CN113106104B

CN113106104B - Rice blast resistance related gene OsNAC29 and application thereof

Info

Publication number: CN113106104B
Application number: CN202110422115.XA
Authority: CN
Inventors: 李生平; 唐定中; 杨德卫; 方建波; 刁志娟
Original assignee: Fujian Agriculture and Forestry University
Current assignee: Fujian Agriculture and Forestry University
Priority date: 2021-04-20
Filing date: 2021-04-20
Publication date: 2023-02-24
Anticipated expiration: 2041-04-20
Also published as: CN113106104A

Abstract

The invention relates to the technical field of plant genetic engineering, in particular to clone, function verification and application of a mediation rice blast resistance related gene OsNAC29, which finds and proves the positive regulation and control effect of the OsNAC29 gene in rice blast resistance reaction for the first time, knocking out OsNAC29 by a genetic engineering method can reduce the resistance of rice to rice blast, and improving the expression level of OsNAC29 can obviously enhance the resistance of rice to rice blast, and field experiments show that the function deletion of OsNAC29 does not influence the agronomic characters of rice, so that the invention can be applied to the genetic improvement of the resistance of rice and other related agricultural crops.

Description

Rice blast resistance related gene OsNAC29 and application thereof

Technical Field

The invention relates to the field of rice disease resistance breeding, in particular to a rice blast resistance related gene OsNAC29 and application thereof.

Background

The rice blast caused by ascomycete Magnaporthe oryzae is a worldwide rice disease, has become a serious obstacle for restricting the continuous stable yield and high yield of rice due to the increasing severity of the damage area and the damage degree, occurs in the whole period of the growth and development of the rice, and is generally divided into seedling blast, leaf blast, panicle blast and grain blast according to the difference of the occurrence part and the occurrence time of the disease. The rice blast infected plant has no strict species specificity, rice is one of the hosts of the rice blast fungus, and the rice blast fungus can also infect other agricultural crops, such as barley, rye, wheat and the like. At present, the prevention and control means for rice blast in agriculture mainly comprises chemical pesticides, but the method easily causes the drug resistance of germs, cannot achieve long-term effective treatment, and the chemical pesticides are not easy to degrade, can cause serious environmental problems and harm human health. Therefore, the deep research and excavation of the broad-spectrum resistance gene of the rice blast can provide gene resources and theoretical basis for the breeding of disease-resistant varieties, and has important effects in agricultural production, rice disease control and agronomic character improvement.

At present, with the rapid development of molecular biology, at least more than 100 rice blast-resistant major resistance genes have been reported, the major resistance gene sites are identified on the other 11 chromosomes except the 3 rd chromosome of rice, and a plurality of rice blast resistance sites are contained on the 6 th, 11 th and 12 th chromosomes of rice, and the resistance genes are clustered at the Pita, pi9 and Pik gene sites. So far, at least 23 rice blast resistant genes, such as Pi37, pit, pi35, pish, pib, pi21, pi36, pi9, pi2, piz-t, pi-d2, pi-d3, pi5, pik, pikm, pik-p, pb1, pi1, pikh, pia, piCO39, pi-ta and Pi54, have been cloned.

In order to resist the invasion of pathogenic bacteria, plants develop a unique immune system in a long evolutionary process, and the unique immune system mainly comprises two levels of immune pathways, namely a PTI (PRR-triggered immunity) reaction triggered by a pathogenic bacteria surface Associated Molecular patterns (PAMPs) and an ETI (Effector-triggered immunity) reaction triggered by a pathogenic bacteria Effector protein (Effector). In the PTI reaction, plants recognize conserved Pathogen surface Associated Molecular patterns (PAMPs) or microorganism Associated Molecular patterns (microMPs) through Pattern Recognition Receptors (PRRs) on cell membranesobe-associated molecular patterns, MAMPs), which in turn initiate a series of immune responses, such as Reactive Oxygen Species (ROS) bursts, ca ²⁺ Influx, mitogen-activated protein Kinase (MAPK) activation, and up-regulation of defense gene expression, among others. The early stage of the experiment shows that OsNAC29 is up-regulated after being induced by rice blast fungi, the rice blast resistance of rice is obviously reduced after the gene is knocked out, and the rice blast resistance of the rice is obviously enhanced by improving the expression level of the gene, so that the OsNAC29 gene has a basis for playing a role in the rice blast disease-resistant reaction.

