CN117568360A - Ula diagram wheat ENHANCED DISEASE SUSCEPTIBILITY 1 analog gene TuEDS1 and application thereof - Google Patents
Ula diagram wheat ENHANCED DISEASE SUSCEPTIBILITY 1 analog gene TuEDS1 and application thereof Download PDFInfo
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Abstract
The invention discloses a similar gene of Ula drawing wheat ENHANCED DISEASE SUSCEPTIBILITY 1TuEDS1And applications thereof. Said Ula diagram wheat ENHANCED DISEASE SUSCEPTIBILITY 1 analog geneTuEDS1The nucleotide sequence of the polypeptide is shown as SEQ ID NO.1, and the amino acid sequence of the encoded protein is shown as SEQ ID NO. 2. Experiments show that the Ula diagram wheat ENHANCED DISEASE SUSCEPTIBILITY 1 analogue geneTuEDS1Not only for wheat stripe rustSexual function geneYrU1Is essential for normal resistance function in riceoseds1Overexpression alone in mutantsTuEDS1Can also obviously improve riceoseds1Resistance to rice blast of the mutant. Thus, the Ula diagram wheat ENHANCED DISEASE SUSCEPTIBILITY 1 analog geneTuEDS1Has stronger application potential in plant resistance breeding.
Description
Technical Field
The invention belongs to the technical field of plant genetic engineering, and in particular relates to a ENHANCED DISEASE SUSCEPTIBILITY 1 similar gene of Ula drawing wheatTuEDS1And the application of the gene in plant resistance breeding.
Background
Common wheat @Triticum aestivum) Is the most important staple food crop worldwide, and up to 55% of population depends on the crop to feed, and wheat diseases such as stripe rust, scab and powdery mildew seriously threaten the wheat production, and greatly damages the yield and quality of the wheat. Wherein the wheat stripe rust is prepared from wheat stripe rust bacteriaPuccinia striiformisf. sp.tritici，Pst) A parasitic fungus disease caused by infecting the leaves and other parts of wheat. The wheat stripe rust can harm wheat and related species plants, once infected and diffused, hypha growing in a host body can compete with plant leaves for nutrients, and a large amount of summer spores generated later can cover the host leaves to prevent photosynthesis of the host plants. Because the application of chemical agents always screens out resistant strains, and pesticide residues pollute grains and ecological environment, and finally, people are retrospectively killed, the cultivation of self-resistant wheat varieties is the most effective, economical and intelligent coping strategy. The cultivation of resistant varieties needs to excavate a large amount of excellent resistance genes and research the resistance mechanism, and the search and research of high-efficiency disease-resistant related genes carried by local wheat varieties and closely related species plants are tightly carried out, so that the cultivation is urgent to meet the requirements of resistant breeding.
Uradil diagram wheat @Triticum urartu) Is the original ancestor of the common wheat A genome, and compared with the donor ancestor of the common wheat B, D genome (namely, aegilops sieboldii and aegilops crudus), the characteristics of the uralensis wheat in plant morphology, wheat ear development and the like are more similar to those of the common wheat. G1812 is a material in Ula diagram wheat whose complete genomic sequence has been sequenced and published, which contains 7 pairs of chromosomes, with a genome size of about 5.6G, and the functional genes have also been substantially annotated complete. PI428309 is also UralWheat material whose genome was not sequenced, but resistance screening found that it had excellent resistance to wheat stripe rust and wheat powdery mildew. By hybridizing PI428309 with the infectious uracratia wheat material G1812 to construct a genetically localized population, and using experimental methods such as map cloning, complementation verification, etc., we have cloned and reported several major, broad-spectrum resistance genes, such as wheat powdery mildew resistance genes, from PI428309Pm60It encodes a typical CC-NBS-LRR (CNL) protein. Whereas in PI428309 another resistance geneYrU1The protein codes a special CNL protein with an ankyrin structural domain at the N end and a WRKY structure at the C end, and the protein endows PI428309 with wheat stripe rust resistance, thereby providing an excellent resistance source for wheat resistance breeding.
