KR20210157192A - Pepper CbNLR09 gene enhanced resistance to anthracnose - Google Patents

Abstract

The present invention relates to CbNLR09 gene derived from Capsicum annuum cultivars resistant to pepper anthracnose. According to the present invention, by using the expression of the CbNLR09 gene, Capsicum annum cultivars with enhanced or improved resistance to pepper anthracnose, which is caused by Colletotrichum spp., can be developed.

Description

CbNLR09 gene derived from pepper to enhance anthrax resistance {Pepper CbNLR09 gene enhanced resistance to anthracnose}

The present invention relates to a CbNLR09 gene derived from red pepper that enhances anthrax resistance and a method for producing pepper with enhanced anthrax resistance using the same.

Due to the rapid increase of the world population over the past 100 years, all mankind on the planet has faced the difficult problem of securing food. production has been increased Among them, the pest and weed control method using chemical pesticides has been recognized as an essential factor in increasing agricultural productivity, and thus, the use of chemical pesticides worldwide is steadily increasing.

However, the continuous use of chemical pesticides made a great contribution to solving the shortage of food, but due to indiscriminate abuse, pesticide poisoning, groundwater contamination, soil contamination, toxicity to livestock, residues in agricultural products, resistance to various pests and weeds It caused side effects such as the appearance of individuals. Meanwhile, efforts to protect the global environment have been made since the declaration of Rio by the UN Conference on Environment and Development in 1992, and with the establishment of the WTO, multilateral negotiations on international trade that linked the environment and trade, that is, the Green Round, started in earnest to provide food to mankind. In addition, since it was necessary to switch to eco-friendly agriculture, the limit on the use of chemical pesticides and the allowable amount of residues in the field of crop protection are being strengthened as part of that.

Red pepper has been deeply rooted in the Korean diet for about 300 years since it was introduced to Korea, and is an important food crop that cannot be omitted from the traditional diet. Red pepper is the most cultivated seasoning vegetable crop among the total vegetable cultivation area in terms of production area, and it is a crop with a high contribution to farm household income. It is increasing by 7.1%.

However, as red peppers are continuously cultivated, sufficient sources of infection are created in the soil, and they are easily infected during the summer rainy season, increasing the damage from pepper diseases. Among them, red pepper anthrax is one of the biggest diseases of red pepper, and it mainly occurs on leaves, stems, and fruits, and more than 10% of peppers are infected at harvest time every year, acting as a direct impediment to the decrease in pepper production.

Chili anthrax is a disease that is a major problem in Korea, Thailand, India, and Indonesia. The fungus that causes pepper anthrax is a fungus, and four species ( C. acutatum , C. capsici , C. gloeosprioides , C. cocodes ) belonging to the genus Colletotrichum spp. have. Among them, C. capsici is reported to be a major problem not only in Korea but also in Thailand, India, and Indonesia, and it is known that the disease is more common in the Red family than in the Green family. The current method for testing anthrax resistance is to harvest red pepper fruits and directly inoculate them with anthrax. In addition to the inconvenience of doing so, there is a problem that proper selection cannot be made if the disease conditions are not maintained well because the degree of disease is affected by the environment.

Republic of Korea Patent Registration No. 10-0952632

An object of the present invention is to provide a CbNLR09 gene derived from pepper that enhances anthrax resistance, a recombinant vector including the gene, and a transformant transformed with the recombinant vector.

In order to solve the above problems, the present invention provides a CbNLR09 protein derived from pepper for anthrax resistance enhancement consisting of the amino acid sequence shown in SEQ ID NO:2.

The anthrax may be caused by a strain of the genus Colletotrichum spp.

The present invention also provides a CbNLR09 gene represented by SEQ ID NO: 1 encoding the CbNLR09 protein.

The present invention also provides a recombinant vector comprising the CbNLR09 gene.

The present invention also provides a transformant transformed with the recombinant vector.

The transformant may be a plant.

The plant may be red pepper.

The present invention also provides a composition for enhancing anthrax resistance comprising the recombinant vector.

The present invention also provides a method for producing a plant having improved anthrax resistance, comprising the step of introducing the recombinant vector into the plant.

The present invention also provides a method for enhancing anthrax resistance of a plant, comprising the step of introducing the recombinant vector into the plant.