Disclosure of Invention

In order to improve the genetic improvement of the resistance of relevant agricultural crops such as rice and the like, an NAC family transcription factor gene OsNAC29 which is infected and induced and expressed by rice blast bacteria is found by analyzing and screening the interaction expression spectrum of the rice and the rice blast bacteria, and the expression level of the gene OsNAC29 is up-regulated by nearly 10 times after the gene OsNAC is induced by the rice blast bacteria, so that the rice blast-resistant gene OsNAC29 is provided and subjected to functional identification, and the positive regulation and control effect of the OsNAC29 gene in the rice blast-resistant reaction is found and proved for the first time; the OsNAC29 gene is knocked out in a rice variety flower 11 (ZH 11) by a genetic engineering method, so that the resistance of rice to rice blast is reduced, compared with a wild type, the scab and magnaporthe grisea growth quantity of an Osnac29 mutant is more, the expression level of rice disease-resistant related genes is reduced, the resistance of rice to rice blast can be obviously enhanced by improving the expression level of the OsNAC29, and the function deletion of the OsNAC29 is found through field experiment statistics and does not influence the agronomic characters of the rice, so that the rice blast-resistant gene OsNAC29 provided by the invention can be applied to the genetic improvement of the resistance of rice and other related agricultural crops.

The invention mainly aims to provide a rice blast resistance gene OsNAC29, the CDS sequence of the OsNAC29 gene is shown as SEQ ID No.1, and the total length of the CDS sequence is 1041bp.

Another objective of the invention is to provide a protein coded by a rice blast-resistant gene OsNAC29, wherein the amino acid sequence of the protein coded by the OsNAC29 gene is shown in SEQ ID NO.2 and codes 346 amino acids.

The invention further aims to provide application of the rice blast resistance related gene OsNAC29 or protein coded by the rice blast resistance related gene OsNAC29 in improving plant disease resistance.

The invention also aims to provide application of the rice blast resistance related gene OsNAC29 or protein coded by the rice blast resistance related gene OsNAC29 in improving the rice blast resistance.

The invention has the beneficial effects that:

(1) According to the invention, various identification methods such as a spray inoculation method, a Punch inoculation method, magnaporthe grisea growth detection and the like are adopted to prove that OsNAC29 gene deletion reduces the disease resistance of rice to rice blast; the disease course related gene detection proves that the OsNAC29 gene deletion causes the expression of the plant disease resistance related gene to be reduced; an OsNAC29 overexpression experiment proves that the overexpression of the OsNAC29 gene can enhance the disease resistance of rice to rice blast, and the OsNAC29 positively regulates the disease resistance of the rice to the rice blast; the results all show that the OsNAC29 gene is an excellent candidate gene for creating molecular design of transgenic rice and crops with enhanced rice blast resistance, and has important theoretical value and wide application prospect.

(2) According to the invention, field experiment statistics is carried out, and main agronomic traits such as plant height, ear length, effective tiller number, grain length and grain width of rice after OsNAC29 gene mutation are investigated, so that OsNAC29 gene deletion does not influence agronomic traits, yield and quality of rice, and can be applied to genetic improvement for improving resistance of related agricultural crops such as rice.