Although cloning and mechanism studies of wheat disease resistance genes have been reported, the current available resistance genes for resistance breeding are very limited, and the immune mechanisms of most known resistance genes are known to be quite superficial. At present, it is generally considered that the innate immunity of plants to pathogenic bacteria mainly comprises two layers, namely an immune response pattern-triggered immunity (PTI) induced by molecular patterns related to identifying pathogenic bacteria or injury and an immune response effector-triggeredimmunity (ETI) induced by effector factors secreted by identifying pathogenic bacteria, and the different layers have mutual synergic and promoting relations, so that a large number of complex resistance related genes are often involved in a plant immune mechanism. Revealing the mechanism of wheat against parasitic pathogenic fungi, and further breeding superior resistant varieties, there is an urgent need to further clone and explore more wheat resistance related genes and to delve into the mechanism by which they participate in immune responses.
Based on the background content, the Ula wheat material PI428309 is inoculated with the wheat stripe rustPstTranscriptome sequencing analysis of the samples before and after the sample found a similar gene of protein ENHANCED DISEASE SUSCEPTIBILITY 1 in Ula map wheat PI428309, which plays a key role in Arabidopsis ETI reactionTuEDS1There was no significant change in transcript levels after inoculation with rust. But through gene silencing experiments, it was confirmed thatTuEDS1For Ural diagramWheat stripe rust resistance gene of wheat material PI428309YrU1It is essential to exert a resistance function and,TuEDS1takes part in YrU-mediated stripe rust resistance. Further experiments on the over-expression also prove that,TuEDS1but also can complement riceoseds1Mutant due to knockoutOsEDS1Whereas the significantly reduced rice blast resistance demonstrated in urapidotia wheatTuEDS1Can also play a disease-resistant function in the background of rice. We have also found that TuEDS1 interacts with the analog TuADR1 of arabidopsis ACTIVATED DISEASE RESISTANCE 1 in uracrat wheat, and that silencing TuADR1 of uracrat wheat material PI428309 also significantly reduces YrU 1-mediated wheat stripe rust resistance in PI428309, and we have also found that TuADR1 interacts with the Coiled-Coil (CC) domain of YrU 1. TuEDS1 and TuADR1 exert resistance function on YrU, and are likely to sense and transmit YrU 1-sensed disease resistance signals, so as to activate downstream immune response. Through researching the disease resistance functions of TuEDS1 and TuADR1 in Ula diagram wheat PI428309, the invention provides a novel high-efficiency resistance related gene for wheat disease resistance breeding practice and also discloses a wheat stripe rust resistance geneYrU1The mechanism of regulating immune response lays a solid foundation.
Disclosure of Invention
The invention aims to provide a ENHANCED DISEASE SUSCEPTIBILITY 1 analogue gene of Ula drawing wheatTuEDS1And the application thereof deepens understanding and exploring the mechanism of regulating and controlling immune response of plant disease-resistant genes, and provides novel and efficient source-resistant genes for crop resistance breeding so as to cultivate excellent resistant varieties.
In order to solve the technical problems, the invention adopts the following technical scheme:
the invention firstly provides a ENHANCED DISEASE SUSCEPTIBILITY 1 analogue gene of Ula drawing wheatTuEDS1Said Urapine wheat ENHANCED DISEASE SUSCEPTIBILITY 1 analog geneTuEDS1The nucleotide sequence of the coding region is shown as SEQ ID NO. 1.
The invention also provides a similar gene of ENHANCED DISEASE SUSCEPTIBILITY 1 of the Ula drawing wheatTuEDS1Encoded proteinsThe protein has an amino acid sequence shown as SEQ ID NO. 2.
The invention also provides a ENHANCED DISEASE SUSCEPTIBILITY 1 analogue gene containing the Ula drawing wheatTuEDS1Is a recombinant vector of (a).