According to the present invention, it was confirmed that anthrax resistance increased when the CbNLR09 gene isolated from anthrax-resistant pepper varieties was overexpressed, and anthrax susceptibility was increased when CbNLR09 expression was suppressed. do.

1 is a result of analyzing CbNLR09 gene expression change after treatment with anthrax.
2 is a schematic diagram of a vector for CbNLR09-overexpression and a tobacco transformant transformed with the overexpression vector.
3 is a result of CbNLR09 gene expression analysis of tobacco overexpression transformants.
4 is a result of anthrax resistance analysis of CbNLR09-tobacco overexpression after anthrax inoculation.
5 is a result of lesion analysis using Image J after inoculation with tobacco overexpressing anthrax.
6 is a result of qRT-PCR analysis of the expression of CbNLR09 and disease resistance genes (NbPR1, NbPR10, and NbPR2) in tobacco overspecies.
7 is a result of analyzing the level of CbNLR09 gene expression after suppressing the expression of the CbNLR09 gene using a virus-induced gene silencing (VIGS) system.
8 is a result of observing lesions after inoculation of anthrax in red pepper fruit after suppressing the expression of CbNLR09 gene with the VIGS system.
9 is an Image J analysis result of lesions after inoculation of anthrax in red pepper fruit after suppressing the expression of CbNLR09 gene with the VIGS system.
10 is a result of qRT-PCR analysis of CaPR1 and CaPR2 expression before and after anthrax inoculation after suppressing the expression of CbNLR09 gene with the VIGS system.
11 is a nucleotide sequence of SEQ ID NO: 1.
12 is an amino acid sequence of SEQ ID NO:2.

The present invention is to identify the role of the CbNLR09 gene involved in anthrax resistance, and to breed pepper varieties with improved anthrax resistance by overexpressing the CbNLR09 gene in this plant (red pepper).

The present invention provides a CbNLR09 protein for promoting anthrax resistance comprising the amino acid sequence shown in SEQ ID NO:2.

In the present invention, "CbNLR09" is a protein consisting of the amino acid sequence shown in SEQ ID NO:2. The amino acid sequence of SEQ ID NO: 2 includes a CC-NB-ARC-LRR structure.

The CbNLR09 protein may be encoded by the nucleotide sequence of SEQ ID NO: 1.

As a result of analyzing the expression of CbNLR09 gene after anthrax treatment in An-S, anthrax susceptible variety, and PBC80, resistant variety, it was confirmed that the expression of CbNLR09 in the resistant variety was higher than that of the susceptible variety, and the CbNLR09 gene was It was confirmed that anthrax resistance was enhanced in the overexpressed tobacco transformant, and it was confirmed that anthrax resistance was reduced in red pepper in which the CbNLR09 gene was suppressed by a virus-induced gene silencing system.

As it was confirmed that the increase in CbNLR09 expression promotes or improves anthrax resistance, the CbNLR09 protein of the present invention has an anthrax resistance enhancing effect.

In the present invention, "anthrax" is a disease of plants caused by infection with anthrax, and lesions appear on fruits and leaves. In the present invention, anthrax may be caused by an infection of the genus Colletotrichum spp.

The genus Coletotricum ( Colletotrichum spp .) The strain includes a Colletotrichum acutatum ( Colletotrichum acutatum ) or Colletotrichum capsici ( Colletotrichum capsici ) strain.

In an embodiment of the present invention, the CbNLR09 gene overexpressing tobacco transformant was inoculated with a Coletotrichum acutatum or Colletotrichum capsici strain, and as a result of analyzing the length of the lesion, tobacco It was confirmed that the anthrax resistance was improved, and it was confirmed that the resistance to Colletotrichum capsici was significantly increased compared to that of Colletotrichum acutatum.

Therefore, the genus Coletotrichum of the present invention ( Colletotrichum spp .) The strain may be a Coletotrichum capsici ( Colletotrichum capsici ).

The present invention also provides a CbNLR09 gene encoding the CbNLR09 protein.

In the present invention, the “CbNLR09” gene includes the nucleotide sequence represented by SEQ ID NO: 1. The CbNLR09 gene was isolated from PBC80, an anthrax resistant pepper variety, and encodes the amino acid sequence of SEQ ID NO:2. The amino acid sequence of SEQ ID NO: 2 includes a CC-NB-ARC-LRR structure.