Drawings

SEQ ID NO 1 of the sequence Listing is the CDS sequence of the OsNAC29 gene cloned in the present invention;

the sequence table SEQ ID NO.2 is the amino acid sequence of the protein coded by the OsNAC29 gene cloned in the invention;

FIG. 1 is a diagram showing the identification of disease-resistant phenotype of plants subjected to OsNAC29 gene knockout in example 1 of the present invention;

FIG. 2 is a map showing the detection of disease-associated disease course genes in Osnac29 and ZH11 in example 2 of the present invention;

FIG. 3 is a graph showing the identification of OsNAC29 overexpression disease-resistant phenotype in example 3 of the present invention;

FIG. 4 is a graph showing the subcellular localization and transcriptional activity analysis of OsNAC29 protein in example 5 of the present invention.

Detailed Description

The invention will be described in further detail with reference to the following drawings and specific embodiments, but the invention is not limited thereto; in the invention, a rice variety ZH11, rice blast fungus Guy11 and ZHong1-GFP are taken as research objects.

Example 1

OsNAC29 gene disease-resistant phenotype identification

(1) Materials and methods

The rice cultivation method comprises the following steps: hulling rice, and soaking in 75% ethanol for 2min. Pouring out ethanol, continuously soaking for 40min by using 50% of pasteurization solution, reversing for several times from top to bottom in the period, operating in a super clean bench, pouring out the sterilization solution, washing the seeds by using sterile water for 5-6 times, pouring out the sterile water, uniformly spreading the seeds in an MS culture medium, generally 15-20 seeds/dish, culturing in a greenhouse at 28 ℃ for one week to transplant the seedlings, placing the seedlings in a long-day incubator, and after one week, transferring the seedlings to the outdoor for continuous culture, wherein the proper illumination and moisture are ensured in the period;

preparing a rice blast fungus spore liquid: placing the stored rice blast fungus filter paper sheets in a CM solid culture medium, culturing at a constant temperature of 28 ℃ to enable the rice blast fungi to grow, cutting the culture medium into small blocks by using a sterile scalpel on a super clean bench after the growth area of the rice blast fungi reaches more than half of the area of a culture dish, transferring the small blocks to a rice bran culture medium for continuous culture, slightly scraping hyphae on the surface of the culture medium by using a sterile scraper on the super clean bench after the hyphae of the rice blast fungi cover more than 3/4 area of the rice bran culture medium, and continuously culturing for 3-5 days by using an illumination culture box to promote spore production; preparing 0.2% Tween-20 solution, pouring into rice bran culture medium, scraping gently with scraper on the rice bran culture medium to elute spores, filtering spores, counting with blood cell counting plate until spore concentration reaches 1 × 10 ⁵ Per ml;

spray inoculation method: the prepared concentration is 1 × 10 ⁵ Uniformly spraying bacterial liquid per ml on rice for 2-3 weeks to ensure that all rice leaves are coated with the bacterial liquid, performing dark treatment in a high-humidity environment for 24 hours, and performing illumination for 12 hours and black for 12 hoursContinuously culturing under dark condition, keeping environment at high humidity, and observing the phenotype after 3-5 days;

a perforation inoculation method comprises the following steps: taking rice seedlings which grow for about 1 month and have leaf width larger than 1cm, and lightly pressing the rice seedlings on the rice leaves by using a puncher to form wounds but cannot be punched through the wounds; taking 10 μ l of 5 × 10 ⁵ Dripping the spore liquid of/ml on the surface of the perforated leaf, sealing the perforated leaf by using an adhesive tape, and paying attention to the fact that the spore liquid does not flow out; placing the inoculated rice in a greenhouse at 26 ℃ for dark treatment for 24h, then placing the rice in 12h of illumination and 12h of dark and high-humidity conditions for continuous culture for 5-8 days, and counting the size of leaf lesions;

and (3) detecting the biomass of the rice blast fungi: respectively selecting equivalent leaves with consistent respective morbidity trends of the mutant and the wild type, extracting rice genome DNA, determining the concentration, taking Ubiquitin of rice and MoPot2 of Magnaporthe grisea as reference genes, and detecting the growth condition of the Magnaporthe grisea in the rice leaves by qPCR (quantitative polymerase chain reaction) at the DNA level.