The invention also provides a ENHANCED DISEASE SUSCEPTIBILITY 1 analogue gene containing the Ula drawing wheatTuEDS1Is a virus-induced gene silencing vector.
The invention also provides a ENHANCED DISEASE SUSCEPTIBILITY 1 similar gene of the Ula drawing wheatTuEDS1The application in constructing rice blast resistant rice.
The invention also provides a ENHANCED DISEASE SUSCEPTIBILITY 1 similar gene of the Ula drawing wheatTuEDS1At interferenceYrU1Use in stripe rust resistance.
The invention has the beneficial effects that:
the invention clones ENHANCED DISEASE SUSCEPTIBILITY 1 analog gene in Ula drawing wheat PI428309TuEDS1It can regulate and control stripe rust resistance gene in Ula drawing wheat PI428309YrU1For a pair ofPstResistance to CYR 33. In addition, the Ula diagram wheat ENHANCED DISEASE SUSCEPTIBILITY 1 analog geneTuEDS1The rice blast resistance function can be exerted in the rice body, which shows that the gene can exert the resistance function in the heterologous plant. It can be seen that ENHANCED DISEASE SUSCEPTIBILITY 1 analog gene derived from Ula diagram wheat PI428309TuEDS1Has the function of wheat stripe rust resistance and can play the function of resistance in heterologous plant rice, and has important application value in the field of plant disease resistance breeding.
Drawings
Fig. 1: sequence comparison (a) and phylogenetic tree analysis (b) of the Ula diagram wheat ENHANCED DISEASE SUSCHEPALITY 1 analog protein TuEDS1 with the Arabidopsis ENHANCED DISEASE SUSCHEPALITY 1 and ENHANCED DISEASE SUSCHEPALITY 1 analog protein in other plants.
Fig. 2: silencing uracratia wheat ENHANCED DISEASE SUSCEPTIBILITY 1 analog genesTuEDS1Resulting in urapidotian wheat PI428309 pairsPst CYR33Is significantly reduced.
(a) Barley leaf spot virus induced silencing in PI428309TuEDS1Post inoculationPstCYR33, phenotype observed 14 days post inoculation;
(b) Detection of samples before inoculation with RumexTuEDS1Expression level (P) indicates extreme significance of the difference<0.01, using t-test);
(c) Biological statistics of the rust bacteria (representing significant differences, P <0.05, with t-test) were performed 14 days after inoculation;
(d) And counting the number of the rust fungus spore piles in a unit area on the leaf blade. Significant differences, P <0.05, using t-test).
Fig. 3: BSMV:GFPand BSMV:TuEDS1inoculation of Rumex strigosus after infection of Ula diagram wheat PI428309Pst CYR33Is a histological observation of fungal growth.
(a) Uralensis wheat PI428309 is BSMV:GFPand BSMV:TuEDS1infection with rust bacteriaPstAfter CYR33, structural morphology changes after the wheat leaf blades grow 2 dpi, 3 dpi and 5 dpi are observed by using Wheat Germ Agglutinin (WGA) staining, and SSV (stomatal inferior sac), IH (infected hypha), HMC (haustorium master cell) and H (haustorium master);
(b) After inoculation of 2 dpi, BSMV was counted:GFPand BSMV:TuEDS1after treatment, the number of Hyphal Branches (HB), haustorium blasts (HMC) and haustorium numbers (H) of the rust;
(c) After inoculating 3 dpi of the rust, counting the length of the rust infection hypha (P < 0.01);
(d) After inoculation with 5 dpi, the hyphal invasion area was counted (P < 0.05).
Fig. 4: there is an interaction between TuEDS1 and the analog TuADR1 of arabidopsis ACTIVATED DISEASE RESISTANCE in urapida wheat.
(a) Through a bimolecular fluorescence complementation experiment, the interaction between TuEDS1 and TuADR1 is found;
(b) The interaction between TuEDS1 and TuADR1 is proved by a firefly enzyme complementation imaging experiment;
(c) The interaction between TuEDS1 and TuADR1 was demonstrated by co-immunoprecipitation experiments.