As a result of comparative analysis of CbNLR09 gene expression after anthrax treatment in An-S, anthrax susceptible variety, and PBC80, resistant variety, in the above-described example, it was confirmed that the expression of CbNLR09 in the resistant variety was higher than that of the susceptible variety, and the CbNLR09 gene It was confirmed that anthrax resistance was enhanced in tobacco transformants overexpressing , and anthrax resistance was reduced in red pepper in which the CbNLR09 gene was suppressed by a virus-induced gene silencing system.

That is, the CbNLR09 gene is involved in anthrax resistance, and it was confirmed that anthrax resistance was enhanced when the CbNLR09 gene was overexpressed. can do it

The present invention also provides a recombinant vector comprising the CbNLR09 gene represented by SEQ ID NO: 1.

The description of the CbNLR09 gene in the present invention is the same as described above.

In the present invention, "recombinant" refers to a cell in which the cell replicates a heterologous nucleic acid, expresses the nucleic acid, or expresses a peptide, a heterologous peptide, or a protein encoded by the heterologous nucleic acid. Recombinant cells can express genes or gene segments not found in the native form of the cell, either in sense or antisense form. In addition, recombinant cells can express genes found in cells in a natural state, but the genes are modified and re-introduced into cells by artificial means.

As used herein, "vector" means a DNA preparation having a DNA sequence operably linked to a suitable regulatory sequence capable of expressing the DNA in a suitable host. A vector may be a plasmid, a phage particle, or simply a potential genomic insert. Upon transformation into an appropriate host, the vector may replicate and function independently of the host genome, or in some cases may be integrated into the genome itself. In particular, "plant transformation vector" refers to a recombinant DNA molecule comprising a desired coding sequence and an appropriate nucleic acid sequence essential for expressing a coding sequence operably linked in a specific plant host organism. Promoters, enhancers, termination signals and polyadenylation signals available in plant cells are known to those skilled in the art. The vector of the present invention is most preferably a vector for plant transformation, but is not limited thereto.

Vectors of the invention can typically be constructed as vectors for cloning or expression. In addition, the vector of the present invention can be constructed using a prokaryotic cell or a eukaryotic cell as a host. For example, when the recombinant vector of the present invention is an expression vector and a prokaryotic cell is used as a host, a strong promoter capable of propagating transcription (eg, pLλ promoter, trp promoter, lac promoter, T7 promoter, tac promoter, etc.) , a ribosome binding site for initiation of translation and a transcription/translation termination sequence.

The vector of the present invention may be prepared using a genetic recombination technique well known in the art, and for site-specific DNA cleavage and ligation, an enzyme generally known in the art may be used.

The present invention also provides a transgenic plant transformed with the recombinant vector.

The transformant of the present invention is characterized in that the CbNLR09 gene is overexpressed by being transformed with a recombinant vector containing the CbNLR09 gene represented by SEQ ID NO: 1.

As it was confirmed that overexpression of the CbNLR09 gene enhances anthrax resistance in the above-described embodiment, the transformant of the present invention may have enhanced or improved anthrax resistance by overexpression of the CbNLR09 gene.

The description of the recombinant vector in the present invention is as described above.

As used herein, the term "transformation" refers to a phenomenon in which DNA enters the cell and changes the genetic trait when DNA, which is a genetic material, is injected into a living cell of another lineage. Or also called type conversion.

In the present invention, "transformation" of a plant with the vector can be performed by transformation techniques known to those skilled in the art.

Specifically, transformation method using Agrobacterium, microprojectile bombardment, electroporation, PEG-mediated fusion, microinjection, liposome-mediated method ( liposome-mediated method, In planta transformation, Vacuum infiltration method, floral meristem dipping method, or Agrobacterium spraying method can be used. have.

In the present invention, the term "plant" is meant to include not only mature plants, but also plant cells, plant tissues, and seeds of plants that can develop into mature plants.