(2) Results and analysis

Referring to fig. 1, fig. 1a shows that the magnaporthe grisea strain Guy11 is inoculated by a spray inoculation method, the inoculation is observed and photographed after 3 days, co39 is an inoculation positive control (Co 39 is a rice blast susceptible variety), ZH11 is a wild type, osnac29-1 and Osnac29-2 are two allelic mutants created by a CRISPR/Cas9 technology, and Bar =1cm; FIG. 1b shows the detection of the growth of Magnaporthe grisea in the spray inoculation method of diseased leaves, and 2-fold is adopted for the calculation of the relative gene expression level ^△△CT A method; FIG. 1c shows the inoculation of Pyricularia oryzae Guy11 by the punch inoculation method, and photographs were taken after 7 days of inoculation, with Bar =1cm; FIG. 1d detection of amount of Magnaporthe grisea in the case of a holing inoculation method, 2-fold calculation of relative gene expression level ^△△CT A method;

from the phenotypes of spray inoculation and punch inoculation, after the function of the OsNAC29 gene is deleted, the lesion area of rice is obviously enlarged and increased (as shown in figures 1a and 1c), and the biomass of rice blast bacteria is increased (as shown in figures 1b and 1d), which indicates that the OsNAC29 gene mutation causes the reduction of the rice blast resistance; wherein, FIG. 1a shows that Osnac29-1 and Osnac29-2 both show reduced resistance to Pyricularia oryzae and increased lesion spots relative to ZH11, FIG. 1c shows that Osnac29-1 and Osnac29-2 have larger plaque areas relative to ZH11, indicating that Osnac29-1 and Osnac29-2 are more susceptible to diseases, FIG. 1b shows that the biomass of Pyricularia oryzae in the mutant is significantly higher than that in the wild type, and FIG. 1d shows that the biomass of Pyricularia oryzae in the mutant is significantly higher than that in the wild type.

Example 2

Detection of expression level of disease-resistant related genes in mutant Osnac29 and wild type ZH11

(1) Materials and methods

Taking rice seedlings growing for 2-3 weeks, taking three seedlings as a group, respectively taking rice leaves of a mutant Osnac29 and a wild type ZH11 inoculated with rice blast fungi at three time periods of 0h,24h and 48h, extracting sample RNA, performing reverse transcription to obtain cDNA, detecting the expression condition of disease-resistant related genes in the ZH11 and the Osnac29 by using real-time fluorescent quantitative PCR, and detecting the accumulation condition of PR1a, PR5, PR6, PR10 and WRKY45 transcripts by using Ubiquitin as an internal reference gene;

(2) Results and analysis

Referring to FIG. 2, FIG. 2a shows the transcription level of PR1a in wild type and Osnac29 mutants detected by real-time quantitative PCR in inoculation Guy11, 24h and 48 h; FIG. 2b is a graph showing the transcription level of PR5 in wild type and Osnac29 mutants at time of inoculation Guy11, 24h and 48h by real-time quantitative PCR; FIG. 2c shows the transcriptional level of PR6 in wild type and Osnac29 mutants using real-time quantitative PCR for detection of inoculum Guy11 h,24h and 48 h; FIG. 2d is a graph showing the transcriptional level of PR10 in wild type and Osnac29 mutants using real-time quantitative PCR for detection of inoculum Guy11 h,24h and 48 h; FIG. 2e is a graph showing the transcription levels of WRKY45 in wild type and Osnac29 mutants using real-time quantitative PCR to detect inoculum Guy11 h,24h and 48 h;

as can be seen from FIG. 2, the expression of all 4 disease course related genes PR1a, PR5, PR6 and PR10 and the expression of the rice blast positive regulatory gene WRKY45 in Osnac29 are down-regulated to a certain extent compared with the wild type, which shows that after the OsNAC29 gene is mutated, the resistance of rice to rice blast is reduced, and the OsNAC29 gene regulates the resistance of rice to rice blast in a positive regulation manner.

