to the glycosyl transferase family. This enzyme is mainly found in plants, but also appears in bacteria. This domain corresponds to the amino acid sequence of SEQ ID NQ: 3, for example, amino acid positions 1 to 545.

Signature of the sucrose synthase family, plants and cyanobacteria (I-PR012820): This signature represents sucrose synthases, an enzyme that, despite its name, uses generally rarely produce sucrose. Saccharose plus UDP (or ADP) becomes D-fructose plus UDP-glucose (or ADP-glucose), which is then available for cell wall (or starch) biosynthesis. The enzyme is homologous to sucrose phosphate synthase, which catalyzes the penultimate step in sucrose synthesis. Sucrose synthase is found until now exclusively in plants and cyanobacteria. This domain corresponds to the amino acid sequence of SEQ ID NQ: 3, for example, from amino acid positions 18 to 794.

These domains can easily be recognized by computer-based searches using, for example, PROSITE profiles (PROSITE is available at www.expansv.ch/prosite). Alternatively, the BLOCKS database and algorithm (blocks.fhcrc.org) can be used to identify these domains. Other databases and algorithms are also available (pFAM: http://www.sanger.ac.uk/SOftware/Pfam/ INTERPRO: http://www.ebi.ac.uk/interpro/; PF numbers quoted above refer to database and pFAM algorithm and IPR numbers to database and INTERPO algorithm).

The present invention provides various amino acid sequences and variant nucleotide sequences (cDNA as well as genomic DNA) for SUsC isoforms isolated from Gossypium hirsutum cultivars (SEQ ID NOs .: 3-4), Gossypium barbadense cv (SEQ ID NOs. : 5 and 6), Gossypium arboretum (SEQ ID NOs: 7 and 8), Gossypium raimondii (SEQ ID NOs: 9 and 10) and Gossypium Iongicaiyx (SEQ ID NOs: 11 and 14).

Other variant nucleotide sequences may be obtained

of other cultivars of Gossypium hirsutum or Gossypium barbadense or parent cotton plants such as Gossypium arboretum, Gossypium herbaceum and Gossypium raimondii and Gossypium Iongicalyx or other Gossypium species, especially Gossypium species comprising a D-type genome such as Gossypium arid davidsonii, Gossypium gossypioides, Gossypium klotzschianum, Gossypium turberi, Gossypium triobum, Gossypium turneri, Gossypium mustelinum, Gossypium tomentosum, Gossypium darwinii. Variant amino acid sequences or nucleotide sequences include modifications of a sequence by the addition, deletion or substitution of amino acids or nucleotides, respectively. PCR amplification of SusC-conducting nucleic acids is possible from other Gossypium species comprising type D genome. Such PCR amplified nucleic acids appear to comprise

a characteristic EcoRI restriction site for nucleic acids encoding Sus type C.

Sucrose synthase C isoform variants can be found in stringent hybridization using the nucleotide sequence of any SEQ ID NO: 1, 4, 6, 8, 10, 12, or 14 thereof or comprising at least about 25 or 50 nucleotides. of SEQ ID NO: 1, 4, 6, 8, 10, 12 or 14 or the complementary nucleotide sequences thereof, as a probe. A particular useful probe may be the nucleotide sequence encoding the N-terminal or C-terminal sequence of SEQ ID Nos: 15 and 16, such as the nucleotide sequences of any one of SEQ ID NQ: 1, 4, 6, 8, 10, 12 or 14 encoding the amino acid sequence of SEQ ID NO: 15 or 16.

"Stringent hybridization conditions" as used herein means that hybridization will generally occur if there is at least 95% and preferably at least 97% sequence identity between the probe and the target sequence. Examples of stringent hybridization conditions are overnight incubation in a solution comprising 50% formalin, 5 χ SSC (15 mM NaC1150, 15 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6). , 5x Denhardt's solution, 10% dextran sulfate, and 20 MiTVhiI of denatured sheared carrier DNA such as salmon sperm DNA, followed by washing of the 0.1 χ SSC hybridization support at approximately 65 ° C, preferably by about 65 ° C. 10 minutes. Other hybridization and wash conditions are well known and are exemplified in Sambrook et al., Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor, NY (1989), particularly chapter 11.

Such variant sequences may also be obtained by DNA amplification using sucrose synthase (SusC) gene specific oligonucleotides as primers such as, but not limited to, oligonucleotides comprising or consisting of about 20 to about 50 consecutive nucleotides of the sequence. of nucleotide of SEQ ID NO: 1, 4, 6, 8, 20, 12 or 14 or their complement. SEQ ID Nos: 17 and 18 show the nucleotide sequences of primers that are particularly suitable as SusC specific primers.

In addition to exploiting natural variation for SusC isoforms, variant sequences may also be obtained by induced generation of variation in vitro or in vivo. Various methods for in vitro induced generation of variant nucleotide sequences are available in the art including, but not limited to, shuffling techniques or direct DNA evolution as described in US Patent 5,605,793, US Patent 5,811,238 and US Patent 5,830,721. Methods for in vivo induced generation of variant nucleotide sequences are also well known in the art and may include exposure of cotton plants or cotton progenitor plants to mutagens such as ionization radiation, SEM, MMS or the like, followed by isolation of the Nucleic acids encoding SusC1 for example, as described elsewhere herein.

Having identified the C-terminal and N-terminal consensus sequences for a SusC sucrose synthase, the inventors identified the following nucleotide sequences in databases encoding amino acid sequences comprising such consensus sequences. Using the N-terminal consensus region (SEQ ID NQ: 16) the following sequences (identified by their Accession Number) were isolated: EM-BL.C0499214 (Gossypium hirsutum cDNA; EMBL: C0499090 ( Gossypium hirsutum) ·, EMBLCP498472 (Gossypium hirsutum cDNA) ·, EMBL: C) 498387 (Gossypium hirsutum cDNA) ·, EMBL: C) 497432 (Gossypium hirsutum cDNA); EMBL: C0497275 (Gossypium hirsutum cDNA); MS-BL: C0496887 (Gossypium hirsutum cDNA); EMBL: C0496593 (Gossypium hirsutum cDNA); EMBL.O0495954 (Gossypium hirsutum cDNA); MS-BL: C0495804 (Gossypium hirsutum cDNA); EMBL: C0495643 (Gossyp / um hirsutum cDNA); EMBLC0495623 (Gossypium hirsutum cDNA); MS-BL: C0495601 (Gossypium hirsutum cDNA); EMBL: C0495511 (Gossypium hirsutum cDNA); EMBL: C0494831 (Gossypium hirsutum cDNA); MS-BL: C0494779 (Gossypium hirsutum cDNA); EMBL: C0494693 (Gossypium hirsutum cDNA); EMBLC0494536 (Gossypium hirsutum cDNA); MS-BLC0494182 (Gossypium hirsutum cDNA); EMBL: C0493935 (Gossypium hirsutum cDNA); EMBL.C0493780 (Gossypium hirsutum cDNA); MS-BL: C0493725 (Gossypium hirsutum cDNA); EMBL.C0493724 (Gossypium hirsutum cDNA); EMBL: C0493589 (Gossypium hirsutum cDNA); MS-BL.-CO493043 (Gossypium hirsutum cDNA); EMBL: C0493007 (Gossypium hirsutum cDNA); EMBL: C0492969 (Gossypium hirsutum cDNA); MS-BL.C0492566 (Gossypium hirsutum cDNA); EMBL: C0490959 (Gossypium hirsutum cDNA); EMBL: BF272227 (Gossypium arboreum cDNA clone GA_Eb0014E15f); EMBL: C0493732 (Gossypium hirsutum cDNA); MS-BL: C0493111 (Gossypium hirsutum cDNA); EMBL: C0491540 (Gossypium hirsutum cDNA); EMBLC0493583 (Gossypium hirsutum cDNA); MS-BL: C0493765 (Gossypium hirsutum cDNA); and EMBLC0498054 (Gossypium hirsutum cDNA). All sequences cited are incorporated herein by reference. Using the C-terminal consensus region (SEQ ID NQ: 15) the following sequences (identified by their accession number) were isolated: EMBL: DT463218 (Gossypium hirsutum cDNA clone GH_CHX13D22-3); EMBLC0499051 (Gossypium hirsutum cDNA); MS-BLC0498850 (Gossypium hirsutum cDNA); EMBL: C0498685 (Gossypium hirsutum cDNA); EMBL: C0498361 (Gossypium hirsutum cDNA); MS-BL: C0498044 (Gossypium hirsutum cDNA); EMBL: C0497702 (Gossypium hirsutum cDNA); EMBLC0497506 (Gossypium hirsutum cDNA); MS-BL: C0497291 (Gossypium hirsutum cDNA); EMBL: C0497151 Gossypium hirsutum cDNA); EMBL: C0496924 (Gossypium hirsutum cDNA); MS-BL: C0496907 (Gossypium hirsutum cDNA); EMBL: C0496748 (Gossypium hirsutum cDNA); EMBL: C0496747 (Gossypium hirsutum cDNA); MS-BL: C0496373 (Gossypium hirsutum cDNA); EMBL: C0496003 (Gossypium hirsutum cDNA); EMBL: C0495974 (Gossypium hirsutum cDNA); MS-BL: C0495570 (Gossypium hirsutum cDNA); EMBL: C0494691 (Gossypium hirsutum cDNA); EMBL: C0493817 (Gossypium hirsutum cDNA); MS-BL: C0493779 (Gossypium hirsutum cDNA); EMBL: C0493750 (Gossypium hirsutum cDNA); EMBL: C0492925 (Gossypium hirsutum cDNA); MS-BL: C0492917 (Gossypium hirsutum cDNA); EMBLC0492529 (Gossypium hirsutum cDNA); EMBL: C0492496 (Gossypium hirsutum cDNA); MS-BL: C0493902 (Gossypium hirsutum cDNA); EMBL: BQ412019 Gossypium

arboreum cDNA clone GA_Ed0053A09r); EMBL: C0497086 (Gossypium

hirsutum cDNA) EMBL: C0494597 (Gossypium hirsutum cDNA); MS-BL: C0494374 (Gossypium hirsutum cDNA); EMBLC0499615 (Gossypium hirsutum cDNA); EMBLC0490812 (Gossypium hirsutum cDNA); MS-BL: C0493178 (Gossypium hirsutum cDNA).

Suitable isoform C sucrose synthase variant forms suitable for the invention may then have an amino acid sequence comprising an amino acid sequence having at least about 60% or about 70% or about 80% or about 90% or about 95% sequence identity with either the N-terminal consensus sequence amino acid sequence (SEQ ID Ne: 16) or the C-terminal consensus sequence (SEQ ID NQ: 15) or both. Variants may have an amino acid sequence having at least about 60% or about 70% or about 80% or about 90% or about 95% sequence identity to the amino acid sequence of any of the above. SusC isoforms of SEQ ID Ng: 2, 3, 5, 7, 9, 11 or 13. Nucleotide sequences encoding such variants may have a nucleotide sequence having at least about 60% or about 70% or about 80%. or about 90% or about 95% sequence identity to the nucleotide sequence of any of the nucleotide sequences of SEQ ID NQ: 1, 4, 6, 8, 10, 12 or 14.

For purposes of the present invention, the "sequence identity" of two related nucleotide or amino acid sequences, expressed as a percentage, refers to the position number in the two optimally aligned sequences that have identical (x100) divided residues. by the number of positions compared. A gap, that is, a position in an alignment where a residue is present in one sequence but not the other, is considered as a position with non-identical residues. The alignment of the two sequences is performed through the Needleman and Wunsch algorithm (Needleman and Wunsch, 1970). The above computer-aided sequence alignment can conveniently be accomplished using standard software programs such as GAP which is part of the Wisconsin Package Version 10.1 (Genetics Computer Group, Madison, Wisconsin, LI) using scoring default matrix with a spawn penalty of 50 and a spine extension penalty of 3.

It will be clear that whenever nucleotide sequences of RNA molecules are defined by reference to the nucleotide sequence of corresponding DNA molecules, the thymine (T) in the nucleotide sequence must be replaced by uracil (U). Whether reference is made to RNA or DNA molecules will be clear from the context of the application. The functional level of sucrose synthase type C isoform or saccharose synthases with similar characteristics can also be increased by in vivo induced sequence variation. For this purpose, methods of induced sequence variation known in the art as described elsewhere herein can be applied to fiber production plants, such as cotton plants and fiber producing plants can be identified that exhibit greater activity. for isoform C sucrose synthase activity (for example, using the enzyme assay described herein) or which exhibit a higher concentration of sucrose synthase C, particularly sucrose isoform C sucrose in the apoplastic fluid or cell wall, particularly during the developmental phase of the sucrose synthase. secondary cell wall formation, and particularly in fiber cell walls or in the apoplastic fluid surrounding such fibers.

For some fiber producing plants, it may be advantageous to modify fiber characteristics by decreasing the functional level of sucrose synthase isoform C or sucrose synthases with similar characteristics. This can again conveniently be accomplished by introducing a gene encoding a silencing RNA.

In one embodiment, the gene encoding the silencing RNA may encode a silencing RNA molecule or an inhibitory RNA molecule capable of reducing expression of an endogenous gene encoding an isoform C sucrose synthase to alter fiber characteristics. Such reduction of expression of a gene encoding an isoform C sucrose synthase should preferably occur by post-transcriptional silencing. However, it will be clear that even when an inhibitory RNA molecule decreases the expression of a SusC gene through post-transcriptional silencing, such an RNA molecule can also perform other functions within a cell, such as guiding DNA methylation of the endogenous SusC gene, again eventually leading to lower expression of this gene. Also, expression of endogenous SusC genes may be reduced by transcriptional silencing, for example, using RNAi or dsRNA directed against the promoter region of the endogenous SusC gene.

Several methods are available in the art for producing a silencing RNA molecule, that is, an RNA molecule that when expressed reduces the expression of a particular gene or group of genes, including so-called "sense" or "RNA" technologies. "antisense".

Then, in one embodiment, the inhibitory RNA molecule encoding the chimeric gene is based on the so-called antisense technology. In other words, the chimeric gene coding region comprises a nucleotide sequence of at least 20 consecutive nucleotides from the nucleotide sequence complement of an endogenous SusC gene. Such a chimeric gene may be constructed by operably linking a DNA fragment comprising at least 20 nucleotides of a SusC gene, which may be isolated or identified as described elsewhere in this application, in reverse orientation to a plant expressible promoter and region of formation. 3 'terminus involved in transcription termination and polyadenylation. It will be clear that there is no need to know the exact nucleotide sequence or the complete nucleotide sequence of such an isolated SusC gene DNA fragment.

In another embodiment, the inhibitory RNA molecule encoding the chimeric gene is based on so-called co-suppression technology. In other words, the chimeric gene coding region comprises a nucleotide sequence of at least 20 consecutive nucleotides from the nucleic acid sequence of a plant endogenous SusC gene. Such a chimeric gene may be constructed by operably linking a DNA fragment comprising at least 20 nucleotides of a SusC gene, which may be isolated or identified as described elsewhere in this application, in direct orientation to a plant expressible promoter and region of formation. 3 'terminus involved in transcription termination and polyadenylation. Again, it will be clear that there is no need to know the exact nucleotide sequence or the complete nucleotide sequence of such a DNA fragment or isolated SusC gene.

The efficiency of the chimeric genes mentioned above in reducing

15

The expression of endogenous SusC genes can be further enhanced by the inclusion of a DNA element that results in the expression of aberrant non-polyadenylated inhibitory RNA molecules, or results in the retention of inhibitory RNA molecules in the nucleus of DNA. cells One such DNA element suitable for this purpose is a region of DNA encoding a self-splicing ribozyme, as described in WO 0001133 (incorporated by reference). Another such DNA element suitable for this purpose is a region of DNA encoding a nuclear localization or jRNA retention signal as described in

PCT / AU03O0292 published as W003O76619 (incorporated by reference).

A convenient and very efficient down-regulation of expression of a gene of interest uses so-called double stranded RNA (dsRNA) or interfering RNA (RNAi) 1 as described, for example, in W09963050 (incorporated herein). of reference). In this technology, an RNA molecule is introduced into a plant cell, whereby the RNA molecule is capable of forming a double stranded RNA region in at least about 19 to about 21 nucleotides, and with the that one strand of this double stranded RNA region is roughly identical in nucleotide sequence to the target gene ("sense region"), while the other strand is roughly identical in nucleus sequence to the complement of the target gene. target gene or sense region ("antisense region"). It is expected that for silencing target gene expression, the nucleotide sequence of the 19 consecutive nucleotide sequences may be mismatched, or the sense and antisense regions may differ by one nucleotide. To achieve the construction of such RNA molecules or chimeric coding genes, use may be made of the vector as described in WO 02059294.

Thus, in one embodiment of the invention, a method for altering the fiber characteristics of a fiber producing plant, such as cotton, is provided comprising the step of introducing a chimeric gene into a cell of the fiber producing plant. where the chimeric gene

20 comprises operably linked DNA elements that follow: (a) a plant expressible promoter, preferably a promoter

plant expressible which preferably controls transcription in fiber cells;

(b) a region of transcribed DNA, which when transcribed gives a

double stranded RNA molecule capable of reducing the expression of an endogenous SusC gene to the fiber producing plant, and the RNA molecule comprising a first and a second DNA region where i) the first RNA region comprises a sequence of nucle-

an otide of at least 19 consecutive nucleotides having at least about 94% sequence identity with the nucleotide sequence of an endogenous SusC gene;

ü) the second RNA region comprises a sequence of

cleotide complementary to at least 19 consecutive nucleotides of the first RNA region;

üi) the first and second RNA regions are capable of matching

Base line to form a double stranded RNA molecule between at least 19 nucleotides of the first and second regions; and (c) a 3 'end region comprising transcription termination and polyadenylation signals functioning in plant cells.

The length of the first and second RNA regions (sense and antisense region) may range from about 19 nucleotides (nt) to a length equaling the length (in nucleotides) of an endogenous SusC gene. The total length of the sense or antisense nucleotide sequence may then be at least 25 nt or at least about 50 nt or at least about 100 nt or at least about 150 nt or at least about 200 nt or at least minus about 500 nt. It is expected that there will be no limit to the total length of the sense or antisense nucleotide sequence. However, for practical reasons (such as, for example, chimeric gene stability) it is expected that the length of the sense or antisense nucleotide sequence should not exceed 5000 nt, particularly it should not exceed 2500 nt and could be limited to about 1000 nt.

It will be understood that the longer the total length of the sense or antisense region, the less stringent the sequence identity needs between these regions and the corresponding sequence in an endogenous SusC gene or complement thereof. Preferably, the nucleic acid of interest should have a sequence identity of at least about 75% with the corresponding target sequence, particularly at least about 80%, more particularly preferably at least about 85%, very particularly about 90%, especially about 95%, more especially about 100%, most especially being identical to the corresponding part of the target sequence or its complement. However, it is preferred that the nucleic acid of interest always includes a sequence of about 19 consecutive nucleotides, particularly about 25 nt, more particularly about 50 nt, especially about 100 nt, most particularly about 250 nt with 100%. sequence identity with the corresponding part of the target nucleic acid. Preferably, for calculating sequence identity and designing the corresponding sense or antisense sequence, the number of gaps should be minimized, particularly for shorter sense sequences. dsRNA encoding chimeric genes according to the invention

they may comprise an intron, such as a heterologous intron, located, for example, in the spacer sequence between the sense and antisense RNA regions according to the disclosure of WO 9963050 (incorporated herein by reference). In another embodiment, the silencing RNA molecule or

Inhibitor RNA may be a microRNA molecule designed, synthesized and / or modulated to target and cleave endogenous SusC genes in a fiber producing plant such as a cotton plant. Several methods have been described for generating and using miRNAs for a specific target gene (including, but not limited to, Schwab et al. (2006, Plant Cell, 18 (5): 1121-1133), W02006 € 44322, W02005O47505, WP 06009836, incorporated herein by reference). Generally, an existing miRNA structure is modified at the target gene recognition portion so that the generated miRNA now guides the RISC complex to cleave the transcribed RNA molecules from the target nucleic acid. MiRNA structures could be modified or synthesized so that miRNA now comprises 21 consecutive nucleotides from the nucleotide sequence of a SusC1 encoding nucleotide sequence such as the sequences represented in the Sequence Listing, and allowing mismatches according to the described rules. here below. Particularly suitable sequences from which 21 consecutive nucleotides may be chosen comprise nucleotide sequences encoding SusC-specific amino acid sequences as described herein.

Thus, in one embodiment, the invention provides a method for down-regulating the expression of one or increased plant resistance to adverse growth conditions, comprising steps a. introduction of a chimeric gene into cells of said plants, the

said chimeric gene comprising the operably linked DNA regions as follows:

i. an expressible plant promoter;

ii. a region of DNA which upon introduction and transcription in a plant cell is processed into miRNA, whereby the miRNA is able to recognize and guide mRNA cleavage of a gene encoding SusC from the plant; and

iii. optionally a 3 'DNA region involved in transcription termination and polyadenylation.

The mentioned miRNA processed DNA region may comprise a nucleotide sequence that is essentially complementary to a nucleotide sequence of at least 21 consecutive nucleotides of a SusC encoding gene, provided that one or more of the mismatches is allowed:

The. A mismatch between the 5 'end nucleotide

miRNA and the corresponding nucleotide sequence in the RNA molecule; B. A mismatch between any of the nucleotides at position 1 to position 9 of the miRNA and the corresponding nucleotide sequence in the RNA molecule;

ç. Three mismatches between any of the nucleotides at position 12 to miRNA position 21 and the corresponding nucleotide sequence in the RNA molecule as long as there are no more than two consecutive mismatches.

Decreasing the functional level of isoform C Sucrose synthase may be advantageous, for example, in fiber cells leading to cotton down fiber to reduce or prevent energy divergence and metabolites for producing downward downward fiber. costs of cotton production.

As used herein, an "endogenous gene" is a naturally occurring gene in the fiber-producing plant species that have been chosen for modulation of fiber characteristics or a gene that naturally occurs in a species from another fiber-producing plant. but can be introduced into the species of the fiber producing plant that has been chosen for modulation of fiber characteristics through conventional generation techniques.

It will be clear that endogenous SusC gene expression can also be down-regulated using chimeric genes as described herein, where the DNA encoding the target specific RNA has a nucleotide sequence of at least 20 consecutive nucleotides selected from the sequences of nucleotide encoding the amino acid sequences of SEQ ID NO: 2, 3, 5, 7, 9, 11 or 13 or their complement, or where the target specific RNA has a nucleotide sequence of at least 20 consecutive nucleotides selected from the following: nucleotide sequences of SEQ ID NO: 1, 4, 6, 8, 10, 12 or 14.

As used herein, the term "promoter" means any DNA that is recognized and linked (directly or indirectly) by a DNA-dependent RNA polymerase during transcription initiation. A promoter includes the transcription initiation site, and binding sites for transcription initiation factors and RNA polymerase, and may comprise a number of other sites (e.g., enhancers) to which gene expression regulatory proteins may bind. .

As used herein, the term "plant expressible promoter" means a DNA sequence that is capable of controlling (initiation) transcription in a plant cell. The chimeric gene promoter described herein may be naturally associated with the coding regions or it may be a heterologous promoter. The term plant expressible promoter includes any plant origin promoter, but also any non-plant origin promoter that is capable of directing transcription in a plant cell, that is, certain promoters of viral or bacterial origin as such. such as CaMV35S, the underground clover virus promoter N5 4 or Ng 7 or T-DNA gene promoters and the like.

The plant expressible promoter may be constitutive or it may initiate transcription of the downstream bound region in a spatial or temporary manner. A plant expressible promoter that controls transcription initiation and maintenance preferably in fiber cells is a promoter that directs transcription of the operably linked DNA region to a higher level in fiber cells and base epidermis cells. than in other cells or tissues of the plant. Such promoters include the cotton promoter of a fiber-specific β-tubulin gene (as described in W00210377), the cotton promoter of a fiber-specific actin gene (as described in W00210413), the promoter of a protein-specific gene. - cotton fiber specific lipid transfer (as described in US5792933), a cotton expansin gene promoter (W09830698) or a cotton chitinase gene promoter (US2003106097) or specific gene promoters described in US6259003 or US6166294. The promoter may also be inducible by chemical compound (generally in combination with a transcriptional activator) as described, for example, in W09321334, US5514578, EP0823480, W098O5789, W001 / 34821 or W00220811.

The methods described here will change the fiber characteristics produced by the modified plants including strength, length, fiber fineness, fiber maturity ratio, immature fiber content, fiber uniformity, and micronaire.

- Fiber "micronaire" (which can be determined by HVI) is a unitless measure that depends on both fiber maturity (or wall thickness determined by secondary wall cellulose content) and fiber diameter.

- Fiber beam strength (via HVI) is expressed in units of (cN4ex). It is the specific strength of the fiber bundle where the

Individual fiber fineness (tex) is calculated from the "micronaire" value.

- Fiber fineness (through AFIS) is expressed as (mTex). It represents the weight in milligrams of one kilometer of fiber. One thousand meters of fiber with a mass of 1 milligram is equal to 1 millitex.

- The fiber maturity ratio (through AFIS) is an ex-

pressure of the degree of cell wall thickening (depending on the cellulose deposition in the secondary cell wall). It is the ratio of fibers with a circularity ratio of 0.5 (or more) divided by the amount of fibers with 0.25 (or less) of circularity. (Thicker wall fibers are less prone to collapse and remain more circular

when drying). The higher the maturity ratio, the more mature the fibers are and the better the fibers are for dyeing.