In the present invention, the plant is not particularly limited, and as an example, food crops including rice, wheat, barley, corn, soybean, potato, wheat, red bean, oat or sorghum; vegetable crops including Arabidopsis thaliana, Chinese cabbage, radish, red pepper, strawberry, tomato, watermelon, cucumber, cabbage, melon, pumpkin, green onion, onion or carrot; special crops including ginseng, tobacco, cotton, sesame, sugar cane, sugar beet, perilla, peanut or rapeseed; fruit trees including apple trees, pear trees, jujube trees, peaches, poplars, grapes, tangerines, persimmons, plums, apricots or bananas; flowers including roses, gladiolus, gerberas, carnations, chrysanthemums, lilies or tulips; and ryegrass, red clover, orchard grass, alpha alpha, tall fescue, or any one selected from the group consisting of forage crops including perennial ryegrass, and specifically may be pepper.

The present invention also provides a composition for enhancing anthrax resistance comprising the recombinant vector.

The description of the recombinant vector and anthrax in the present invention is the same as described above.

The composition can enhance anthrax resistance of plants by transforming plants with a recombinant vector including the CbNLR09 gene represented by SEQ ID NO: 1 as an active ingredient.

The present invention also provides a method for producing a plant having improved anthrax resistance, comprising the step of introducing the recombinant vector into the plant.

The description of the recombinant vector, plant, and anthrax in the present invention is the same as described above.

The step of introducing in the present invention is to transform the plant with the CbNLR09 gene recombinant vector, and can be performed by transformation techniques known to those skilled in the art.

In the plant produced by the production method of the present invention, the expression of the CbNLR09 gene or a protein thereof is increased in the plant by introducing a recombinant vector containing the CbNLR09 gene involved in anthrax resistance, and the anthrax resistance is enhanced.

The present invention also provides a method for enhancing anthrax resistance of a plant, comprising the step of introducing the recombinant vector into the plant.

The step of introducing in the present invention is to transform the plant with the CbNLR09 gene recombination vector, and can be performed by transformation techniques known to those skilled in the art.

Plants produced by the production method of the present invention can enhance anthrax resistance by introducing a recombinant vector containing the CbNLR09 gene involved in anthrax resistance.

In the present invention, the description of anthrax and plants is the same as described above.

Hereinafter, the configuration and effects of the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.

1. Isolation of anthrax resistance gene

The CbNLR09 gene isolated from the anthrax-resistant pepper variety PBC80 consists of the nucleotide sequence of SEQ ID NO: 1 (2,748 bp, FIG. 11), and was inferred to encode 915 amino acids (SEQ ID NO: 2, FIG. 12), CC-NB - It has an ARC-LRR structure.

To analyze the effect of the CbNLR09 gene on anthrax resistance, CbNLR09 gene expression was analyzed in red pepper fruits 0 to 48 hours after anthrax-resistant (PBC80) and susceptible (An-S) peppers were treated with anthrax. As a result of the analysis, it was confirmed that CbNLR09 gene expression was induced approximately 1.8 times higher in the resistant variety (35.09) than in the susceptible variety (19.53) by anthrax treatment.

2. Construction of CbNLR09 gene overexpression vector and tobacco transformant

A vector for overexpression, pEarleyGate201/CbNLR09, was prepared using the Gateway system on the pEarleyGate201 vector containing the 35S promoter and bar marker (FIG. 2). Tobacco transformants were prepared by transforming the prepared pEarleyGate201/CbNLR09 vector into tobacco.

3. CbNLR09 gene expression analysis and anthrax resistance function assay in tobacco transformants

The expression of the CbNLR09 gene was analyzed by performing RT-PCR on the tobacco transformant prepared above. As a result of the analysis, CbNLR09 expression was not confirmed in the control Nb, but CbNLR09 expression was confirmed in the overexpressing tobacco transformant.

To verify the resistance of CbNLR09-overexpressing protoplasts , two types of anthrax bacteria, Colletotrichum acutatum (Ca) and Colletotrichum capsici (Cc), a dominant bacteria in Southeast Asia, were inoculated into tobacco and their resistance to anthrax was analyzed.

4 is a result of anthrax resistance analysis of CbNLR09-tobacco overexpression after anthrax inoculation. As a result of lesion length investigation 7 days after inoculation of CbNLR09 overexpression with anthrax ( C. acutatum or C. capsici ), it was confirmed that resistance was increased compared to tobacco as a control ( FIGS. 4 and 5 ).

Specifically, as a result of analyzing the lesions caused by anthrax, the disease resistance to the C. acutatum strain showed a significant difference in some (8-4, 15-3) overexpressing organisms, but the remaining overexpressing organisms (1-5, 2- 4, 3-5), it was confirmed that there was no significant difference between the control group and the lesion.