Example 3

OsNAC29 overexpression for improving rice blast resistance of rice

(1) Materials and methods

And (3) detecting the expression level of the OsNAC29 overexpression plant: respectively extracting RNA from three strains of ZH11, OENAC29-1 and OENAC29-2, performing reverse transcription to obtain cDNA, detecting the expression level of OsNAC29 in an over-expression plant by using real-time fluorescence quantitative PCR, and detecting the material growth cycle of the over-expression level detection material of OsNAC29 for 1 month;

identification of rice blast resistance of OsNAC29 over-expression plants: the wild rice material ZH11, the over-expression material OENAC29-1, OENAC29-2 and the inoculation control Co39, the rice cultivation and inoculation method is the same as the example 1, the growth cycle of the inoculation material is 2-3 weeks, the morbidity phenotype is observed after 3-5 days of spraying inoculation, and the biomass of the rice blast fungi is quantitatively analyzed;

(2) Results and analysis

Referring to FIG. 3, FIG. 3a shows identification of OsNAC29 expression level in OsNAC29 overexpression plants OENAC29, wherein three repeats of OENAC29-1 are abbreviated as OE-1 (1), OE-1 (2) and OE-1 (3), and three repeats of OENAC29-2 are abbreviated as OE-2 (1), OE-2 (2) and OE-2 (3), wherein rice seedlings with 1 month of growth cycle are used as materials, ZH11 is used as a negative control, RNA samples are extracted, and reverse transcription is performed to cDNA for qRT-PCR analysis; FIG. 3b is a photograph of a spray inoculated rice blast fungus strain Guy11 taken 3 days after inoculation with Bar =1cm; FIG. 3c is a photograph of a 7-day inoculated punch inoculation method inoculated with Pyricularia oryzae strain Guy11, with Bar =1cm;

as can be seen from FIG. 3, the expression level of OsNAC29 in the over-expressed plants is obviously up-regulated by a multiple compared with the wild type, and is between 6 and 12 times on average (FIG. 3 a), and the over-expressed material meets the experimental requirements; the results of the spray inoculation experiments show that the lesion area of the OsNAC29 over-expression plants is reduced compared with that of control groups after inoculation, and bacterial biomass detection of diseased leaves also shows that the growth amount of Magnaporthe grisea in the over-expression plants is less than that of wild type (see 3b,3 c), which shows that the over-expression of OsNAC29 can improve the rice blast resistance of rice, and the phenotype further proves that OsNAC29 is regulating and controlling the rice blast resistance of rice.

Example 4

Statistical analysis of agronomic characters of Osnac29 mutant

(1) Materials and methods

Respectively sowing 100 seeds on a wild ZH11 and a mutant Osnac29 for germination, raising seedlings in a paddy field after the seeds germinate for 3 days, transplanting seedlings in the paddy field after the seedlings grow for 25 to 30 days, keeping the row spacing and the plant spacing basically consistent, photographing the plants in a heading stage, and then selecting 20 plants of the ZH11 and the Osnac29-1 with consistent growth vigor for statistics of agronomic characters in a mature stage, wherein the agronomic character indexes comprise the plant height, the spike length, the effective tillering number, the average grain number of each spike, the grain length, the grain width and the like of the rice;

(2) Results and analysis

Statistical results show that the Osnac29 mutant and ZH11 have no difference from wild type in main agronomic traits including plant height, panicle length, effective tiller number, grain length, grain width and the like (as shown in Table 1), which indicates that the function deletion of OsNAC29 does not influence the growth and development of rice;

TABLE 1 statistical survey of agronomic traits for Osnac29 mutants

Traits	ZH11	Osnac29-1
			Plant height(cm)	99.82±1.86	102.67±1.53
Panicle length(cm)	23.15±1.32	23.73±1.70
			Number of effective panicle	9.20±1.04	9.00±1.00
Spikelets per panicle	144.46±4.26	144.9±4.03
			Grain length of 10seeds(cm)	7.92±0.11	7.85±0.09
Grain width of 10seeds(cm)	3.62±0.08	3.53±0.06