Fig. 5: in riceoseds1Overexpression in mutantsTuEDS1Resistance of the mutant to rice blast is enhanced.
（a）TuEDS1T 0 Identifying target proteins of transgenic plants;
b)TuEDS1-GFP-1、TuEDS1-GFP-5strain T 1 Identifying TuEDS1 protein of the transgenic plant;
(b) For wild ZH11 and mutantoseds1Rice plantTuEDS1T 1 Substituted over-expression strainTuEDS1-1#、TuEDS1-2#After spraying and inoculating the Pyricularia oryzae strain Guy for 11 weeks, observing the disease spots, and finding that the number of the over-expression plant disease spots is obviously smaller than that of riceoseds1A mutant;
(c) When the punched fungus-grafting Guy11 shows fusiform lesions on most ZH11 leaves after 8 days, the size of the lesions is observed, and the over-expression plants are found to be relative to riceoseds1The mutant lesions were significantly smaller;
(d) Extracting DNA of leaf blade at disease position after perforation and inoculation, detecting fungus biomass in leaf blade by fluorescent quantitative PCR, finding out over-expressionTuEDS1Rice of (2) relative to mutantoseds1The fungal biomass in the leaves was significantly less, the difference was significant (meaning there was a significant difference, P)<0.05, using t-test);
(e) Statistics of disease spot area of disease leaf after perforation and inoculation shows that over-expression is foundTuEDS1Rice plants of (2) relative to mutantsoseds1The area of lesions is significantly reduced (P represents significant difference, P)<0.05, using t-test).
Detailed Description
The following examples are intended to illustrate, but not limit the scope of the invention.
The invention discovers that the Ula diagram wheat ENHANCED DISEASE SUSCEPTIBILITY 1 analogue geneTuEDS1Expression level after inoculation with Leptosphaeria gracilisNo significant change occurs, but byTuEDS1Silencing and heterologous stable over-expression are performed, and the invention proves thatTuEDS1Plays an important role in wheat stripe rust resistance and rice blast resistance. For the convenience of the reader to understand the contents and objects of the present invention, specific technical implementation steps of the present invention will be described in detail by the following examples. The technical means described in the examples are conventional means familiar to those skilled in the art. The disease resistance or susceptibility in the following examples is referred to as stripe rust, rice blast; the stripe rust is wheat stripe rust and is prepared from wheat stripe rust bacteriaPuccinia striiformisf. sp.tritici，Pst) The strain CY33 of the wheat rust bacteria which is popular in northwest China is specifically caused by the fact that the wheat rust bacteria without special marks are hereinafterPstCYR 33). The rice blast is rice blast, and the rice blast bacteria is [ ], rice blast bacteria [ ], rice blastMagnaporthe oryzae) Infection of rice. In the following, if there is no specific identification, pyricularia oryzae will specifically be referred to as Pyricularia oryzae strain GUY11. The rice ZH11 is the middle flower 11 of the japonica rice variety,oseds1is ENHANCED DISEASE SUSCEPTIBILITY 1 analog gene of rice ZH11 by gene editingOsEDS1And the resulting mutant.