On the other hand, for the C. capsici strain, it was confirmed that all of the overexpressing forms showed significantly fewer lesions compared to the control group.

That is, it was confirmed that overexpression of the CbNLR09 gene enhances anthrax resistance by C. acutatum or C. capsici , and has a better anthrax resistance enhancement effect on C. capsici.

4. Expression analysis of disease resistance-associated gene markers in CbNLR09 gene overexpressing transformants

Next, the expression changes of the disease resistance marker genes NbPR1, NbPR2, and NbPR10 in tobacco transformants overexpressing the CbNLR09 gene were analyzed by qRT-PCR. As a result of the analysis, it was confirmed that the expression of CbNLR09 was increased in the CbNLR09 overexpressing tobacco transformant, and at the same time the expression of NbPR1, NbPR2 and NbPR10 genes was increased ( FIG. 6 ).

5. Analysis of the Effect of Gene Suppression on Anthrax Resistance

Tobacco ( Nicotiana benthamiana ) It was confirmed that overexpression of the CbNLR09 gene promotes anthrax resistance. After suppressing CbNLR09 gene expression using virus-induced gene silencing (VIGS) in red pepper, anthrax resistance was analyzed. did.

After suppressing CbNLR09 gene expression in anthrax-sensitive pepper (An-S) and resistant pepper (PB80), anthrax ( C. acutatum K1) was inoculated, and symptoms were observed 6 days later.

CbNLR09 gene expression was suppressed in pepper using the pTRV system, and CbNLR09 gene expression was confirmed in each pepper after 8 days. As a result of the analysis, it was confirmed that the expression of the CbNLR09 gene was reduced by the pTRV system ( FIG. 7 ).

Next, CbNLR09 gene expression suppressed pepper was inoculated with anthrax (C. acutatum K1) and the disease was observed 6 days later. As a result, in contrast to the overexpressed tobacco transformant, it was sensitive to anthrax compared to the control pepper whose expression was not suppressed. It was confirmed that the deterioration (FIG. 8). After 8 days, the lesions were analyzed using Image J, and as a result, it was confirmed that suppression of CbNLR09 gene expression increased the sensitivity of red pepper fruit to anthrax ( FIG. 9 ).

In addition, the expression of the anthrax resistance marker genes (CaPR1 and CaPR2) was analyzed using qRT-PCR before and after anthrax inoculation 8 days after CbNLR09 gene expression was suppressed using the pTRV system. As a result, the expression of CaPR1 and CaPR2 decreased. was confirmed (FIG. 10).

That is, it was confirmed that the CbNLR09 overexpressing tobacco transformant isolated from the pepper anthracnose resistant variety PBC80 was effective in enhancing anthrax resistance by regulating the expression of tobacco disease resistance marker genes. It was confirmed that the red pepper fruit became more sensitive to anthrax and the expression of disease resistance marker genes was reduced.

