AU744673B2 - Gene associated with disease resistance in plants - Google Patents

Description

WO 99/36542 PCT/SG98/00004 1 GENE ASSOCIATED WITH DISEASE RESISTANCE IN PLANTS Background of the Invention Field of the Invention Bacterial, fungal and viral infections of plants grown for food and fiber cause substantial economic losses to farmers and consumers. For example, rice blast is the most economically devastating disease of cultivated rice, caused by the filamentous fungus Magnaporthe grisea (Ou, 1985). (A bibliography is provided at the end of the written description.) It occurs in most rice growing areas worldwide, costs farmers $5 billion annually (Moffat, 1994). The disease reduces rice yield significantly, particularly in the temperate flooded and tropical upland rice ecosystem. The use of resistant cultivars is the most economical and effective method of controlling the disease. With the advent of transgenic plant technology, it is possible to identify natural host defense mechanisms and to transfer genes associated with these mechanisms to or control expression of such genes in commercial cultivars. It is hoped that expression of such genes will confer disease resistance to the transgenic plants.

SUBSTITUTE SHEET (RULE 26) WO 99/36542 PCT/SG98/00004 2 Background Art Over the last decades, much has been learned about the genetics of resistance to the blast fungus. While the molecular mechanism of host defenses to this pathogen is mostly unknown, blast fungus is believed to infect rice plants in a manner typical of other foliar pathogens. Infection by M.grisea is initiated when a conidium lands on a leaf surface. In a drop of water, a conidium produces a germ tube that grows and differentiates a specialized infection structure called an appressorium that adheres tightly to the plant surface (Bourett and Howard, 1990). The specialized cell generates enormous turgor pressure that is used to penetrate the underlying plant surface (Howard, 1994) The penetration into plant cells by pathogen invasion may damage the cell structure and activate genes responsive to wounding.

In plants, two mitogen-activated protein ("MAP") kinases involved in defense response to wounding have been identified (Usami et al., 1995; Bogre et al., 1997). Usami et al., (1995) reported a MAP kinase that is induced by wounding leaves from a variety of plant species including dicotyledonous and monocotyledonous plants. Another MAP kinase in alfalfa, p44MMK4, was activated by wounding. After wounding, the activity of p44MMK4 rose within 1 minute but decreased to basal levels within 30 minutes. It has been demonstrated that a MAP kinase, PMK1, plays a role in appresorrium formation and infectious growth in rice blast fungus M.grisea (Xu and Hamer, 1996).

SUBSTITUTE SHEET (RULE 26) WO 99/36542 PCT/SG98/00004 3 The MAP kinase signaling cascade is one of the major pathways involved in transducing extracellular stimuli into intracellular responses in mammals and yeasts (Shyy and Chien, 1997, Gabay et al., 1997, Samejima et al., 1997). MAP kinase is a specific class of serine/threonine protein kinases and has been implicated in a wide variety of physiological processes, such as cell growth, differentiation, oncogenesis and response to environmental stresses (Herskowitz, 1995, Cohen, 1997). In mammals, MAP kinases or extracellular signal regulated kinases ("ERKs") were originally identified as transducers of mitogens (substances that induce proliferation). Later, MAP kinases were also shown to be involved with signaling hormones, neurotransmitters and signals for differentiation(Marshall, 1994). At present, MAP kinase pathways are best understood in yeast and animals and several distinct MAP kinase pathways have been identified (Ruis and Schuller, 1995). The basic module of a MAP kinase cascade is a specific set of three functionally interlinked kinases. The activation of MAP kinases is brought about by upstream (i.e.

earlier in the reaction sequence) kinases through phosphorylation of the conserved threonine and tyrosine residues that are located close to kinase domain VIII in all MAP kinases (Marshall, 1994; Hirt, 1997). These dual-specificity MAP kinase kinases (MAPKKs) can only catalyse the activation of specific MAP kinase and can not substitute for each other. The MAPKKs are themselves activated by phosphorylation through upstream kinases that either belong to the class of MAPKK kinases (MAKKKs), or are raf and mos proteins (Marshall, 1994; Hirt, 1997).

SUBSTITUTE SHEET (RULE 26) WO 99/36542 PCT/SG98/00004 4 In plants, several genes encoding MAP kinases have been identified from alfalfa (Jonak et al., 1993; 1995), Arabidopsis (Mizoguchi et al., 1994), pea (Stafstrom et al., 1993), petunia (Decroocq-Ferrant et al., 1995), tobacco (Wilson et al., 1993) and parsley (Ligterink et al., 1997). Similar to mammalian kinases, AtMAPK1 and AtMAPK2 are shown to be involved in cell proliferation (Jonak et al., 1993, Mizoguchi et al., 1994). Several stress-induced MAP kinases have also been identified in plants which are responsive to cold, heat, wounding, drought and mechanical stresses (Bogre et al., 1997, Jonak et al., 1996; Seo et al., 1995, Ligterink et al., 1997; Zhang and Klessig, 1997).

The 48 kD MAP kinase, ERMK, is rapidly activated upon high-affinity binding of a fungal elicitor to a plasma membrane receptor in parsley cells (Ligterink et al., 1997). The activated ERMK is translocated into the nucleus where it may be involved in the transcriptional activation of defense genes. Recently, a MAP kinase, p48 SIP, is identified to be activated in tobacco cells by salicylic acid (SA) treatment which is an endogenous signal for the activation of several plant defense response (Zhang and Klessig, 1997).

These studies suggest that MAP kinases are an important component in the signal transduction pathway of plant defense to pathogen infection. Ligterink et al. (1997) and Zhang and Klessig, (1997) have found a elicitor-responsive MAP kinase in parsley suspension cells and a SA-activated MAP kinase in tobacco suspension cells respectively. However, no evidence was found that MAP kinase is activated by natural pathogen infection in plant species. Accordingly, a need exists for the identification of MAP kinase genes associated SUBSTITUTE SHEET (RULE 26) WO 99/36542 PCT/SG98/00004 with such defense mechanisms and means for expressing such genes in host plants (or regulating their expression) to confer disease resistance.

Summary of the Invention In accordance with this invention, a novel MAP kinase gene and protein that it encodes have been discovered. Based on sequence analysis, this novel gene is a new member of the MAP kinase gene family which encodes a 519 amino acid 59 kD protein. It is designated as BIMKI for blast induced MAP kinase. BIMK1 was strongly induced by rice blast fungus M.grisea and is postulated to be involved in the defense response of rice to blast infection.

