Histone demethylase gene HOVUSG0811000 related to powdery mildew resistance and application of histone demethylase gene HOVUSG0811000

Abstract

The invention provides a histone demethylase gene HOVUSG0811000 related to powdery mildew resistance and an application of the histone demethylase gene HOVUSG0811000 related to powdery mildew resistance. The nucleotide sequence of the HOVUSG0811000 is as shown in SEQ ID NO. 1 (sequence identifier number 1). It is found for the first time that HOVUSG0811000 improves the resistance of highland barley to powdery mildew by regulating phenol amine metabolism, and the HOVUSG0811000 has a good application prospect in construction of powdery mildew resistant plant varieties.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of genetic engineering, and particularly relates to a HOVUSG0811000 gene and its use for resisting powdery mildew. BACKGROUND

[0002] Hordeum vulgare L.var.nudum, belonging to the family of Poaceae and the genus of Hordeum, is also known as naked barley, and is an important raw material for beer, medicine and health products, and is famous for its rich nutritional value and unique taste and flavor. However, naked barley is often attacked by powdery mildew. In recent years, due to the influence of global warming and other factors, the degree of damage caused by powdery mildew has shown a clear trend of aggravation, and has become a major disease that limits the high yield and high quality of naked barley. Therefore, it is of great significance to analyze the molecular mechanism of naked barley resistance to powdery mildew for carrying out disease-resistant molecular breeding research, and provides valuable gene resources for breeding powdery mildew-resistant varieties. SUMMARY

[0003] In the previous work, it was found that after the inoculation of naked barley with powdery mildew, the histone H3K27me3 modification level of the key gene HvTBT1 in the phenylamine synthesis pathway was significantly decreased. Correlation analysis showed that a demethylation modification protein HOVUSG0811000 and the key compounds and synthesis genes of the aromatic phenylamine metabolic pathway had extremely significant co-expression. This suggests that during the process of resisting powdery stress, HOVUSG0811000 may participate in the regulation of the resistance of naked barley to powdery mildew by directly regulating the gene expression of the aromatic phenylamine synthesis pathway, which provides an important clue for exploring the accumulation process of aromatic phenylamine and its function in resisting powdery mildew.

[0004] The present application provides a gene, and the nucleotide sequence of the gene is shown in SEQ ID NO. 1.

[0005] The present application also provides a recombinant vector, which comprises the nucleotide sequence shown in SEQ ID NO. 1.

[0006] Further, the recombinant vector is a recombinant PC186 vector.

[0007] The present application also provides a recombinant bacterium comprising the recombinant vector.

[0008] Further, the recombinant bacterium is a recombinant Agrobacterium.

[0009] The present application provides a method for constructing a plant resistant to powdery mildew, and the gene fragment shown in SEQ ID NO. 1 is introduced into the plant.

[0010] Further, the method for transforming the plant is one of Agrobacterium method, gene gun method, electroporation method, PEG-mediated method, liposome method and calcium phosphate-DNA co-precipitation method.

[0011] Further, the plant is tobacco or barley.

[0012] The application also provides the use of the above-mentioned gene, recombinant vector and / or recombinant bacteria in preparing a plant variety resistant to powdery mildew.

[0013] Further, the plant is tobacco or barley.

[0014] The present application has the following beneficial effects: the present application discovers a new gene HOVUSG0811000 in highland barley, which positively regulates the resistance of highland barley to powdery mildew. In order to verify that HOVUSG0811000 can positively regulate the resistance of highland barley to powdery mildew, the present application constructs three different transgenic strains of HOVUSG0811000: HOVUSG0811000 overexpression strain (named HOVUSG0811000-OE), HOVUSG0811000 gene editing strain (named HOVUSG0811000-Crisp#), and HOVUSG0811000 gene silencing VIGS strain (named HOVUSG0811000-VIGS). Further analysis found that, compared with WT, the histone H3K27me3 level of the key gene HvTBT1 in the phenylamine synthesis pathway in the HOVUSG0811000 gene overexpression strain was significantly reduced, while it was significantly increased in the HOVUSG0811000 gene editing strain and the gene silencing VIGS strain; at the same time, the expression of HvTBT1 was the highest in the high expression strain and the lowest in the silencing strain. After inoculation with powdery mildew Bgh, the HOVUSG0811000 gene editing strain and the HOVUSG0811000 gene silencing VIGS strain were highly susceptible to powdery mildew, and compared with the wild type, a large number of visible conidia were produced on the leaves, while the resistance of the HOVUSG0811000 gene overexpression strain to powdery mildew was significantly enhanced. At the same time, the microcolony index (MI) on the leaves of each strain was calculated to evaluate the resistance of different transgenic strains to powdery mildew. The results showed that the MI of the HOVUSG0811000 gene editing strain and the HOVUSG0811000 gene silencing VIGS strain was 31.2% and 29.1% respectively, which was significantly higher than that of the wild type strain (MI was 16.7%). While the conidial colonization on the leaves of the HOVUSG0811000 gene overexpression strain was significantly inhibited, and the MI was only 8.6%. In addition, after NAB and NBT staining of the leaves of different transgenic strains at 2dpi, the DAB and DBT staining in the HOVUSG0811000 gene overexpression strain was significantly deeper than that of the wild type, while the staining degree of the HOVUSG0811000 gene editing strain and the HOVUSG0811000 gene silencing VIGS strain was the shallowest, indicating that H2O2 and superoxide anion free radicals (O 2- ·) accumulated significantly in the HOVUSG0811000 overexpression strain, suggesting that the related defense response was activated. In summary, the related experimental results of the overexpression strain, the gene editing strain and the gene silencing VIGS strain strongly suggest that HOVUSG0811000 can positively regulate the resistance of highland barley to powdery mildew.