Example 1 Ula drawing wheat Material PI428309 ENHANCED DISEASE SUSCEPTIBILITY 1 analog GeneTuEDS1Is cloned from (A)
With reference to the genomic sequence published by urapidotian wheat G1812, it was found that a similar gene of the protein ENHANCED DISEASE susceptitiflity 1, which plays a key role in the arabidopsis ETI reaction, in urapidotian wheat G1812 was tug1812G0500002322.01 and the encoded protein was tug1812G0500002322.01.p01. Comparing the transcriptome database of Ula drawing wheat PI428309 to obtain the allele of TuG1812G0500002322.01 in Ula drawing wheat PI428309TuEDS1Is a sequence of mRNA of (A) in a host cell. To obtain the true presence in urapidotian wheat PI428309TuEDS1Designing specific primer for full-length geneTuEDS1-F andTuEDS1-R) obtained by reverse transcription of Promega kit and high-fidelity amplification of KOD-Fx DNA polymerase (TOYOBO)TuEDS1Is a gene sequence of (a). Obtaining by using online coding region prediction softwareTuEDS1The coding region sequence of the polypeptide is shown as SEQ ID NO.1, and the amino acid sequence of the coding protein is shown as SEQ ID NO. 2. Specific primersTuEDS1-F andTuEDS1the nucleotide sequence of R is as follows:
TuEDS1-F：5’- TGCGAGCTCAAGCATCAAAGGCTA-3’，
TuEDS1-R：5’- TGGCTACCTCATCAAGCGACG-3’。
inoculating wheat stripe rust after growth of Ural diagram wheat PI428309 to one leaf and one heart periodPstCY33, inoculated with rust on wheat at two time points, 0 h and 36 h, respectively, after inoculationPstSamples of uralensis wheat leaves after CY33 were taken and transcriptome sequenced. Based on the results of transcriptome analysis, vaccinationPstAfter the period of CY33, the process is completed,TuEDS1does not significantly change the transcript level of (a).
TuG1812G0500002322.01.P01 is identical to the amino acid sequence of TuEDS1, but has very low sequence similarity to Arabidopsis ENHANCED DISEASE SUSCEPTIBILITY 1. FIG. 1 shows the amino acid sequence comparison of Arabidopsis ENHANCED DISEASE SUSCEPTIBILITY 1 with TuEDS1 and other Gramineae ENHANCED DISEASESUSCEPTIBILITY 1-like proteins, and their phylogenetic relationship was analyzed.
Example 2TuEDS1Regulation in urapidoti wheat PI428309YrU1Mediated stripe rust resistance
RNA in leaves of Ula drawing wheat PI428309 is extracted and reversely transcribed into cDNA as a template, and a primer gamma-TuEDS1F and gamma-)TuEDS1-R amplification and recovery of PCR product. The plasmid with Barley stripemosaic virus RNA gamma (BSMV) is digested with restriction enzyme NheI. The recovered PCR product was ligated with linearized plasmid using Vazyme ClonExpress: II One Step Cloning Kit C112 homologous recombination kit to construct silencing vector BSMV:TuEDS1and vector BSMV incorporating GFP fragment was constructed in the same way:GFPas a control. The experimental procedure described in the reference [ Holzberg S, brosio P, gross C, pogue GP. 2002. Barley stripe mosaic virus-induced genesilencing in a monocot Plant J30:315-327 ], at Ula map wheat PI428309 ]In one-leaf one-heart period, infecting plant leaves with the constructed recombinant barley mosaic virus, and then detecting the fourth leaf of the infected plantTuEDS1Detecting the expression level and inoculating the wheat stripe rust CY33 to detect a resistance phenotype; the experiment selects urapidotian wheat material G1812 infected with the wheat stripe rust strain CY33 as a negative control and urapidotian wheat PI428309 (CK) which is not subjected to virus treatment as a positive control. Primer gamma-TuEDS1F and gamma-)TuEDS1The sequence of R is as follows:
γ-TuEDS1-F：5’-TTTTTTTTTTTTTTAGCTAGCTCCCCGTCTTGTGCAGTTCT-3’，
γ-TuEDS1-R：5’-GATTCTTCTTCCGTTGCTAGCCGCTCTTCCCGTCGCTG-3’。
experimental results show (fig. 2), relative to BSMV:GFPprocessing, in BSMV:TuEDS1after being infected with urapidotian wheat PI428309, the urapidotian wheat PI428309 can be certainly reducedTuEDS1Is a factor (B) of the expression level of (C). After inoculating wheat stripe rust, it was found that silencingTuEDS1Can obviously weaken the stripe rust resistance of the urapidoti wheat PI 428309. This experiment strongly demonstratesTuEDS1Involved in regulating and controlling urapidoti wheat PI428309YrU1Mediated stripe rust resistance response.