In one aspect, the invention relates to the deoxyribonucleic acid that comprises the novel MAP kinase gene, its messenger ribonucleic acid ("mRNA) transcript and the protein that it encodes. In related aspects, the invention involves expression vectors that contain the novel gene operably linked to a plant active promoter and to plant cells and plants that have been transformed with such vectors.

In a further aspect, the invention concerns a method for conferring disease resistance in plants, particularly monocot plants such as rice, wheat, maize, barley and asparagus, which comprises genetically modifying the plant to effect expression of the novel MAP kinase gene.

Brief Description of the Drawings SUBSTITUTE SHEET (RULE 26) II WO 99/36542 PCT/SG98/00004 6 Figure 1 is an autoradiogram of a Southern hybridization analysis of restriction enzyme digested rice genomic DNA using labeled BIMK1 cDNA as a probe.

Figure 2 is an autoradiogram of a Northern analysis of total RNA (50 mg) isolated from rice leaf tissue at different time points after inoculation with M.grisea using labeled BIMK1 cDNA as a probe.

Detailed Description of the Invention The gene encoding a MAP kinase, identified as BIMK1 has been identified for rice, cloned and sequenced. The sequence of the full-length clone, including 5' and 3' untranslated regions, is provided in SEQ ID NO:1. The region from nucleotide 13 through nucleotide 1569 encodes the 519 amino acid 59kD protein whose sequence is shown in SEQ ID NO:2. The BIMK1 gene was isolated from rice infected with the rice blast pathogen, Magnaporthe grisea.

The invention provides an isolated DNA having substantially the sequence spanning nucleotides 13 through 1569 of SEQ ID NO:1. The invention further provides isolated mRNA complementary to the deoxyribonucleic acid having substantially the sequence spanning nucleotides 13 through 1569 of SEQ ID NO:1.

The invention also provides an isolated protein having substantially the sequence shown in SEQ ID NO:2.

"Isolated" as used herein, means that the nucleic acid or protein is in an environment different from its natural environment. For example, it may be cloned in a cloning or expression vector, it may reside in a bacterial cell, it may be associated with other means for transformation of plants or plant cells or it may SUBSTITUTE SHEET (RULE 26) WO 99/36542 PCT/SG98/00004 7 reside in a plant with which it is not naturally associated. As used herein, the term "substantially the sequence" means a sequence that is predominantly that of the identified sequence, provided that the nucleic acid or protein retains the kinase functions of the native molecule. Thus, conservative substitutions, deletions and additions that do not significantly reduce the function of the protein are contemplated.

Probes, primers, antisense molecules and other nucleic acid molecules that are complementary to regions of the BIMKl gene will be useful for its amplification and analysis, regulation of its expression and the like. Accordingly, the invention provides DNA or RNA molecules that are capable of hybridizing to the DNA molecules described above (or their complements) under stringent hybridization conditions. Such conditions are well known in the art and include those conditions under which stable hybrids will form when there is at least about 75%, preferably at least about 80%, most preferably at least about 100% homology between the DNA or RNA molecule and the corresponding region of the target DNA.

The DNA can be incorporated in plant or bacterial cells using conventional recombinant DNA technologies.

Generally, such techniques involve inserting the DNA into an expression vector which contains the necessary elements for the transcription and translation of the inserted protein coding sequences and one or more marker sequences to facilitate selection of transformed cells or plants.

A number of plant-active promoters are known in the art and may be used to effect expression of the nucleic acid sequences disclosed herein. Suitable SUBSTITUTE SHEET (RULE 26) WO 99/36542 PCT/SG98/00004 8 promoters include, for example, the nos promotor, the small subunit chlorophyll A/B binding polypeptide, the promotor of cauliflower mosaic virus, and promoters naturally associated with MAP kinase genes, such as BIMK1 in plants. SEQ ID NO: 6 provides the sequence of the 5' untranslated region upstream of the BIMK1 coding sequence. This region contains the putative promoter for this gene. SEQ ID NO: 6 overlaps the 5' end of the BIMK1 coding region, the ATG start codon appearing at position 1378-80. A "TATA" box appears at positions 1302-1306 of the sequence. In addition to directing expression of the MAP kinase DNA described herein, this promoter has general utility as a plant-active promoter, particularly for effecting expression of transgenes in monocotylodonous plants, such as rice.

Once the isolated DNA of the present invention has been cloned into an expression vector, it may be introduced into a plant cell using conventional transformation procedure-s. The term "plant cell" is intended to encompass any cell derived from a plant including undifferentiated tissues such as callus and suspension cultures, as well as plant seeds, pollen or plant embryos. Plant tissues suitable transformation include leaf tissues, root tissues, meristems, protoplasts, hypocotyls, cotyledons, scutellum, shoot apex, root, immature embryo, pollen, and anther.

One technique for transforming plants is by contacting tissue of such plants with an inoculum of a bacterium transformed with a vector comprising DNA in accordance with the present invention. Generally, this procedure involves inoculating the plant tissue with a suspension of bacteria and incubating the tissue for 48 SUBSTITUTE SHEET (RULE 26) WO 99/36542 PCT/SG98/00004 9 to 72 hours on regeneration medium without antibiotics at 25-280 C.

Bacteria from the genus Agrobacterium can be utilized advantageously to transform plant cells.

Suitable species of such bacteria include Agrobacterium tumefaciens and Agrobacterium rhizogens. Agrobacterium tumefaciens strains LBA4404 or EHA105) is particularly useful due to its well-known ability to transform plants.

Another approach to transforming plant cells with the nucleic acid of this invention involves propelling inert or biologically active particles into plant cells. This technique is disclosed in U.S. Pat. Nos.

4,945,050, 5,036,006 and 5,100,792 all to Sanford et.

al., which are hereby incorporated by reference.

Generally, this procedure involves propelling inert or biologically active particles at the cells under conditions effective to penetrate the outer surface of the cell and to be incorporated within the interior thereof. When inert particles are utilized, the vector can be introduced into the cell by coating the particles with the vector comprising the isolated DNA of this invention. Biologically active particles dried yeast cells, dried bacterium or a bacteriophage, each containing DNA sought to be introduced) can also be propelled into a plant cell tissue.