- Immature fiber content ("% IFC" through AFIS) is the percentage of fibers with a maturity of less than 0.25. The lower the% of IFC, the more suitable the fiber for dyeing.

- Several different units are used as indicators of

fiber length, shows values for three of these as now described. Upper mean ("UHM" through HIVI) is the average length of the longest half of the fibers (weight basis).

- The fiber uniformity index ("Ul" through HVI) expressed

hurry the ratio of the average value (Average Length) to the Length

Upper Middle. It is a measure of fiber length spread within a population; if all fibers have the same length Ul would be 100%. Short Fiber Content ("% SFC1 through HVI) is the percentage of fibers less than [1.27 cm] [1/2]" in length on a weight basis. HVI is thought to measure Short Fiber Content as determined through genetics only since the measurement does not impose additional potential fiber break stress.

Other fiber length indicators include weight-based length ("L (p)" [cm] [in] via AFIS) is the average fiber length calculated on a weight basis. Length based on number ("L (n) M [cm] [in] through AFIS) is the average fiber length calculated by number. Length" L5% (n) "[cm] [in .] (through AFIS) is the 5% span Iength or the 5% extended length of the fibers when they are parallel and randomly distributed. "L2.5% (n)" [cm] [in] (across (AFIS) is the 2.5% span Iength, or the 2.5% extended length of the fibers when they are parallel or randomly distributed. "UQL (p)" (in AFIS) is the com - upper quartile length of fiber by weight or the length exceeded by 25% of fiber by weight Finally, "SFC (n)" [cm] [in] and "SFC (p)" [in] (by AFIS) is the percentage of fibers of less than 1.27 cm (0.50 inch) in length based on number and weight, respectively. In contrast to HVI, AFIS beats the fibers before taking these measurements, which has the potential to cause fiber breakdown. AFIS SFC values are a good indication of fiber characteristics after normal processing.

It will be clear that the methods and means described herein for altering fiber characteristics in fiber producing plants such as cotton plants may be combined with other methods for altering fiber characteristics as known in the art.

In one embodiment of the invention, the methods and means of the present application are combined with those described in W02005 / 017157 (incorporated herein by reference) or W001 / 17333. The combined expression of the genes is expected to result in a synergistic effect on fiber length and / or strength increase. The methods and means of the present application may also be combined with other methods and means for modifying fiber characteristics as described, for example, in PCT / EP2006 / 005853 or W098 / 00549.

Chimeric genes may be introduced through subsequent transformations into cells of a plant or may be combined into cells of a plant by crossing plants comprising one chimeric gene each.

As the onset of secondary cell wall synthesis in fibers, so that cotton fibers overlap the fiber elongation phase, it may be further advantageous to delay the onset of sucrose C isoform expression to a later stage, thereby prolonging the production. fiber stretching phase. This delay in SusC gene expression may still be combined with increased expression of SusC gene expression at a later stage. One way to achieve this r-step would be to "exchange" the endogenous SusC gene (s) for a SusC gene whose expression is activated in a change later than the endogenous SusC genes, for example, by reducing expression of the endogenous SusC gene as described herein and introducing the SusC coding region under the control of a promoter that is activated at a later developmental stage of secondary cell wall synthesis, such as as, for example, the E6 promoter (John, ME, Keller, G., Proc. Nati Acad. Sci. USA 1996, 93: 12768-12773). The chimeric SusC coding region under control of a promoter expressed at a later stage in secondary cell wall synthesis may be introduced ectopically, or the endogenous SusC gene may be introduced by homologous recombination techniques, as described, for example, in PCTVEP2006 / 003086. As the SusC gene of Gossypium barbadense cultivars, such as variedades varieties, appears to be expressed at a later stage than the SusC gene of G. hirsutum varieties, "exchange" of the SusC gene in G. hirsutum varieties by SusC genes from G. barbadense growers are expected to prolong the fiber elongation phase and lead to longer cotton fibers. The invention also encompasses the chimeric genes described herein, as well as plants, seeds, tissues comprising such chimeric genes and fibers produced from such plants.

Methods for transforming plants are well known in the art. Methods for turning cotton plants are also well known in the art. Agrobacterium-mediated transformation of cotton has been described, for example, in US 5,004,863 or US 6,483,013 and cotton transformation by particle bombardment is reported, for example, in WO 92/15675.

Chimeric genes according to the invention may be introduced into plants in a stable or transient manner using methods well known in the art. Chimeric genes may be introduced into plants or may be generated within the plant cell as described, for example, in EP 1339859.

Chimeric genes can be introduced by transformation into cotton plants from which embryogenic callus can be derived, such as Coker 312, Coker310, Coker 5Acala SJ-5, GSC25110, FIBERMAX 819, Siokra 1-3, T25, GSA75, Acala SJ2, Acala SJ4, Acala SJ5, Acala SJ-C1, Acala B1644, Acala B1654-26, Acala B1654-43, Acala B3991, Acala GC356, Acala GC510, Acala GAM1, Acala C1, Acala Royale, Aeala Maxxa, Acala Prema , Acala B638, Acala B1810, Acala B2724, Acala B4894, Acala B5002, Non-Acala Siokra picker variety, stripper FC2017, Coker 315, STONEVILLE 506, STONEVILLE 825, DP50, DP61, DP90, DP77, DES119, McN235 , HBX87, HBX191, HBX107, FC 3027, CHEMBRED A1, CHEMBRED A2, CHEMBRED A3, CHEMBRED A4, CHEMBRED B1, CHEMBRED B2, CHEMBRED B3, CHEMBRED C2, CHEMBRED C3, CHEMBRED C4, PAYMASTER 145, HS, HS SICA- PIMA S6 GOLD BLANK ΡΙΜΑ, FIBERMAX FM5013, FIBERMAX FM5015, FIBERMAX FM5017, FIBERMAX FM989, FIBERMAX FM832, FIBERMAX FM966, FIBERMAX FM958, FIBERMAX FM989, FIBERMAX FM958, FIBERMAX FM832, FIBERMAX FM991, FIBERMAX FM819, FIBERMAX FM960, FIBERMAX FM960, FIBERMAX FM981, FIBERMAX FM5035, FIBERMAX FM175, FIBERMAX FM935 FM5024 and plants with genotypes derived from them. "Cotton" as used herein includes Gossypium hirsutum or Gossypium barbadense. "Cotton progenitor plants" include Gossypium arboreum, Gossypium herbaceum and Gossypium raimondii and Gossypium longicaiyx.

However, the methods and means of the present invention may also be employed for other plant species such as hemp, jute, lignin and woody plants, including, but not limited to, Pinus spp., Pulpus spp., Picea spp. , Eucalyptus spp., Etc.

The obtained transformed plant can be used in a conventional generation scheme to produce more transformed plants with the same characteristics or to introduce the chimeric gene according to the invention into other varieties of the same or related plant species, or in hybrid plants. Seeds obtained from transformed plants contain the chimeric genes of the invention as a stable genomic insert and are also encompassed by the invention.

In another embodiment, the method is provided for identifying allelic variations of proteins involved in fiber characteristics in a different genotype or variety population of a particular plant species, preferably a fiber producing plant species, which are correlated or alone. or in combination with the quantity and / or quality of fiber production. This method includes the following steps:

(a) provision of a population of different varieties or genotypes

of a particular plant species or interbreeding plant species comprising different allelic forms of the nucleotide sequences encoding sucrose synthase isoform C, such as nucleotide sequences encoding SEQ ID NO: 1, 4, 6, 8, 10, 12 or 14. Different allelic forms can be identified using the methods described elsewhere in this application. Preferably, a segregating population is pro- vided, where different combinations of allelic variations of SusC proteins are present. Methods for producing segregation populations are well known in the plant generation technique.

b) determination of parameters related to fiber characteristics for each individual of the population;

c) determining the presence of a particular allelic form of the SusC-encoding nucleotide sequences for each individual in the population; and

d) correlating the occurrence of particular fiber trait with the presence of a particular allelic form of the aforementioned nucleotide sequence or a particular combination of such allelic forms.

The resulting information can be used to accelerate generation program varieties with particular fiber characteristics and dryness resistance by determining the presence or absence of allelic forms using conventional molecular biology techniques.

Allelic forms of the SusC gene associated with particular fiber characteristics can also be identified, isolated and introduced into plants, such as cotton plants, whereby the expression of endogenous SusC has been reduced or eliminated. Such reduction of expression of endogenous SusC genes may conveniently be accomplished by post-transcriptional or transcriptional silencing as described herein, or may be achieved by inactivating, as well as deleting, endogenous SusC genes. Introduction of allelic forms can be accomplished through generation techniques or through transformation with the isolated genes.

In another embodiment of the invention, antibodies raised against the novel type of sucrose synthase genes are provided, particularly antibodies recognizing SusC proteins having the amino acid sequences of SEQ ID Ng: 2, 3, 5, 7, 9, 11 or 13.

As used herein, "comprising" shall be construed as specifying the presence of the declared features, integers, steps or components as referred to, but does not preclude the presence or addition of one or more of the features, integers, steps or components, or groups thereof. So, for example, a nucleic acid or protein comprising a nucleotide or amino acid sequence may comprise more nucleotides or amino acids than those actually quoted, that is, being embedded in a larger nucleic acid or protein. A chimeric gene comprising a DNA1 region that is functionally or structurally defined may comprise additional DNA regions, etc.

The following non-limiting examples describe the identification of a new type of sucrose synthase involved in secondary plant cell wall synthesis and chimeric genes for altering fiber characteristics in fiber producing plants such as cotton and uses. of the same. Unless otherwise stated in the Examples, all recombinant DNA techniques are performed according to standard protocols as described in Sambrook et al. (1989) Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor Laboratory Press, NY and in Volumes 1 and 2 of Ausubel et al. (1994) Current Protocols in Molecular Biology, Current Protocols, USA. Standard materials and methods for plant molecular work are described in Plant Molecular Bio Laby Labfax (1993) by R.D.D. Croy, jointly published by BIOS Scientific Publication Ltd. (UK) and Blackwell Scientific Publications, UK.

In the description and examples, reference is made to the following sequences represented in the sequence listing: SEQ ID N-: 1: CDNA nucleotide sequence encoding sucrose cotton type C synthase.

SEQ ID NO: 2: Amino acid sequence of cDNA encoding sucrose cotton type C synthase.

SEQ ID NO: 3: SusC amino acid sequence of a Gossypium hirsutum cultivar.

SEQ ID NQ: 4: SusC genomic DNA nucleotide sequence from Gossypium hirsutum cultivar.

SEQ ID N9: 5: SusC amino acid sequence of Gossypium barbadense cultivar.

SEQ ID Ns: 6: SusC genomic DNA nucleotide sequence of Gossypium barbadense cultivar encoding the SUSc protein of SEQ ID N9: 7. SEQ ID NO: 7: SusC amino acid sequence of Gossypium arboreum. SEQ ID Ng: 8: Gossypium arboretum SusC genomic DNA nucleotide sequence.

SEQ ID NO: 9: Gossypium raimondii SusC amino acid sequence. SEQ ID NO: 10: Gossypium raimondii SusC genomic DNA nucleotide sequence.

SEQ ID NQ: 11: SusC amino acid sequence of Gossypium Iongicalyx subtype 1

SEQ ID NQ: 12: Gossypium Iongicalyx subtype 1 SusC genomic DNA nucleotide sequence

SEQ ID NQ: 13: Gossypium Iongicalyx Subtype 2 SusC Amino Acid Sequence

SEQ ID N-: 14: Gossypium Iongicalyx subtype 2 SusC genomic DNA nucleotide sequence

SEQ ID NQ: 15: Amino acid sequence of SusC C-terminal consensus sequence

SEQ ID Ns: 16: Amino Acid Sequence of SusC N-terminal Consensus Sequence

SEQ ID NO: 17: Oligonucleotide sequence used as SUS C-specific primer (5'UTR)

SEQ ID NO: 18: Oligonucleotide sequence used as SUS C-specific primer (3'UTR). Examples

Example 1. Identification of type C sucrose synthases in Gossypium species

SusC Identification. It has been shown that there are at least 3 classes of Sus genes expressed on cotton fiber during development, called types A, B and C. Types A / D are homologous to the previously reported sequence (Accession number U73588). and type B shows strong sequence similarity to the published sequence. The type C sequences provided in this document are novel and show differential expression between stretching phase and secondary cell wall. Type C appears to be the isoform that appears at later stages in fiber development. SusC is only 76% identical to type A and B proteins at the amino acid level. Using RTPCR and Northern analysis it has been shown that the developmental expression profile of type C transcript precedes secondary cell wall synthesis but shows a more commensurate pattern with a role in this phase of fiber development while AeB types are expressed very early in elongation. Fiber The type C gene is also strongly expressed in stem tissue. cDNA was isolated from fiber tissue at different stages of

its development: 5, 10, 15, 20, 40 days after anthesis (dpa). Libraries were made from these cDNAs and DNA was isolated. This DNA was then used to characterize susyC expression using 9 ng of each library DNA as the starting material for a PCR reaction using the SusC-type specific primer of SEQ ID Nos: 17 and 18. of the amplified fragment is 2550 bp for cDNA and 3353 bp for genomic DNA. Appropriate controls were activated to validate the semiquantitative character of this assay. Figure 2 shows the results, and also allows us to conclude that SusC mRNA is relatively more abundant at 20-40 dpa.

Protein fractionation. Fiber protein extracts at elongation stage (8-11 DAF) and secondary cell wall synthesis stage (15-25 DAF) were fractionated to provide soluble (cytosolid) and microsomal (plasma membrane, cytoskeleton and tonoplast). The microsomal fraction was further purified to enrich plasma membrane proteins. This PM fraction no longer has any cytoskeletal or cell wall contamination as judged by western blotting against actin. Also included was another cell fraction comprising intact fiber apoplastic proteins obtained by gentle washing with isotonic buffer including protease inhibitors and EG-TA. These protein preparations were activated on SDS-PAGE and Sus proteins identified by western blotting. The gel fragments were excised and subjected to tryptic digestion followed by tandem mass spectrometry to identify the present Sus form. Examples of signature peptides used to identify the SusC isoform are shown in Tables 1A and 1B. The predicted molecular weight of SusC is 90.3 kDa as opposed to 92.4 for type A / D and B proteins allowing for separation of form C from S / PAGE isoforms. This separation was confirmed by analysis of these 2 bands by peptide MS-MS (the larger band contains A and B signature peptides while the smaller band alone has unique type C peptides).

Table 1A: SusC-Specific Peptides Detected in Apoplast

apoplast 8d apoplast 17d apoplast 25d No type C detected 9.20e + 005 (R) KAEEYLTPLSSDTPYSV FEK () 1.45e + 006 (R) IQDVNSLQHALR () 8.17e + 005 2.37e006 (K) LLTLTG VYG FSK (H) RLS SDTPYSVFEK () 1.32e + 006 2.16e006 (K) YTWQIYSEK () (K) NLTGLVEFYAK (N) 6.90e + 005 (R) STQEAVVSSPLVALAI R () 7.45e + 006 (K) LQG LDITPR (I) 3.64e + 006 ( K) YTWQIYSEK () 5.53e + 006 (R) VVDGIDVFDPK (F) 3.34e + 006 (K) LLTLTGVYGFSK (H) 4.17e + 005 (K) D TLGQYESHIAF-TLPGLYR (V) 3.50e + 005 (K) LFVE L) Table 1B: SusC specific peptides detected in microsomal fraction

microsome 8d microsome 13d microsome 20d microsome 25d No type C detected No type C detected 6.06E + 006 (R) KAEEYLTPLSSDTPY SVFEK () 3.94E + 006 (R) KAEEYLTPLS SDTPYSVFEK () 7.64E + 006 (K) LLTLTGVYGF SK (H) 1.13E + 006 (K) LFVEE MPVAE-YLR (L) 2.31 E + 006 (R) IQ DVNSLQHALR () 3.89E + 006 (R) VVDGIDVFDPK (F) 6.70E + 006 (R) ) VVDGIDVFDPK (F 3.68E + 006 (K) LLTLTGVYGF SK (H) 1.82E + 006 (R) YIEmLHAWmYNNR (V) 3.97E + 005 (K) SGFNIDPYNGDL AAETLANFFEK (C) 3.19E + 006 (K) YT E + 006 (K) NLTGLV EF-YAK (N) 1.99E + 006 (R) STQEA VVSSPLVAL AIR () 1.26E + 006 (R) STQEAVV SSPL-VALA R (1.05E + 006 (R) LGESLATHPQQAK (S) 2.05 E + 006 (K) NLTGLVEFYAK (N) 5.72E + 006 (K) LFVEE m PVAE-YLR (L) 3.01 E + 006 (K) D TLGQYESHIA FTLPGLYR (V) 8.25E + 005 (K) DTLGQYESHIAFTLP GLYR (V) 2.18E + 006 (R) IQ DVNSL-QHALR () 5.25E + 006 (K) LQG LDITPR (I) 8.02E + 005 (R) YIEmLH AWmYNNR (V) 3.28E + 006 (K) NLTGLVEFYAK (N) 3.60E +005 (K) TKYPDSDIN WK (Q 9.91E + 005 (K) SIGNGmDFLNR (H) 5.35E + 005 (R) L GESLATHPQQ AK ( S) 5.59E + 005 (R) ALEEELLHR (F) 2.77E + 006 (K) LFVEE mPVAE-YLR (L)

Table 2 presents a summary of the spec. and SDSPAGE / Western. Of particular note is the low abundance of Sus-C protein from soluble, microsomal and PM fractions. This seems paradoxical considering the high levels of transcript present in the final stages of fiber development. End-developmental apoplastic washes describe that type C protein is present at high levels (relative to A) in western blots, the inverse of observations for cytosolic, microsomal, and PM fractions. Sus-C is highly abundant at 25 DAF and could not be detected at 8 DAF by peptide MS-MS. In contrast, Sus-A possibly Sus-B are ubiquitous during development (in all fractions). Sus-A is the major, and possibly the only isoform in PM fractions and active as an 85 kDa protein in SDS-

PAGE.

Table 2. Summary of spec data Mass and SDSPAGBWestern

microsomal 8 dpa (upper band) Susy A + B microsomal 8 dpa (lower band) Susy A or A + B microsomal 15 dpa Susy A + C or A + B + C plasma membrane 8 dpa Susy A or A + B plasma membrane 15 dpa Susy A or A + B soluble 8 dpa Susy A or A soluble B 15 dpa Susy A + B apoplast 8 dpa (upper range) Susy A + B (very low amount) 8 dpa (lower range) Susy A + B (predominant range) apoplast 25 dpa (upper range) Susy A + B + C * (very low amount) 25 dpa apoplast (lower range) Susy C (predominant range)

The presence of C in this sample is most likely due to the large lower band contamination that corresponds to SusC.

The number of peptides compares with Sus-C in the apoplast fraction at 25 DAF and the western signal indicates that this isoform is abundant, and only predominantly present in the 25 DAF apoplast. Interestingly, SusA and B are present as the dominant forms in the 8 DAF fiber apoplast. These results imply a major role for Sus in the cotton fiber cell wall and the correlation of Sus-C expression at the RNA level with secondary wall formation and protein abundance at 25 DAF suggest that Sus -C is the main cell wall isoform in the secondary wall synthesis stage. Immuno-gold EM work clearly shows Sus signal on the external secondary cell wall. This is entirely in accordance with the presence of Sus-C protein in the cell wall stage apoplast. Example 2. SusC isoform analysis and identification of differences between different sucrose synthase isoforms in cotton and related species

Phosphorylation sites. A serine phosphorylation site situated at the N-end of the protein has been well characterized in the literature, with RXXS as a consensus sequence. Phosphorylation of this serine specific amino acid in Zea mays (with the following positions: 12RXXS15) appears to be related to the release of the susy enzyme from the plasma membrane (Winter, H, et al., 1997 - FEBS Letters 420, 151 Hardin, SC et al., 2004, Plant Physiology 134, 1427-1438). Mutation of this same serine amino acid at position 8RXXS11 makes mung bean sucrose synthase more efficient in using sucrose to make UDPglycosis (= Km higher) (Nakai T. et al., 1999, Proc. Natl. Acad. Sci ., USA 96, 14-18). Phosphorylation at this specific site has also been characterized in soybean and appears to be related to the loss in hydrophobicity and then putatively to the weakening of plasma membrane binding (Zhang, XQ et al., 1999, Archives of Biochemistry and Biophysics 371, 70-82). This site of phosphorylation has also been characterized in other plant species: (sycamore) (Pozueta-Romero, J. et al., 2004 - Physoiogia Piantarum 122, p. 275), Japanese pear (Tanase, K. et al., 2002). -Physoiogia Piantarum 114, p. 21) and Gossypium hirsutum (upiand cotton) (Haigler, CH et al. - 2001 - Piant Molecular Biology 47, 29-51). Conclusions from these studies indicate that phosphorylation has an influence on the Km value of the enzyme.

A similar consensus sequence can be identified in all sucrose synthase G proteins isolated with the following amino acid positions: 27RXXS30. This phosphorylation site is then situated further into the protein. A similar signature can be found on other known plant sucrose synthase genes: L19762, AY205302, AY205085, AJ537575, M18745, U2487, X75332, Y76091, X82504, AJ131999. Most of these genes have both 8RXXS11 and 27RXXS30 phosphorylation sites. L19762 and AJ131999 proteins only have the latter.

Another serine phosphorylation site was characterized in the literature: Ser170 de susl (maize). This site was correlated with susy protein labeling for proteolysis (Hardin, S.C. et al., 2003, The Plant Journal, 35, 588-603). This amino acid Ser is also present in SusC. Only two sucrose synthases do not contain this Serine residue: Atsus5 and AB018561.

Domains and protein, functional sites and specific motifs present in SusC (using the Inter Pro Scan program). SusC contains two domains called sucrose synthase (from amino acids 1 to 545) and glycosyl transferase (from amino acids 549 to 737 respectively) and a family region called sucrose synthase (from amino acids 18 to 794). UDP Binding Site: The following sequence represents the Uridine binding region on sucrose synthase C proteins: 270VVIMTPHGYFAQDNVLGYPDTGGQVVYILDQVRALEEELLHRFKLQGLDI TPRILVITRL329. This sequence shows a very high homology in all known plant sucrose synthases.

Genomic structure. A comparison was made at the genomic level of the SusyC gene with the other isolated cotton susy genes (Figure 3). It will be immediately clear that the susyC gene has a particular genomic structure by the fact that it has fewer introns compared to the other susy genes of Gossypium spp. The exon structure of the SusyC gene intron is also considerably different from the Arabidopsis susy genes.

Hydrophobicity. The hydrophobicity plot of susyC was analyzed by comparing it to other known plant sucrose synthases. N-terminal extremity appears to be hydrophobic, a feature not to be found in most plant sucrose synthases. The following saccharose synthases were more similar to SusC at this level: SUS6 (Arabidopsis thaliana), mung bean sucrose synthase (D10266), eucalyptus sucrose synthase (DD014303) and poplar sucrose synthase (AY341026).

Actin binding region. A putative actin binding region has been described in the literature and appears to be present in the sucrose syntheses (Winter, H. et al., 1998 - FEBS Ietters 430, 205-208. Winter, H. and Hu- ber, SC - 2000). - Critical Reviews in Plant Sciences 19 (1), 31-67). This region binds specifically to F-actin. The putative SusyC actin binding site has the following amino acid sequence: 383KDVAAEITKEFQ394. The amino acids highlighted in bold are those that are specific to SusyC. The consensus sequence for this site between different plant sucrose synthases is: [^ E, D] -D- [V, A] - [A, G, S, T] -XE- [L, V, I] - [ T, S1A1M] - [K, R, G, M, L] -E- [F *, L, M] - [Q, N].

Interestingly, 383K (= lys) has a basic side chain whereas amino acid residues (E (= glu) and D (= asp) have acidic side chains.) As K is unique to susyC this may indicate that this specific protein does not bind to the F-actin filaments In addition, the amino acid F (= Phe), unique to susyC, is rougher than its counterparts L and M from the other plant sucrose synthases. .

SusC protein-specific amino acid sequences. By aligning the different plant sucrose synthases it became apparent that some regions of the susyC gene were very different and specific. These regions are located respectively at the N- and C-terminal ends of the susyC gene and have the following amino acid sequence:

A (N-terminal region) = 36HKSQKLLSVLDKEAGNQALDGMVV59 B (C-terminal region) = 765AYQEQRGRKRYIEMLHAWMYNNRVKT790

The N-terminal region was analyzed by examining its predicted 2D structure. For this purpose the following programs were used: HCA (hydrophobic cluster analysis) and JPRED. Results from both programs were aligned and compared with the other Gossypium hirsutum sucrose synthases (Figure 4). It is clear from this analysis that susC at this site is absolutely unique in structure. Different phosphorylation sites are also indicated confirming the unique characteristic of the susyC gene. A similar analysis was performed on the C-terminal region (Figure 5). Again, the structure of SusC was different from the structure of the other cotton sacsase in this region.

SusC N-terminal and C-terminal specific regions were used to identify EST sequences in databases. The results are summarized in table 3 below. Eggs Wall Wall Other Species Other Total (-3 to 0) Cell Cell Gossy Species Primary Maria Plant Species (0 to 15 (20 to 40 dpa) dpa) Region N - 0 0 35 1 (= 7-10 dpa) 0 36 terminal Region C- 1 0 30 1 (= 7-10 dpa) 0 32 terminal

Table: number of ESTs extracted from some specific tissues and plant species.