[0015] Obviously, according to the above content of the present application, according to the ordinary technical knowledge and common practice in the art, other various forms of modifications, replacements or changes can be made without departing from the above basic technical idea of the present application.

[0016] The above content of the present application will be further explained in detail by the following specific embodiments in the form of examples. However, it should not be understood that the above subject matter of the present application is limited to the following examples. Any technology realized based on the above content of the present application belongs to the scope of the present application. BRIEF DESCRIPTION OF DRAWINGS

[0017] Figure 1 .HOVUSG0811000 overexpression strain amplification (left) and qRT-PCR identification (right) results.

[0018] Figure 2 .HOVUSG0811000 gene editing strain identification results.

[0019] Figure 3 .HOVUSG0811000 gene silencing VIGS strain identification results.

[0020] Figure 4 .HOVUSG0811000 different transgenic strain powdery mildew resistance identification results. A, 8dpi, the growth of leaf molecular spores of HOVUSG0811000 editing strain, VIGS strain, overexpression strain and wild type strain (2-week-old seedlings start to do infection treatment) was observed, and the microcolony formation index (MI) on the leaf was counted (**P<0.01; Student's t-test). The scale is 3mm. B, 8dpi, the growth state of powdery mildew on the leaves of different strains was observed by trypan blue staining. C, 2 days after inoculation of powdery mildew, the leaves of different strains were stained with 3,3'-diaminobenzidine (DAB) to observe the accumulation of H2O2. D, 2dpi, the leaves of different strains were stained with nitroblue tetrazolium (NBT) to detect the accumulation of superoxide anion free radicals (O 2 -·) accumulation. The scale is 50μm.

[0021] Figure 5Effect of HOVUSG0811000 on the key synthetic genes HvTBT1 and HvTBT2 of phenylamine pathway. A-B, RT-qPCR was used to determine the expression of HvTBT1 and HvTBT2 in different transgenic lines of HOVUSG0811000 and WT line after inoculation with E.graminis. C, Schematic diagram of gene structure for ChIP assay. The primer sets (P1, P2) used in ChIP assay are shown relative to the transcription start sites (TSSs) and ATG positions. D, The enrichment of HOVUSG0811000 binding on HvTBT1 and HvTBT2 sites was determined by ChIP assay. P1 and P2 represent TSS and coding region, respectively. TUA4 (a-tubulin) is a negative control for HOVUSG0811000 and histone methylation enrichment. E, The level of histone H3K27me3 modification on HvTBT1 in different transgenic lines after inoculation with E.graminis was determined by ChIP assay. F-G, Analysis of HOVUSG0811000 regulation on HvTBT1 transcriptional activity. DETAILED DESCRIPTION

[0022] The raw materials and equipment used in the present application are known products, which can be obtained by purchasing commercially available products.

[0023] The wild type highland barley used in the specific embodiment of the present application is highland barley receptor C1, which shows a susceptible type to powdery mildew.

[0024] Example 1: Construction of HOVUSG0811000 different transgenic lines

[0025] 1) HOVUSG0811000 screening of candidate genes

[0026] Applicants previously identified a MeJA-responsive histone demethylase HOVUSG0811000 based on the transcriptome, proteome and extensive targeted metabolome databases of highly resistant (K69) and susceptible highland barley varieties (G72) before and after E.graminis infection; metabolites and synthetic genes involved in the aromatic phenylamine pathway to resist E.graminis infection have a highly significant correlation with HOVUSG0811000.