<110> REPUBLIC OF KOREA (MANAGEMENT : RURAL DEVELOPMENT ADMINISTRATION) Pepper & Breeding Institutec <120> Pepper CbNLR09 gene enhanced resistance to anthracnose <130> DP20200132 <160> 2 <170> KoPatentIn 3.0 <210> 1 <211> 2748 <212> DNA <213> Artificial Sequence <220> <223> CbNLR09 from PBC80 <400> 1 atggcagatg tagcattaca acttgctgtg gagatattgg ttccgataat aaaggagact 60 tggaagttga ttgatagtgg aaaagaagat tctgcaaatg aactactggg agaacttaat 120 cgtttaaagg ccttcttgca ggatgctgca aaatatcgtc aaagcaacag tgaacagtgg 180 aaaaactttg tcaaggaggt ccaagttatg gtatatgaag ctgagggtct cattgataag 240 ctcatggttg agatgaagct gcaccaaaag aaaaataaat ttgaacagat cgtggatctt 300 aaatatcaca aaacccttaa ggatcttgta gaagacatca aagcaattct tggaaaggtg 360 aagaagattc gcgaagcaaa tctgcaggct ttccaggcaa agccaatcct tgattatcag 420 ccggaaattg ttgcccccgg cacacaggat actttattgg aagaaaacaa agttgtcggt 480 tttgatgagg aagcaaaagt agtgatcaag cgacttgtta aaggaaccaa agatttagat 540 gttatccctg tggtggggtt gcctggacta gggaaaacca ctctggcaat aaaaatctct 600 aaagatccgc aagtttccta tgattttttc ctgaccattt gggtgagcgt aggcccgcaa 660 tccaaactga agggggtctt ccttagtatt ctgaaagcgt tcaagaaaca gactacagaa 720 tatcaagaca tggatgtgaa ggaattgcca aagataatat gtgaattcat tgacaaaggc 780 ggcaaatgtc tcattgtatt ggatgacgtg tggacaacag acgttgtgga tgctgtcatg 840 aatgttttcc ccgaaaaaag caaaggccac cgtatcatga tcaccactcg tgatggacgt 900 attggtagat atgccaatgc taatcctcac atgttgaaat ttctggaaat ggaggaaagt 960 ttccaattgt tggtaaatag agtttttggc agtaatattc gaaggtgtcc tgaagagtta 1020 atagaacatg gggaaagcat cgcaaaacaa tgttttggag tgccacttgc agttgtggta 1080 attgcaggag ctctaagagg acgcaccagc aaaagtgact ggaaaatggt tgaggacaat 1140 gtaaaacacc ttataaataa agatgatgac ccaaaaagct gcttgaaatt tgtggaaatg 1200 agttatgttt atctacccga agagatgaag gcgtgtttcc tatattgtgg tgcctttcca 1260 caaggctttg aaattcctgc ttggaagttg atccgcttat ggatttccga gggattgata 1320 aactccaact taacaggaag ccccgaggat atagcagagt attacttgaa cgaccttatc 1380 aacaggaact tagtaatagt agtgcagaaa agggctaatg gtcaagtaaa aacatgccgt 1440 attcatgaca tgttacacca gttttgcaag atagaggcta ataacgaagg tcttttccac 1500 gaagtatgtg aaaaaacaga tcaggctggt ctttctatac cagatctaga tagttcgcgt 1560 cacttgtgta ttccactctc tcttttgaaa gcttttatct ccaccgaacc atatgctgag 1620 catgttagat cattcttgtg tttttccaca aaacaaaaag aaagtgagaa gcttagcaac 1680 attcaacaac tccacaaagc ctttccactg gtcagggtct tggatgttga atcccttgaa 1740 tttagtttca gcaagtattt cagagagcta tatcatttga ggtacatttc catctcagtt 1800 gaaagcagtg tccttccgac gttcttcggt aaattttgga atttacaaac tcttataatt 1860 tatacaaagg catccaccat tgaaattaaa gctgaaatat ggaacatgct acggttgagg 1920 catgtgcaca ccaatgtccc tgcaaaattg ccatcctcta ctacccacac agtagatgaa 1980 tcttctcgcc tacaaacttt gtctaaagtt gcaccagaaa gttgcagaga agatgcgctt 2040 gcgcgggctt gtaatctcag aaaactaact attcaaggga aaatggctga ttttcttgaa 2100 attaacatgg gtgggttcaa caactttcaa aagctaaagt gcctggagca attgaaactg 2160 ctgaatgata atgtagacag atccatgagc ggagttcttc accttcctcc cgcattcccc 2220 aaatttctat gcaaactgaa gaagttaact ttgtcaaata caaggtttgc ttggagcgag 2280 gcgagtagat tggggcagtt