Example 3. Cotton Overexpression of SusC

Using standard recombinant DNA techniques, the following DNA elements are operably linked:

• a specific cotton fiber promoter as described in W02004 / 066571

• a DNA region encoding a sucrose synthase C having the nucleotide sequence of SEQ ID N-1

• a terminator region 3 'nos.

This chimeric gene is introduced into a T-DNA vector along with a selectable bar gene. The T-DNA vector is introduced into Agrobacterium tumefaciens and is used to produce transgenic cotton plants as described in US 6,483,013.

Transgenic cotton plants comprising the chimeric gene are analyzed for increased SusC1 expression particularly in fibers, and the fibers obtained from these plants are analyzed for fiber strength, fiber length, fiber maturity ratio, immature fiber content, fiber uniformity and micronaire. Example 4. Down-regulation of SusC isoform expression in

cotton Using standard recombinant DNA techniques, the following DNA elements are operably linked:

• a CaMV 35S promoter region

• a sense RNA coding region corresponding to the nucleotide sequence of SEQ ID Ng 1 encoding the specific SusC N-terminal sequence of SEQ ID Ne 18 or the specific SusC C-terminal sequence of SEQ ID Ne 17.

• An antisense RNA coding region corresponding to the nucleotide sequence complement of the antisense RNA region.

sense.

• a terminator region 3 'nos.

This chimeric gene is introduced into a T-DNA vector along with a selectable bar gene. The T-DNA vector is introduced into Agrobacterium tumefaciens and is used to produce transgenic cotton plants as described in US 6,483,013.

Transgenic cotton plants comprising the chimeric gene are analyzed for lower SusC1 expression particularly in fibers, and the fibers obtained from these plants are analyzed for fiber strength, fiber length, fiber maturity ratio, immature fiber content. , fiber uniformity and micronaire.

Example 5. Overexpression of SusC isoform in transgenic Arabidopsis thaliana

Using standard recombinant DNA techniques, the following DNA elements have been operably linked: · a CaMV35S promoter region

• the SusC encoding DNA region of SEQ ID Ne: 2 (SEQ ID N6: 1 from nt84ant2474)

• a 3 'terminator region in the

In addition, the following DNA element was operably linked to the stem-specific promoter of the alfalfa S2A gene (Abraham).

Hams et al., 1995, Plant Mo. , 27: 413-528) which results in vascular expression and trichome in a range of plant species • The SusC-encoding DNA region of SEQ ID NQ: 2 (SEQ ID Ns: 1 from nt 84 to nt 2474) .

• A terminator region 3 'nos.

These chimeric genes were introduced into a T-DNA vector along with a selectable bar gene. The T-DNA vector was introduced into A-grobacterium tumefaciens and used to produce transgenic A. thaliana using a floral dip method. Plants were regenerated and phenotypes recorded in the T2 generation with several interesting phenotypes routinely observed in transgenic plants obtaining one of the transgenes described above.

Many of the transgenic strains for both constructs show marked root branching in culture, including extensive lateral root growth. Additionally observed were loss of apical dominance, multiple floral branches, loss of rosette symmetry and stiffness, leaf breakage and floral philotaxis, and delayed flowering time.

Increased branching and loss of apical dominance are thought to be due to reduced cell elongation in meristematic zones due to premature secondary thickening of cell walls.

A large proportion of transgenic lineages with any of the chimeric genes showed a phenotypic effect on tritic structures. Larger branching and thick pedestals are usually seen (Figures 6C and D) and trichome density appears to be higher.

Stems and floral branches of Sus-C transgenic lineages usually show extreme thickening and are bifurcated, possibly fasciated (Figures 6A and B).

Three methods were used to relate phenotype to transgene expression:

1) RT-PCR for Sus-C and the endogenous Sus gene

2) Western blotting using Sus-C specific antibody (see Figure 7)

3) Sus activity test

Transgenic lines showing high levels of 35S-Sus-C expression correlated with more severe phenotypes Sequence Listing <110> Bayer BioScience N.V.

Commonwealth Scientific and Industrial Research

Organization

Ruan, Yong-Ling

Furbank, Robert

Brill, Lizz

Van Thournhout, Michel

Arioli, Antonio

<120> Identification of a Type of Sucrose Synthase and Use in Fiber Modification

<130> BCS 06-2008 WO1 <150> EP 06015433.3 <151> 2006-07-25 <150> US 60/834066 <151> 2006-07-28 <160> 18

<170> PatentIn version 3.3 <210> 1 <211> 2635 <212> DNA

<213> Gossypium hirsutum <220>

<221> CDS

<222> (84) .. (2474)

<400> 1

acgcggggtg tctttgattt ttatccatat cctaattctt catacattcc ttcccttctg 60

ccatttcagg aacccaaaaa aca atg gct tca till agt gtt tgt gag cgt ttg 113

Met Ala Ser Ile Ser Val Cys Glu Arg Leu 10

ggt gaa tet cta gct act cat cca cag cag gea aag tet by ttg tca 161

Gly Glu Ser Leu Wing Thr His Pro Gln Gln Wing Lys Ser Ile Leu Ser 20 25

agg att gaa age ct gga aag ggt att cat aag tet

Arg Ile Glu Being Read Gly Lys Gly Ile His Lys Being Read Gln Lys Leu 35 40

tcg gtt ctc gat aaa gag gcc gga aat caa gea ctt gat ggg atg gtg 257

Ser Val Leu Asp Lys Glu Wing Gly Asn Gln Wing Leu Asp Gly Met Val

45 50 55

gtg ggg gtc ctc agg tcc act cag gaa gct gta gtg tcg tet cca ttg 305

Val Gly Val Leu Arg Be Thr Gln Glu Wing Val Val Ser Be Pro Leu 60 65 70

gtt gcc ctt gcc att cgt tca gct cct gga gtt tgg gag tac att gct 353

Val Wing Leu Wing Ile Arg Ser Wing Pro Gly Val Trp Glu Tyr Ile Wing 75 80 85 90

gtg gag gtc caa aag ctt tt gtg gag gaa atg ccc gtt gct gag tat 401

Val Glu Val Gln Lys Leu Phe Val Glu Glu Met Pro Val Wing Glu Tyr

95 100 105

cta cgg ttg aag gaa gaa ctt gtt gat gga age tcc aat ggc gag ttt 449

Leu Arg Leu Lys Glu Glu Leu Val Asp Gly Ser As Asn Gly Glu Phe

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atg ttg gaa ttg gac tt ggt gea ttc aat aat tet gtt cct cgt cca 497

Met Leu Glu Leu Asp Phe Gly Ala Phe Asn Asn Ser Val Pro Arg Pro

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tet ctt tca aag tcc att ggt aat ggc atg gac ttc ctc aac cgc cac 545

Be Read Be Lys Be Ile Gly Asn Gly Met Asp Phe Read Asn Arg His

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ctt tet gcc aag cta ttt caa gac aag gag aac ttg aac ttg ttg ctt 593

Leu Ser Ala Lys Leu Phe Gln Asp Lys Glu Asn Leu Asn Leu Leu Leu 155 160 165 170

gaa ttt ctc caa att cac tgc cag aag gga aag ggt atg ctg ttg aat 641

Glu Phe Leu Gln Ile His Cys Gln Lys Gly Lys Gly Met Leu Leu Asn

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25 Asp Arg Ile

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Val Wing Asn

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Gln Gly Leu 315

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Gln Asp Val Asn Ser 190

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ttt ctg ggg att ggt Phe Leu Gly Ile Gly 225

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cct gtc gca ctt gaa Pro Val Wing Leu Glu 255

gtt gta until atg act Val Val Ile Met Thr 270

gga tac cct gac ac Gly Tyr Pro Asp Thr 290

cgt gcc ttg gag gaa Arg Wing Leu Glu Glu 305

gac att acc cca cgt Asp Ile Thr Pro Arg 320

gta gga aca tgc act Val Gly Thr Thr Cys 335

tat tet gat att ctt Tyr Ser Asp Ile Leu 350

Read Gln His Wing Leu 195

tcg gat acc ccg tac Ser Asp Thr Pro Tyr 215

ttg gag aag gga tgg Leu Glu Lys Gly Trp 230

cat ctt cta ttg gat His Leu Leu Leu Asp 245

age ttc ctt gga aga Ser Phe Leu Gly Arg 260

ccc cat gga tac tt pro Gly Tyr Phe 275

ggt ggt cag gtt gtt Gly Gly Gln Val Val 295

gag ttg ctt cat cgt Glu Leu Leu His Arg 310

until cta gtc att act Ile Leu Val Ile Thr 325

ggt cag cgt ctt gag Gly Gln Arg Leu Glu 340

cgg gta ccc ttc aga Arg Val Pro Phe Arg 355

Arg Lys Wing 200

tca gtt ttc 737

To be Val Phe

ggt gat aat 785

Gly Asp Asn

ctc ctt cag 833

Leu Leu Gln 250

until cca ttg 881

Ile Pro Leu 265

gcc caa gac 929

Gln Wing Asp 280

tat till tta 977

Tyr Ile Leu

ttc aag ctg 1025

Phe Lys Leu

cgg ctc ctc 1073

Arg Leu Leu 330

aaa gtg tat 1121

Lys Val Tyr 345

act gag aag 1169

Thr Glu Lys

360 10

15

gga att gta cgt cca tgg till tca cga ttc aaa gtc tgg cct tac ctg 1217

Gly Ile Val Arg Pro Trp Ile Ser Arg Phe Lys Val Trp Pro Tyr Leu

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gag act tac act aag gat gtt gea gct gag until acc aaa gag ttc cag 1265

Glu Thr Tyr Thr Lys Asp Val Wing Wing Glu Ile Thr Lys Glu Phe Gln

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ggc aag cct gat ctg att ggg aac tac agt gat gga aac att gtt 1313

Gly Lys Pro Asp Leu Ile Val Gly Asn Tyr Ser Asp Gly Asn Ile Val 395 400 405 410

gt tet tta ttg gea cat cat aag ttt gat gtt aca cag tgc act att gct 1361

Ala Ser Leu Leu Wing His Lys Phe Asp Val Thr Gln Cys Thr Ile Wing

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cgt gea ctc gag aag acg aaa tac ccg gat tca gac att aac tgg aaa 1409

Arg Wing Read Glu Lys Thr Lys Tyr Pro Asp Ser Asp Ile Asn Trp Lys

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cag ctt gag gat aag tat ttc tcc tgt cag ttt act gct gat ctt 1457

Gln Leu Glu Asp Lys Tyr His Phe Ser Cys Gln Phe Thr Wing Asp Leu

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att gct atg aac cat act gat ttt till till acc age acc ttc caa gag 1505

Ile Ala Met Asn His Thr Asp Phe Ile Ile Thr Be Thr Phe Gln Glu

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att gct gga age aag gac act ctc ggc caa tac gag agt cac att gct 1553

Ile Wing Gly Be Lys Asp Thr Read Gly Gln Tyr Glu Be His Ile Wing 475 480 485 490

ttc act ctt cg ggg ctc tac cgc gtt gtt gat ggg until gat gtt ttt 1601 Phe Thr Leu Pro Gly Leu Tyr Arg Val Val Asp Gly Ile Asp Val Phe

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gat ccc aaa ttc aat att gtc tcc cct ggt gct gat atg age ata tac 1649

Asp Pro Lys Phe Asn Ile Val Ser Pro Gly Wing Asp Met Ser Ile Tyr

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ttc cct tac acg gaa gag aag cgg agg ttg aag aag ttc cac ccg gag 1697

Phe Pro Tyr Thr Glu Glu Lys Arg Arg Read Le Lys Lys Phe His Pro Glu

20

30 att gaa gag Ile Glu Glu 540

gta cta aaa Val Leu Lys 555

gac cga gp Asp Arg Val

10

age cgg ctg Ser Arg Leu

aga aag gaa 1 5 Arg Lys Glu 605

tat gaa ctt Tyr Glu Leu 620

tcg tcc cag Ser Ser Gln 635

tgc gac acc Cys Asp Thr

25

ggc ttg act Gly Leu Thr

aca tgt tac Thr Cys Tyr 685

ttc aac until

530

ctt ctt tac age cct Leu Leu Tyr Ser Pro 545

gac cgc aac cc cc Asp Arg Asn Lys Pro 560

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agg gaa ctg gtt aac Arg Glu Leu Val Asn 590

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till gaa aaa tac aag Ile Glu Lys Tyr Lys 625

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999 ggc cct gct gag Gly Gly Pro Wing Glu 690

gat cct tat aac ggt

48

535

gtt gag aat aca gag Val Glu Asn Thr Glu 550

att ctg ttt acc atg Ile Leu Phe Thr Met 565

ctc gta gag ttc tat Leu Val Glu Phe Tyr 580

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gaa aag gct gaa atg Glu Lys Ala Glu Met 615

ttg aat gga caa ttc Read Asn Gly Gln Phe 630

aat ggt gaa ctc tat Asn Gly Glu Leu Tyr 645

cag cct cct tac tac Gln Pro Ile Tyr 660

acc tgt gga cca cca Thr Cys Gly Leu Pro 675

att ata gtt cac gga Ile Ile Val His Gly 695

gat ttg gct gcc gag

cac tta tgt 1745

His Leu Cys

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Wing Lys Asn 585

gga gat agg 1889 Gly Asp Arg 600

aag aag atg 1937

Lys Lys Met

agg tgg ata 1985 Arg Trp Ile

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gag gct ttt 2081

Glu Wing Phe 665

aca ttt gea 2129 Thr Phe Ala 680

aaa tcg ggg 2177

Lys Ser Gly

acc ctt gcc 2225 10

Phe Asn Ile Asp Pro Tyr Asn Gly Asp Leu Wing Wing Wing Glu Thr Leu Wing

700 705 710

aat ttc ttg gag aag tgc aaa gcg gat cca tct tat tgg gat gag until 2273

Asn Phe Phe Glu Lys Cys Lys Wing Asp Pro Ser Tyr Trp Asp Glu Ile

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tcc cag gga ggg ttg aaa cgc ata cag gag aag tat acg tgg cag att 2321

Be Gln Gly Gly Read Lys Arg Ile Gln Glu Lys Tyr Thr Trp Gln Ile

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tac tcc gag aag cta ttg act ctc acc gga gt tat ggc ttt tcg aaa 23 69 Tyr Ser Glu Lys Leu Leu Thr Leu Thr Gly Val Tyr Gly Phe Ser Lys

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cat gta gct tac cag gag caa cgt ggg cgc aag cgt tac att gaa atg 2417 His Val Tyr Gln Glu Arg Gly Arg Lys Tyr Ile Glu Met

765 770 775

ttg cat gea tgg atg tat aac aat cgg gtc aag act gtt cca cta gct 2465 Leu His Ala Trp Met Tyr Asn Asn Arg Val Lys Thr Val Pro Leu Ala 780 785 790

gtt gag taa agaaaaagca aaaggctgct tggaaaccaa ggaatttcca 2514

Val glu

795

agaatgattg ctattgaagt tttatttttg cttctttttt tcaatttcgt aaatgtttaa 2574 tttcaacttt ggggtttccc cttggaatgt tttgaactct gggtttggtc ccattgattg 2634 c

2635

<210> 2 <211> 796 <212> PRT

<213> Gossypium hirsutum <400> 2

15

20

30

Met Ala Ser Ile Ser Val Cys Glu Arg Leu Gly Glu Ser Leu Wing Thr 10 I5

His Pro Gln Gln Wing Lys Be Ile Leu Be Arg Ile Glu Be Leu Gly 25 30 Lys Gly Ile His Lys Be Gln Lys Leu Leu Be Val Leu Asp Lys Glu

40 45

Gly Wing Asn Gln Wing Read Asp Gly Met Val Val Gly Val Leu Arg Ser

50 55 60

Thr Gln Glu Wing Val Val Ser Be Pro Leu Val Wing Leu Wing Ile Arg 65 70 75 80

Ser Pro Wing Gly Val Trp Glu Tyr Ile Wing Val Glu Val Gln Lys Leu

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Phe Val Glu Glu Met Pro Val Glu Wing Tyr Leu Arg Leu Lys Glu Glu

100 105 110

Read Val Asp Gly Be Ser Asn Gly Glu Phe Met Read Le Glu Read Le Asp Phe

115 120 125

Gly Ala Phe Asn Asn Be Val Pro Arg Pro Be Read Lys Be Ile

130 135 140

Gly Asn Gly Met Asp Phe He Read Asn Arg His He Read Be Wing Lys Leu Phe 145 150 155 160

Gln Asp Lys Glu Asn Leu Asn Leu Leu Leu Leu Glu Phe Leu Gln Ile His

165 170 175

Cys Gln Lys Gly Lys Gly Met Leu Read Asn Asp Arg Ile Gln Asp Val

180 185 190

Asn Be Read Gln His Wing Read Arg Lys Wing Glu Glu Tyr Leu Thr Pro

195 200 205

Read Be Ser Asp Thr Pro Tyr Be Val Phe Glu Lys Arg Phe Read Gly

210 215 220

Ile Gly Leu Glu Lys Gly Trp Gly Asp Asn Asn Wing Glu His Val Leu Glu 225 230 235 240

Met Ile His Leu Leu Leu Asp Leu Leu Gln Wing Pro Asp Pro Val Wing

245 250 255

Read Glu Ser Phe Read Gly Arg Ile Pro Read Val Wing Asn Val Val Ile

260 265 270

Met Thr Pro His Gly Tyr Phe Wing Gln Asp Asn Val Leu Gly Tyr Pro 275 280 285 Asp Thr Gly Gly Gln Val Val Tyr Ile Leu Asp Gln Val Arg Wing Alu Leu

290 295 300

Glu Glu Glu Leu Read His Arg Phe Lys Leu Gln Gly Leu Asp Ile Thr 305 310 315 320

Pro Arg Ile Leu Val Ile Thr Arg Leu Leu Pro Asp Wing Val Gly Thr

325 330 335

Thr Cys Gly Gln Arg Read Le Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp

340 345 350

Ile Leu Arg Val Pro Phe Arg Thr Glu Lys Gly Ile Val Arg Pro Trp

355 360 365

Ile Ser Arg Phe Lys Val Trp Pro Tyr Leu Glu Thr Tyr Thr Lys Asp

370 375 380

Val Wing Wing Glu Ile Thr Lys Glu Phe Gln Gly Lys Pro Asp Leu Ile 385 390 395 400

Val Gly Asn Tyr Be Asp Gly Asn Ile Val Wing Be Read Leu Wing His

405 410 415

Lys Phe Asp Val Thr Gln Cys Thr Ile Wing Arg Wing Leu Glu Lys Thr

420 425 430

Lys Tyr Pro Asp Be Asp Ile Asn Trp Lys Gln Read Glu Asp Lys Tyr

435 440 445

His Phe Ser Cys Gln Phe Thr Wing Asp Read Ile Wing Met Asn His Thr

450 455 460

Asp Phe Ile Ile Thr Be Thr Phe Gln Glu Ile Wing Gly Ser Lys Asp 465 470 475 480

Thr Leu Gly Gln Tyr Glu Being His Ile Wing Phe Thr Leu Pro Gly Leu

485 490 495

Tyr Arg Val Val Asp Gly Ile Val Val Asp

500 505 510

Val Ser Pro Gly Wing Asp Met Ile Tyr Phe Pro Tyr Thr Glu Glu

515 520 525

Lys Arg Arg Leu Lys Lys Phe His Pro Glu Ile Glu Glu Leu Leu Tyr 530 535 540 Ser Pro Val Glu Asn Thr Glu His Leu Cys Val Leu Lys Asp Arg Asn 545 550 555 560

Lys Pro Ile Leu Phe Thr Met Wing Arg Leu Asp Arg Val Lys Asn Leu

565 570 575

Thr Gly Leu Val Glu Phe Tyr Ala Lys Asn Ser Arg Leu Arg Glu Leu

580 585 590

Val Asn Leu Val Val Val Gly Gly Asp Arg Arg Lys Glu Ser Lys Asp

595 600 605

Read Glu Glu Lys Wing Glu Met Lys Lys Met Tyr Glu Read Le Ile Glu Lys

610 615 620

Tyr Lys Read Asn Gly Gln Phe Arg Trp Ile Be Ser Gln Met Asn Arg 625 630 635 640

Val Arg Asn Gly Glu Read Tyr Arg Tyr Ile Cys Asp Thr Lys Gly Wing

645 650 655

Phe Val Gln Pro Ile Tyr Glu Wing Phe Gly Leu Thr Val Val Glu

660 665 670

Met Thr Cly Wing Gly Leu Pro Thr Phe

675 680 685

Glu Wing Ile Ile Val His Gly Lys Ser Gly Phe Asn Ile Asp Pro Tyr

690 695 700

Asn Gly Asp Read Wing Wing Glu Thr Read Wing Wing Asn Phe Phe Glu Lys Cys 705 710 715 720

Lys Wing Asp Pro Being Tyr Trp Asp Glu Ile Being Gln Gly Gly Leu Lys

725 730 735

Arg Ile Gln Lys Tyr Thr Trp Gln Ile Tyr Be Glu Lys Leu Leu

740 745 750

Thr Leu Thr Gly Val Tyr Gly Phe Ser Lys His Val Wing Tyr Gln Glu

755 760 765

Gln Arg Gly Arg Lys Arg Tyr Ile Glu Met Read His Wing Trp Met Tyr

770 775 780

Asn Asn Arg Val Lys Thr Val Pro Leu Wing Val Glu 785 790 795 <210> 3 <211> 796 <212> PRT

<213> Gossypium hirsutum <4 00> 3

Met Wing Ser Ile Ser Val Cys Glu Arg Leu Gly Glu Ser Leu Wing Thr 10 15

His Pro Gln Gln Wing Lys Be Ile Read G Be Arg Ile Glu Be Read Gly

25 30

Lys Gly Ile His Lys Be Gln Lys Leu Read Le Be Val Leu Asp Lys Glu

40 45

Wing Gly Asn Gln Wing Leu Asp Gly Met Val Val Glu Val Leu Arg Ser

50 55 60

Thr Gln Glu Wing Val Val Ser Be Pro Leu Val Wing Leu Wing Ile Arg 65 70 75 80

Ser Pro Wing Gly Val Trp Glu Tyr Ile Wing Val Glu Val Gln Lys Leu

85 90 95

Phe Val Glu Glu Met Pro Val Glu Wing Tyr Leu Arg Leu Lys Glu Glu

100 105 110

Read Val Asp Gly Be Ser Asn Gly Glu Phe Met Read Le Glu Read Le Asp Phe

115 120 125

Gly Ala Phe Asn Asn Be Val Pro Arg Pro Be Read Lys Be Ile

130 135 140

Gly Asn Gly Met Asp Phe He Read Asn Arg His He Read Be Wing Lys Leu Phe 145 150 155 160

Gln Asp Lys Glu Asn Leu Asn Leu Leu Leu Leu Glu Phe Leu Gln Ile His

165 170 175

Cys Gln Lys Gly Lys Gly Met Leu Read Asn Asp Arg Ile Gln Asp Val

180 185 190

Asn Be Read Gln His Wing Read Arg Lys Wing Glu Glu Tyr Leu Thr Pro

195 200 205

Read Being Ser Asp Thr Pro Tyr Being Val Phe Glu Lys Arg Phe Leu Gly 210

Ile Gly Leu Glu Lys 225

Met Ile His Leu Leu 245

Read Glu Ser Phe Leu 260

Met Thr Pro His Gly 275

Asp Thr Gly Gly Gln 290

Glu Glu Glu Leu Leu 305

Pro Arg Ile Leu Val 325

Thr Cys Gly Gln Arg 340

Ile Leu Arg Val Pro 355

Ile Ser Arg Phe Lys 370

Val Wing Glu Ile Wing 385

Val Gly Asn Tyr Ser 405

Lys Phe Asp Val Thr 420

Lys Tyr Pro Asp Ser 435

His Phe Ser Cys Gln 450

Asp Phe Ile Ile Thr

215

Gly Trp Gly Asp Asn Wing 230 235

Leu Asp Leu Leu Gln Wing 250

Gly Arg Ile Pro Leu Val 265

Tyr Phe Ala Gln Asp Asn 280

Val Val Tyr Ile Leu Asp 295

His Arg Phe Lys Leu Gln 310 315

Ile Thr Arg Leu Leu Pro 330

Read Glu Lys Val Tyr Gly 345

Phe Arg Thr Glu Lys Gly 360

Val Trp Pro Tyr Leu Glu 375

Thr Lys Glu Phe Gln Gly 390 395

Asp Gly Asn Ile Val Wing 410

Gln Cys Thr Ile Wing His 425

Asp Ile Asn Trp Lys Gln 440

Phe Thr Wing Asp Leu Ile 455

Ser Thr Phe Gln Glu Ile

220

Glu His Val Leu Glu 240

Pro Asp Pro Val Wing 255

Wing Asn Val Val Ile 270

Val Leu Gly Tyr Pro 285

Gln Val Arg Wing Leu 300

Gly Leu Asp Ile Thr 320

Asp Wing Val Gly Thr 335

Thr Lys Tyr Ser Asp 350

Ile Val Arg Pro Trp 365

Thr Tyr Thr Lys Asp 380

Lys Pro Asp Leu Ile 400

Ser Leu Leu Wing His 415

Wing Read Glu Lys Thr 430

Read Glu Asp Lys Tyr 445

Met Wing Asn His Thr 460

Gly Ser Lys Wing Asp 465 470 475 480

Thr Leu Gly Gln Tyr Glu Being His Ile Wing Phe Thr Leu Pro Gly Leu

485 490 495

Tyr Arg Val Val Asp Gly Ile Val Val Asp

500 505 510

Val Ser Pro Gly Wing Asp Met Ile Tyr Phe Pro Tyr Thr Glu Glu

515 520 525

Lys Arg Arg Read Lys Lys Phe His Pro Glu Ile Glu Glu Read Leu Tyr

530 535 540

Ser Pro Val Glu Asn Thr Glu His Leu Cys Val Leu Lys Asp Arg Asn 545 550 555 560

Lys Pro Ile Leu Phe Thr Met Wing Arg Leu Asp Arg Val Lys Asn Leu

565 570 575

Thr Gly Leu Val Glu Phe Tyr Ala Lys Asn Ser Arg Leu Arg Glu Leu

580 585 590

Val Asn Leu Val Val Val Gly Gly Asp Arg Arg Lys Glu Ser Lys Asp

595 600 605

Read Glu Glu Lys Wing Glu Met Lys Lys Met Tyr Glu Read Le Ile Glu Lys

610 615 620

Tyr Lys Read Asn Gly Gln Phe Arg Trp Ile Be Ser Gln Met Asn Arg 625 630 635 640

Val Arg Asn Gly Glu Read Tyr Arg Tyr Ile Cys Asp Thr Lys Gly Wing

645 650 655

Phe Val Gln Pro Ile Tyr Glu Wing Phe Gly Leu Thr Val Val Glu

660 665 670

Met Thr Cly Wing Gly Leu Pro Thr Phe

675 680 685

Glu Wing Ile Ile Val His Gly Lys Ser Gly Phe Asn Ile Asp Pro Tyr

690 695 700

Asn Gly Asp Read Wing Wing Glu Thr Read Wing Wing Asn Phe Phe Glu Lys Cys 705 710 715 720