Another method of transforming plant cells is the electroporation method. This method involves mixing the protoplasts and the desired DNA and forming holes in the cell membranes by electric pulse so as to introduce the DNA into the cells, thereby transforming the cells. This method currently has high SUBSTmTTE SHEET (RULE 26) WO 99/36542 PCT/SG98/00004 reproducibility and various genes have been introduced into monocotyledons, especially rice plants by this method (Toriyama et. al., 1988, Shimamoto et al., 1989 and Rhodes et al., 1988).

Similar to the electroporation method is a method in which the desired gene and protoplasts are mixed and the mixture is treated with polyethylene glycol thereby introducing the gene into the protoplasts. This method is different from the electroporation method in that PEG is used instead of an electric pulse (Zhang W. et. al., 1988, Datta et al., 1990 and Christou et al., 1991).

Other methods include 1) culturing seeds or embryos with nucleic acids (Topfer R. et al., 1989, Ledoux et al., 1974) 2) treatment of pollen tube, (Luo et al., 1988) 3) liposome method (Caboche, 1990 and Gad et al., 1990) and 4) the microinjection method (Neuhaus G. et al., 1987).

Known methods for regenerating plants from transformed plant cells may be used in preparing transgenic plants of the present invention. Generally, explants, callus tissues or suspension cultures can be exposed to the appropriate chemical environment cytokinin and auxin) so the newly grown cells can differentiate and give rise to embryos which then regenerate into roots and shoots.

The isolated DNA of the present invention is believed to be useful in enhancing resistance to disease-causing pathogens in both monocotyledonous plants ("monocots"), and dicotyledonous plants ("dicots"). It is preferred for use with commercially important monocots, such as rice, wheat, barley, maize and asparagus.

SUBSTITUTE SHEET (RULE 26) WO 99/36542 PCT/SG98/00004 11 In plants in which the BIMK1 gene naturally resides, enhanced disease resistance may be achieved by controlling expression of the endogenous gene, rather than transforming the plant with a vector containing the gene. Such control may be achieved, for example, by modifying or replacing endogenous promoters, enhancers or other control signals that regulate expression of the gene, for example, to achieve enhanced expression or programmed expression.

The predicted protein sequence of BIMK1 carries all 11 conserved domains for the catalytic function of serine/threonine protein kinase. The expression of BIMK1 was rapidly induced as early as 4 hours after inoculation with M.grisea, evincing the involvement of BIMK1 in the defense response to the blast fungus.

Several stress-induced MAP kinases have been identified in dicots. As shown in Table 1 below, the protein sequences of these genes showed 70-75% homology. For example, Parsley ERMK and tobacco SIMK have 74.4% protein identity. However, as shown in Table 1, BIMK1 only has about 50% identity with these two stress-related MAP kinases isolated from dicot plants. This suggests the divergence of MAP kinases in monocot and dicot plant species. In addition to sequence differences, BIMK1 is about 500 bp longer than all cloned MAP kinase genes. The 3' region of the gene contains a domain similar with ADH genes in animals.

The function of this domain in the defense response to blast infection isunknown.

The invention is further illustrated by the following examples, which are not intended to be limiting.

SUBSTITUTE SHEET (RULE 26) WO 99/36542 PCT/SG98/00004 12

EXAMPLES

Materials and Methods Rice plants and blast inoculation The resistant isogenic line C101A51 carrying the Pi-2 gene and the susceptible cultivar C039 were used in the experiment. Three week-old rice plants were inoculated with a Philippine isolate P06-6 of M.grisea.

After inoculation, plants were kept in dark in a dew chamber for 24 hours at 260 C. Then, inoculated plants were move into a growth chamber in 10 hours light with 14 hours dark at 25-260 C for 7 days. Leaf tissue was harvested from both cultivars at 0, 4, 8, 12, 24, 48.

72 hours after inoculation.

RNA isolation, cDNA synthesis and RT-PCR RNeasy mini kit (Qiagen, Germeny) was used to isolate total RNA from 150-200 mg rice leaf tissue.

Poly(A)+ RNA fractionated from total RNA using Qiagen Oligotex Spin Column was used as a template in a reverse transcriptase-mediated polymerase chain reaction (RT-PCR). Two primers, CF9-RT and CF9-Rev, were designed based on the DNA sequence of the cloned gene Cf-9, a tomato resistance gene to the leaf mould fungus Cladosporium fulvum (Jones et al., 1994). The primer sequence of CF9-RT is 5'-AAAAGCACAAGTTGCTGC-3' (SEQ ID NO:3) which is the DNA sequence 217-235 bp after the start codon. The sequence of CF9-Rev is 5'TAACGTCTATCGACTTCT-3' (SEQ. ID NO:4) which is the reverse strand sequence of Cf-9 from 1408 to 1426 bp after the start codon. RT-PCR was conducted following protocols provided by the manufacturer

(GIBCO-BRL,

SUBSTITUTE SHEET (RULE 26) WO 99/36542 PCT/SG98/00004 13 Life-Technology, USA). The amplified cDNAs were then separated in 1.2% agarose gel.

Cloning and DNA sequencing Specific bands were cloned into pGEM-T vector (Promega, USA). Clones were sequenced using the ABI PRISM 377 DNA sequencer (Perkin-Elmer, CA, USA). The sequence was analyzed with softwares DNAstar and Sequencher BAC library screening and subcloning Protocols for BAC filter preparation and screening were as described Wang et al. (1995). Hybridization and washing conditions were the same as described in Hoheisel et al., (1993).

Southern hybridization Rice genomic DNA was isolated as described by Dellporta et al. (1984).- DNA was digested with restriction enzymes and separated in 0.8% agarose gel, and then transferred onto Hybond-N+ membrane (Amersham, UK). Probes were labeled using megaprimer labelling kit (Amersham, UK). Rapid hybridization solution (Clonetech, USA) was used.

SUBSTTUTE SHEET (RULE 26) WO 99/36542 PCT/SG98/00004 14 Northern hybridization Total RNA used in the Northern blot analysis was isolated using a Trizol total RNA isolation reagent (GIBCO-BRL, Life-Technology, USA). Fifty micrograms of total RNA per lane was separated in 1.0% agarose gel and transferred onto Hybond-N+ membrane (Amersham, UK) using NorthernMax kit (Ambion, USA) following the manufacturer's instruction. Northern hybridization was carried out same as Southern hybridization described above.