[0027] The sequence of HOVUSG0811000 gene is (SEQ ID NO. 1):

[0028] ATGAAAAGGGAGAACATGGCTGTGGAGGTAACAACAAACACCAAGAGGCCCAGTCTAGCTGAGGGGCATAGTACCAAAGCATTAAAAGGTGTAGATGATGTGTCCCACCCCTCACCAACTACAGTGGTTCCCCGTGTTAGCTTTAGGATTGCCGATAGAGCAAACAAGGTGAAAACAGAGATGACGGAAGAGGATGATGACCCTGCTTGCCGTCCGAAGCCAAAGGCCACTTCCCATTCGCGAACAGGGCCACCAAAGAAGATTGAGGCTGAAACAAAGAAGCAGATGAAGACGAGCAAAGGAGACAGCAAGATGATGGCTCCAGCTGCTCCAAAGGATGATGAGGAGCACCCATCTGCTGCCAAGGGGGGGGGCCTTCTGTGGGCCCTGCCACTAAGCAGGAGTTGTCTGGGCGTTAGAACCGATGCCGAGGCCGTTCGTGCAGATGGTCGGCGGGAGGCCGCCTGCGCGAGCGCCCTTGCTGAGAGGGAGGCACTCACTAACCGTTGTGAACAAGTGGAGGCCGGCTTCAGGGTCCTTCGGGACGCCTCCAAGGCCCGCGAACCGAAGCTCGCCAACTGTGACTCTGAGCTAGCGAGGGTCGCGGTGGAGCAGATCACGGAGCATGGGCGCATGGAGGCGCTTCAGCGCGAGGTGGTTGAGGCTTGGGACGCCCACGAGAAGTAG.

[0029] 2) HOVUSG0811000 was overexpressed in the barley receptor C1, and the HOVUSG0811000 overexpression strain (named HOVUSG0811000-OE) was constructed

[0030] First, the overexpression vector PC186-HOVUSG0811000 was successfully constructed. Next, Agrobacterium tumefaciens suspension was prepared. Glycerol-containing suspension was streaked onto MG / L plates and incubated in the dark at 28°C for two days. Colonies were then picked from the plates and inoculated into MG / L liquid medium, incubated overnight at 28°C and 220 rpm. Next, barley recipient material C1 seeds (2-2.5 mm in size) were selected, disinfected sequentially with alcohol and sodium hypochlorite, rinsed with sterile water, and the embryos were peeled off. The embryos were then immersed in an Agrobacterium tumefaciens suspension for 5 minutes. If co-transformation was to be performed, equal volumes of the two bacterial suspensions were mixed before immersion. The embryos were then placed on co-culture medium and incubated in the dark at 23°C for 2 days. After co-culture, the hypocotyl of the embryos was removed, and the remaining embryos were placed on a culture medium and incubated in the dark at 25°C for 5 days. Two rounds of selection culture were then performed. First, the embryos were transferred to a selection medium and incubated in the dark at 25°C for 2 weeks, then transferred to a new selection medium and incubated under the same conditions for 2 weeks. The selected callus tissue was transferred to a transitional medium, covered with paper towels, and cultured for 2 weeks at 25°C under 16 hours of light and 8 hours of darkness. Afterward, the callus tissue was transferred to a differentiation medium and cultured for 2 weeks under the same light conditions. Once regenerated shoots differentiated, they were transferred to a rooting medium and cultured for another 2 weeks. Finally, the rooted regenerated plants were transplanted into planting tubes and cultured in a greenhouse. The overexpression of HOVUSG0811000 was confirmed by gene amplification and qRT-PCR. Figure 1 ).

[0031] The PC186 vector sequence is as follows (SEQ ID NO.2):