Lys Wing Asp Pro Be Tyr Trp Asp Glu Ile Be Gln Gly Gly Leu Lys 735 Lys Leu Leu 750

Tyr Gln Glu Trp Met Tyr

<210> 4

<211> 3353

<212> DNA

<213> Gossypium hirsutum <220>

<221> misc_feature

<222> (32). . (34)

<223> initiation codon <220>

<221> exon

<222> (32) .. (111)

<223> exonl <220>

<221> exon

<222> (197) .. (314)

<223> exon2 <220>

<221> exon

<222> (394) .. (545)

<223> exon3 <220>

<221> exon

<222> (625) .. (817)

725

Arg Ile Gln Glu Lys Tyr 740

Thr Leu Thr Gly Val Tyr 755

Gln Arg Gly Arg Lys Arg 770

Asn Asn Arg Val Lys Thr 785 790

730

Thr Trp Gln Ile Tyr Ser Glu 745

Gly Phe Ser Lys His Val Wing 760 765

Tyr Ile Glu Met Read His Wing 775 780

Val Pro Leu Wing Val Glu 795 <223> exon4 <220>

<221> exon <222> (897) .. (1328)

<223> exon5 <220>

<221> exon <222> (1399). . (1689)

<223> exon6 <220>

<221> exon

<222> (1783). . (1949)

<223> exon7 <220>

<221> exon

<222> (2050) .. (2274)

<223> exon8

<220>

<221> exon

<222> (2358). . (3060)

<223> exon9 <220>

<221> exon

<222> (3169). . (3195)

<223> exonlO <220>

<221> misc_feature

<222> (3196) .. (3171)

<223> Termination Codon

<400> 4

cccttctgcc atttcaggaa cccaaaaaac atg gct tca till agt gtt tgt 52

Met Ala Ser Ile Ser Val Cys 1 5

gag cgt ttg ggt gaa tct cta gct act cat cca cag cag gca aag tct 100

Glu Arg Read Gly Glu Be Read Ala Thr His Pro Gln Gln Ala Lys Ser 15 20

until ttg tca ag gtataatgta aataagtgct gccttcaaat ttctggtctt 151

Ile Leu Ser Arg 25

tcttcgttat tttttattac attgtttata tatgtcatat tgcag g att gaa age 206

Ile Glu Ser

30

ctc gga aag ggt att cat aag tct caa aag ctg ctc tcg gtt ctc gat 254

Leu Gly Lys Gly Ile His Lys Be Gln Lys Leu Read Le Be Val Leu Asp

40 45

aaa gag gcc gga aat caa gca ct gat ggg atg gtg gtg gag gtc ctc 3 02

1 5 Lys Glu Wing Gly Asn Gln Wing Read Asp Gly Met Val Val Glu Val Leu 50 55 60

agg tcc act cag gtcacaataa aacaaaccct ttctttggtt atgtttcatg 354

Arg Ser Thr Gln 65

atgagaatat tggactgata tgttaccgtt atctaatag gaa gct gta gtg tcg 4 08

Glu Wing Val Val Ser 70

tct cca ttg gtt gcc ctt gcc att cgt

Ser Pro Leu Wing Val Leu Wing Ile Arg Ser Pro Wing Gly Val Trp Glu

75 80 85

tac att gct gtg gag gtc caa aag ctt ttt gtg gag gaa atg ccc gtt 504

Tyr Ile Wing Val Glu Val Gln Lys Leu Phe Val Glu Glu Met Pro Val

90 95 100

gct gag tat cta cgg ttg aag gaa gaa ct gt gat gga ag 545

Glu Wing Tyr Leu Arg Leu Lys Glu Glu Leu Val Asp Gly Ser 105 110 115

gtaggtagtt ttctcttaca gctcctggtt tatattatgt ttgagtttat gttgaacaat 605 10

15

gctcaatgat gtggttcag c tcc aat ggc gag ttt atg ttg gaa ttg gac 655

Ser Asn Gly Glu Phe Met Leu Glu Leu Asp 120 125

tt ggt gca ttc aat aat tct gtt cct cgt cca tct ctt tca aag tcc 703

Phe Gly Ala Phe Asn Asn Be Val Pro Arg Pro Be Read Be Lys Be

130 135 140

att ggt aat ggc atg gac ttc ctc aac cgc cac ctt tct gcc aag cta 751

Ile Gly Asn Gly Met Asp Phe Leu Asn Arg His Leu To Be Ala Lys Leu

145 150 155

ttt caa gac aag gag aac ttg aac ttg ttg ct gaa ttt ctc caa att 799

Phe Gln Asp Lys Glu Asn Leu Asn Leu Leu Leu Leu Glu Phe Leu Gln Ile 160 165 170 175

cac tgc cag aag gga aag gtaaaagttt cactttcact atttgttttt 847

His Cys Gln Lys Gly Lys 180

cctttctgta tacatcaaaa cttaagaatc gttcaatgtt atgccacag ggt atg ctg 905

Gly Met Leu

ttg aat gac aga until hunting gat gtg aat tcc ctc hunting cat gca tta agg 953

Leu Asn Asp Arg Ile Gln Asp Val Asn Ser Leu Gln His Wing Leu Arg 185 190 195 200

aag gcc gag gag ctg act cct cta tcc tcg gat acc ccg tac tca 1001

Lys Wing Glu Glu Tyr Leu Thr Pro Leu Ser Ser Asp Thr Pro Tyr Ser

205 210 215

gt ttc gag aaa agg ttt ctg ggg att ggt ttg gag aag gga tgg ggt 1049

Val Phe Glu Lys Arg Phe Leu Gly Ile Gly Leu Glu Lys Gly Trp Gly

220 225 230

gat aat gct gag cat gtt ctt gag atg until cat ctt cta ttg gat ctc 1097

Asp Asn Glu Wing His Val Leu Glu Met Ile His Leu Leu Leu Asp Leu 235 240 245

ctt cag gca cct gat cct gtc gca ct gaa age ttc ctt gga aga until 1145

Read Gln Wing Pro Asp Pro Val Wing Read Glu Ser Phe Leu Gly Arg Ile

20

25 10

250 255 260

cca ttg gtc gcc aat gtt gta until atg act ccc cat gga tac ttt gcc 1193

Pro Leu Val Wing Asn Val Val Ile Met Thr Pro His Gly Tyr Phe Wing 265 270 275 280

caa gac aat gtt ttg gga tac cct gac aca ggt ggt cag gtt gtt tat 1241

Gln Asp Asn Val Leu Gly Tyr Pro Asp Thr Gly Gly Gln Val Val Tyr

285 290 295

till tta gat caa gtt cgt gcc ttg gag gaa gag ttg ctt cat cgt ttc 1289

Xle Leu Asp Gln Val Arg Wing Leu Glu Glu Glu Leu Read His Arg Phe

300 305 310

aag ctg caa gga ctc gac att acc cca cgt until cta gtc gtgagtatct 1338

Lys Leu Gln Gly Leu Asp Ile Thr Pro Arg Ile Leu Val 315 320 325

gaatgttaat tcagtttcga tcattatgtt taatgcttaa tttggctgtt tgtgtaccag 1398 att act cgg ctc cct gat gct gta gga aca act tgc ggt cag cgt 1446 Ile Thr Arg Leu Pro Asp Thr Gly Val Gly

330 335 340

ct gag aaa gtg tat gga acc aag tat tet gat att ctt cgg gta ccc 1494

Leu Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp Ile Leu Arg Val Pro 345 350 355

ttc aga act gag aag gga att gta cgt cca tgg until tca cga ttc aaa 1542

Phe Arg Thr Glu Lys Gly Ile Val Arg Pro Trp Ile Be Arg Phe Lys

360 365 370

gtc tgg cct tac ctg gag act tac act aag gat gtt gea gct gag until 1590

Val Trp Pro Tyr Leu Glu Thr Tyr Thr Lys Asp Val Wing Wing Glu Ile 375 380 385

acc aaa gag ttc cag ggc aag cct gat ctg att gtt ggg aac tac agt 1638

Thr Lys Glu Phe Gln Gly Lys Pro Asp Ile Ile Val Gly Asn Tyr Ser 390 395 400 405

gat gga aac att gtt gct tet tta ttg gea cat aag ttt gat gtt aca 1686

Asp Gly Asn Ile Val Wing Be Read Leu Wing His Lys Phe Asp Val Thr 410 415 420

30 10

cag gtttgttaaa taccaattaa ttgaacaagt tccttgctgt attttgatgt 1739

Gln

tccccgtgca atctaaaaac ttggttttca atgttttatg cag tgc act att gct 1794

Cys Thr Ile Wing 425

cat gca ctc gag aag ag aaa tac ccg gat tca gac att aac tgg aaa 1842

His Wing Read Glu Lys Thr Lys Tyr Pro Asp Ser Asp Ile Asn Trp Lys

430 435 440

cag ctt gag gat aag tat ttc tcc tgt cag ttt act gct gat ctt 1890

Gln Leu Glu Asp Lys Tyr His Phe Ser Cys Gln Phe Thr Wing Asp Leu

445 450 455

att gct atg aac cat act gat ttt until till acc age acc ttc caa gag 1938

Ile Ala Met Asn His Thr Asp Phe Ile Ile Thr Be Thr Phe Gln Glu

460 465 470

att gct gga ag gtaatctatc tcccacttta ctacatttgg ttcccttgta 1989

Ile Wing Gly Ser

475

cgttgtaaat gttgccacct atttgagtcc ctaacaatgt tttcattcca tgattttcag 2049 c aag gac act ctc gg ca ca ca tac gag ttc act ctt cca 2098 Lys Asp Thr Leu Gly Gln Tyr Glu Being His Ileu

ggg ctc tac cgc gtt gtt gat ggg till gat gtt ttt gat ccc aaa ttc 2146

Gly Leu Tyr Arg Val Val Asp Gly Ile Val Val Asp Phe Asp Pro Lys Phe 495 500 505 510

aat att gtc tcc cct ggt gct gat atg act ata tac ttc cct tac acg 2194

Asn Ile Val Ser Pro Gly Wing Asp Met Ser Ile Tyr Phe Pro Tyr Thr

515 520 525

gag gag ag cgg agg ttg aag aag ttc cac ccg gag att gaa gag ctt 2242

Glu Glu Lys Arg Arg Read Lys Lys Phe His Pro Glu Ile Glu Glu Leu 530 535 540

ct tac age cct gt gt gt aa gag aa gag cac tt gtgagatttt ggtcctattt 2294 10

15

Leu Tyr Ser Pro Val Glu Asn Thr Glu His Leu

545 550

ttacaccctt ttcactagat tttgtgttca gtgttaacaa tgtcttaaat tgttgtctca 2354 cag a tgt cta aaa gac cgc aac ag ccg att ctg ttt acc atg gca 2403 Cys Val Leu Lys Asp L5 As5 Arg 5

agg ttg gac cga gtg aag aac tta act ggg ctc gta gag ttc tat gcc 2451

Arg Leu Asp Arg Val Lys Asn Leu Thr Gly Leu Val Glu Phe Tyr Wing

570 575 580

aag aac acts cgg ctg agg gaa ctg gtt aac ttg gtt gta gta ggt gga 2499

Lys Asn Be Arg Leu Arg Glu Leu Val Asn Leu Val Val Val Gly Gly 585 590 595 600

gat agg aag gaa tec aag gac tta gaa gaa aag gct gaa atg aag 2547

Asp Arg Arg Lys Glu Be Lys Asp Leu Glu Glu Lys Wing Glu Met Lys

605 610 615

aag atg tat gaa ctt until gaa aaa tac aag ttg aat gga caa ttc aga 2595

Lys Met Tyr Glu Leu Ile Glu Lys Tyr Lys Leu Asn Gly Gln Phe Arg

620 625 630

tgg ata tcg tcc cag atg aac cga gtg aga aat ggt gaa ctc tat cgt 2643

Trp Ile Ser Be Gln Met Asn Arg Val Arg Asn Gly Glu Leu Tyr Arg 635 640 645

tat att tgc gac acc aga gga gca ttc gtt cag cct cct ata tac gag 2691

Tyr Ile Cys Asp Thr Lys Gly Phe Val Gln Pro Pro Ile Tyr Glu 650 655 660

gct ttt ggc ttg act gtt gtt gag gca atg acc tgt gga cta cca aca 2739 Phe Gly Wing Leu Thr Val Val Glu Wing Met Thr Cys Wing Gly Leu Pro Thr 665 670 675 680

ttt gca aca tgt tac ggg ggc cct gct gag att ata gtt cac gga aaa 2787

Phe Wing Thr Cys Tyr Gly Gly Pro Glu Wing Ile Ile Val His Gly Lys 685 690 695

tcg ggg ttc aac till gat cct tat aac ggt gat ttg gct gcc gag acc 2835

Ser Gly Phe Asn Ile Asp Pro Tyr Asn Gly Asp Read Wing Wing Glu Thr 10

700 705 710

ctt gcc aat ttc ttc gag aag tgc aaa gcg gat cca tct tat tgg gat 2883

Leu Wing Asn Phe Phe Glu Lys Cys Lys Wing Asp Pro To Be Tyr Trp Asp

715 720 725

gag till tcc cag gga ggg ttg aaa cgc ata cag gag aag tat aca tgg 2931

Glu Ile Being Gln Gly Gly Leu Lys Arg Ile Gln Glu Lys Tyr Thr Trp

730 735 740

cag att tac tcc gag aag cta ttg act ctc acc gga gt tat ggc ttt 2979

Gln Ile Tyr Ser Glu Lys Leu Leu Thr Leu Thr Gly Val Tyr Gly Phe 745 750 755 760

tcg aaa cat gta gct tac cag gag caa cgt ggg cgc aag cgt tac att 3027

Ser Lys His Val Tyr Wing Gln Glu Gln Arg Gly Arg Lys Arg Tyr Ile

765 770 775

catg atg ttg cat gea tgg atg tat aac aat cgg gtaaaacaat catgtctatt 3080 Glu Met Leu His Ala Trp Met Tyr Asn Asn Arg 780 785

attattatta catcaaacat cattttactc tttcattgaa ctgaaatttt catgtttctg 3140 aatgtttgtt taattgtgtg ctttacag gtc aag act gtt cca cta gct gtt 3192

Val Lys Thr Val Pro Leu Wing Val 790 795

gag taaagaaaaa gcaaaaggct gcttggaaac caaggaattt ccaagaatga 3245

Glu

20

25

ttgctattga agttttattt ttgcttcttt ttttcaattt cgtaaatgtt taatttcaac 3305 tttggggttt ccccttggaa tgttttgaac tctgggtttg gtcccatt 3353

<210> 5 <211> 796 <212> PRT

<213> Gossypium barbadense

<400> 5

Met Ala Ser Ile Be Val Cys Glu Arg Leu Gly Glu Ser Leu Wing Thr 10 15 His Pro Gln Gln Wing Lys Ser Ile Leu Be Arg Ile Glu Ser Leu Gly

25 30

Lys Gly Ile His Lys Be Gln Lys Leu Read Le Be Val Leu Asp Lys Glu

40 45

Wing Gly Asn Gln Wing Leu Asp Gly Met Val Val Glu Val Leu Arg Ser

50 55 60

Thr Gln Glu Wing Val Val Ser Be Pro Leu Val Wing Leu Wing Ile Arg 65 70 75 80

Ser Pro Wing Gly Val Trp Glu Tyr Ile Wing Val Glu Val Gln Lys Leu

85 90 95

Phe Val Glu Glu Met Pro Val Glu Wing Tyr Leu Arg Leu Lys Glu Glu

100 105 110

Read Val Asp Gly Be Ser Asn Gly Glu Phe Met Read Le Glu Read Le Asp Phe

115 120 125

Gly Ala Phe Asn Asn Be Val Pro Arg Pro Be Read Lys Be Ile

130 135 140

Gly Asn Gly Met Asp Phe He Read Asn Arg His He Read Be Wing Lys Leu Phe 145 150 155 160

Gln Asp Lys Glu Being Leu Asn Leu Leu Leu Leu Glu Phe Leu Gln Ile His

165 170 175

Cys Gln Lys Gly Lys Gly Met Leu Read Asn Asp Arg Ile Gln Asp Val

180 185 190

Asn Be Read Gln His Wing Read Arg Lys Wing Glu Glu Tyr Leu Thr Pro

195 200 205

Read Be Ser Asp Thr Pro Tyr Be Val Phe Glu Lys Arg Phe Read Gly

210 215 220

Ile Gly Leu Glu Lys Gly Trp Gly Asp Asn Asn Wing Glu His Val Leu Glu 225 230 235 240

Met Ile His Leu Leu Leu Asp Leu Leu Gln Wing Pro Asp Pro Val Wing

245 250 255

Leu Glu Ser Phe Leu Gly Arg Ile Pro Leu Val Wing Asn Val Val Ile 260 265 270 Met Thr Pro His Gly Tyr Phe Wing Gln Tyr Asri Val Leu Gly Tyr Pro

275 280 285

Asp Thr Gly Gly Gln Val Val Tyr Ile Leu Asp Gln Val Arg Wing Alu

290 295 300

Glu Glu Glu Leu Read His Arg Phe Lys Leu Gln Gly Leu Asp Ile Thr 305 310 315 320

Pro Arg Ile Leu Val Ile Thr Arg Leu Leu Pro Asp Wing Val Gly Thr

325 330 335

Thr Cys Gly Gln Arg Read Le Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp

340 345 350

Ile Leu Arg Val Pro Phe Arg Thr Glu Lys Gly Ile Val Arg Pro Trp

355 360 365

Ile Ser Arg Phe Lys Val Trp Pro Tyr Leu Glu Thr Tyr Thr Lys Asp

370 375 380

Val Wing Wing Glu Ile Thr Lys Glu Phe Gln Gly Lys Pro Asp Leu Ile 385 390 395 400

Val Gly Asn Tyr Be Asp Gly Asn Ile Val Wing Be Read Leu Wing His

405 410 415

Lys Phe Asp Val Thr Gln Cys Thr Ile Wing His Wing Leu Glu Lys Thr

420 425 430

Lys Tyr Pro Asp Be Asp Ile Asn Trp Lys Gln Read Glu Asp Lys Tyr

435 440 445

His Phe Ser Cys Gln Phe Thr Wing Asp Read Ile Wing Met Asn His Thr

450 455 460

Asp Phe Ile Ile Thr Be Thr Phe Gln Glu Ile Wing Gly Ser Lys Asp 465 470 475 480

Thr Leu Gly Gln Tyr Glu Being His Ile Wing Phe Thr Leu Pro Gly Leu

485 490 495

Tyr Arg Val Val Asp Gly Ile Val Val Asp

500 505 510

Val Ser Pro Gly Wing Asp Met Ser Ile Tyr Phe Pro Tyr Thr Glu Glu 515 520 525 Lys Arg Arg Read Lys Lys Lys Phe His Pro Glu Ile Glu Read Leu Tyr

530 535 540

Ser Pro Val Glu Asn Thr Glu His Leu Cys Val Leu Lys Asp Arg Asn 545 550 555 560

Lys Pro Ile Leu Phe Thr Met Wing Arg Leu Asp Arg Val Lys Asn Leu

565 570 575

Thr Gly Leu Val Glu Phe Tyr Ala Lys Asn Ser Arg Leu Arg Glu Leu

580 585 590

Val Asn Leu Val Val Val Gly Gly Asp Arg Arg Lys Glu Ser Lys Asp

595 600 605

Read Glu Glu Lys Wing Glu Met Lys Lys Met Tyr Glu Read Le Ile Glu Lys

610 615 620

Tyr Lys Read Asn Gly Gln Phe Arg Trp Ile Be Ser Gln Met Asn Arg 625 630 635 640

Val Arg Asn Gly Glu Read Tyr Arg Tyr Ile Cys Asp Thr Lys Gly Wing

645 650 655

Phe Val Gln Pro Wing Ile Tyr Glu Wing Phe Gly Leu Thr Val Val Glu

660 665 670

Met Thr Cly Wing Gly Leu Pro Thr Phe

675 680 685

Glu Wing Ile Ile Val His Gly Lys Ser Gly Phe Asn Ile Asp Pro Tyr

690 695 700

Asn Gly Asp Read Wing Wing Glu Thr Read Wing Wing Asn Phe Phe Glu Lys Cys 705 710 715 720

Lys Wing Asp Pro Being Tyr Trp Asp Glu Ile Being Gln Gly Gly Leu Lys

725 730 735

Arg Ile Gln Lys Tyr Thr Trp Gln Ile Tyr Be Glu Lys Leu Leu

740 745 750

Thr Leu Thr Gly Val Tyr Gly Phe Ser Lys His Val Wing Tyr Gln Glu

755 760 765

Gn Arg Gly Arg Lys Arg Tyr Ile Glu Met Leu His Wing Trp Met Tyr 770 775 780 Asn Asn Arg Val Lys Thr Val

<210> 6 <211> 3352 <212> DNA

<213> Gossypium barbadense <220>

<221> misc_feature <222> (32) .. (34) <223> initiation codon <220>

<221> exon <222> (32) .. (111)

<223> exonl <220>

<221> exon <222> (197) .. (314)

<223> exon2 <220>

<221> exon

<222> (394) .. (545)

<223> exon3 <22 0>

<221> exon

<222> (625) .. (817)

<223> exon4 <220>

<221> exon

<222> (897) .. (1328)

223> exon5 220>

221> exon <222> (1399) .. (1689) <223> exon6 <220>

<221> exon <222> (1783) .. (1949) <223> exon7 <220>

<221> exon <222> (2050) .. (2274) <223> exon8 <220>

<221> exon

<222> (2357) .. (3059)

<223> exon9

<220>

<221> exon <222> (3168) .. (3194) <223> exonlO <22 0>

<221> misc_feature <222> (3195) .. (3197) <223> Termination Codon <4 00> 6

cccttctgcc atttcaggaa cccaaaaaac a atg gct tca till agt gtt tgt

Met Ala Ser Ile Ser Val Cys 1 5

gag cgt ttg ggt gaa tet tta gct act cat cca cag cag gea aag tet Glu Arg Leu Gly Glu Ser Leu Ala Thr His Pro Gln Ala Lys Ser

15 20

until ttg tca ag gtataatgta aataagtgct gccttcaaat ttctggtctt Ile Leu Ser Arg 25 tcttcgttat tttttattac attgtttata tatgtcatat tgcag g att gaa age 206

Ile Glu Ser 30

ctc gga aag ggt att cat cat aag tet caa aag ctg ctc tcg gtt ctc gat 254

Leu Gly Lys Gly Ile His Lys Being Gln Lys Leu Leu Being Val Leu Asp 40 45

aaa gag gcc gga aat caa gea ctt gat ggg atg gtg gtg gag gtc ctc 302

Lys Glu Wing Gly Asn Gln Wing Read Asp Gly Met Val Val Glu Val Leu 50 55 60

agg tcc act cag gtcacaataa aacaaaccct ttctttggtt atgtttcatg 354

Arg Ser Thr Gln 65

atgagaatat tggactgata tgttaccgtt atctaatag gaa gct gta gtg tcg 408

Glu Wing Val Val Ser

grandfather

tet cca ttg gtt gcc ctt gcc att cgt tca gct cct gga gtt tgg gag 456

Ser Pro Leu Wing Val Leu Wing Ile Arg Ser Pro Wing Gly Val Trp Glu

75 80 85

tac att gct gtg gag gtc caa aag ctt ttt gtg gag gaa atg ccc gtt 504

Tyr Ile Val Glu Wing Val Gln Lys Leu Phe Val Glu Glu Met Pro Val 90 95 100

gct gag tat cta cgg ttg aag gaa gaa ct gt gat gga ag 545

Glu Wing Tyr Leu Arg Leu Lys Glu Glu Leu Val Asp Gly Ser 105 110 115

gtaggtagtt ttctcttaca gctcctggtt tatgttatgt ttgagtttat gttgaacaat 605 gctcaatgat gtggttcag c tcc aat ggc gag ttt atg ttg gaa ttg gac 655