Example 1 Isolation of a cDNA fragment induced after blast infection Total RNA was isolated from leaf tissue inoculated with isolate P06-6 8 hours after inoculation. Purified mRNA was used as template in the first strand cDNA synthesis. When primers CF9-RT and CF9-Rev were used in RT-PCR, four bands were amplified in both C101A51 (compatible) and C039 (incompatible) post-inoculation (data not shown). These cDNA fragments were then cloned into the pGEM-T vector. Clones with different insert sizes were sequenced. A database search revealed that the clone with 350 bp insert is highly homologous to mammalian and yeast MAP kinases.

Example 2 Isolation of genomic clones from rice BAC library To clone the full-length genomic fragment of this gene, a rice BAC library of cultivar IR64 (Yang et al., 1997) was screened using the 350 bp cDNA fragment SUBSTITUTE SHEET (RULE 26) WO 99/36542 PCT/SG98/00004 described in Example 1 as a probe. Four positive BAC clones (3-07, 17-H21, 43-H15 and 43-F5) were identified from the whole BAC library. The miniprepared DNA of the three BAC clones was digested with 3 different enzymes to check if they are overlapping clones in a chromosomal region. Based on the restriction patterns, it was found that these three clones were overlapping clones. Thus, one BAC clone (3-07) was chosen and subcloned into pBluescript-SK (Strategene, USA). The recombinant clone which hybridized with the 350 bp cDNA fragment (Ml, 4.5 kb) was identified and used for sequencing. Based on a comparison with known MAP kinase genes, it was found that it contains the region of the gene including the putative promoter and part of coding region (about 400 bp).

Example 3 Isolation of a full-length cDNA using RT-PCR To isolate a full length cDNA from rice, a primer containing sequence spanning the start codon ATG(5'-AACACAGTGGAAATGGAGTTCTTCA-3') SEQ ID NO:5 was designed based on the genomic DNA sequence. RT-PCR was performed using this primer and a oligo-dT primer (Life-Technologies, USA). From the cDNA prepared from the infected leaves of C101A51 (8 hours after inoculation), a 2.0 kb PCR product was obtained. This PCR product was cloned into pGEM-T vector and sequenced. The sequence is shown in SEQ ID NO:1. It contains a 1557 bp open reading frame corresponding to 519 amino acids (SEQ ID NO:2). This gene was designated BIMK1 for blast induced MAP kinase. This amino acid sequence was compared to the sequence of SUBSTTUTE SHEET (RULE 26) WO 99/36542 PCT/SG98/00004 16 several MAP kinases isolated from a variety of organisms. As shown in Table 1, the sequences are significantly homologous. In section A of the Table, multiple alignment of the deduced amino acid sequence (N-terminal) of BIMK1 with other members of MAP kinases from other organisms is shown. The amino acid sequence of BIMK1 is compared to that of MsERK (Duerr et al., 1993) from Medicago sativa, WIPK (Seo et al., 1995) from tobacco, ATMPK (Mizuguchi et al., 1994) from Arabidopsis, ERK2 (Owaki el al., 1992) from human, ERM (Ligterink et al., 1997) from parsley. Bold type represents amino acid residues that match the EIMKl.

Gaps were induced to maximize alignment. The conserved TXY (in BIMK1, is an aspartic acid while in most MAP kinase it is a glutamic acid) phosphorylation motif for MAP kinase is indicated by asterix. The 11 MAP kinase subdomains are labeled in Roman numerals (Hanks et al., 1988). The M.grisea BIMK1 gene contains all 11 highly conserved subdomains which are present in all known MAP kinases in mammals and plants.

Interestingly, BIMK1 also contains 50 amino acids homologous to mammalian alcohol dehydrogenase (ADH) in its C-terminal. Section B of the Table shows multiple alignment of the deduced amino acid sequence (C-terminal) of BIMK1 with other ADH genes in animals and plants. ADH is present in many organisms that metabolize ethanol, including human, in an oxidoreductase reaction with NAD+/NADH as an essential co-factor.

Example 4 SUBSTTrUTE SHEET (RULE 26) 17 BIMK1 is conserved in rice genome and mapped to a region clustering blast resistance genes DNAs of C101A51 and C039 were digested the restriction enzymes BamHI, EcoRI and HindIII. Southern hybridization was carried out as described in the section of Materials and Methods using the cDNA fragment of BIMK1 as probe. No polymorphism was detected between resistant and susceptible lines for three enzymes (Figure 1).

Similar results have been obtained using DNAs of 4 other cultivars (data not shown). These result indicated that BIMK1 is conserved among rice cultivars. BIMK1 has been mapped on rice chromosome 12 between makers RG341 and RG574, a region clusterring rice blast resistance genes Pi-4(t) and Pi-6(t).

Example BIMKI was induced by rice blast fungus Total RNA was isolated from rice leaf tissue collected at 20 different timepoints after inoculation. The blot was hybridized Susing BIMK1 cDNA fragment as probe labelled with 32p. It was found S* that BIMK1 was highly induced as early as 4 hours after inoculation. The expression of the gene BIMK1 was reduced 24 hours after inoculation (Figure The induction level of BIMK1 in both S 25 resistant (C101A51) and susceptible (C039) lines was very similar (Figure Since C101A51 and Co39 have the same genetic background except C101A51 carries a rice blast resistance gene, Pi-2, it is suggested that BIMK1 was induced independently from Pi-2 and is involved in a general defense pathway to blast.

In the claims which follow and in the preceding description of the invention, except where the context requires otherwise due to express language or necessary implication, the word "comprising", or grammatical variations thereof, is used in the sense of "including", i.e. the features specified may be associated with further features in various embodiments of the invention.

It is to be understood that a reference herein to a prior art document does not constitute an admission that the document forms 17a part of the common general knowledge in the art in Australia or in any other country.

a a a. a a a.

Table 1

(A)

BIMK1 MsERK1 WI PR A TM PK 1 ERK2 E RM BIMK1 Ms ERR]

WIPR

ATMPR].