Inoculating Rumex tricuspidatum, sampling at 2 dpi, 3 dpi, and 5 dpi, staining the fungal structure in the leaf with wheat germ lectin (WGA), and counting germination condition, hypha length, and hypha infection area of Rumex tricuspidatum spores. The experimental results showed (fig. 3), BSMV 2 days after inoculation:TuEDS1treatment and control BSMV:GFPin contrast, the numbers of HB (hyphal branching), HMC (haustorium blasts) did not change significantly, but the number of H (haustorium) increased significantly; 3 days after inoculation, BSMV:TuEDS1treatment and control BSMV:GFPcompared with the prior art, the germination length of hypha is obviously increased; 5 days after inoculation, BSMV:TuEDS1treatment and control BSMV:GFPcompared with the prior art, the hypha infection area is obviously increased. The above-described results illustrate that,TuEDS1down-regulation of transcript levels favors the development of the stripe rust,TuEDS1plays an important role in the stripe rust resistance of urapidotian wheat PI 428309.
Example 3 interaction between TuEDS1 and the analog TuADR1 of ACTIVATED DISEASE RESISTANCE 1 in Ula diagram wheat PI428309
Studies have reported that ACTIVATED DISEASE RESISTANCE (ADR 1) and ENHANCED DISEASE subscore (EDS 1) in arabidopsis, in combination with other disease-associated proteins, form a complex of cascade defense signals that together regulate plant immunity in PTI and ETI. In urapidotia wheat PI428309, whether TuEDS1 and TuADR1 have an interaction relationship and whether they have a possibility of forming a disease-resistant complex or not, we have performed the following experiment.
First, verification was performed by a bimolecular fluorescence complementation experiment (BiFC). Will build up YFP C TuEDS1 and YFP N The TuADR1 fusion expression vector was transformed into Agrobacterium GV3101, and subsequently the Agrobacterium mixture containing both vectors was injected into tobacco leaves with a single vector and empty mixture co-injection as negative control. After normal cultivation in the greenhouse of 48h, the tobacco leaves after injection were taken and observed under a laser confocal microscope. As shown in fig. 4a, yellow fluorescence was observed in the epidermal cells of the tobacco leaf of the experimental group, while no yellow fluorescence was observed in the control group, indicating that TuEDS1 interacted with TuADR1 in the bimolecular fluorescence complementation experiment.
Second, interaction was verified using a firefly complementary imaging experiment (Luciferase complementation imaging assay). The constructed TuEDS1-nLuc and TuADR1-cLuc fusion expression vector was transformed into Agrobacterium GV3101, and then both Agrobacterium were co-injected into tobacco leaves, and TuEDS1-nLuc and YFP-cLuc, tuADR1-cLuc and YFP-nLuc were injected on the same tobacco leaves as negative controls. After 48 to h is injected, tobacco leaves are taken, a layer of luciferase substrate is uniformly coated on the back surfaces of the leaves, and after 5 minutes of dark treatment, the fluorescence intensity is observed by a plant living body imager. As shown in FIG. 4b, the regions where TuEDS1-nLuc and TuADR1-cLuc were co-injected were able to detect fluorescent signals, whereas the negative control region did not detect fluorescent signals, indicating that TuEDS1 and TuADR1 also interacted in luciferase complementary imaging experiments.
Finally, the interaction was verified by Co-immunoprecipitation (Co-IP). The fusion expression vectors TuADR1-GFP and TuEDS1-HA are respectively transformed into agrobacterium GV3101, and the agrobacterium tumefaciens GV3101 and TuEDS1-HA are mixed and injected into tobacco leaves, and the TuEDS1-HA and empty GFP are mixed and injected to serve as negative controls. Total proteins were extracted 48h after injection, incubated with GFP beads at 4deg.C for 2 h to bind, and washing buffer was used to remove non-specifically bound proteins, followed by western blot detection. As shown in fig. 4c, it was found that bands of TuEDS1-HA could be detected with anti-HA antibodies in the experimental group after GFP beads incubation and not in the negative control, indicating interaction of TuEDS1 and TuADR1 in the Co-IP experiment.