Ser Asn Gly Glu Phe Met Leu Glu Leu Asp 120 125

ttt ggt gea ttc aat aat tet gtt cct cgt cca tet ctt tca aag tcc Phe Gly Ala Phe Asn Asn Ser Val Pro Arg Pro Ser Read Lys Ser 130 135 140

att ggt aat ggc atg gac ttc ctc aac cgc cac ctt tet gcc aag cta

703

751 Ile Gly Asn Gly Met Asp Phe Leu Asn Arg His Leu Ser Ala Lys Leu

145 150 155

tt caa gac aag gag age ttg aac ttg ttg ct gaa ttt ctc caa att 799

Phe Gln Asp Lys Glu Being Read Asn Read Leu Read Leu Glu

cac tgc cag aag gga aag gtaaaagttt cactttcact atttgttttt 847

His Cys Gln Lys Gly Lys 180

ccttctgta tacatcaaaa cttaagaatc gttcaatgtt atgccacag ggt atg ctg 905 Gly Met Leu

ttg aat gac aga until hunting gat gtg aat tcc ctc hunting cat gea tta agg 953

Leu Asn Asp Arg Ile Gln Asp Val Asn Ser Leu Gln His Wing Leu Arg

185 190 195 200

aag gcc gag gag ctg act cct cta tcc tcg gat acc ccg tac tca 1001

Lys Wing Glu Glu Tyr Leu Thr Pro Leu Ser Ser Asp Thr Pro Tyr Ser

205 210 215

gt ttc gag aaa agg ttt ctg ggg att ggt ttg gag aag gga tgg ggt

Val Phe Glu Lys Arg Phe Read Gly Ile Gly Read Glu Lys Gly Trp Gly

220 225 230

gat aat gct gag cat gtt ctt gag atg until cat ctt cta ttg gat ctc 1097

Asp Asn Alu Glu His Val Leu Glu Met Ile His Leu Leu Leu Asp Leu

235 240 245

ct cag gea cct gat cct gtc gea

Read Gln Wing Pro Asp Pro Val Wing Read Glu Ser Phe Leu Gly Arg Ile 250 255 260

cca ttg gtc gcc aat gtt gta until atg act ccc cat gga tac ttt gcc 1193

Pro Leu Val Wing Asn Val Val Ile Met Thr Pro His Gly Tyr Phe Wing 265 270 275 280

caa tac aat gtt ttg gga tac cct gac aca ggt ggt cag gtt gtt tat 1241 Gln Tyr Asn Val Leu Gly Tyr Pro Asp Thr Gly Gly Gln Val Val Tyr 285 290 295 10

15

20

25

till tta gat caa gtt cgt gcc ttg gag gaa gag ttg ctt cat cgt ttc 1289 Ile Leu Asp Gln Val Arg Wing Leu Glu Glu Glu Leu Leu His Arg Phe

300 305 310

aag ctg caa gga ctc gac att acc cca cgC until cta gtc gtgagtatct 1338

Lys Leu Gln Gly Leu Asp Ile Thr Pro Arg Ile Leu Val 315 320 325

gaatgttaat tcagtttcga tcattatgtt taatgcttaa tttggctgtt tgtgtaccag 1398 att act cgg etc ctc cct gat gct gta aca act tgc ggt cag cgt 1446

Ile Thr Arg Read Leu Pro Asp Wing Val Gly Thr Thr Cys Gly Gln Arg 330 335 340

ctt gag aaa gtg tat gga acc aag tat tet gat att ctt cgg gta ccc 1494 Leu Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp Ile Leu Arg Val Pro

345 350 355

ttc aga act gag aag gga att gta cgt cca tgg until tca cga ttc aaa 1542

Phe Arg Thr Glu Lys Gly Ile Val Arg Pro Trp Ile Be Arg Phe Lys

360 355 370

gtc tgg cct tac ctg gag act tac act aag gat gtt gea gct gag until 1590 Val Trp Pro Tyr Leu Glu Thr Tyr Thr Lys Asp Val Wing Ala Glu Ile 375 380 385

acc aaa gag ttc cag ggc aag cct gat ctg att gtt ggg aac tac agt 163 8

Thr Lys Glu Phe Gln Gly Lys Pro Asp Read Ile Val Gly Asn Tyr Ser

390 395 400 405

gat gga aac att gtt gct tet tta ttg gea cat aag ttt gat gtt aca 1686

Asp Gly Asn Ile Val Wing Be Read Leu Wing His Lys Phe Asp Val Thr

410 415 420

cag gtttgttaaa taccaattaa ttgaacaagt tccttgctgt attttgatgt 1739

Gln

30

tccccgtgca atctaaaaac ttggttttca atgttttatg cag tgc act att gct 1794

Cys Thr Ile Wing 425

cat gea ctc gag aag ag aaa tac ccg gat tca gac att aac tgg aaa 1842 15

His Wing Read Glu Lys Thr Lys Tyr Pro Asp Ser Asp Ile Asn Trp Lys

430 435 440

cag ctt gag gat aag tat ttc tcc tgt cag ttt act gct gat ctt 1890

Gln Leu Glu Asp Lys Tyr His Phe Ser Cys Gln Phe Thr Wing Asp Leu 445 450 455

att gct atg aac cat act gat ttt until till acc age acc ttc caa gag 1938

Ile Ala Met Asn His Thr Asp Phe Ile Ile Thr Be Thr Phe Gln Glu

460 465 470

att gct gga ag gtaatctatc tcccacttta ctacatttgg ttcccttgta 1989

Ile Wing Gly Ser 475

cgttgtaaat gttgccacct atttgagtcc ctaacaatgt tttcattcca tgattttcag 2049 c aag gac act ctc gg ca ca ca tac gag ttc act ctt cca 2098 Lys Asp Thr Leu Gly Gln Tyr Glu Being His Ileu

ggg ctc tac cgc gtt gtt gat ggg till gat gtt ttt gat ccc aaa ttc 2146

Gly Leu Tyr Arg Val Val Asp Gly Ile Val Val Asp Phe Asp Pro Lys Phe 495 500 505 510

aat att gtc tcc cct ggt gct gat atg act ata tac ttc cct tac acg 2194

Asn Ile Val Ser Pro Gly Wing Asp Met Ser Ile Tyr Phe Pro Tyr Thr

515 520 525

gag gag ag cgg agg ttg aag aag ttc cac ccg gag att gaa gag ctt 2242

Glu Glu Lys Arg Arg Leu Lys Lys Phe His Pro Glu Ile Glu Glu Leu

530 535 540

ct tac age cct gt gag aat aca gag cac tt gtgagatttt ggtcctattt 2294 Leu Tyr Ser Pro Val Glu Asn Thr Glu His Leu

545 550

ttacaccctt tcactagatt ttgtgttcag tgttaacaat gtcttaaatt gttgtctcac 2354 ag a tgt gta cta aaa gac aag ccg att ctg ttt acc atg gea 2402 Cys Val Leu Lys Asp Arg Asn Lys Pro Ile Leu

555 560 565

agg ttg gac cga gtg aag aac tta act ggg ctc

20

25 Arg Read Asp Arg Val Lys Asn Read Thr Gly Read Val Glu Phe Tyr Wing

570 575 580

aag aac acts cgg ctg agg gaa ctg gtt aac ttg gtt gta gta ggt gga 2498

Lys Asn Be Arg Leu Arg Glu Leu Val Asn Leu Val Val Val Gly Gly 585 590 595 600

gat agg aga aag gaa tcc aag gac tta gaa gaa aag gct gaa atg aag 2546

Asp Arg Arg Lys Glu Be Lys Asp Leu Glu Glu Lys Wing Glu Met Lys

605 610 615

aag atg tat gaa ctt till gaa aaa tac aag ttg aat gga caa ttc aga 2594

Lys Met Tyr Glu Leu Ile Glu Lys Tyr Lys Leu Asn Gly Gln Phe Arg 620 625 630

tgg ata tcg tcc cag atg aac cga gtg aga aat ggt gaa ctc tat cgt 2,642

Trp Ile Ser Be Gln Met Asn Arg Val Arg Asn Gly Glu Leu Tyr Arg 635 640 645

tat att tgc gac acc aag gga gea ttc gtt cag cct gct ata tac gag 2690 Tyr Ile Cys Asp Thr Lys Gly Ala Phe Val Gln Pro Ala Ile Tyr Glu

650 655 660

gct ttt ggc ttg act gtt gtt gag gea atg acc tgt gga cta cca aca 2738

Phe Gly Leu Wing Thr Val Val Glu Wing Met Thr Cys Wing Gly Leu Thr Thr 665 670 675 680

ttt gea aca tgt tac ggg ggc cct gct gag att ata gtt cac gga aaa 2786

Phe Wing Thr Cys Tyr Gly Gly Pro Glu Wing Ile Ile Val His Gly Lys

685 690 695

tcg ggg ttc aac until gat cct tat aac ggt gat ttg gct gcc gag acc 2834

Ser Gly Phe Asn Ile Asp Pro Tyr Asn Gly Asp Read Wing Wing Wing Glu Thr 700 705 710

ctt gcc aat ttc ttc gag aag tgc aaa gcg gat cca tet tat tgg gat 2882

Leu Wing Asn Phe Phe Glu Lys Cys Lys Wing Asp Pro To Be Tyr Trp Asp

715 720 725

gag till tcc cag gga ggg ttg aaa cgc ata cag gag aag tat aca tgg 2930

Glu Ile Being Gln Gly Gly Leu Lys Arg Ile Gln Glu Lys Tyr Thr Trp 730 735 740

30 10

cag att tac tcc gag aag cta ttg act ctc acc gga gt tat ggc ttt 2978

Gln Ile Tyr Ser Glu Lys Leu Leu Thr Leu Thr Gly Val Tyr Gly Phe 745 750 755 760

tcg aaa cat gta gct tac cag gag caa cgt ggg cgc aag cgt tac att 3026

Ser Lys His Val Tyr Wing Gln Glu Gln Arg Gly Arg Lys Arg Tyr Ile

765 770 775

gaa atg ttg cat gca tgg atg tat aac aat cgg gtaaaacaat catgtctatt 3079 Glu Met Leu His Wing Trp Met Tyr Asn Asn Arg

780 785

attattatta catcaaacat cattttactc tttcattgaa ctgaaaattt catgtttctg 3139 aatgtttgtt taattgtgtg ctttacag gtc aag act gtt cca cta gct gtt 3191

Val Lys Thr Val Pro Leu Wing Val 790 795

gag taaagaaaaa gcaaaaggct gcttggaaac caaggaattt ccaagaatga 3244

Glu

ttgctattga agttttattt ttgcttcttt ttttcaattt ggtaaatgtt taatttcaac 3304 tttggggttt ccccttggaa tgttttgaac tctgggtttg gtcccatt 3352

<210> 7

<211> 796

<212> PRT

<213> Gossypium arboreum <4 00> 7

Met Ala Ser Ile Ser Val Cys Glu Arg Leu Gly Glu Ser Leu Ala Thr

1 5 10 15

His Pro Gln Gln Wing Lys Be Ile Read G Be Arg Ile Glu Be Read Gly

25 30

Lys Gly Ile His Lys Be Gln Lys Leu Read Le Be Val Leu Asp Lys Glu

40 45

Wing Gly Asn Gln Wing Leu Asp Gly Met Val Val Glu Val Leu Arg Ser

50 55 60

Thr Gln Glu Wing Val Val Ser Be Pro Leu Val Wing Leu Wing Ile Arg 65 70 75 80

Ser Pro Wing Gly Val Trp Glu Tyr Ile Wing Val Glu Val Gln Lys Leu

85 90 95

Phe Val Glu Glu Met Pro Val Glu Wing Tyr Leu Arg Leu Lys Glu Glu

100 105 110

Read Val Asp Gly Be Ser Asn Gly Glu Phe Met Read Le Glu Read Le Asp Phe

115 120 125

Gly Ala Phe Asn Asn Be Val Pro Arg Pro Be Read Lys Be Ile

130 135 140

Gly Asn Gly Met Asp Phe He Read Asn Arg His He Read Be Wing Lys Leu Phe 145 150 155 160

Gln Asp Lys Glu Asn Leu Be Leu Leu Leu Leu Glu Phe Leu Gln Ile His

165 170 175

Cys Gln Lys Gly Lys Gly Met Leu Read Asn Asp Arg Ile Gln Asp Val

180 185 190

Asn Be Read Gln His Wing Read Arg Lys Wing Glu Glu Tyr Leu Thr Pro

195 200 205

Leu Thr Be Asp Thr Pro Tyr Be Val Phe Glu Lys Arg Phe Arg Glu

210 215 220

Ile Gly Leu Glu Lys Gly Trp Gly Asp Asn Asn Wing Glu His Val Leu Glu 225 230 235 240

Met Ile His Leu Leu Leu Asp Leu Leu Gln Wing Pro Asp Pro Val Wing

245 250 255

Leu Glu Ser Phe Leu Gly Thr Ile Pro Leu Val Wing Asn Val Val Ile

260 265 270

Met Thr Pro His Gly Tyr Phe Ala Gln Asp Asn Val Leu Gly Tyr Pro

275 280 285

Asp Thr Gly Gly Gln Val Val Tyr Ile Leu Asp Gln Val Arg Wing Alu

290 295 300

Glu Glu Glu Leu Read His Arg Phe Lys Leu Gln Gly Leu Asp Ile Thr 305 310 315 320

Pro Arg Ile Leu Val Ile Thr Arg Leu Leu Pro Asp Wing Val Gly Thr 325 330 335

Thr Cys Gly Gln Arg Read Le Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp

340 345 350

Ile Leu Arg Val Pro Phe Arg Thr Glu Lys Gly Ile Val Arg Pro Trp

355 360 365

Ile Ser Arg Phe Lys Val Trp Pro Tyr Leu Glu Thr Tyr Thr Lys Asp

370 375 380

Val Wing Glu Wing Val Thr Lys Glu Phe Gln Gly Lys Pro Asp Leu Ile 385 390 395 400

Val Gly Asn Tyr Ser Asp Gly Asn Ile Val Wing Ser Leu Leu Wing Leu

405 410 415

Lys Phe Asp Val Thr Gln Cys Thr Ile Wing His Wing Leu Glu Lys Thr

420 425 430

Lys Tyr Pro Asp Be Asp Ile Asn Trp Lys Gln Read Glu Asp Lys Tyr

435 440 445

His Phe Ser Cys Gln Phe Thr Wing Asp Read Ile Wing Met Asn His Thr

450 455 460

Asp Phe Ile Ile Thr Be Thr Phe Gln Glu Ile Wing Gly Ser Lys Asp 465 470 475 480

Thr Leu Gly Gln Tyr Glu Being His Ile Wing Phe Thr Leu Pro Gly Leu

485 490 495

Tyr Arg Val Val Asp Gly Ile Val Val Asp

500 505 510

Val Ser Pro Gly Wing Asp Met Ile Tyr Phe Pro Tyr Thr Glu Glu

515 520 525

Lys Arg Arg Read Lys Lys Phe His Pro Glu Ile Glu Glu Read Leu Tyr

530 535 540

Ser Pro Val Glu Asn Thr Glu His Leu Cys Val Leu Lys Asp Arg Asn 545 550 555 560

Lys Pro Val Leu Phe Thr Met Wing Arg Leu Asp Arg Val Lys Asn Leu

565 570 575

Thr Gly Leu Val Glu Phe Tyr Ala Lys Asn Ser Arg Leu Arg Glu Leu 580 585 590

Val Asn Leu Val Val Val Gly Gly Asp Arg Arg Lys Glu Ser Lys Asp

595 600 605

Read Glu Glu Lys Wing Glu Met Lys Lys Met Tyr Glu Read Le Ile Glu Lys

610 615 620

Tyr Lys Read Asn Gly Gln Read Arg Trp Ile Be Ser Gln Met Asn Arg 625 630 635 640

Val Arg Asn Gly Glu Read Tyr Arg Tyr Ile Cys Asp Thr Lys Gly Wing

645 650 655

Phe Val Gln Pro Wing Ile Tyr Glu Wing Phe Gly Leu Thr Val Val Glu

660 665 670

Met Thr Cly Wing Gly Leu Pro Thr Phe

675 680 685

Glu Wing Ile Ile Val His Gly Lys Ser Gly Phe Asn Ile Asp Pro Tyr

690 695 700

Asn Gly Asp Read Wing Wing Glu Thr Read Wing Wing Asn Phe Phe Glu Lys Cys 705 710 715 720

Lys Wing Asp Pro Being Tyr Trp Asp Glu Ile Being Gln Gly Gly Leu Lys

725 730 735

Arg Ile Gln Lys Tyr Thr Trp Gln Ile Tyr Be Glu Lys Leu Leu

740 745 750

Thr Leu Thr Gly Val Tyr Gly Phe Ser Lys His Val Wing Tyr Gln Glu

755 760 765

Gln Arg Gly Arg Lys Arg Tyr Ile Glu Met Read His Wing Read Met Tyr

770 775 780

Asn Asn Arg Val Lys Thr Val Pro Leu Wing Val Glu 785 790 795

<210> 8 <211> 3352 <212> DNA

<213> Gossypium arboreum <220> <221> <222> <223> <220> <221> <222> <220> <221> <222> <223> <220> <221> <222> <223> <220> <221> <222> <223> <22 0> <221> <222> <220> <221> <222> <223> <220> <221> <222>

<223> <220>

exon

(32). . (111) exonl

misc_feature (32) .. (34) ATG starcodon

exon

(197) .. (314) exon2

exon

(390) .. (541) exon3

exon

(621) .. (813) exon4

exon

(893) .. (1324) exon5

exon

(1395) .. (1685) exon6

exon

(1779). . (1945) exon7 10

<221> exon <222> (2046) .. (2270) <223> exon8 <220>

<2 21> exon <222> (2355) .. (3057) <223> exon 9 <220>

<221> exon <222> (3162) .. (3188) <223> exonlO <220>

<221> misc_feature <222> (3189). . (3191) <223> Termination Codon <400> 8

cccttctgcc atttcaggaa ccaaaaaagc a atg gct tca till agt gtc tgt 52

Met Ala Ser Ile Ser Val Cys 1 5

gag cgt ttg ggt gaa tet cta gct act cat cca cag cag gea aag tet 100

Glu Arg Read Gly Glu Be Read Ala Thr His Pro Gln Gln Ala Lys Ser

15 20

until ttg tca ag gtataatgta aataagtgct gccttcaaag ttctggtctt 151

Ile Read Ser Arg

25

tctttgttat tttttgttac attgtttata tatgtcatat tgcag g att gaa age 206

Ile Glu Ser 30

ct gga aag ggt att cat cat aag tet caa aag ctg etc tca gtt ctc gat 254

Leu Gly Lys Gly Ile His Lys Be Gln Lys Leu Read Le Be Val Leu Asp

40 45

aaa gag gcc gga aat caa gea ctt gat ggg atg gtg gta gag gtc ctc 302 15

Lys Glu Wing Gly Asn Gln Wing Leu Asp Gly Met Val Val Glu Val Leu

50 55 60

agg tcc act cag gtcacaataa aacaaaccct ttctttggtt atgtatgatg 3 54

Arg Ser Thr Gln

65

agaatattgg actgatatgt taccattatc tatag gaa gct gta gtg tcg tct 407

Glu Wing Val Val Ser Ser 70

cca ttg gtt gcc ctt gcc att cgt tca gct cct gga gtt tgg gag tac 455

Pro Leu Val Wing Leu Wing Ile Arg Ser Wing Pro Gly Val Trp Glu Tyr 75 80 85

att gct gtg gag gtc caa aag ctt ttt gtg gag gaa atg ccc gtt gct 503

Ile Wing Val Glu Val Gln Lys Leu Phe Val Glu Glu Met Pro Val Wing

90 95 100

gag tat cta cgg ttg aag gaa gaa ct gt gat gga ag gtaggtagtt 551

Glu Tyr Leu Arg Leu Lys Glu Glu Leu Val Asp Gly Ser 105 110 115

ttctcttaca gctcctggtt tatgttatgt ttgagtttat gttgaacaat gctcaatgat 611 gtggttcag c tcc aat ggc gag ttt atg ttg gaa ttg gac ttt ggt gca 660

Ser Asn Gly Glu Phe Met Leu Glu Leu Asp Phe Gly Wing 120 125 130

ttc aat aat tct gtt cct cgt cca tct ctt tca aag tcc att ggt aat 708

Phe Asn Asn Be Val Pro Arg Pro Be Read Be Lys Be Ile Gly Asn

135 140 145

ggc atg gac ttc ctc aac cgc cac ctt tct gcc aag cta ttt caa gac 756

Gly Met Asp Phe Read Asn Arg His Read Be Ala Lys Read Phe Gln Asp

150 155 160

aag gag aac ttg age ttg ttg ct gaa tt ctc caa att cac tgc cag 804

Lys Glu Asn Leu Be Leu Leu Leu Glu Phe Leu Gln Ile His Cys Gln

165 170 175

aag gga aag gtaaaagttt cactttcact atttgttttt cctttctgta 853

Lys Gly Lys

20

25

30 180

tacatcaaaa cttaagaatc gttcaatgtt atgccacag ggt atg ctg ttg aat 907

Gly Met Leu Leu Asn 185

gac aga till caa gat gtg aat tcc ctc caa cat gea ttg agg aag gcc 955

Asp Arg Ile Gln Asp Val Asn Be Read Gln His Wing Read Le Arg Lys Wing

190 195 200

gag gag tat ctg act cct cta acg tcg gat acc ccg tac tcg gtt ttc 1003

Glu Glu Tyr Leu Thr Pro Leu Thr Be Asp Thr Pro Tyr Be Val Phe

205 210 215

gag aaa agg ttt cgg gag att ggt ttg gag aag gga tgg ggt gat aat 1051

Glu Lys Arg Phe Arg Glu Ile Gly Leu Glu Lys Gly Trp Gly Asp Asn

220 225 230

gct gag cat gtt ctt gag atg until cat ctt cta ttg gat ctc ctt cag 1099

Glu Wing His Val Leu Glu Met Ile His Leu Leu Leu Asp Leu Leu Gln 235 240 245 250

gt cct gat cct gtc gea gt ct gaa age ttc ct gga aca till cca ttg 1147

Pro Wing Asp Pro Val Wing Wing Glu Ser Phe Leu Gly Thr Ile Pro Wing 255 260 265

gtc gcc aat gtt gta until atg act ccc cat gga tac ttt gcc caa gac 1195 Val Ala Asn Val Val Ile Met Thr Pro His Gly Tyr Phe Ala Gln Asp

270 275 280

aat gtt ttg gga tac cct gac aca ggt ggt cag gtt gtt tat till tta 1243

Asn Val Leu Gly Tyr Pro Asp Thr Gly Gly Gln Val Val Tyr Ile Leu 285 290 295

gat caa gtt cgt gcc ttg gag gaa gag ttg ctt cat cgt

Asp Gln Val Arg Wing Leu Glu Glu Glu Leu Read His Arg Phe Lys Leu

300 305 310

caa gga ctc gac att acc cca cga till cta gtc gtgagtatct gaatattaat 1344 Gln Gly Leu Asp Ile Thr Pro Arg Ile Leu Val 315 320 325

tcggtttcga tcattatgtt taatgcttaa tttggctgtt tatgtaccag att act 1400

30 Ile Thr

cgg ctc ctc cct gat gct gta gga aca act tgc ggt cag cgt ctt gag 1448

Arg Leu Leu Pro Asp Wing Val Gly Thr Thr Cys Gly Gln Arg Leu Glu 330 335 340

aaa gtg tat gga acc aag tat tct gat att ctt cgg gta ccc ttc aga 1496

Lys Tyr Val Gly Thr Lys Tyr Ser Asp Ile Read Arg Val Pro Phe Arg

345 350 355

act gag aag gga att gta cgt cca tgg until tca cga ttc aaa gtc tgg 1544

Glu Thr Lys Gly Ile Val Arg Pro Trp Ile Be Arg Phe Lys Val Trp 360 365 370 375

cct tac ctg gag act tac act aag gat gtt gea gct gag gtc acc aaa 1592

Pro Tyr Leu Glu Thr Tyr Thr Lys Asp Val Wing Glu Wing Val Thr Lys 380 385 390

gag ttc cag ggc aaa cct gat ctg att gtt ggg aac tac agt gat gga 1640 Glu Phe Gln Gly Lys Pro Asp Leu Ile Val Gly Asn Tyr Ser Asp Gly

395 400 405

aac att gtt gt tt tt tt ttg gea ctg aag ttt gat gtt aca cag 1685

Asn Ile Val Wing Be Read Leu Wing Leu Lys Phe Asp Val Thr Gln 410 415 420

gtttgttaaa tgccaattaa ttgaacaagt tcctttctgt attttgattt ttcccgtgca 1745 atctaaaaac ttggttttca atgttttatg cag tgc act att gct cat gea ctt 1799