ERR2

RM

MEGGGAP PADT VMS D -t 1AD

-MEFFT

AAPAPPQMGIENIPA VLSHGGRFIQYNIFG ANMGAGGGQFPDFPS VLTHGGQYVQFDIFG MATLVDPPN GIRNEGK-HYSMWQ MA AAAAAGP EMVRG

EYGEASQ-YQIQ-EV

NI EEVTAKYKPP IMP

NFFEITTRYRPPIMP

TLFEIDTKYMP-IRP

QVFDVGPRYTN-LSY

I

IGKGSYGVVAAAVDT

IGKGAYGIVCSAHNS

IGRGAYGIVCSVLNT

IGRGAYGVVCSSVNS

IGEGAYGMVCSAYDN

II

RTGERVAIKI N DVF

ETNEHVAVKKIANAF

ELNEMVAVIKCIANAF

DTNE KVAIKKI HNVY

LNKVRVAIKKIS-PF

ETNEMVAVKKIANAF MANPGDGQYTDFPA IQTHGGQFIQYNIFG NLFQVTRRYRPPIMP IGRGAYGIVCSIMNT

III

EHVSDATRILREIKL

DNRIDAKRTLRE IKL DI YMDAKRTLREIKaJ

ENRIDALRTLRELKL

EHQTYCQRTLREI11( DN YMDARRTLRE IKI

LRLLRHPDIAEIKHI

LRHMDHENVVAI RD I

LRHLDHENVIGLRDV

LRHLRHENVIALKDV

LLRFRMEN IIGIN DI

LRHLDHENVIAITDV

VII

KICDFGLARAS FNDA

KICDFGLARVTSET-

KICDFGLARPNIEN-

KICDFGLARASNTKG

KICDFGLARVADPDH

KICDFGLARHNTDDE

MLPPSRREFQD IYVV V PPPQRE VFNDVY IA I PPPLRREFSDVYIA MMPI HKMSFKDVYLV

IRAPTIEQMKDVYIV

I PPPLRREFTDVYIA VIii] PSAI FWTDYVATRWY

DFMTEYVVTRWY

-ENMTEYVVTRWY

-QFMTEYVVTRWY

DHTGFLTEYVATRWY

FMTEYVVTRWY

FELME SDLHQV IRAN YELMDTDLHQI IRSN TELMDTDLHQI IRSN YELMDTDLHQI IKSS QDLM4ETDLYKLLRT- TELMDTDLHQI IRSN RAPE IMWLI FSKYTP

RAPELL-LNSSDYTA

RAPELL-LNSSDYTA

PAPELL-LCCDNYGT

RAPE IM-LNS KGYTK

RAPELL-LNSSDYTV

DDLTPEHYQFFLYQL

QALS EEHCQYFLYQI

QGLSEDHCQYFMYQL

QRLSNDHCQYFLFQL

QHLSNDHICYFLYQI

QGLS EEIICQYFLYQL Ix

AIDIWSIGCIFAELL

AID VWS VGCIFMELM

AIDVWSVGCIFMELM

S IDVWSVGCIFAELL

SIDIWSVGCILAEML

AIDIWSVGCIYMELM

-VI

LRALKYIHAANVFHR

LRGLKYIHSANVLHR

LRGLKYIHSANVLHR

LRGLKYIHSANILHR

LRGLKYIHSANVLHR

LRGLKYIHSANIIHR

TGRPLFPGKNVVHQL

DRKPLFPGRDHVHQL

NRKPLFGGKDHVHQI

GRKPIFQGTECLNQL

SNRPIFPGKHYLDQL

NRKPLFPGKDHVHQM

138 180 168 157 146 164 228 265 253 243 235 249 BIMK1 MsERR].

WIPK

ATM PR1 ERR2

ERM

BIMR1 MsERKl

WIPR

ATMPK1 ERR2 E RM DLKPKb4ILANS DCKL

DLKPSNLLLNANCDL

DLKPSNLLVNANCDL

DLKPGNLLVNANC DL DLKPSNLLLNTTC DL

DLKPSNILLNANCDL

x DI ITDLLGTPSSETL

RLLMELIGTPSEDDL

RLLTELLGTPTEADL

KLIVNI IGSQREEDL

NHILGILGSPSQEDL

RLLTELLGSPTEADL

SRIRNEKARRYLSTM

GE'L-NENARRYIRQL

GFLQNEDARRYIRQL

EFIVNPKAKRYIRSL

NCIINLKARNYLLSL

GFVRNEDAKRFILQL

DDVRELIYRE ILEYH

EQMKELIYREALAFN

EQIKDMIYQEALSLN

EMIREMIWNEMLHYH

EKLKELI FEETARFQ EQI KDMIYQEALAFN

RKKHAVPFSQKFRNT

PPYRRQSFQEKFPHV

PQH PRQQLAEVFPHV

PYSPGMSLSRLYPCA

PHKNRVPWNRLFPNA

P RH PRQ PLRQLY PQ V DPLALRLLE RLLAFD H PEA].DLVEKMLTFD NPLAI DLVDKMLTFD HVLAI DLLQKMLVFD

DSKALDLLDKMLTFN

HPLAI DLI DRMLTFD xi

PKDRPSAEEALADPY

PRKRITVEDALAHPY

PTRRI TVEEALDHPY PS KR].SASEALQHPY PH KRI.EVEQAI.AHPY PSKRI TVEEALAHPY FASLANVEREPS RH P

LTSLHDISDEP--VC

LAKLHDAGDEP--IC

MAPLYDPNANP-- PA LEQYYDPSDEP- -IA LARLHDIADEP-- IC 318 352 341 331 323 237 B IMR 1 Ms ERR].

WIPR

ATMPR].

ERR2 E RM I SKLEFEFERRKLTK MT PFSFDFEQHALTE

PVPFSFDFEQQGIGE

QVPI DLDVDED-LRE

EAPFRFDMELDDLPK

TKPS FE FE TAHLG E

PQMLQEYMKG

PEYQQ

PEYA

PQASTLNTEL

PG YRS

PDCA

358 (519) 387 375 370 358 371 0 Table 1 (continued)

(B)

BIMKl-rice 445 SAGQNGVTSTDLSSRSYLKSAS-ISASKCVAVKDNKEPEDDYISEEM-EGSVDGLFQVF-J..QFLV 509 ADH-rabbit 214 AGASRIIAVDINKDKFPK-AKEVGATECINPQDYKKPIQEVIQE-ISDGGVDFSF-VIGRLDTVTV 277 ADH-horse 212 AAGAARIIGVDINKDKFAK-AKEVGATECVNPQDYKKPIQEVLTE-MSNGVDFS.E-VIGRLDTMp/ 275 ADH-hurnan 161 AAGAARIIAVDINKDKFAK-AKELGATECINPQDYKKPIQEVLKE-MTDGGVDFSFE-VIGRLDTMM 224 00 WO 99/36542 PCT/SG98/00004 Bibliography Bogre, Meskiene, Barker, Heberle-Bors, Huskisson NS, Hirt, (1997). Wounding induces the rapid and transient activation of a specific kinase pathway. Plant Cell, 9, 75-83.