Example 4 heterologous overexpressionTuEDS1Can enhance rice blast resistance of rice
To exploreTuEDS1Whether or not the rice can be supplementedoseds1Resistance to Pyricularia oryzae lost by the mutant we regulated the 35S promoterTuEDS1(GFP tag fused to C-terminus) into Rice mutantoseds1And exploring the role of the pestilence in rice disease resistance by means of perforation and spray inoculation.
The sequence similarity of both TuEDS1 and OsEDS1 was 80.03% compared to the amino acid sequence (see fig. 1 a), and it was speculated that EDS1 evolved relatively conservatively in rice and wheat.
In the pair T 0 After TuEDS1-GFP protein identification of the plants over-expressed in the generation (see FIG. 5 a), we selectedTuEDS1 -1 # AndTuEDS1 -2 # the two lines were subjected to subsequent rice inoculation experiments (see FIGS. 5b, c). Flower 11 (ZH 11) is respectively centered by the experimental methods of spraying and punching inoculation,oseds1mutant and overexpressing strainsTuEDS1 -1 # 、TuEDS1 -2 # （T 1 Generation), the positive plants in the strain are inoculated with rice blast fungus Guy11, the disease leaf blades are taken 7 days after inoculation to observe the phenotype, and the biological statistics and the disease leaf area statistical analysis are carried out on the sampling leaf blades, and the result is shown in a graph b, c, d, e of fig. 5. After the rice blast fungus is inoculated, the rice blast fungus,TuEDS1over-expressed plants compared to the original mutantsoseds1The phenotype with more disease resistance is shown, and the biomass statistics and the disease spot area are obviously lower than those of mutantsoseds1(after t-test, P<0.05 And the disease resistance phenotype of ZH11 is basically oneSo that.
Description of the above Experimental resultsTuEDS1AndOsEDS1has a certain similarity in not only amino acid sequence, but alsoTuEDS1Can be used in paddy riceoseds1The mutant plays a disease-resistant function to compensateoseds1The rice blast resistance caused by the deficiency of OsEDS1 is reduced.
The foregoing has been described in some detail by way of illustration of the general principles and specific embodiments of the invention. Certain modifications and derivatives of this invention will be readily apparent to those skilled in the art, and it is therefore intended to be within the scope of this invention as claimed herein without departing from the essential characteristics thereof.
Claims (5)
1. Ula diagram wheat ENHANCED DISEASE SUSCEPTIBILITY 1 analog geneTuEDS1The method is characterized in that: said Ula diagram wheat ENHANCED DISEASE SUSCEPTIBILITY 1 analog geneTuEDS1The nucleotide sequence of (2) is shown as SEQ ID NO. 1.
2. A method for preparing a plant-derived substance from the plant-derived substance of the Ula-leaf wheat ENHANCED DISEASE SUSCEPTIBILITY 1 as defined in claim 1TuEDS1The encoded protein is characterized in that: the amino acid sequence of the protein is shown as SEQ ID NO. 2.
3. A composition comprising urarel as defined in claim 1 wheat ENHANCED DISEASE SUSCEPTIBILITY 1 analog geneTuEDS1Is a recombinant vector of (a).
4. Overexpression of the Ula drawing wheat ENHANCED DISEASE SUSCEPTIBILITY 1 analog Gene according to claim 1TuEDS1In improving riceoseds1Use of mutant rice blast resistance.
5. Silencing the Urapine wheat ENHANCED DISEASE SUSCEPTIBILITY 1 analog Gene of claim 1TuEDS1Use of YrU of uracrat wheat PI428309 for attenuating resistance to wheat stripe rust mediated by YrU.
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