Cys Thr Ile Wing His Wing Leu 425

gag aag acg aaa tac ccg gat tca gac att aac tgg aaa caa ctt gag 1847 Glu Lys Thr Lys Tyr Pro Asp Ser Asp Ile Asn Trp Lys Gln Leu Glu 430 435 440 445

gat aag tat cac ttc tcc tgt cag ttt act gct gat ctt att gct atg 1895

Asp Lys Tyr His Phe Be Cys Gln Phe Thr Wing Asp Leu Ile Wing Met 450 455 460

aac cat act gat ttt until till acc age acc ttc caa gag att gct gga 1943

Asn His Thr Asp Phe Ile Ile Thr Be Thr Phe Gln Glu Ile Wing Gly 10

465 470 475

ag gtaatctatc tcccacttta ctacatttgg ttcccttgta cgttgtaaat 1995

To be

gttgccacct atttgagtcc ctaacaatgt ttccattcca tgattttcag c aag gac 2052

Lys Asp 480

act ctc ggc caa tac gag agt cac up to gct ttc act ctt cca ggg ctc 2100

Thr Leu Gly Gln Tyr Glu Being His Ile Wing Phe Thr Leu Pro Gly Leu

485 490 495

tac cgc gtt gtt gat ggg until gat gtt ttt gat ccc aaa ttc aat att 2148

Tyr Arg Val Val Asp Gly Ile Val Val Asp

500 505 510

gtc tcc cet ggt gct gat atg age atta tac ttc cct tac acg gaa gag 2196

Val Ser Pro Gly Wing Asp Met Ile Tyr Phe Pro Tyr Thr Glu Glu 515 520 525

aag cga agg ttg aag aag ttc cac ccg gag att gaa gag ctt tac 2244

Lys Arg Arg Read Lys Lys Phe His Pro Glu Ile Glu Glu Read Leu Tyr

530 535 540

age cct gtt gag aa aca gag cac tt gtaagatttt ggtcctagtt 2290

Ser Pro Val Glu Asn Thr Glu His Leu 545 550

ttacacccgt tttcaccgga ttttgtgttc agtgttaaca atgtcttaaa ttgttgtctc 2350 acag a tgt gta cta aaa gac aac aag ccg gtt ctg ttt acc atg 2397

Cys Val Leu Lys Asp Arg Asn Lys Pro Val Leu Phe Thr Met 555 560 565

gea agg ttg gac cga gtg aag aac tta act ggg ctc gta gag ttc tat 2445

Wing Arg Read Asp Arg Val Lys Asn Read Thr Thr Gly Read Val Glu Phe Tyr

570 575 580

gcc aag aac age cgg ctg agg gaa ctg gtt aac ttg gtt gta gta ggt 2493

Wing Lys Asn Ser Arg Leu Arg Glu Leu Val Asn Leu Val Val Val Gly 585 590 595

20

25

30 10

15

gga gat agg aga aag gaa tcc aag gac tta gaa gaa aag gct gaa atg 2541

Gly Asp Arg Arg Lys Glu Be Lys Asp Leu Glu Glu Lys Wing Glu Met 600 605 610 615

aag aag atg tat gaa ctt until gaa aaa tac aag ttg aat gga caa ttg 2589

Lys Lys Met Tyr Glu Leu Ile Glu Lys Tyr Lys Leu Asn Gly Gln Leu

620 625 630

aga tgg ata tcg tcc cag atg aac cgt gtg aga aat ggt gaa ctc tat 2637

Arg Trp Ile Being Gln Met Asn Arg Val Arg Asn Gly Glu Leu Tyr

635 640 645

cgt tat att tgc gac acc aga gga gea ttc gtt cag cct gct ata tac 2685

Arg Tyr Ile Cys Asp Thr Lys Gly Phe Val Gln Pro Wing Ile Tyr

650 655 660

gg gt tt gg tt gt act gtt gt gt gt atg acc tgt gga cta cca 2733

Glu Wing Phe Gly Leu Thr Val Val Glu Wing Met Thr Cys Gly Leu Pro

665 670 675

aca ttt gea aca tgt tac ggg ggc cct gct gag att ata gtt cac gga

Thr Phe Wing Thr Cys Tyr Gly Gly Pro Wing Glu Ile Ile Val His Gly 680 685 690 695

aaa tcg ggg ttc aac until gac cct tat aac ggt gat ttg gct gcc gag 2829

Lys Ser Gly Phe Asn Ile Asp Pro Tyr Asn Gly Asp Read Wing Wing Glu

700 705 710

acc ctt gcc aat ttc ttc gag aaa tgc aaa gcg gat cca tet tat tgg 2877

Thr Leu Wing Asn Phe Phe Glu Lys Cys Lys Wing Asp Pro Be Tyr Trp

715 720 725

gat gag till tcc cag gga ggg ttg aaa cgc ata cag gag aag tat aca 2925

Asp Glu Ile Be Gln Gly Gly Leu Lys Arg Ile Gln Glu Lys Tyr Thr

730 735 740

tgg cag att tac tcc gag aag cta ttg act ctc acc gga gtt tat ggc 2973

Trp Gln Ile Tyr Be Glu Lys Leu Leu Thr Leu Thr Gly Val Tyr Gly

745 750 755

ttt tcg aaa cat gta cgt tac cag gag caa cgt ggg cgc aag cgt tac 3021

Phe Ser Lys His Val Tyr Wing Gln Glu Gln Arg Gly Arg Lys Arg Tyr

25

30 10

760 765 770 775

att gaa atg ttg cat gca ctg atg tat aac cgg gtaaaacagt 3067

Ile Glu Met Leu His Wing Leu Met Tyr Asn Asn Arg

780 785

catgtctatt attattatta catcaaacat cattttactc tttcattgaa ctgaaaattt 3127 catatttctg aatgtttaat tgtgtgcttt acag gtc aag act gtt cct cta gct 3182

Val Lys Thr Val Pro Leu Wing 790

gtt gag taaagaaaaa gcaaaaagct gcttggaaac taaggaattt ccaagaatga 3238

Val Glu 795

ctgctgttga agttttattt ttgcttttat tatttttttt ttcaattggt aaatgtttaa 3298 tttcaacttt gggtttcccc ttggaatgtt tcgaactctg ggtttggtcc catt 3352

<210> 9

<211> 796

<212> PRT

<213> Gossypium raimondii <4 00> 9

Met Ala Ser Ile Ser Val Cys Glu Arg Leu Gly Glu Ser Leu Ala Thr

1 5 10 15

His Pro Gln Gln Wing Lys Be Ile Read G Be Arg Ile Glu Be Read Gly

25 30

Lys Gly Ile His Lys Be Gln Lys Leu Read Le Be Val Leu Asp Lys Glu 40 45

Gly Wing Asn Gln Wing Read Asp Gly Met Val Val Glu Val Leu Arg Ser 50 55 60

Thr Gln Glu Wing Val Val Ser Be Pro Leu Val Wing Leu Wing Ile Arg 65 70 75 80

Ser Pro Wing Gly Val Trp Glu Tyr Ile Wing Leu Glu Phe Gln Lys Leu

85 90 95

Phe Val Glu Glu Met Pro Val Glu Wing Tyr Leu Arg Leu Lys Glu Glu 100 105 110 Leu Val Asp Gly Be Ser Asn Gly Glu Phe Met Leu Glu Leu Asp Phe

115 120 125

Gly Ala Phe Asn Asn Be Val Pro Arg Pro Be Read Lys Be Ile

130 135 140

Gly Asn Gly Met Asp Phe He Read Asn Arg His He Read Be Wing Lys Leu Phe 145 150 155 160

Gln Asp Lys Glu Asn Leu Asn Leu Leu Leu Leu Glu Phe Leu Gln Ile Arg

165 170 175

Cys Gln Lys Gly Lys Gly Met Leu Read Asn Asp Arg Ile Gln Asp Val

180 185 190

Asn Be Read Gln His Wing Read Arg Lys Wing Glu Glu Tyr Leu Thr Pro

195 200 205

Read Be Ser Asp Thr Pro Tyr Be Val Phe Glu Lys Arg Phe Read Gly

210 215 220

Ile Gly Leu Glu Lys Gly Trp Gly Asp Asn Asn Wing Glu His Val Leu Glu 225 230 235 240

Met Ile His Leu Leu Leu Asp Leu Leu Gln Wing Pro Asp Pro Val Wing

245 250 255

Read Glu Ser Phe Read Gly Arg Ile Pro Read Val Wing Asn Val Val Ile

260 265 270

Met Thr Pro His Gly Tyr Phe Ala Gln Asp Asn Val Leu Gly Tyr Pro

275 280 285

Asp Thr Gly Gly Gln Val Val Tyr Ile Leu Asp Gln Val Arg Wing Alu

290 295 300

Glu Glu Glu Leu Read His Arg Phe Lys Leu Gln Gly Leu Asp Ile Thr 305 310 315 320

Pro Arg Ile Leu Val Ile Thr Arg Leu Leu Pro Asp Wing Val Gly Thr

325 330 335

Thr Cys Gly Gln Arg Read Le Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp

340 345 350

Ile Leu Arg Val Pro Phe Arg Glu Thr Lys Gly Ile Val Arg Pro Trp 355 360 365 Ile Be Arg Phe Lys Val Trp Pro Tyr Leu Glu Thr Tyr Thr Lys Asp

370 375 380

Val Wing Wing Glu Ile Thr Lys Glu Phe Gln Gly Lys Pro Asp Leu Ile 385 390 395 400

Val Gly Asn Tyr Be Asp Gly Asn Ile Val Wing Be Read Leu Wing His

405 410 415

Lys Phe Asp Val Thr Gln Cys Ser Ile Wing His Wing Leu Glu Lys Thr

420 425 430

Lys Tyr Pro Asp Be Asp Ile Asn Trp Lys Gln Read Glu Asp Lys Tyr

435 440 445

His Phe Ser Cys Gln Phe Thr Wing Asp Read Ile Wing Met Asn His Thr

450 455 460

Asp Phe Ile Ile Thr Be Thr Phe Gln Glu Ile Wing Gly Ser Lys Asp 465 470 475 480

Thr Leu Gly Gln Tyr Glu Being His Ile Wing Phe Thr Leu Pro Gly Leu

485 490 495

Tyr Arg Val Val Asp Gly Ile Val Val Asp

500 505 510

Val Ser Pro Gly Wing Asp Met Ile Tyr Phe Pro Tyr Thr Glu Glu

515 520 525

Lys Arg Arg Read Lys Lys Phe His Pro Glu Ile Glu Glu Read Leu Tyr

530 535 540

Ser Pro Val Glu Asn Thr Glu His Leu Cys Val Leu Lys Asp Arg Asn 545 550 555 560

Lys Pro Ile Leu Phe Thr Met Wing Arg Leu Asp Arg Val Lys Asn Leu

565 570 575

Thr Gly Leu Val Glu Phe Tyr Ala Lys Asn Ser Arg Leu Arg Glu Leu

580 585 590

Val Asn Leu Val Val Val Gly Gly Asp Arg Arg Lys Glu Ser Lys Asp

595 600 605

Leu Glu Glu Lys Wing Glu Met Lys Lys Met Tyr Glu Leu Ile Glu Lys 610 615 620 Tyr Lys Read Asn Gly Gln Phe Arg Trp Ile Be Ser Gln Met Asn Arg 625 630 635 640

Val Arg Asn Gly Glu Read Tyr Arg Tyr Ile Cys Asp Thr Lys Gly Wing

645 650 655

Phe Val Gln Pro Wing Ile Tyr Glu Wing Phe Gly Leu Thr Val Val Glu

660 665 670

Met Thr Cly Wing Gly Leu Pro Thr Phe

675 680 685

Glu Wing Ile Ile Val His Gly Lys Ser Gly Phe Asn Ile Asp Pro Tyr

690 695 700

Asn Gly Asp Read Wing Wing Glu Thr Read Wing Wing Asn Phe Phe Glu Lys Cys 705 710 715 720

Lys Wing Asp Pro Being Tyr Trp Asp Glu Ile Being Gln Gly Gly Leu Lys

725 730 735

Arg Ile Gln Lys Tyr Thr Trp Gln Ile Tyr Be Glu Lys Leu Leu

740 745 750

Thr Leu Thr Gly Val Tyr Gly Phe Ser Lys His Val Wing Tyr Gln Glu

755 760 765

Gln Arg Gly Arg Lys Arg Tyr Ile Glu Met Read His Wing Read Met Tyr

770 775 780

Asn Asn Arg Val Lys Thr Wing Pro Leu Wing Val Glu 785 790 795

<210> 10 <211> 3354 <212> DNA

<213> Gossypium raimondii <220>

<221> exon <222> (32) .. (111) <223> exonl <220>

<221> misc_feature <222> (32). . (34) <223> initiation codon <22 0>

<221> exon

<222> (197) .. (314)

<223> exon 2 <220>

<221> exon

<222> (394). . (545)

<223> exon 3 <220>

<221> exon

<222> (625). . (817)

<223> exon4 <220>

<221> exon

<222> (897) .. (1328)

<223> exon 5 <220>

<221> exon

<222> (1399) .. (1689)

<223> exon 6 <22 0>

<221> exon

<222> (1783) .. (1949)

<223> exon 7 <220>

<221> exon

<222> (2050) .. (2274)

<223> exon 8 <220>

<221> exon 15

<222> (2358) .. (3060) <223> exon 9 <220>

<221> exon <222> (3169) .. (3195) <223> exon 10 <220>

<221> misc_feature <222> (3196) .. (3198) <223> Termination Codon <400> 10

cccttctgcc atttcaggaa cctaaaaaac atg gct tca till agt gtt tgt 52

Met Ala Ser Ile Ser Val Cys 1 5

gag cgt ttg ggt gaa tet cta gct act cat cca cag cag gea aag tet 100

Glu Arg Read Gly Glu Be Read Ala Thr His Pro Gln Gln Ala Lys Ser

15 20

until ttg tca ag gtataatgta aataagtgct gccttcaaat ttctggtctt 151

Ile Leu Ser Arg 25

tctttgttat tttttattac attgtttata tatgtcatat tgcag g att gaa age 206

Ile Glu Ser 30

gg aag ggt att cat cat aag tet caa aag ctg etc tcg gtt ctc gat 254

Leu Gly Lys Gly Ile His Lys Be Gln Lys Leu Read Le Be Val Leu Asp

40 45

aaa gag gcc gga aat caa gea ctt gat ggg atg gtg gtg gag gtc ctc 302

Lys Glu Wing Gly Asn Gln Wing Leu Asp Gly Met Val Val Glu Val Leu

50 55 60

agg tcc act cag gtcacaataa aacaaaccct ttctttggtt atgtttcatg 354

Arg Ser Thr Gln 65

20

30 15

atgagaatat tggactgata tgttaccgct atctaatag gaa gct gta gtg tcg 408

Glu Wing Val Val Ser 70

tct cca ttg gtt gcc ctt gcc att cgt

Ser Pro Leu Wing Val Leu Wing Ile Arg Ser Pro Wing Gly Val Trp Glu 75 80 85

tac att gct ctg gag ttc caa aag ctt tt gtg gag gaa atg ccc gtt 504

Tyr Ile Wing Read Glu Phe Gln Lys Read Phe Val Glu Glu Met Pro Val 90 95 100

gct gag tat cta cgg ttg aag gaa gaa ct gt gat gga ag 545

Glu Wing Tyr Leu Arg Leu Lys Glu Glu Leu Val Asp Gly Ser

105 110 115

gtaggtagtt ttctcttaca gctcctggtt tatattatgt ttgagtttat gttgaacaat 605 gctcaatgat gtggttcag c tcc aat ggc gag ttt atg ttg gaa ttg gac 655

Ser Asn Gly Glu Phe Met Leu Glu Leu Asp 120 125

tt ggt gca ttc aat aat tct gtt cct cgt cca tct ctt tca aag tcc 703

Phe Gly Ala Phe Asn Asn Be Val Pro Arg Pro Be Read Be Lys Be

130 135 140

att ggt aat ggc atg gac ttc ctc aac cgc cac ctt tct gcc aag cta 751

Ile Gly Asn Gly Met Asp Phe Leu Asn Arg His Leu To Be Ala Lys Leu

145 150 155

ttt caa gac aag gag aac ttg aac ttg ttg ct gaa ttt ctc caa att 799

Phe Gln Asp Lys Glu Asn Leu Asn Leu Leu Leu Leu Glu Phe Leu Gln Ile 160 165 170 175

cgc tgc cag aag gga aag gtaaaagttt cactttcact atttgttttt 847

Arg Cys Gln Lys Gly Lys 180

cctttctgta tacatcaaaa cttaagaatc gttcaatgtt atgccacag ggt atg ctg 905

Gly Met Leu

20

ttg aat gac aga until hunting gat gtg aat tcc ctc hunting cat gca ttg agg

953 Leu Asn Asp Arg Ile Gln Asp Val Asn Ser Leu Gln His Wing Leu Arg 185 190 195 200

aag gcc gag gag tat ctg act cct cta tcg tcg gat acc ccg tac tca 1001

Lys Wing Glu Glu Tyr Leu Thr Pro Leu Ser Ser Asp Thr Pro Tyr Ser 205 210 215

gt ttc gag aaa agg ttt ctg ggg att ggt ttg gag aag gga tgg ggc 1049 Val Phe Glu Lys Arg Phe Leu Gly Ile Gly Leu Glu Lys Gly Trp Gly

220 225 230

gat aat gct gag cat gtt ctt gag atg until cat ctt cta ttg gat ctc 1097

Asp Asn Glu Wing His Val Leu Glu Met Ile His Leu Leu Leu Asp Leu 235 240 245

ct cag gea cct gat cct gtc gea

Read Gln Wing Pro Asp Pro Val Wing Read Glu Ser Phe Leu Gly Arg Ile 250 255 260

cca ttg gtc gcc aat gtt gta until atg act ccc cat gga tac ttt gcc 1193 Pro Leu Val Ala Asn Val Val Ile Met Thr Pro His Gly Tyr Phe Ala 265 270 275 280

caa gac aat gtt ttg gga tac cct gac aca ggt ggt cag gtt gtt tat 1241

Gln Asp Asn Val Leu Gly Tyr Pro Asp Thr Gly Gly Gln Val Val Tyr 285 290 295

till tta gat caa gtt cgt gcc ttg gag gaa gag ttg ctt cat cgt ttc 1289

Ile Leu Asp Gln Val Arg Wing Leu Glu Glu Glu Leu Read His Arg Phe

300 305 310

aag ctg caa gga ctc gac att acc cca cgt until cta gtc gtgagtatct 1338

Lys Leu Gln Gly Leu Asp Ile Thr Pro Arg Ile Leu Val 315 320 325

gaatgttaat tcagtttcga tcattatgtt taatgcttaa tttggctgtt tgtgtaccag 1398 att act cgg ctc cct gat gct gta gga aca act tgc gga cag cgt 1446

Ile Thr Arg Read Leu Pro Asp Wing Val Gly Thr Thr Cys Gly Gln Arg 330 335 340

ct gag aaa gtg tat gga acc aag tat tet gat att ctt cgg gta ccc 1494

Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp Ile Leu Arg Val Pro 10

15

345 350

ttc aga act gag aag gga att gta cgt cca tgg until

Phe Arg Thr Glu Lys Gly Ile Val Arg Pro Trp Ile 360 365

gcc tgg cct tac ctg gag act tac act aag gat gtt Val Trp Pro Tyr Leu Glu Thr Tyr Lys Asp Val 375 380 385

acc aaa gag ttc cag ggc aag cct gat ctg att gtt Thr Lys Glu Phe Gln Gly Lys Pro Asp

gat gga aac att gtt gct tet tta ttg gea cat aag Asp Gly Asn Ile Val Ala Ser Leu Leu Ala His Lys

410 415

cag gtttgttaaa tgccaattaa ttgaacaagt tccttgctgt Gln

355

tca cga ttc aaa 1542

To be Arg Phe Lys

370

gea gct gag until 1590 Ala Ala Glu Ile

ggg aac tac agt 1638 Gly Asn Tyr Ser 405

ttt gat gtt aca 1686 Phe Asp Val Thr 420

attttgatgt 1739

20

25

30

tcctcgtgca atctaaaaac ttggttttca atgttttatg cag tgc tet att gct 1794

Cys Ser Ile Wing 425

cat gea etc gag aag aca aaa tac ccg gat tca gac att aac tgg aaa 1842

His Wing Read Glu Lys Thr Lys Tyr Pro Asp Ser Asp Ile Asn Trp Lys

43O 435 440

cag ctt gag gat aag tat ttc tcc tgt cag ttt act gct gat ctt 1890

Gln Leu Glu Asp Lys Tyr His Phe Ser Cys Gln Phe Thr Wing Asp Leu

445 450 455

att gct atg aac cat act gat ttt till till acc act acc ttc caa gag 1938 Ile Ala Met Asn His Thr Asp Phe Ile Ile Thr Be Thr Phe Gln Glu 460 465 47Q

att gct gga ag gtaatctatc tcccacttta ctacatttgg ttcccttgta 1989

Ile Wing Gly Ser

475

cgttgtaaat gttgccacct atttgagtcc ctaacaatgt tttcattcca tgattttcag 2049 c aag gac act ctc gg ca ca ca tac gag ttc act ctt cca 2098 Lys Asp Thr Leu Gly Gln Tyr Glu Ser His Ileu 480

ggt ctc tac cgc gtt gtt gat ggg till gat gtt ttt gat ccc aaa ttc 2146

Gly Leu Tyr Arg Val Val Asp Gly Ile Val Val Asp Phe Asp Pro Lys Phe 495 500 505 510

aat att gtc tcc cct ggt gct gat atg act ata tac ttc cct tac acg 2194

Asn Ile Val Ser Pro Gly Wing Asp Met Ser Ile Tyr Phe Pro Tyr Thr 515 520 525

ga gag aag cgg agg ttg aag aag ttc cac ccg gag att gaa gag ctt 2242 Glu Glu Lys Arg Arg Leu Lys Lys Phe His Pro Glu Ile Glu Glu Leu

530 535 540

ct tac age cct gt gag aat aca gag cac tt gtgagatttt ggtcctattt 2294 Leu Tyr Ser Pro Val Glu Asn Thr Glu His Leu 545 550

ttacaccctt ttcactagat tttgtgttca gtgttaacaa tgtcttaaat tgttgtctca 2354 cag a tgt cta aaa gac cgc aac ag ccg att ctg ttt acc atg gea 2403 Cys Val Leu Lys Asp Arg Asn P5 555

agg ttg gac cga gtg aag aac tta act ggg ctc gta gag ttc tat gcc 2451 Arg Leu Asp Arg Val Lys Asn Leu Thr Gly Leu Val Glu Phe Tyr Ala

570 575 580

aag aac acts cgg ctg agg gaa ctg gtt aac ttg gtt gta gta ggt gga 2499

Lys Asn Be Arg Leu Arg Glu Leu Val Asn Leu Val Val Val Gly Gly 585 590 595 600

gat agg aga aag gaa tcc aag gac tta gaa gaa aag gct gaa atg aag 2547

Asp Arg Arg Lys Glu Be Lys Asp Leu Glu Glu Lys Wing Glu Met Lys

605 610 615

aag atg tat gaa ctt until gaa aaa tac aag ttg aat gga caa ttc aga 2595

Lys Met Tyr Glu Leu Ile Glu Lys Tyr Lys Leu Asn Gly Gln Phe Arg 620 625 630

tgg ata tcg tcc cag atg aac cgt gtg aga aat ggt gaa ctc tat cgt 2643 Trp Ile Ser Ser Gln Met Asn Arg Val Asn Gly Glu Leu Tyr Arg

635 640 645

tat att tgc gac acc aga gga gca ttc gtt cag cct gct ata tac gag 2691

Tyr Ile Cys Asp Thr Lys Gly Wing Phe Val Gln Pro Ile Wing Tyr Glu 650 655 660

gct ttt ggc ttg act gtt gtt gag gca atg acc tgt gga cta cca aca 273 9 Phe Gly Wing Leu Thr Val Val Glu Wing Met Thr Cys Wing Gly Leu Pro Thr 665 670 675 680

ttt gca aca tgc tac ggg ggc cct gct gag att ata gtt cac gga aaa 2787

Phe Wing Thr Cys Tyr Gly Gly Pro Glu Wing Ile Ile Val His Gly Lys

685 690 695

tcg ggg ttc aac till gat cct tat aac ggt gat ttg gct gcc gag acc 2835

Ser Gly Phe Asn Ile Asp Pro Tyr Asn Gly Asp Read Wing Wing Wing Glu Thr 700 705 710

1 5 ctt gcc aat ttc ttc gag aag tgc aaa gcg gat cca tet tat tgg gat 2883 Leu Asa Phe Phe Glu Lys Cys Lys Ala Asp Pro Ser Tyr Trp Asp

715 720 725

gag till tcc cag gga ggg ttg aaa cgc ata cag gag aag tat aca tgg 2931 Glu Ile Ser Gln Gly Gly Leu Lys Arg Ile Gln Glu Lys Tyr Thr Trp 730 735 740

cag att tac tcc gag aag cta ttg act ctc acc gga gt tat ggc ttt 2979

Gln Ile Tyr Ser Glu Lys Leu Leu Thr Leu Thr Gly Val Tyr Gly Phe 745 750 755 760

tcg aaa cat gta gct tac cag gag caa cgt ggg cgc aag cgt tac att 3027

Ser Lys His Val Tyr Wing Gln Glu Gln Arg Gly Arg Lys Arg Tyr Ile

765 770 775

gaa atg ttg cat gca ttg atg tat aac aat cgg gtaaaacaat catgtctatt 3080 Glu Met Leu His Ala Leu Met Tyr Asn Asn Arg 780 785

attattatta catcaaacat cattttactc tttcattgaa ctgaaaattt catgtttctg 3140 aatgtttgtt taattgtgtg ttttacag gtc aag act gct cca gct gtt 3192