Caboche (1990). Physiol. Plant. 79:173-176).

Christou et al. (1991) Bio/Technology 9:957-962 Cohen P. (1997). The search for physiological substrates of MAP and SAP kinases in mammalian cells.

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76:835-840 SUBSITUTE SHEET (RULE 26) WO 99/36542 PCT/SG98/00004 24 SEQUENCE LISTING GENERAL INFORhMATION: APPLIC(ANT: Inst itute for molecular Agrobioloqy (except for t S) He, Chaozu (for US) Wang, Guo-Liang (for US) (ii) TITLE OF INVENTION: Gene Associated with Disease Resistance in Plants fi)NUMBER OF SEQUENCES: 6 INFORMATION FOR SEQ ID NO:l: SEQUENCE CHARACTERISTICS: LENGTH: 1957 base pairs TYPE: nucleic acid STRANDEDNESS: double TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

(vi) ORIGINAL

SOURCE:

ORGANISM: Oryza sativa STRAIN: Cl01A5i (vii) IMMEDIATE SOURCE: CLONE: BIMKI (xi) SEQUENCE DESCRIPTION: SEQ ID NO:l: AACACAGTGG AAATGGAGTT CTTCACTGAG TATGGAGAAG

GAAGTCATTG

GAGCGGGTTG

ATATTGCGTG

CACATTATGC

ATGGAATCAG

CAGTTTTTCC

CATCGCGATC

GATTTCGGAC

TATGTTGCAAu ACTCCTGCAp,

CCACTATTTC

ACTCCATCT

ACCATGCGGA

GCTCTTCGTC

GAAGCTTTGG

CATCCAATCT

GCAAAGGAJ\G

CGATCAAGAA

AGATCAAGCT

TTCCCCCTTC

ATCTCCATCA

TGTACCAACT

TAAAGCCCA

TTGCCCGAGC

CGAGGTGGTA

TTGATATTTG

CTGGGAAGAA

CAGAAACCTT

AAAALACATGC

TGCTAGAGCG

CTGATCCGTA

CAAAAuCTTGA

TTATGGAGTA

GATCAATGAT

CCTTCGTCTG

TCGAAGGGAG

AGTCATCAGA

TCTTCGTGCT

GAATATACTG

ATCATTCAAT

CCGAGCACCT

GAGTATTGGG

TGTTGTGCAC

ATCCAGGATT

TGTCCCCTTC

'rTTACTGGCA

CTTCGCAAGT

GTTTGAATTc

GTTGCTGCTG

GTGTTTGAGC

CTCCGTCACC

TTCCAAGATA

GCGAACGATG

CTCAAGTACA

GCAAACTCAG

GATGCCCCTT

GALATTATGTG

TGCATATTTG

CA.ATTAGATA

CGAAATGAGA

TCTCAGAAGT

TTTGATCCTA

CTTGCTAATG

GAGAGACGGA

CAAGCCAGTA

CACTAGATAC

ATGTATCAGA

CAGACATAGC

TTTATGTTGT

ACCTCACCCC

TCCATGCAGC

ACTGCAAATT

CAGCAATAT'r

GCTCATTTTT

CTGAACTTCT

TTATAACAGA

AGGCCAGGAG

TCCGCAATAC

AAGATCGGCC

TGGAACGTGA

AGCTGACAAA

CCAGATCCA

CCGCACGGGT

CGCTACGCGC

TGAGATCAAA

TTTTGAGCTC

GGAGCACTAC

TAATGTATTT

GAAAATATGT

'rTGGACGGATI

CTCCAAATAC

CACTGGGAGA

TCTTCTTGGA

ATACTTGAGC

TGACCCCTTG

TTCAGCTGAA

GCCCTCAAGA

AGATGATGTT

120 180 240 300 360 420 480 5410 600 660 720 780 8 960 1020 SUBSTITUTE SHEET (RULE 26) WO 99/36542 PCT/SG98/00004 AGAGAATTAA TTTATCGAGA GATTTTGGAG TATCACCCAC AGATGCTGCA AGAGTATATG

AAAGGTGGAG

TTTGCACACC

CATGCTTCTT

ACCAATGACC

TCACAAGATG

AGTTCGAGGA

GACAATAAAG

TTGTTTGAAC

AAGATTTTGT

TTGGACTTTG

GCTCATCCAT

CATGGATTAA

AATGAATAGC

TAGTCATATC

TCCATTCCTC

AGGAGATTAG

TTGAGGAGAA

TACCGAGGGA

AAGAGAGGAG

CACAACAACA

GCTATCTGAA

AACCAGAGGA

AAGTTTTCAG

GAGGCGCACC

GACAATGCAA

GTTCACATAT

TGTATTATCC

AAGCAGCCAG

CATGCTTTTT

CTTCCTCTAT

CTACAGCAAA

GAGAGTAGGT

TGCAGATTCC

TGGATCTGCT

GAGTGCAAGC

TGATTACATC

GATGCAATTC

AAATGCTGAT

GTATGCAACA

TCTTCTTGCC

CTCTGATGTA

CTTGTGCATC

TTGTAATGGT

C CAAGTGGGG

GGAGAAAGAG

GTATCAAAGG

GTTGCCCGCA

GGCCAAAATG

ATTAGTGCTT

TCTGAAGAAA

CTAGTGCACA

AATTTCCA-AG

GCCAGCCCGA

ATTGTGCTGT

ACACTAGATT

ATGTGGGCAT

ATATGAAACA

CTGTGTT

TTGATCGCTT

GTTCTCCACT

ATGGTTATAA

CTACAGTAAG

GTGTGACATC

CCAAGTGTGT

TGGAAGGGTC

ACGATGACGA

CAGGATGCTG

GATGATTGGC

CTGTCACTAC

AGTTCATCTG

GTTCATTTTC

GTTTATCAGT

CAAACGACAG

GCAGAGGAAG

CCAACAAAAC

CCCTCCAATG

CACAGACTTG

CGCTGTCAAG

GGTCGATGGA

TGATCAGTGC

CACTGCAAGT

ATCTTCTTAT

AGGACCCCTG

TCCATGGAGG

CAGTGAGATC

GAGACTGTGG

1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1957 TTTGAAGAAC TCCATTTCCA INFORMATION FOR SEQ ID NO:2: SEQUENCE CHARACTERISTICS: LENGTH: 519 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: protein- (iii) HYPOTHETICAL:

YES

(iv) ANTI-SENSE:

NO

(vi) ORIGINAL SOURCE: ORGANISM: Oryza sativa STRAIN: (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: Met Giu Phe Phe Thr Gin Tyr Gly Glu Ala Ser Gin Tyr 10 Giu Val Ile Gly Lys Gly Ser Tyr Gly Val Val Ala Ala 25 Thr Arg Thr Gly Giu Arg Val Ala Ile Lys Lys Ile Asn 40 45 Glu His Val Ser Asp Ala Thr Arg Ile Leu Arg Giu Ile 5560 Arg Leu Len Arg His Pro Asp Ile Ala Giu Ile Lys His 70 75 Pro Pro Ser Arg Arg Gin Phe Gin Asp Ile Tyr Val Val 90 Gin Ile Gin Ala Val Asp Asp Val Phe Lys Len Len Ile Met Phe SUBSTITIJT SHEET (RULE 26) WO 99/36542 PCT/SG98/00004 26 met Giu Ser Asp Leu His Gin Val Ile Arg Ala Asn Asp Asp Leu Thr 100 105 110 Pro Giu His Tyr Gin Phe Phe Leu Tyr Gin Leu Leu Arg Ala Leu Lys 115 120 125 Tyr Ile His Ala Ala Asn Val Phe His Arg Asp Leu Lys Pro Lys Asn 130 135 140 Ile Leu Ala Asn Ser Asp Cys Lys Leu Lys Ile Cys Asp Phe- Giy Leu 145 150 155 160 Ala Arq Ala Ser Phe Asn Asp Ala Pro Ser Ala Ile Phe Trp Thr Asp 165 170 175 Tyr Val Ala Thr Arg Trp Tyr Arg Ala Pro Glu Leu Cys Gly Ser Phe 180 185 190 Phe Ser Lys Tyr Thr Pro Ala Ile Asp Ile Trp Ser Ile Gly Cys Ile 195 200 205 Phe Ala Giu Leu Leu Thr Gly Arg Pro Leu Phe Pro Gly Lys Asn Val 210 215 220 Val His Gin Leu Asp Ile Ile Thr Asp Leu Leu Giy Thr Pro Ser Ser 225 230 235 240 Giu Thr Leu Ser Arg Ile Arg Asn Giu Lys Ala Arg Arg Tyr Leu Ser 245 250 255 Thr Met Arg Lys Lys His Aia Val Pro Phe Ser Gin Lys Phe Arg Asn 260 265 270 Thr Asp Pro Leu Aia Leu Arg Leu Leu Giu Arg Leu Leu Aia Phe Asp 275 280 285 Pro Lys Asp Arg Pro Ser Ala Giu Giu Ala Leu Ala Asp Pro Tyr Phe 290 295 300 Ala Ser Leu Ala Asn Val Giu Arg Giu Pro Ser Arg His Pro Ile Ser 305 310 315 320 Lys Leu Giu Phe Giu Phe Giu Arg Arg Lys Leu Thr Lys Asp Asp Val 325 330 335 Arg Giu Leu Ile Tyr Arg Giu Ile Leu Giu Tyr His Pro Gin Met Leu 340 345 350 Gin Giu Tyr Met Lys Gly Gly Glu Gin Ile Ser Phe Leu Tyr Pro Ser 355 360 365 Gly Val Asp Arg Phe Lys Arg Gin Phe Ala His Leu Glu Giu Asn Tyr 370 375 380 Ser Lys Gly Giu Arg Gly Ser Pro Leu Gin Arg Lys His Ala Ser Leu 385 390 395 400 Pro Arg Glu Arg Val Giy Val Ser Lys Asp Gly Tyr Asn Gin Gin Asn 405 410 415 Thr Asn Asp Gin Glu Arg Ser Ala Asp Ser Val Ala Arg Thr Thr Vai 420 425 430 Ser Pro Pro Met Ser Gin Asp Ala Gin Gin His Gly Ser Ala Gly Gin 435 440 445 Asn Gly Val Thr Ser Thr Asp Leu Ser Ser Arg Ser Tyr Leu Lys Ser 450 455 460 Ala Ser Ile Ser Ala Ser Lys Cys Val Ala Val Lys Asp Asn Lys Giu 465 470 475 480 SUBSTITUTE SHEET (RULE 26) WO 99/36542 PCT/SG98/00004 27 Pro Glu Asp Asp Tyr Ile Ser Glu Glu Met Glu Gly Ser Val Asp Gly 485 490 495 Leu Phe Glu Gin Val Phe Arg Met Gin Phe Leu Val His Asn Asp Asp 500 505 510 Asp Asp Gin Cys Lys Ile Leu 515 INFORMATION FOR SEQ ID NO:3: SEQUENCE CHARACTERISTICS: LENGTH: 18 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid DESCRIPTION: /desc "Synthetic DNA" (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: AAAAGCACAA GTTGCTGC INFORMATION FOR SEQ ID NO:4: SEQUENCE CHARACTERISTICS: LENGTH: 18 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid DESCRIPTION: /desc "Synthetic DNA" (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE:

NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:4: TAACGTCTAT CGACTTCT INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 25 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid DESCRIPTION: /desc "Synthetic DNA" (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID SUBSTITUTE SHEET (RULE 26) I I WO 99/36542 28 AACACAGTGG AAATGGAGTT CTTCA INFORMATION FOR SEQ ID NO:6: SEQUENCE CHARACTERISTICS: LENGTH: 1678 base pairs TYPE: nucleic acid STRANDEDNESS: double TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE:

NO

(vi) ORIGINAL SOURCE: ORGANISM: Oryza sativa STRAIN: ClOIA51 (vii) IMMEDIATE SOURCE: CLONE: BIMK1 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6: PCT/SG98/00004