[0032] GTGCGGTTTCATGCTTGTTCCTCTTGGCGTTCATTCTCGGCGGCCGCCA

[0033] GGGCGTCGGCCTCGGTCAATGCGTCCTCACGGAAGGCACCGCGCCGCC

[0034] TGGCCTCGGTGGGCGTCACTTCCTCGCTGCGCTCAAGTGCGCGGTACA

[0035] GGGTCGAGCGATGCACGCCAAGCAGTGCAGCCGCCTCTTTCACGGTGC

[0036] GGCCTTCCTGGTCGATCAGCTCGCGGGCGTGCGCGATCTGTGCCGGGG

[0037] TGAGGGTAGGGCGGGGGCCAAACTTCACGCCTCGGGCCTTGGCGGCCT

[0038] CGCGCCCGCTCCGGGTGCGGTCGATGATTAGGGAACGCTCGAACTCGG

[0039] CAATGCCGGCGAACACGGTCAACACCATGCGGCCGGCCGGCGTGGTGG

[0040] TGTCGGCCCACGGCTCTGCCAGGCTACGCAGGCCCGCGCCGGCCTCCT

[0041] GGATGCGCTCGGCAATGTCCAGTAGGTCGCGGGTGCTGCGGGCCAGGC

[0042] GGTCTAGCCTGGTCACTGTCACAACGTCGCCAGGGCGTAGGTGGTCAA

[0043] GCATCCTGGCCAGCTCCGGGCGGTCGCGCCTGGTGCCGGTGATCTTCTC

[0044] GGAAAACAGCTTGGTGCAGCCGGCCGCGTGCAGTTCGGCCCGTTGGTT

[0045] GGTCAAGTCCTGGTCGTCGGTGCTGACGCGGGCATAGCCCAGCAGGCC

[0046] AGCGGCGGCGCTCTTGTTCATGGCGTAATGTCTCCGGTTCTAGTCGCAA

[0047] GTATTCTACTTTATGCGACTAAAACACGCGACAAGAAAACGCCAGGAA

[0048] AAGGGCAGGGCGGCAGCCTGTCGCGTAACTTAGGACTTGTGCGACATG

[0049] TCGTTTTCAGAAGACGGCTGCACTGAACGTCAGAAGCCGACTGCACTA

[0050] TGTGGTTGGCATGCACATACAAATGGACGAACGGATAAACCTTTTCACG

[0051] TGTGGTTGGCATGCACATACAAATGGACGAACGGATAAACCTTTTCACG

[0052] TGTGGTTGGCATGCACATACAAATGGACGAACGGATAAACCTTTTCACG

[0053] TGTGGTTGGCATGCACATACAAATGGACGAACGGATAAACCTTTTCACG

[0054] TGTGGTTGGCATGCACATACAAATGGACGAACGGATAAACCTTTTCACG

[0055] TGTGGTTGGCATGCACATACAAATGGACGAACGGATAAACCTTTTCACG

[0056] TGTGGTTGGCATGCACATACAAATGGACGAACGGATAAACCTTTTCACG

[0057] TGTGGTTGGCATGCACATACAAATGGACGAACGGATAAACCTTTTCACG

[0058] TGTGGTTGGCATGCACATACAAATGGACGAACGGATAAACCTTTTCACG

[0059] TGTGGTTGGCATGCACATACAAATGGACGAACGGATAAACCTTTTCACG

[0060] TGTGGTTGGCATGCACATACAAATGGACGAACGGATAAACCTTTTCACG

[0061] TGTGGTTGGCATGCACATACAAATGGACGAACGGATAAACCTTTTCACG

[0062] TGTGGTTGGCATGCACATACAAATGGACGAACGGATAAACCTTTTCACG

[0063] GAGTATTTTGACAACAGGACTCTACAGTTTTATCTTTTTAGTGTGCATGT

[0064] GTTCTCCTTTTTTTTTGCAAATAGCTTCACCTATATAATACTTCATCCATT

[0065] TTATTAGTACATCCATTTAGGGTTTAGGGTTAATGGTTTTTATAGACTAAT

[0066] TTTTTTAGTACATCTATTTTATTCTATTTTAGCCTCTAAATTAAGAAAACT

[0067] AAAACTCTATTTTAGTTTTTTTATTTAATAATTTAGATATAAAATAGAATA

[0068] AAATAAAGTGACTAAAAATTAAACAAATACCCTTTAAGAAATTAAAAAA

[0069] ACTAAGGAAACATTTTTCTTGTTTCGAGTAGATAATGCCAGCCTGTTAA

[0070] ACGCCGTCGACGAGTCTAACGGACACCAACCAGCGAACCAGCAGCGT

[0071] CGCGTCGGGCCAAGCGAAGCAGACGGCACGGCATCTCTGTCGCTGCCT

[0072] CTGGACCCCTCTCGAGAGTTCCGCTCCACCGTTGGACTTGCTCCGCTGT

[0073] CGGCATCCAGAAATTGCGTGGCGGAGCGGCAGACGTGAGCCGGCACG