Val Lys Thr Wing Pro Read Wing Val 790 795

gag taaagaaaaa gcaaaaggct gcttggaaac taaggaattt ccaagaatga 3245

Glu

ttactattga agttttattt ttgcttcttt tttttcaatt tggtaaatgt ttaatttcaa 3305 ctttggggtt tccccttgga atgttttgaa ctctgggttt ggtcccatt 3354

<210> 11 <211> 796 <212> PRT <213> Gossypium Iongicalyx <4 00> 11

Met Wing Ser Ile Ser Val Cys Glu Arg Leu Gly Glu Ser Leu Wing Thr 10 15

His Pro Gln Gln Wing Lys Be Ile Read G Be Arg Ile Glu Be Read Gly

20 25 30

Lys Gly Ile His Lys Be Gln Lys Leu Read Le Be Val Leu Asp Lys Glu

40 45

Gly Wing Asn Gln Wing Read Asp Gly Met Val Val Glu Val Leu Arg Ser 50 55 60

Thr Gln Glu Wing Val Val Ser Be Pro Leu Val Wing Leu Wing Ile Arg 65 70 75 80

Ser Pro Wing Gly Val Trp Glu Tyr Ile Wing Val Glu Val Gln Lys Leu

85 90 95

Phe Val Glu Glu Met Pro Val Glu Wing Tyr Leu Arg Leu Lys Glu Glu 100 105 110

Read Val Asp Gly Be Ser Lys Gly Glu Phe Met Read Le Glu Read Le Asp Phe

115 120 125

Gly Wing Phe Asn Asn Be Val Pro Arg Pro Be Read Lys Be Ile 130 135 140

Gly Asn Gly Met Asp Phe He Read Asn Arg His He Read Be Wing Lys Leu Phe 145 150 155 160

Gln Asp Lys Glu Asn Leu Asn Leu Leu Leu Leu Glu Phe Leu Gln Ile His 165 170 175

Cys Gln Lys Gly Lys Gly Met Leu Read Asn Asp Arg Ile Gln Asp Val

180 185 190

Asn Be Read Gln His Wing Read Arg Lys Wing Glu Glu Tyr Leu Thr Pro

195 200 205

Leu Thr Be Asp Thr Pro Tyr Be Val Phe Glu Lys Arg Phe Arg Glu

210 215 220

Ile Gly Leu Glu Lys Gly Trp Gly Asp Asn Asn Wing Glu His Val Leu Glu 225 230 235 240

Met Ile His Leu Leu Leu Asp Leu Leu Gln Wing Pro Asp Pro Val Wing

245 250 255

Read Glu Ser Phe Read Gly Arg Ile Pro Read Val Wing Asn Val Val Ile

260 265 270

Met Thr Pro His Gly Tyr Phe Ala Gln Asp Asn Val Leu Gly Tyr Pro

275 280 285

Asp Thr Gly Gly Gln Val Val Tyr Ile Leu Asp Gln Val Arg Wing Alu

290 295 300

Glu Glu Glu Leu Read His Arg Phe Lys Leu Gln Gly Leu Asp Ile Thr 305 310 315 320

Pro Arg Ile Leu Val Ile Thr Arg Leu Leu Pro Asp Wing Val Gly Thr

325 330 335

Thr Cys Gly Gln Arg Read Le Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp

340 345 350

Ile Leu Arg Val Pro Phe Arg Thr Glu Lys Gly Ile Val Arg Pro Trp

355 360 365

Ile Ser Arg Phe Lys Val Trp Pro Tyr Leu Glu Thr Tyr Thr Lys Asp

370 375 380

Val Wing Wing Glu Ile Thr Lys Glu Phe Gln Gly Lys Pro Asp Leu Ile 385 390 395 400

Val Gly Asn Tyr Be Asp Gly Asn Ile Val Wing Be Read Leu Wing His

405 410 415

Lys Phe Asp Val Thr Gln Cys Thr Ile Wing His Wing Leu Glu Lys Thr 420 425 430

Lys Tyr Pro Asp Be Asp Ile Asn Trp Lys Gln Read Glu Asp Lys Tyr

435 440 445

His Phe Ser Cys Gln Phe Thr Wing Asp Read Ile Wing Met Asn His Thr

450 455 460

Asp Phe Val Ile Thr Be Thr Phe Gln Glu Ile Wing Gly Ser Lys Asp 465 470 475 480

Thr Leu Gly Gln Tyr Glu Being His Ile Wing Phe Thr Leu Pro Gly Leu

485 490 495

Tyr Arg Val Val Asp Gly Ile Val Val Asp

500 505 510

Val Ser Pro Gly Wing Asp Met Ile Tyr Phe Pro Tyr Thr Glu Glu

515 520 525

Lys Arg Arg Read Lys Lys Phe His Pro Glu Ile Glu Glu Read Leu Tyr

530 535 540

Ser Pro Val Glu Asn Thr Glu His Leu Cys Val Leu Lys Asp Arg Asn 545 550 555 560

Lys Pro Val Leu Phe Thr Met Wing Arg Leu Asp Arg Val Lys Asn Leu

565 570 575

Thr Gly Leu Val Glu Phe Tyr Ala Lys Asn Ser Arg Leu Arg Glu Leu

580 585 590

Val Asn Leu Val Val Val Gly Gly Asp Arg Arg Lys Glu Ser Lys Asp

595 600 605

Read Glu Glu Lys Wing Glu Met Lys Lys Met Tyr Glu Read Le Ile Glu Lys

610 615 620

Tyr Lys Read Asn Gly Gln Phe Arg Trp Ile Be Ser Gln Met Asn Arg 625 630 635 640

Val Arg Asn Gly Glu Read Tyr Arg Tyr Ile Cys Asp Thr Lys Gly Wing

645 650 655

Phe Val Gln Pro Wing Ile Tyr Glu Wing Phe Gly Leu Thr Val Val Glu

660 665 670

Wing Met Thr Cys Gly Leu Pro Thr Phe Wing Thr Cys Tyr Gly Gly Pro 675 680 685 Wing Glu Ile Ile Val

69O 695 700

Asn Gly Asp Read Wing Wing Glu Thr Read Wing Wing Asn Phe Phe Glu Lys Cys 705 ΊΟ 715 720

Lys Wing Asp Pro Being Tyr Trp Asp Glu Ile Being Gln Gly Gly Leu Lys

725 730 735

Arg Ile Gln Lys Tyr Thr Trp Gln Ile Tyr Be Glu Lys Leu Leu

740 745 750

Thr Leu Thr Gly Val Tyr Gly Phe Ser Lys His Val Wing Tyr Gln Glu 755 760 765

Gln Arg Gly Arg Lys Arg Tyr Ile Glu Met Read His Wing Read Met Tyr 770 775 780

Asn Asn Arg Val Lys Thr Val Pro Leu Wing Val Glu 785 790 795

<210> 12 <211> 3353 <212> DNA

<213> Gossypium longicalyx <220>

<221> exon <222> (32) .. (111) <223> exon 1 <220>

<221> misc_feature <222> (32) .. (34) <223> ATG initiation codon <220>

<221> exon <222> (195) .. (312) 223> exon 2 220> <221> exon <222> (392) .. (543) <223> exon 3 <220>

<221> exon <222> (623) .. (815) <223> exon 4 <220>

<221> exon <222> (891) .. (1322) <223> exon 5 <220>

<221> exon <222> (1393) .. (1683) <223> exon 6 <22 0>

<221> exon <222> (1777) .. (1943) <223> exon 7 <220>

<221> exon <222> (2044) .. (2268) <223> exon 8 <220>

<221> exon <222> (2353) .. (3055) <223> exon 9 <220>

<221> exon <222> (3160) .. (3186) <223> exon 10 <220> 10

15

<221> misc_feature <222> (3187) .. (3189) <223> Termination Codon <400> 12

cccttctgcc atttcaggaa cctaaaaaac a atg gct tca till agt gtc tgt 52

Met Ala Ser Ile Ser Val Cys 1 5

gag cgt ttg ggt gaa tet cta gct act cat cca cag cag gea aag tet 100

Glu Arg Read Gly Glu Be Read Ala Thr His Pro Gln Gln Ala Lys Ser

15 20

until ttg tca ag gtataatgta aataagtgct gccttcaaag ttctggtctt 151

Ile Leu Ser Arg 25

tctttgttat tttttattac attgtttata tgtcatattg cag g att gaa age ctt 207

Ile Glu Ser Leu 30

gga aag ggt att cat cat aag tet caa aag ctg etc tcg gtt ctc gat aaa 255

Gly Lys Gly Ile His Lys Be Gln Lys Read Leu Read Val Leu Asp Lys

40 45

gag gcc gga aa caa gea ctt gat ggg atg gtg gtg gag gtc ctc agg 3 03

Glu Wing Gly Asn Gln Wing Leu Asp Gly Met Val Val Glu Val Leu Arg

50 55 60

tcc act cag gtcacaataa aacaaaccct ttctttggtt atgtttcatg 352

Ser Thr Gln 65

atgagaatat tggactgata tgttaccatt atctaatag gaa gct gta gtg tcg 406

Glu Wing Val Val Ser 70

tet cca ttg gtt gcc ctt gcc att cgt tca gct cct gga gtt tgg gag 454

Ser Pro Leu Wing Val Leu Wing Ile Arg Ser Pro Wing Gly Val Trp Glu 75 80 85

tac att gct gtg gag gtc caa aag ctt tt gtg gag gaa atg ccc gtt 502

20

25 10

15

Tyr Ile Wing Val Glu Val Gln Lys Leu Phe Val Glu Glu Met Pro Val

90 95 100

gct gag tat cta cgg ttg aag gaa gaa ctt gtt gac gga ag 543

Glu Wing Tyr Leu Arg Leu Lys Glu Glu Leu Val Asp Gly Ser

105 110 115

gtaggtagtt ttctcttaca gcCcctggtt tatgttatgt ttgagtttat gttgaacaat 603 gctcaatgat gtggttcag c tcc aag ggc gag ttt atg ttg gaa ttg gac 653

Ser Lys Gly Glu Phe Met Leu Glu Leu Asp 120 125

ttt ggt gca ttc aat aat tct gtt cct cgt cca tct ctt tca aag tcc 701

Phe Gly Ala Phe Asn Asn Be Val Pro Arg Pro Be Read Be Lys Be

130 135 140

att ggt aat ggc atg gac ttc ctc aac cgc cac ctt tct gcc aag cta 749

Ile Gly Asn Gly Met Asp Phe Leu Asn Arg His Leu To Be Ala Lys Leu

145 150 155

ttt caa gac aag gag aac ttg aac ttg ttg ct gaa ttt ctc caa att 797

Phe Gln Asp Lys Glu Asn Leu Asn Leu Leu Leu Leu Glu Phe Leu Gln Ile 160 165 170 175

cac tgc cag aag gga aag gtaaaagttt cactttcact atttgttttt 845

His Cys Gln Lys Gly Lys

180

cctttataca tcaaaactca agaatcgttc aatgttatgc cacag ggt atg ctg ttg 902

Gly Met Leu Leu 185

aat gac aga till cat gat gtg aat tcc ctc caa cat gca ttg agg aag 950

Asn Asp Arg Ile Gln Asp Val Asn Being Read Gln His Wing Read Arg Lys

190 195 200

gcc gag gag tat ctg act ccc cta acg tcg gat acc ccg tac tcg gtt 998

Glu Glu Wing Tyr Leu Thr Pro Leu Thr Be Asp Thr Pro Tyr Be Val

205 210 215

ttc gag aaa agg ttt cgg gag att ggt ttg gag aag gga tgg ggt gat 1046

Phe Glu Lys Arg Phe Arg Glu Ile Gly Leu Glu Lys Gly Trp Gly Asp 10

15

20

25

30

220 225 230

aat gct gag cat gtt ctt gag atg until cat ctt cta ttg gat ctc ctt 1094

Asn Wing Glu His Val Leu Glu Met Ile His Leu Leu Leu Asp Leu Leu

235 240 245

cag gea cct gat cct gtc gea ct gaa age ttc ctt gga aga till cca 1142 Gln Ala Pro Asp Pro Val Leu Glu Ser Phe Leu Gly Arg Ile Pro 250 255 260 260 265

ttg gtc gcc aat gtt gta till atg act ccc cat gga tac ttt gct caa 1190 Leu Val Ala Asn Val Val Ile Met Thr Pro His Gly Tyr Phe Ala Gln

270 275 280

gac aat gtt ttg gga tac cct gac aca ggt ggt cag gtt gtt tat till 1238 Asp Asn Val Leu Gly Tyr Pro Asp Thr Gly Gly Gln Val Val Tyr Ile

285 290 295

tta gat caa gtt cgt gcc ttg gag gaa gag ttg ctt cat cgt ttc aag 1286

Leu Asp Gln Val Arg Wing Leu Glu Glu Glu Leu Read His Arg Phe Lys

300 305 3io

ctg caa gga etc gac att acc cca cgt till cta gtc gtgagtatct 1332

Read Gln Gly Read Asp Ile Thr Pro Arg Ile Read Val 315 320 325

gaatgttaat tcggtttcga tcattatgtt taatgcttaa tttggctgtt tatgtaccag 1392 att act cgg ctc cct gat gct gta gga aca act tgc ggt cag cgt 1440

Ile Thr Arg Read Leu Pro Asp Wing Val Gly Thr Thr Cys Gly Gln Arg

330 335 340

ct gaa aaa gtg tat gga acc aag tat tet gat att ctt cgg gta ccc 1488

Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp Ile

345 350 355

ttc aga act gag aag gga att gta cgt cca tgg until tca cga ttc aaa 1536

Phe Arg Thr Glu Lys Gly Ile Val Arg Pro Trp Ile Be Arg Phe Lys

360 365 370

gtc tgg cct tac ctg gag act tac act aag gat gtt gea gct gag until 1584

Val Trp Pro Tyr Leu Glu Thr Tyr Thr Lys Asp Val Wing Wing Glu Ile 375 380 385 acc aaa gag ttc cag ggc aag cct gat ctg att gtt ggg aac tac agt 1632

Thr Lys Glu Phe Gln Gly Lys Pro Asp Read Ile Val Gly Asn Tyr Ser

390 395 400 405

gat generates aac att gtt gct tet tta ttg gea cat aag ttt gat gtt aca 1680

Asp Gly Asn Ile Val Wing Be Read Leu Wing His Lys Phe Asp Val Thr

410 415 420

cag gtttgttaaa tgccaattaa ttgaacaagt tccttgctgt attttgatgt 1733

Gln

10

15

20

25

30

ttcccgtgca atctaaaaac ttggttttca atgttttatg cag tgc act att gct 1788

Cys Thr Ile Wing 425

cat gea ctt gag aag ag aaa tac ccg gat tca gac att aac tgg aaa 1836

His Wing Read Glu Lys Thr Lys Tyr Pro Asp Ser Asp Ile Asn Trp Lys

430 435 440

cag ctt gag gat aag tat ttc tcc tgt cag ttt act gct gat ctt 1884

Gln Leu Glu Asp Lys Tyr His Phe Ser Cys Gln Phe Thr Wing Asp Leu

445 450 455

att gct atg aac cat act gat ttt gtc until acc age acc ttc caa gag 1932

Ile Ala Met Asn His Thr Asp Phe Val Ile Thr Be Thr Phe Gln Glu

460 465 470

att gct gga ag gtaatctatc tcccacttta ctaaatttgg ttcccttgta 1983

Ile Wing Gly Ser 475

cgttgtaaat gttgccacct atttgagtcc ctaacaatgt ttccattcca tgattttcag 2043 c aag gac act ctc gg ca ca ca tac gag ttc act ctt cca 2092 Lys Asp Thr Leu Gly Gln Tyr Glu Ser His Thr

ggg ctc tac cgc gtt gtt gat ggg till gat gtt ttt gat ccc aaa ttc 2140

Gly Leu Tyr Arg Val Val Asp Gly Ile Val Val Asp Phe Asp Pro Lys Phe 495 500 505 510

aat att gtc tcc cct ggt gct gat atg act ata tac ttc cct tac acg 2188 Asn Ile Val Ser Pro Gly Ala Asp Met Ser Ile Tyr Phe Pro Tyr Thr

515 520 525

gag gag aag cga agg ttg aag aag ttc cac ccg gag att gaa gag ctt 2236

Glu Glu Lys Arg Arg Read Lys Lys Phe His Pro Glu Ile Glu Glu Leu 530 535 540

ct tac age cct gtt gag aat aca gag cac tt gtgggatttt ggtcctattt 22 88 Leu Tyr Ser Pro Val Glu Asn Thr Glu His Leu

545 550

ttacaccctt tttcactaga ttttgtgttc agtgttaaca atgtcttgaa ttgttgtctc 2348 acag a tgt gta cta aaa gac ag aag ccg gtt ctg ttt acc atg 2395

Cys Val Leu Lys Asp Arg Asn Lys Pro Val Leu Phe Thr Met 555 560 565

gea agg ttg gac cga gtg aag aac tta act ggg ctc gta gag ttc tat 2443 Arg Wing Leu Asp Arg Val Lys Asn Leu Thr Gly Leu Val Glu Phe Tyr 570 575 580

gcc aag aac age cgg ctg agg gaa ctg gtt aac ttg gtt gta gta ggt 2491

Wing Lys Asn Ser Arg Leu Arg Glu Leu Val Asn Leu Val Val Val Gly

585 590 595

gga gat agg aga aag gaa tcc aag gac tta gaa gaa aag gct gaa atg 2539

Gly Asp Arg Arg Lys Glu Be Lys Asp Leu Glu Glu Lys Wing Glu Met 600 605 610 615

aag aag atg tat gaa ctt until gaa aaa tac aag ttg aat gga caa ttc 2587

Lys Lys Met Tyr Glu Read Ile Glu Lys Tyr Lys Read Asn Gly Gln Phe 620 625 630

aga tgg ata tcg tcc cag atg aac cgt gtg aga aat ggt gaa ctc tat 263 5 Arg Trp Ile Ser Ser Gln Met Asn Arg Val Arg Asn Gly Glu Leu Tyr

635 640 645

cgt tat att tgc gac acc aag gga gea ttc gtt

Arg Tyr Ile Cys Asp Thr Lys Gly Phe Val Gln Pro Wing Ile Tyr 650 655 660

gg gt tt gg tt gt gt gt gt gt gt cta cca 2731

Glu Wing Phe Gly Leu Thr Val Val Glu Wing Met Thr Cys Gly Leu Pro 10

15

20

25

30

665 670 675

aca ttt gca aca tgt tac ggg ggc cct gct gag att ata gtt cac gga 2779

Thr Phe Wing Thr Cys Tyr Gly Gly Pro Wing Glu Ile Ile Val His Gly 680 685 690 695

aaa tcg ggg ttc aac until gac cct tat aac ggt gat ttg gct gcc gag 2827

Lys Ser Gly Phe Asn Ile Asp Pro Tyr Asn Gly Asp Read Wing Wing Glu

700 705 710

acc ctc gcc aat ttc ttc gag aaa tgc aaa gcg gat cca tet tat tgg 2875

Thr Leu Wing Asn Phe Phe Glu Lys Cys Lys Wing Asp Pro Be Tyr Trp

715 720 725

gat gag till tcc cag ggg ttg aaa cgc ata cag gag aag tat aca 2923

Asp Glu Ile Be Gln Gly Gly Leu Lys Arg Ile Gln Glu Lys Tyr Thr

730 735 740

tgg cag att tac tcc gag aag cta ttg act ctc acc gga gtt tat ggc 2971

Trp Gln Ile Tyr Be Glu Lys Leu Leu Thr Leu Thr Gly Val Tyr Gly

745 750 755

ttt tcg aaa cat gta cgt tac cag gag caa cgt ggg cgc aag cgt tac 3019

Phe Ser Lys His Val Tyr Wing Gln Glu Gln Arg Gly Arg Lys Arg Tyr 760 765 770 775

att gaa atg ttg cat gca ttg atg tat aac aat cgg gtaaaacaat 3 065

Ile Glu Met Leu His Wing Leu Met Tyr Asn Asn Arg

780 785

catgtctatt attattatta catcaaacat cattttactc tttcattgaa ctgaaaattt 3125 catgtttctg aatgtttaat tgtgtgcttt acag gtc aag act gtt cct cta gct 3180

Val Lys Thr Val Pro Leu Wing 790

gtt gag taaagaaaca gcaaaaagct gcttggaaac caaggaattt ccaagaatga 3236

Val glu

795

ttgctattga agttttattt ttgcttttct tttttttttt ttttttcaat ttggtaaatg 3296 tttaatttca actttggggt ttccccttgg aatgttttga actctgggtt tggtccc 3353

<210> 13 <211> 796 <212> PRT

<213> Gossypium Iongicalyx <400> 13

Met Wing Ser Ile Ser Val Cys Glu Arg Leu Gly Glu Ser Leu Wing Thr 10 15

His Pro Gln Gln Wing Lys Be Ile Read G Be Arg Ile Glu Be Read Gly

25 30

Lys Gly Ile His Lys Be Gln Lys Leu Read Le Be Val Leu Asp Lys Glu

40 45

Wing Gly Asn Gln Wing Leu Asp Gly Met Val Val Glu Val Leu Arg Ser

50 55 60

Thr Gln Glu Wing Val Val Ser Be Pro Leu Val Wing Leu Wing Ile Arg 65 70 75 80

Ser Pro Wing Gly Val Trp Glu Tyr Ile Wing Val Glu Val Gln Lys Leu

85 90 95

Phe Val Glu Glu Met Pro Val Glu Wing Tyr Leu Arg Leu Lys Glu Glu

100 105 110

Read Val Asp Gly Be Ser Asn Gly Glu Phe Met Read Le Glu Read Le Asp Phe

115 120 125

Gly Ala Phe Asn Asn Be Val Pro Arg Pro Be Read Lys Be Ile

130 135 140

Gly Asn Gly Met Asp Phe He Read Asn Arg His He Read Be Wing Lys Leu Phe 145 150 155 160

Gln Asp Lys Glu Asn Leu Asn Leu Leu Leu Leu Glu Phe Leu Gln Ile His

165 170 175

Cys Gln Lys Gly Lys Gly Met Leu Read Asn Asp Arg Ile Gln Asp Val

180 185 190

Asn Be Read Gln His Wing Read Arg Lys Wing Glu Glu Tyr Leu Thr Pro

195 200 205

Leu Be Ser Asp Thr Pro Tyr Be Val Phe Glu Lys Arg Phe Leu Gly 210 215 220 Ile Gly Leu Glu Lys Gly Trp Gly Asp Asn Wing Glu His Val Leu Glu 225 230 235 240

Met Ile His Leu Leu Leu Asp Leu Leu Gln Wing Pro Asp Pro Val Wing

245 250 255

Read Glu Ser Phe Read Gly Arg Ile Pro Read Val Wing Asn Val Val Ile

260 265 270

Met Thr Pro His Gly Tyr Phe Ala Gln Asp Asn Val Leu Gly Tyr Pro

275 280 285

Asp Thr Gly Gly Gln Val Val Tyr Ile Leu Asp Gln Val Arg Wing Alu

29O 295 300

Glu Glu Glu Read His Read His Arg Be Lys Read His Gln Gly Read Asp Ile Thr 305 310 315 320

Pro Arg Ile Leu Val Ile Thr Arg Leu Leu Pro Asp Wing Val Gly Thr

32S 330 335

Thr Cys Gly Gln Arg Read Le Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp

340 345 350

Ile Leu Arg Val Pro Phe Arg Thr Glu Lys Gly Ile Val Arg Pro Trp

355 360 365

Ile Ser Arg Phe Lys Val Trp Pro Tyr Leu Glu Thr Tyr Thr Lys Asp

370 375 380

Val Wing Wing Glu Ile Thr Lys Glu Phe Gln Gly Lys Pro Asp Leu Ile 385 390 395 400

Val Gly Asn Tyr Be Asp Gly Asn Ile Val Wing Be Read Leu Wing His

405 410 415

Lys Phe Asp Val Thr Gln Cys Thr Ile Wing His Wing Leu Glu Lys Thr

420 425 430

Lys Tyr Pro Asp Be Asp Ile Asn Trp Lys Gln Read Glu Asp Lys Tyr

435 440 445

His Phe Ser Cys Gln Phe Thr Wing Asp Read Ile Wing Met Asn His Thr

450 455 460

Asp Phe Ile Ile Thr Be Thr Phe Gln Glu Ile Wing Gly Be Lys Asp 465 470 475 480 Thr Read Gly Gln Tyr Glu Be His Ile Wing