ggaatgcctt gaggtcctaa agctaaaaga aaatgcattt 2340 agtgggacaa catgggattc agagattgga ggttttaacc aactaaaggt attatggatt 2400 gaaagggcag accttaaaac ttggaaggtc gcaaatcttc aatttcaaag acttcagtgt 2460 cttgttgtta aatcctgcga tgagcttgag gctgtaccaa ttgagttggc tgatgtacgt 2520 acccttcaag aaatgacgct ggagcacaca aaaaaggcta tcaaatctgc gaatgctatc 2580 aaatgcagga agcaagagat gcatcgtgag cacatgcaga aagctgcaga aagcaaatct 2640 ggaacagaaa tcgaaggcaa gaaaaaggtg agtgaagata tggagagttt caaattcgag 2700 ctcactatat tcccccctga aacggctgat tgcaacccca aacattga 2748 <210> 2 <211> 915 <212> PRT <213> Artificial Sequence <220> <223> CbNLR09 from PBC80 <400> 2 Met Ala Asp Val Ala Leu Gln Leu Ala Val Glu Ile Leu Val Pro Ile 1 5 10 15 Ile Lys Glu Thr Trp Lys Leu Ile Asp Ser Gly Lys Glu Asp Ser Ala 20 25 30 Asn Glu Leu Leu Gly Glu Leu Asn Arg Leu Lys Ala Phe Leu Gln Asp 35 40 45 Ala Ala Lys Tyr Arg Gln Ser Asn Ser Glu Gln Trp Lys Asn Phe Val 50 55 60 Lys Glu Val Gln Val Met Val Tyr Glu Ala Glu Gly Leu Ile Asp Lys 65 70 75 80 Leu Met Val Glu Met Lys Leu His Gln Lys Lys Asn Lys Phe Glu Gln 85 90 95 Ile Val Asp Leu Lys Tyr His Lys Thr Leu Lys Asp Leu Val Glu Asp 100 105 110 Ile Lys Ala Ile Leu Gly Lys Val Lys Lys Ile Arg Glu Ala Asn Leu 115 120 125 Gln Ala Phe Gln Ala Lys Pro Ile Leu Asp Tyr Gln Pro Glu Ile Val 130 135 140 Ala Pro Gly Thr Gln Asp Thr Leu Leu Glu Glu Asn Lys Val Val Gly 145 150 155 160 Phe Asp Glu Glu Ala Lys Val Val Ile Lys Arg Leu Val Lys Gly Thr 165 170 175 Lys Asp Leu Asp Val Ile Pro Val Val Gly Leu Pro Gly Leu Gly Lys 180 185 190 Thr Thr Leu Ala Ile Lys Ile Ser Lys Asp Pro Gln Val Ser Tyr Asp 195 200 205 Phe Phe Leu Thr Ile Trp Val Ser Val Gly Pro Gln Ser Lys Leu Lys 210 215 220 Gly Val Phe Leu Ser Ile Leu Lys Ala Phe Lys Lys Gln Thr Thr Glu 225 230 235 240 Tyr Gln Asp Met Asp Val Lys Glu Leu Pro Lys Ile Ile Cys Glu Phe 245 250 255 Ile Asp Lys Gly Gly Lys Cys Leu Ile Val Leu Asp Asp Val Trp Thr 260 265 270 Thr Asp Val Val Asp Ala Val Met Asn Val Phe Pro Glu Lys Ser Lys 275 280 285 Gly His Arg Ile Met Ile Thr Thr Arg Asp Gly Arg Ile Gly Arg Tyr 290 295 300 Ala Asn Ala Asn Pro His Met Leu Lys Phe Leu Glu Met Glu Glu Ser 305 310 315 320 Phe Gln Leu Leu Val Asn Arg Val Phe Gly Ser Asn Ile Arg Arg Cys 325 330 335 Pro Glu Glu Leu Ile Glu His Gly Glu Ser Ile Ala Lys Gln Cys Phe 340 345 350 Gly Val Pro Leu Ala Val Val Val Ile Ala Gly Ala Leu Arg Gly Arg 355 360 365 Thr Ser Lys Ser Asp Trp Lys Met Val Glu Asp Asn Val Lys His Leu 370 375 380 Ile Asn Lys Asp Asp Asp Pro Lys Ser Cys Leu Lys Phe Val Glu Met 385 390 395 400 Ser Tyr Val Tyr Leu Pro Glu Glu Met Lys Ala Cys Phe Leu Tyr Cys 405 410 415 Gly Ala Phe Pro Gln Gly Phe Glu Ile Pro Ala Trp Lys Leu Ile Arg 420 425 430 Leu Trp Ile Ser Glu Gly Leu Ile Asn Ser Asn Leu Thr Gly Ser Pro 435 440 445 Glu Asp Ile Ala Glu Tyr Tyr Leu Asn Asp Leu Ile Asn Arg Asn Leu 450 455 460 Val Ile Val Val Gln Lys Arg Ala Asn Gly Gln Val Lys Thr Cys Arg 465 470 475 480 Ile His Asp Met Leu His Gln Phe Cys Lys Ile Glu Ala Asn Asn Glu 485 490 495 Gly Leu Phe His Glu Val