Example 5

Subcellular localization and transcriptional Activity assay for OsNAC29

(1) Materials and methods

Subcellular localization: cloning CDS sequence of OsNAC29 gene, connecting to pCAMBIA2300-35S-GFP vector by infusion method to obtain pCAMBIA2300-35S-OsNAC29-GFP plasmid, extracting the obtained plasmid and pCAMBIA2300-35S-GFP empty vector plasmid in a large amount by cesium chloride density gradient centrifugation method (plasmid concentration >1 mug/mul), transforming rice protoplast, and observing subcellular localization of cells under laser confocal microscope.

Analysis of transcriptional activation Activity: CDS sequences corresponding to the full length (1-347 aa), the N-terminal (1-138 aa), the C-terminal 1 (OsNAC 29-C1, 139-253 aa) and the C-terminal 2 (OsNAC 29-C2, 254-347 aa) of OsNAC29 were amplified respectively, ligated to pGBKT7 vector by the infusion method, transformed into yeast strain Y2H Gold, grown at SD/-Trp, and then verified for activation activity on SD/-Trp/X-gal and SD/-Trp/X-gal/AbA media, with OsNAC066 as a positive control.

(2) Results and analysis

Referring to fig. 4, fig. 4a is a subcellular localization of OsNAC29 and fig. 4b is an OsNAC29 transcriptional activity assay; FIG. 4a shows that OsNAC29 specifically localizes to the nucleus, and FIG. 4b shows that OsNAC29 has transcription activation activity, and the activation region is located in 254-347 amino acids range at the C-terminal of OsNAC 29.

The above description is only an embodiment of the present invention, and is not intended to limit the scope of the present invention, and all equivalent structures or equivalent processes performed by the present invention or directly or indirectly applied to other related technical fields are also included in the scope of the present invention.