GTAATTTTTT

CTGCATCCAT

GCTTTGTCGC

AAGAACGCGA

ACTGGTAGGG

CCACCGCCGC

CTCCGACCTC

CGTGCCCAAG

TAGTGTGAAG

TTTCAGGCTT

ATCTTCGTGG

GATCTGTGCA

TTTCTTCTTT

TCATCCTGGC

TTAAGTTTCT

TCCATTTCTT

GGAGCTGTTC

CAAGTTGTCC

AGGTTTGACT

GTCGCGTCTT

CTATGTACTG

ATGGTTTTGT

TTTTTAGTTA

CCCCATCACC

CCATCCATCC

GGGAACGCGG

GAGCTTGGAA

AGGGGGATGG

ACGGCGTCCG

GAGGTCGCCG

CGCAAGATGC

TGTGCTTTGC

TCTCGTGATC

CACCTGAATT

AAGTGCAATA

CGATGGTTGC

TGTTGACCTG

TTCTTTCTTT

AGAGGAGTGC

TCAACAATTG

ACTTGTGGTT

TGGTCTGGCC

GCACATGGTA

AAAATGTCAA

TTATAGGGCT

GCTTAATATT

ACCACCACCA

ATTACTCGCC

GAAGAAAGGT

GGCGAGCTGC

GGGGAGGGGG

GCTCCAACCA

ACGACCCGGA

CCTCTCCCCG

TTTGCTTCGT

CCTTGATTCG

ACTTGCATAC

CAGCTCAAGT

TTGTAGAGCG

GTTATTCCAG

CTCTGTTTGC

AATTGCAGCA

GTCCCATCAT

TTGGATGATC

TATGTATATC

TGGAGCAGGT

TTTTTGTTAG

ATTCTACTAC

TTCTTGCAAA

CCATCGCTTT

GAAGACTTCG

CCGAGCTTGG

GGTGGTGTAG

CACGCTCGTC

GTCGTCCAAC

TCTCGTCGCC

TCGAGAGCCA

TTTCTTTTCA

TGGCCACGAG

TGTACAATAT

GCAGGTAAAG

ATAGTTGCTT

TGATCTATGA

TTAGTTCTGA

CTACTATGCA

CCTGTCATAC

TCATCAGAAT

TCTGGTACGG

GTCCTTTCAT

GTTGGTCATG

TGP.AGAAGTT

ATTGTGATCT

CTTCATCTTC

CGCGGGGAGA

AAGGAGAAGA

CTAGCCAATG

GACGGATTCC

GCCGGCGAGG

CTCCGCTCCA

CAAGAAGGTG

GTTTGGGGGT

GGGTTCTTAG

CATTATTTCT

CTTGCGTGTT

GTAAACTGCC

AACGATCGAT

AATTACTTGC

AAAAGCTTGT

TGATCCTTAG

CGGCTACTAA

ACTGTTTCTA

TTGCGA.ATAA

ATATTCCCAG

TTATAACCAG

TGTAGAACAC

GCCTTCTGGT

GAGGAGGCAA

AGAAGTAGCC

CCGGCGGGGA

GCCGCATCTT

AGGCCGCCTC

TCCGCATCCG

AGGAGGTGCC

GAAATGAAAG

ACAAGATCAG

TTTTTTCTAT

CTATCCAATC

ATCCGATTCG

CTCGTAAAAC

TCCTGCATGC

GCCCTCTTTT

GAGCTCATAC

TTAGTACTCC

TTGGGAACAA

ACCTACATGT

GGAAAAGATC

CCACTCTGTA

AGTGGAAATG

120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 SUBSITUJTE SHEET (RULE 26) WO 99/36542 WO 9936542PCT/SG98/00004 29 GAGTTCTTCA CTGAGTATGG A GAAGCAAGC CAGTACAGCC AGTACCAGAT CCAGGAAGTC 1440 ATTGGCAAAG GAAGTTATGG AGTAGTTGCT GCTGCAGTAG ATACCCGCAC GGGTGAGCGG 1500 GTTGCGATCA AGAAATCAAT GATGTGTTTG AGCATGTATC AGACGCTACG CGCATATTGC 1560 GTGAGATCAA GCTCCTTCGT CTGCTCCGTC ACCCAGACAT AGCTGAGATC AAACACATTA 1620 TGCTTCCCCC TTCTCGAAGG GAGTTCCAAG ATATTTATGT TGTTTTTGAG CTCATGGAA 1679 SUBS1TnW SHEET (RULE 26)

Claims (23)

1. An isolated deoxyribonucleic acid comprising a nucleic acid sequence that encodes a protein encoded substantially by the sequence from about position 13 through about position 1569 of SEQ ID NO.:1.

2. The deoxyribonucleic acid of claim 1 operably linked to a plant-active promoter.

3. An expression vector capable of transforming a plant cell which contains the deoxyribonucleic acid of claim 1 operably linked to a promoter that is active in said plant.

4. A plant cell transformed with the vector of claim 3. A plant containing a plant cell of claim 4.

6. A seed of the plant of claim

7. The expression vector of claim 3, wherein the plant is a monocot.

8. The plant cell of claim 4, wherein the plant is a monocot.

9. The plant of claim 5, wherein the plant is a monocot.

10. The seed of claim 6, wherein the plant is a monocot. oe

11. The plant cell of claim 4, wherein the plant is rice, oooo 20 wheat, maize, barley or asparagus.

12. The plant of claim 5, wherein the plant is rice, S. wheat, maize, barley or asparagus.

13. The plant of claim 5, wherein the plant is rice.

14. A seed of the plant of claim 12 or 13. ooo S* 25 15. An isolated deoxyribonucleic acid comprising the oe•• sequence from about position 13 through about position oooo 1569 of SEQ. ID. NO.:1.

16. Isolated messenger RNA complementary to the 30deoxyribonucleic acid of claim 1 or ooooo 31

17. An isolated deoxyribonucleic acid molecule or a ribonucleic acid molecule that hybridizes to the deoxyribonucleic acid of claim 1 or 15 or its complement under stringent hybridization conditions.

18. An isolated protein comprising substantially the amino acid sequence of SEQ ID. NO.:2.

19. A method for conferring disease resistance to a plant which comprises genetically modifying the plant to cause or regulate the expression of the deoxyribonucleic acid of claim 1 or The method of claim 19, wherein the plant is a monocot.

21. The method of claim 19, wherein the plant is rice, wheat, maize, barley or asparagus.

22. The method of claim 21, wherein the plant is rice.

23. An isolated plant promoter having a nucleotide sequence substantially contained in SEQ ID NO:6.

24. An isolated nucleic acid molecule that comprises the o-sequence shown in SEQ ID. NO.:1, substantially as described herein with reference to the Examples. o 20 25. An isolated protein encoded by the molecule of claim 24.

26. A cell comprising a nucleic acid molecule of claim 24 or o00o a protein according to claim

27. A plant, or tissue thereof, comprising a cell according ee to claim 26. •oeo° Dated this 8th day of January 2002 ee:INSTITUTE OF MOLECULAR AGROBIOLOGY oeo• By their Patent Attorneys GRIFFITH HACK