485 490 495

Cys Arg Val Val Asp Gly Ile Asp Val Val Phe Asp Pro

500 505 510

Val Ser Pro Gly Wing Asp Met Ile Tyr Phe Pro Tyr Thr Glu Glu

515 520 525

Lys Arg Arg Read Lys Lys Phe His Pro Glu Ile Glu Glu Read Leu Tyr

530 535 540

Ser Pro Val Glu Asn Thr Glu His Leu Cys Val Leu Lys Asp Arg Asn 545 550 555 560

Lys Pro Ile Leu Phe Thr Met Wing Arg Leu Asp Arg Val Lys Asn Leu

565 570 575

Thr Gly Leu Val Glu Phe Tyr Ala Lys Asn Ser Arg Leu Arg Glu Leu

580 585 590

Val Asn Leu Val Val Val Gly Gly Asp Arg Arg Lys Glu Ser Lys Asp

595 600 605

Read Glu Glu Lys Wing Glu Met Lys Lys Met Tyr Glu Read Le Ile Glu Lys

610 615 620

Tyr Lys Read Asn Gly Gln Phe Arg Trp Ile Be Ser Gln Met Asn Arg 625 630 635 640

Val Arg Asn Gly Glu Read Tyr Arg Tyr Ile Cys Asp Thr Lys Gly Wing

645 650 655

Phe Val Gln Pro Ile Tyr Glu Wing Phe Gly Leu Thr Val Val Glu

660 665 670

Met Thr Cly Wing Gly Leu Pro Thr Phe

675 680 685

Glu Wing Ile Ile Val His Gly Lys Be Gly Phe Asn

690 695 700

Asn Gly Asp Read Wing Wing Glu Thr Read Wing Wing Asn Phe Phe Glu Lys Cys 705 710 715 720

Lys Wing Asp Pro Be Tyr Trp Asp Glu Ile Be Gln Gly Gly Leu Lys 725 730 735 Arg Ile Gln Glu 740

Thr Leu Thr Gly 755

Gln Arg Gly Arg 770

Asn Asn Arg Val 785

<210> 14 <211> 3353 <212> DNA

<213> Gossypium longicalyx <220>

<221> exon <222> (32). . (111) <223> exon 1 <220>

<221> misc_feature <222> (32). . (34) <223> initiation codon <220>

<221> exon <222> (197). . (314) <223> exon 2 <220>

<221> exon <222> (394) .. (545) <223> exon 3 <220>

<221> exon <222> (625) .. (817) <223> exon 4

Lys Tyr Thr Trp Gln Ile Tyr Ser Glu Lys Leu Leu

745 750

Val Tyr Gly Phe Ser Lys His Val Wing Tyr Gln Glu

760 765

Lys Arg Tyr Ile Glu Met Read His Wing Trp Met Tyr

775 780

Lys Thr Val Pro Leu Val Glu Wing 790 795 <220>

<221> exon

<222> (897) .. (1328)

<223> exon 5

<220>

<221> exon <222> (1399) .. (1689)

<223> exon 6 <220>

<221> exon

<222> (1783) .. (1949)

<223> exon 7 <220>

<221> exon

<222> (2050) .. (2274)

<223> exon 8 <220>

<221> exon

<222> (2358) .. (3060)

<223> exon 9 <220>

<221> exon

<222> (3169) .. (3195)

<223> exon 10

<220>

<221> misc_feature <222> (3196) .. (3198) <223> Termination Codon <4 00> 14

cccttctgcc atttcaggaa cccaaaaaac a atg gct tca till agt gtt tgt

Met Wing Ser Ile Ser Val Cys 1 5 10

15

20

25

30

gag cgt ttg ggt gaa tct cta gct act cac cca cag cag gca aag tct 100

Glu Arg Read Gly Glu Be Read Ala Thr His Pro Gln Gln Ala Lys Ser

15 20

until ttg tca ag gtataatgta aataagtgct gccttcaaat ttctggtctt 151

Ile Leu Ser Arg 25

tcttcgttat tttttattac attgtttata tatgtcatat tgcag g att gaa age 206

Ile Glu Ser 30

ctc gga aag ggt att cat aag tct caa aag ctg ctc tcg gtt ctc gat 254

Leu Gly Lys Gly Ile His Lys Be Gln Lys Leu Read Le Be Val Leu Asp

40 45

aaa gag gcc gga aat caa gca ct gat ggg atg gtg gtg gag gtc ctc 3 02

Lys Glu Wing Gly Asn Gln Wing Leu Asp Gly Met Val Val Glu Val Leu

50 55 60

agg tcc act cag gtcacaataa aacaaaccct ttctttggtt atgtttcatg 354

Arg Ser Thr Gln 65

atgagaatat tggactgata tgttaccgtt atctaatag gaa gct gta gtg tcg 408

Glu Wing Val Val Ser 70

tct cca ttg gtt gcc ctt gcc att cgt

Ser Pro Leu Wing Val Leu Wing Ile Arg Ser Pro Wing Gly Val Trp Glu

75 80 85

tac att gct gtg gag gtc caa aag ctt ttt gtg gag gaa atg ccc gtt 504

Tyr Ile Wing Val Glu Val Gln Lys Leu Phe Val Glu Glu Met Pro Val

90 95 100

gct gag tat cta cgg ttg aag gaa gaa ct gt gat gga ag 545

Glu Wing Tyr Leu Arg Leu Lys Glu Glu Leu Val Asp Gly Ser

105 110 115

gtaggtagtt ttctcttaca gctcctggtt tatattatgt ttgagtttat gttgaacaat 605 gctcaatgat gtggttcag c tc aat ggc gag ttt atg ttg gaa ttg gac 655 10

15

30

Ser Asn Gly Glu Phe Met Leu Glu Leu Asp 120 125

ttt ggt gca ttc aat aat tct gtt ccc cgt cca

Phe Gly Ala Phe Asn Asn Be Val Pro Arg Pro Be Read Be Lys Be

130 135 140

att ggt aat ggc atg gac ttc ctc aac cgc cac ctt tct gcc aag cta 751

Ile Gly Asn Gly Met Asp Phe Leu Asn Arg His Leu To Be Ala Lys Leu

145 150 155

ttt caa gac aag gag aac ttg aac ttg ttg ct gaa ttt ctc caa att 799

Phe Gln Asp Lys Glu Asn Leu Asn Leu Leu Leu Leu Glu Phe Leu Gln Ile 160 165 170 175

cac tgc cag aag gga aag gtaaaagttt cactttcact atttgttttt 847

His Cys Gln Lys Gly Lys 180

cctttctgta tacatcaaaa cttaagaatc gttcaatgtt atgccacag ggt atg ctg 905

Gly Met Leu

ttg aat gac aga until hunting gat gtg aat tcc ctc hunting cat gca tta agg 953

Leu Asn Asp Arg Ile Gln Asp Val Asn Ser Leu Gln His Wing Leu Arg 185 190 195 200

aag gcc gag gag ctg act cct cta tcc tcg gat acc ccg tac tca 1001

Lys Wing Glu Glu Tyr Leu Thr Pro Leu Ser Ser Asp Thr Pro Tyr Ser

205 210 215

gt ttc gag aaa agg ttt ctg ggg att ggt ttg gag aag gga tgg ggt 1049

Val Phe Glu Lys Arg Phe Leu Gly Ile Gly Leu Glu Lys Gly Trp Gly 220 225 230

gat aat gct gag cat gtt ctt gag atg until cat ctt cta ttg gat ctc 1097

Asp Asn Alu Glu His Val Leu Glu Met Ile His Leu Leu Leu Asp Leu

235 240 245

ctt cag gca cct gat cct gtc gca ct gaa age ttc ctt gga aga until 1145

Leu Gln Ala Pro Asp Pro Val Wing Leu Glu Ser Phe Leu Gly Arg Ile 250 255 260 260 cca ttg gtc gcc aat gtt gta until atg act ccc cat gga tac ttt gcc 1193

Pro Leu Val Wing Asn Val Val Ile Met Thr Pro His Gly Tyr Phe Wing 265 270 275 280

caa gac aat gtt ttg gga tac cct gac aca ggt ggt cag gtt gtt tat 1241

Gln Asp Asn Val Leu Gly Tyr Pro Asp Thr Gly Gly Gln Val Val Tyr 285 290 295

till tta gat caa gtt cgt gcc ttg gag gaa gag ttg ctt cat cgt tcc 1289

Ile Leu Asp Gln Val Arg Ward Leu Glu Glu Glu

aag ctg caa gga ctc gac att acc cca cgt until cta gtc gtgagtatct 1338

Lys Leu Gln Gly Leu Asp Ile Thr Pro Arg Ile Leu Val 315 320 325

gaatgttaat tcagtttcga tcattatgtt taatgcttaa tttggctgtt tgtgtaccag 1398 att act cgg ctc cct gat gct gta gga aca act tgc ggt cag cgt 1446

Ile Thr Arg Read Leu Pro Asp Wing Val Gly Thr Thr Cys Gly Gln Arg

330 335 340

ct gag aaa gtg tat gga acc aag tat tet gat att ctt cgg gta ccc 1494

Leu Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp Ile Leu Arg Val Pro 345 350 355

ttc aga act gag aag gga att gta cgt cca tgg until tca cga ttc aaa 1542

Phe Arg Thr Glu Lys Gly Ile Val Arg Pro Trp Ile Be Arg Phe Lys

360 365 370

gtc tgg cct tac ctg gag act tac act aag gat gtt gea gct gag until 1590

Val Trp Pro Tyr Leu Glu Thr Tyr Thr Lys Asp Val Wing Wing Glu Ile 375 380 385

acc aaa gag ttc cag ggc aag cct gat ctg att gtt ggg aac tac agt 1638

Thr Lys Glu Phe Gln Gly Lys Pro Asp Ile Ile Val Gly Asn Tyr Ser 390 395 400 405

gat gga aac att gtt gct tet tta ttg gea cat aag ttt gat gtt aca 1686

Asp Gly Asn Ile Val Wing Be Read Leu Wing His Lys Phe Asp Val Thr

410 415 420

cag gtttgttaaa taccaattaa ttgaacaagt tccttgctgt attttgatgt 1739 15

Gln

tccccgtgca atctaaaaac ttggttttca atgttttatg cag tgc act att gct 1794

Cys Thr Ile Wing

425

cat gca ctc gag aag ag aaa tac ccg gat tca gac att aac tgg aaa 1842

His Wing Read Glu Lys Thr Lys Tyr Pro Asp Ser Asp Ile Asn Trp Lys

430 435 440

cag ctt gag gat aag tat ttc tcc tgt cag ttt act gct gat ctt 1890

1 0 Gln Leu Glu Asp Lys Tyr His Phe Ser Cys Gln Phe Thr Wing Asp Leu 445 450 455

att gct atg aac cat act gat ttt until till acc age acc ttc caa gag 1938

Ile Ala Met Asn His Thr Asp Phe Ile Ile Thr Be Thr Phe Gln Glu

460 465 470

att gct gga ag gtaatctatc tcccacttta ctacatttgg ttcccttgta 1989

Ile Wing Gly Ser 475

cgttgtaaat gttgccacct atttgagtcc ctaacaatgt tttcattcca tgattttcag 2049 c aag gac act ctc gg ca ca ca tac gag ttc act ctt cca 2098 Lys Asp Thr Leu Gly Gln Tyr Glu Being His Ileu

ggg ctc tgc cgc gt gt gat ggg until gat gtt ttt gat ccc aaa ttc

Gly Leu Cys Arg Val Val Asp Gly Ile Val Val Asp Phe Asp Pro Lys Phe 495 500 505 510

aat att gtc tcc cct ggt gct gat atg act ata tac ttc cct tac acg 2194

Asn Ile Val Ser Pro Gly Wing Asp Met Ser Ile Tyr Phe Pro Tyr Thr

515 520 525

gag gag ag cgg agg ttg aag aag ttc cac ccg gag att gaa gag ctt 2242

Glu Glu Lys Arg Arg Read Lys Lys Phe His Pro Glu Ile Glu Glu Leu 530 535 540

ct tac age cct gt gag aat aca gag cac tt gtgagatttt ggttctattt 2294 Leu Tyr Ser Pro Val Glu Asn Thr Glu

20

25 10

15

20

25

30

545 550

ttacaccctt ttcactagat tttgtgttca gtgttaacaa tgtcttaaat tgttgtctca 2354 cag a tgt cta aaa gac cgc aac ag ccg att ctg ttt acc atg gca 2403 Cys Val Leu Lys Asp L5 As5 Arg 5

agg ttg gac cga gtg aag aac tta act ggg etc gta gag ttc tat gcc 2451

Arg Leu Asp Arg Val Lys Asn Leu Thr Gly Leu Val Glu Phe Tyr Wing

570 575 580

aag aac acts cgg ctg agg gaa ctg gtt aac ttg gtt gta gta ggt gga 2499

Lys Asn Be Arg Leu Arg Glu Leu Val Asn Leu Val Val Val Gly Gly 585 590 595 600

gat agg aga aag gaa tcc aag gac tta gaa gaa aag gct gaa atg aag 2547

Asp Arg Arg Lys Glu Be Lys Asp Leu Glu Glu Lys Wing Glu Met Lys

605 610 615

aag atg tat gaa ctt until gaa aaa tac aag ttg aat gga caa ttc aga 2595

Lys Met Tyr Glu Leu Ile Glu Lys Tyr Lys Leu Asn Gly Gln Phe Arg

620 625 630

tgg ata tcg tcc cag atg aac cga gtg aga aat ggt gaa ctc tat cgt 2643

Trp Ile Being Gln Met Asn Arg Val Arg Asn Gly Glu Leu Tyr Arg

635 640 645

tat att tgc gac acc aga gga gca ttc gtt cag cct cct ata tac gag 2691

Tyr Ile Cys Asp Thr Lys Gly Phe Val Gln Pro Pro Ile Tyr Glu

650 655 660

gt ttt ggc ttg act gtt gtt gag gca atg acc tgt gga cta cca aca 2739

Phe Gly Leu Wing Thr Val Val Glu Wing Met Thr Cys Wing Gly Leu Thr Thr 665 670 675 680

ttt gca aca tgt tac ggg ggc cct gct gag att ata gtt cac gga aaa 2787

Phe Wing Thr Cys Tyr Gly Gly Pro Glu Wing Ile Ile Val His Gly Lys

685 690 695

tcg ggg ttc aac until gat cct tgt aac ggt gat ttg gct gcc acc 2835

Ser Gly Phe Asn Ile Asp Pro Cys Asn Gly Asp Leu Wing Wing Wing Glu Thr 700 705 710 ctt gcc aat ttc ttc gag aag tgc aaa gat cca tct tat tgg gat 2883

Leu Wing Asn Phe Phe Glu Lys Cys Lys Wing Asp Pro To Be Tyr Trp Asp

715 720 725

gag till tcc cag gga ggg ttg aaa cgc ata cag gag aag tat aca tgg 2931

Glu Ile Being Gln Gly Gly Leu Lys Arg Ile Gln Glu Lys Tyr Thr Trp 730 735 740

cag att tac tcc gag aag cta ttg act ctc acc gga gt tat ggc ttt 2979

Gln Ile Tyr Ser Glu Lys Leu Leu Thr Leu Thr Gly Val Tyr Gly Phe 745 750 755 760

tcg aaa cat gta gct tac cag gag caa cgt ggg cgc aag cgt tac att 3027

Ser Lys His Val Tyr Wing Gln Glu Gln Arg Gly Arg Lys Arg Tyr Ile

765 770 775

gaa atg ttg cat gea tgg atg tat aac aat cgg

780 785

attattatta catcaaacat cattttactc tttcattgaa ctgaaatttt catgtttctg 3140 aatgtttgtt taattgtgtg ctttacag gtc aag act gtt cca cta gct gtt 3192

Val Lys Thr Val Pro Leu Wing Val 790 795

gag taaagaaaaa gcaaaaggct gcttggaaac caaggaattt ccaagaatga 3245

Glu

ttgctattga agttttattt ttgcttcttt ttttcaattt cgtaaatgtt taatttcaac 3305 tttggggttt ccccttggaa tgttttgaac tctgggtttg gtcccatt 3353

<210> 15

<211> 26

<212> PRT

<213> Artificial <220>

<223> C-terminal consensus SusC sequence

<400> 15

Wing Tyr Gln Glu Gln Arg Gly Arg Lys Arg Tyr Ile Glu Met

Trp Wing Met Tyr Asn Asn Arg Val Lys Thr 25

<210> 16 <211> 24 <212> PRT <213> Artificial <220>

<223> N-terminal consensus SusC sequence <400> 16

His Lys Be Gln Lys Leu Read Le Be Val Leu Asp Lys Glu Wing Gly Asn 10 15

Gln Wing Read Asp Gly Met Val Val Val 20

<210> 17 <211> 22 <212> DNA <213> Artificial <220>

<223> SusC specific PCR primer 1 <400> 17

cccttctgcc atttcaggaa cc <210> 18 <211> 22 <212> DNA <213> Artificial <220>

<223> SusC specific PCR primer 1 <400> 18

aatgggacca aacccagagt tc

Claims (34)

Isolated sucrose synthase protein having an amino acid sequence comprising an amino acid sequence selected from: a. an amino acid sequence having at least 80% sequence homology to the amino acid sequence according to any claim 2, 3, 5, 7, 9, 11 or 13; B. an amino acid sequence comprising the amino acid sequence according to any SEQ ID NO: 2, 3, 5, 7, 9, 11 or 13; ç. an amino acid sequence located at the amino-terminal part of said protein, said amino acid sequence comprising at least about 60% sequence identity with the amino acid sequence of SEQ ID NO: 16; or d. an amino acid sequence located at the carbon-terminal part of said protein, said amino acid sequence comprising at least about 60% sequence identity with the amino acid sequence of SEQ ID NO: 15. An isolated sucrose synthase protein according to claim 1, wherein said protein comprises a hydrophobic N-terminal sequence. An isolated sucrose synthase protein according to claim 1 or claim 2, wherein said protein further comprises any of the following amino acid sequences: a. the amino acid sequence of SEQ ID NO: 3 from amino acid 383 to amino acid 394 b. the amino acid sequence of SEQ ID NO: 3 from amino acid 270 to amino acid 329 c. the amino acid sequence of SEQ ID NO: 3 from amino acid 549 to amino acid 737 d. the amino acid sequence of SEQ ID NO: 3 from amino acid 1 to amino acid 545; or e. the amino acid sequence of SEQ ID NO: 3 from amino acid 18 to amino acid 794. An antibody recognizing the isolated protein according to any one of claims 1 to 3. An isolated DNA molecule or nucleic acid encoding protein sucrose synthase as defined in any one of claims 1 to 3. An isolated DNA molecule or nucleic acid according to claim 5 comprising the nucleotide sequence of any SEQ ID NO: 1, 4, 6, 8, 10, 12 or 14. 7. Expression cassette comprising the operably linked DNA molecules that follow: a. a plant expressible promoter b. a DNA encoding a sucrose synthase according to any one of claims 1 to 3 or an isolated DNA according to claim 6; and optionally c. a transcription termination region and polyadenylation. Expression cassette according to claim 7, wherein the plant-expressible promoter preferably controls transcription in fiber cells of a fiber producing plant. Expression cassette comprising the operably linked DNA molecules that follow: a. a plant expressible promoter b. a DNA which when transcribed results in an RNA molecule said RNA molecule comprising a nucleotide sequence selected from: i. a nucleotide sequence of at least 19 consecutive nucleotides having at least about 94% sequence identity with the nucleotide sequence of a gene encoding endogenous saccharose synthase isoform C or the complement of said nucleotide sequence of a gene encoding endogenous sucrose synthase isoform C; ii. a nucleotide sequence of at least 19 consecutive nucleotides having at least about 94% sequence identity to a nucleotide sequence according to claim 5 or claim 6; and optionally iii. a transcription termination region and polyadenylation. Expression cassette comprising the operably linked DNA molecules that follow: a. a plant expressible promoter, preferably a plant expressible promoter that preferably controls transcription in fiber cells; B. a DNA, which when transcribed gives a double stranded RNA molecule capable of reducing the expression of an endogenous SusC gene to the fiber producing plant, and the RNA molecule comprising first and second RNA regions in which it went first. RNA region comprises a nucleic acid sequence of at least 19 consecutive nucleotides having at least about 94% sequence identity to the nucleotide sequence of an endogenous SusC gene or nucleotide sequence according to claim 5 or claim. 6; ii. the second RNA region comprising a nucleotide sequence complementary to at least 19 consecutive nucleotides of the first RNA region; iii. the first and second RNA regions are capable of base pairing to form a double stranded RNA molecule between at least 19 consecutive nucleotides of the first and second regions; and c. a 3 'end region comprising transcription termination and polyadenylation signals functioning in plant cells. An expression cassette according to claim 9 or claim 10, wherein the plant expressible promoter preferably controls the transcription in fiber cells of a fiber producing plant. 12. Expression cassette comprising the operably linked DNA regions that follow: a. an expressible plant promoter; B. a region of DNA that when introduced and transcribed into a plant cell is processed into miRNA, whereby miR-NA is able to recognize and guide mRNA cleavage of a gene encoding plant endogenous SusC; and c. optionally a 3 'DNA region involved in transcription termination and polyadenylation. The expression cassette of claim 12, wherein said DNA region comprises a nucleotide sequence that is essentially complementary to a nucleotide sequence of at least 21 consecutive nucleotides of a SusC1 coding gene such as a SusC gene comprising the nucleotide sequence of any SEQ ID Ne: 1, 4, 6, 8, 10, 12 or 14, provided that one or more of the mismatches is allowed: b. A mismatch between the nucleotide at the 5 'end of the miRNA and the corresponding nucleotide sequence in the RNA molecule; ç. A mismatch between any of the nucleotides at position 1 to position 9 of the miRNA and the corresponding nucleotide sequence in the RNA molecule; or d. Three mismatches between any of nucleotides at position 12 to miRNA position 21 and the corresponding nucleotide sequence in the RNA molecule as long as there are no more than two consecutive mismatches. 14. Plant cell comprising a heterologous plant expressible promoter operably linked to a selected DNA molecule from the following DNA molecules: a. a DNA encoding a sucrose synthase according to any one of claims 1 to 3; B. a DNA according to claim 6; ç. a DNA which when transcribed results in an RNA molecule said RNA molecule comprising a nucleotide sequence of at least 19 consecutive nucleotides having at least about 94% sequence identity to the nucleotide sequence of a gene encoding the endogenous sucrose synthase C isoform or the complement of said nucleotide sequence of a gene encoding an endogenous sucrose synthase C isoform; d. a DNA which when transcribed results in an RNA molecule said RNA molecule comprising a nucleotide sequence a nucleotide sequence of at least 19 consecutive nucleotides having at least about 94% sequence identity with a sequence of nucleotide according to claim 5 or claim 6; and. a DNA, which when transcribed gives a double stranded RNA molecule capable of reducing the expression of an endogenous SusC gene to the fiber producing plant, and the RNA molecule comprising first and second RNA regions in which i. The first RNA region comprises a nucleotide sequence of at least 19 consecutive nucleotides having at least about 94% sequence identity to the nucleotide sequence of an endogenous SusC gene or the nucleotide sequence of the nucleotide sequence. claim 5 or claim 6; ii. the second RNA region comprises a nucleotide sequence complementary to at least 19 consecutive nucleotides from the first RNA region; and iii. the first and second RNA regions are capable of base pairing to form a double stranded RNA molecule between at least 19 consecutive nucleotides of the first and second regions; A DNA that when transcribed into a plant cell is processed into a miRNA, whereby the miRNA is able to recognize and guide the mRNA cleavage of a gene encoding endogenous plant SusC. The plant cell of claim 14, wherein said DNA molecule operably linked to said plant expressible promoter is stably integrated into the genome, preferably the nuclear genome of said plant cell. A plant cell according to claim 14 or claim 15, wherein said cell is from a fiber producing plant. Plant cell according to claim 16, wherein said cell is cotton. Plant comprising the plant cell as defined in any one of claims 14 to 17 or consisting essentially of plant cells as defined in any one of claims 14 to 17. A plant according to claim 18 which is a fiber producing plant. Plant according to claim 18, which is a cotton plant. A plant progeny seed or plant as defined in any one of claims 18 to 20 comprising a plant cell as defined in claim 14. Plant fibers according to claim 20 or 21. 23. Method for modifying the fiber characteristics of a fiber producing plant, said method comprising modifying the functional level of a sucrose synthase isoform C or a sucrose synthase having similar characteristics in cells or cell walls or fluid. apoplastic of said plant. The method of claim 23, wherein the functional level of an isoform C sucrose synthase or sucrose synthase with similar characteristics is increased. The method of claim 24, wherein the functional level of an isoform C sucrose synthase is increased by providing plants with an expression cassette as defined in claim 7 or claim 8. The method of claim 24, wherein the functional level of a sucrose synthase similar to isoform C sucrose synthase is increased by providing the plants with an expression cassette comprising a plant expressible promoter operably linked to a DNA region. encoding a sucrose synthase having a hydrophobic N-terminal region. The method of claim 23, wherein the functional level of an isoform C sucrose synthase or sucrose synthase with similar characteristics is decreased. The method of claim 27, wherein the functional level of an isoform C sucrose synthase is decreased by providing plants with an expression cassette as defined in any one of claims 9 to 13. A method according to any one of claims 23 to 28, wherein one of the following fiber characteristics has been modified: strength, length, fiber fineness, fiber maturity ratio, immature fiber content, fiber uniformity or ". micronaire ". Use of a protein as defined in any one of claims 1 to 3 to modify the characteristics of a fiber in a fiber producing plant. Use of a nucleic acid as defined in claim 5 or claim 6 to modify the characteristics of a fiber in a fiber producing plant. Fibers obtained by the method as defined in any one of claims 23 to 29. A fabric or yarn made of fibers as defined in claim 22 or claim 32. A method for increasing the concentration of UDP-glucose and fructose in the apoplast of a plant cell comprising providing said plant cell with an expression cassette as defined in claim 7 or 8.