Cys Glu Lys Thr Asp Gln Ala Gly Leu Ser 500 505 510 Ile Pro Asp Leu Asp Ser Ser Arg His Leu Cys Ile Pro Leu Ser Leu 515 520 525 Leu Lys Ala Phe Ile Ser Thr Glu Pro Tyr Ala Glu His Val Arg Ser 530 535 540 Phe Leu Cys Phe Ser Thr Lys Gln Lys Glu Ser Glu Lys Leu Ser Asn 545 550 555 560 Ile Gln Gln Leu His Lys Ala Phe Pro Leu Val Arg Val Leu Asp Val 565 570 575 Glu Ser Leu Glu Phe Ser Phe Ser Lys Tyr Phe Arg Glu Leu Tyr His 580 585 590 Leu Arg Tyr Ile Ser Ile Ser Val Glu Ser Ser Val Leu Pro Thr Phe 595 600 605 Phe Gly Lys Phe Trp Asn Leu Gln Thr Leu Ile Ile Tyr Thr Lys Ala 610 615 620 Ser Thr Ile Glu Ile Lys Ala Glu Ile Trp Asn Met Leu Arg Leu Arg 625 630 635 640 His Val His Thr Asn Val Pro Ala Lys Leu Pro Ser Ser Thr Thr His 645 650 655 Thr Val Asp Glu Ser Ser Arg Leu Gln Thr Leu Ser Lys Val Ala Pro 660 665 670 Glu Ser Cys Arg Glu Asp Ala Leu Ala Arg Ala Cys Asn Leu Arg Lys 675 680 685 Leu Thr Ile Gln Gly Lys Met Ala Asp Phe Leu Glu Ile Asn Met Gly 690 695 700 Gly Phe Asn Asn Phe Gln Lys Leu Lys Cys Leu Glu Gln Leu Lys Leu 705 710 715 720 Leu Asn Asp Asn Val Asp Arg Ser Met Ser Gly Val Leu His Leu Pro 725 730 735 Pro Ala Phe Pro Lys Phe Leu Cys Lys Leu Lys Lys Leu Thr Leu Ser 740 745 750 Asn Thr Arg Phe Ala Trp Ser Glu Ala Ser Arg Leu Gly Gln Leu Glu 755 760 765 Cys Leu Glu Val Leu Lys Leu Lys Glu Asn Ala Phe Ser Gly Thr Thr 770 775 780 Trp Asp Ser Glu Ile Gly Gly Phe Asn Gln Leu Lys Val Leu Trp Ile 785 790 795 800 Glu Arg Ala Asp Leu Lys Thr Trp Lys Val Ala Asn Leu Gln Phe Gln 805 810 815 Arg Leu Gln Cys Leu Val Val Lys Ser Cys Asp Glu Leu Glu Ala Val 820 825 830 Pro Ile Glu Leu Ala Asp Val Arg Thr Leu Gln Glu Met Thr Leu Glu 835 840 845 His Thr Lys Lys Ala Ile Lys Ser Ala Asn Ala Ile Lys Cys Arg Lys 850 855 860 Gln Glu Met His Arg Glu His Met Gln Lys Ala Ala Glu Ser Lys Ser 865 870 875 880 Gly Thr Glu Ile Glu Gly Lys Lys Lys Val Ser Glu Asp Met Glu Ser 885 890 895 Phe Lys Phe Glu Leu Thr Ile Phe Pro Pro Glu Thr Ala Asp Cys Asn 900 905 910 Pro Lys His 915

Claims (10)

CbNLR09 protein for promoting anthrax resistance, consisting of the amino acid sequence shown in SEQ ID NO:2. The method of claim 1,
The anthrax is Colletotrichum spp . The protein is caused by the strain. The CbNLR09 gene represented by SEQ ID NO: 1 encoding the CbNLR09 protein of claim 1. A recombinant vector comprising the gene of claim 3 . A transformant transformed with the recombinant vector of claim 4. 6. The method of claim 5,
The transformant is a transformant that is a plant. 7. The method of claim 6,
The transformant is that the plant is red pepper. A composition for enhancing anthrax resistance comprising the recombinant vector of claim 4. A method for producing a plant having improved anthrax resistance, comprising the step of introducing the recombinant vector of claim 4 into the plant. A method for enhancing anthrax resistance of a plant comprising the step of introducing the recombinant vector of claim 4 into the plant.

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