Sequence listing

<110> Fujian agriculture and forestry university

<120> rice blast resistance related gene OsNAC29 and application thereof

<160> 2

<170> SIPOSequenceListing 1.0

<210> 1

<211> 1041

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 1

atggagatga cgatgtcgtc ggcggcgacg tcgctgccgc cggggttccg gttccacccg 60

acggacgagg agctgatcct gcactacctc cgcagccgcg ccaccgccgg gcagtgcccc 120

gtccccatca tcgccgacgt cgacatctac aagttcgatc catgggacct gccatcgaag 180

gcggtgtacg gggagagtga gtggtatttc ttcagcccgc gagaccgcaa gtaccccaac 240

ggaatccggc cgaaccgcgc cgccgggtcg gggtactgga aggcgacggg aaccgacaag 300

cccatccacg acagcgccac cggcgagagc gtcggcgtca agaaggccct cgtcttctac 360

cgcggccgcc ctcccaaggg caccaagacc agctggatca tgcacgagta ccgcctcgcc 420

gccgaccctc tcgccgccgc cgcaaacacc tacaagccct cctcctcctc ccgattccgc 480

aacgtctcca tgaggctgga cgactgggtg ctctgccgga tctacaagaa gtccggccag 540

gcgtcgccga tgatgccgcc gctcgccgcc gactacgacc acgacgagcc gtccggagtc 600

cttgacgacg cctacagctt ctacgcgccg ccgatgatca gcaccacgct catccccaag 660

ctccccaaga tcccctccat ctccgagctc ttcgacgagc acgcgctcgc ccagatcttc 720

gacgccgccg ccgacccgcc ggccgaccac catcagcatg ccctcgccgt ccacccctcc 780

ctgaaccagc tcctcggcgt cggcgacaac ttcctcgcgg agtgctaccc gtcgacggcg 840

tccacggcca ccgttgccgg cggcaagcgc aaggcgagcc cggccggaga ctacgccggc 900

ggcggccaca cgccggcgaa gaggctcaac ggctcatgct tcgacgtggc gccgcagtcc 960

gtggtgggcg gcttgcaagc gacgccgtcg tcagtcctcg ccggactcaa ccaccagatg 1020

cttcctcctc agctattctg a 1041

<210> 2

<211> 346

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 2

Met Glu Met Thr Met Ser Ser Ala Ala Thr Ser Leu Pro Pro Gly Phe

1 5 10 15

Arg Phe His Pro Thr Asp Glu Glu Leu Ile Leu His Tyr Leu Arg Ser

20 25 30

Arg Ala Thr Ala Gly Gln Cys Pro Val Pro Ile Ile Ala Asp Val Asp

35 40 45

Ile Tyr Lys Phe Asp Pro Trp Asp Leu Pro Ser Lys Ala Val Tyr Gly

50 55 60

Glu Ser Glu Trp Tyr Phe Phe Ser Pro Arg Asp Arg Lys Tyr Pro Asn

65 70 75 80

Gly Ile Arg Pro Asn Arg Ala Ala Gly Ser Gly Tyr Trp Lys Ala Thr

85 90 95

Gly Thr Asp Lys Pro Ile His Asp Ser Ala Thr Gly Glu Ser Val Gly

100 105 110

Val Lys Lys Ala Leu Val Phe Tyr Arg Gly Arg Pro Pro Lys Gly Thr

115 120 125

Lys Thr Ser Trp Ile Met His Glu Tyr Arg Leu Ala Ala Asp Pro Leu

130 135 140

Ala Ala Ala Ala Asn Thr Tyr Lys Pro Ser Ser Ser Ser Arg Phe Arg

145 150 155 160

Asn Val Ser Met Arg Leu Asp Asp Trp Val Leu Cys Arg Ile Tyr Lys

165 170 175

Lys Ser Gly Gln Ala Ser Pro Met Met Pro Pro Leu Ala Ala Asp Tyr

180 185 190

Asp His Asp Glu Pro Ser Gly Val Leu Asp Asp Ala Tyr Ser Phe Tyr

195 200 205

Ala Pro Pro Met Ile Ser Thr Thr Leu Ile Pro Lys Leu Pro Lys Ile

210 215 220

Pro Ser Ile Ser Glu Leu Phe Asp Glu His Ala Leu Ala Gln Ile Phe

225 230 235 240

Asp Ala Ala Ala Asp Pro Pro Ala Asp His His Gln His Ala Leu Ala

245 250 255

Val His Pro Ser Leu Asn Gln Leu Leu Gly Val Gly Asp Asn Phe Leu

260 265 270

Ala Glu Cys Tyr Pro Ser Thr Ala Ser Thr Ala Thr Val Ala Gly Gly

275 280 285

Lys Arg Lys Ala Ser Pro Ala Gly Asp Tyr Ala Gly Gly Gly His Thr

290 295 300

Pro Ala Lys Arg Leu Asn Gly Ser Cys Phe Asp Val Ala Pro Gln Ser

305 310 315 320

Val Val Gly Gly Leu Gln Ala Thr Pro Ser Ser Val Leu Ala Gly Leu

325 330 335

Asn His Gln Met Leu Pro Pro Gln Leu Phe

340 345

Claims

1. Rice blast resistance related geneOsNAC29Or related genesOsNAC29The application of the coded protein in improving the disease resistance of rice blast is characterized in that: the above-mentionedOsNAC29The CDS sequence of the gene is shown as SEQ ID NO.1, and the total length of the CDS sequence is 1041bp; the above-mentionedOsNAC29The protein amino acid sequence of the gene code is shown as SEQ ID NO.2, 346 amino acids are coded, wherein, the related gene of the rice blast resistanceOsNAC29Or related genesOsNAC29The encoded protein is overexpressed in rice.