[0074] GCAGGCGGCCTCCTCCTCCTCTCACGGCAcCGGCAGCTACGGGGGATTC

[0075] CTTTCCCACCGCTCCTTCGCTTTCCCTTCCTCGCCCGCCGTAATAAATAG

[0076] ACACCCCCTCCACACCCTCTTTCCCCAACCTCGTGTTGTTCGGAGCGCA

[0077] CACACACACAACCAGATCTCCCCCAAATCCACCCGTCGGCACCTCCGC

[0078] TTCAAGGTACGCCGCTCGTCCTCCCCCCCCCCCCCTCTCTACCTTCTCTA

[0079] GATCGGCGTTCCGGTCCATGGTTAGGGCCCGGTAGTTCTACTTCTGTTC

[0080] ATGTTTGTGTTAGATCCGTGTTTGTGTTAGATCCGTGCTGCTAGCGTTCG

[0081] TACACGGATGCGACCTGTACGTCAGACACGTTCTGATTGCTAACTTGCC

[0082] AGTGTTTCTCTTTGGGGAATCCTGGGATGGCTCTAGCCGTTCCGCAGAC

[0083] GGGATCGATTTCATGATTTTTTTTGTTTCGTTGCATAGGGTTTGGTTTGC

[0084] CCTTTTCCTTTATTTCAATATATGCCGTGCACTTGTTTGTCGGGTCATCTT

[0085] TTCATGCTTTTTTTTGTCTTGGTTGTGATGATGTGGTCTGGTTGGGCGGT

[0086] CGTTCTAGATCGGAGTAGAATTCTGTTTCAAACTACCTGGTGGATTTATT

[0087] AATTTTGGATCTGTATGTGTGTGCCATACATATTCATAGTTACGAATTGAA

[0088] GATGATGGATGGAAATATCGATCTAGGATAGGTATACATGTTGATGCGGG

[0089] TTTTACTGATGCATATACAGAGATGCTTTTTGTTCGCTTGGTTGTGATGA

[0090] TGTGGTGTGGTTGGGCGGTCGTTCATTCGTTCTAGATCGGAGTAGAATA

[0091] CTGTTTCAAACTACCTGGTGTATTTATTAATTTTGGAACTGTATGTGTGT

[0092] GTCATACATCTTCATAGTTACGAGTTTAAGATGGATGGAAATATCGATCT

[0093] AGGATAGGTATACATGTTGATGTGGGTTTTACTGATGCATATACATGATG

[0094] GCATATGCAGCATCTATTCATATGCTCTAACCTTGAGTACCTATCTATTAT

[0095] AATAAACAAGTATGTTTTATAATTATTTTGATCTTGATATACTTGGATGAT

[0096] GGCATATGCAGCAGCTATATGTGGATTTTTTTAGCCCTGCCTTCATACGC

[0097] TATTTATTTGCTTGGTACTGTTTCTTTTGTCGATGCTCACCCTGTTGTTTG

[0098] GTGTTACTTCTGCAGGTCGACTCTAGAGGATCTTAGATCACAAGTTTGT

[0099] ACAAAGTGGTGATCTAGAGGGAGCTCGAATTTCCCCGATCGTTCAAAC

[0100] ATTTGGCAATAAAGTTTCTTAAGATTGAATCCTGTTGCCGGTCTTGCGAT

[0101] GATTATCATATAATTTCTGTTGAATTACGTTAAGCATGTAATAATTAACAT

[0102] GTAATGCATGACGTTATTTATGAGATGGGTTTTTATGATTAGAGTCCCGC

[0103] AATTATACATTTAATACGCGATAGAAAACAAAATATAGCGCGCAAACTA

[0104] GGATAAATTATCGCGCGCGGTGTCATCTATGTTACTAGATCGGGAATTAG

[0105] CTTCATCAACGCAAGACATGGTAAGCTTGCATGCCTGCAGGTCCCCAGA

[0106] TTAGCCTTTTCAATTTCAGAAAGAATGCTAACCCACAGATGGTTAGAGA

[0107] GGCTTACGCAGCAGGTCTCATCAAGACGATCTACCCGAGCAATAATCTC

[0108] CAGGAAATCAAATACCTTCCCAAGAAGGTTAAAGATGCAGTCAAAAGA

[0109] TTCAGGACTAACTGCATCAAGAACACAGAGAAAGATATATTTCTCAAGA

[0110] TCAGAAGTACTATTCCAGTATGGACGATTCAAGGCTTGCTTCACAAACC

[0111] AAGGCAAGTAATAGAGATTGGAGTCTCTAAAAAGGTAGTTCCCACTGA

[0112] ATCAAAGGCCATGGAGTCAAAGATTCAAATAGAGGACCTAACAGAACT

[0113] CGCCGTAAAGACTGGCGAACAGTTCATACAGAGTCTCTTACGACTCAAT

[0114] GACAAGAAGAAAATCTTCGTCAACATGGTGGAGCACGACACACTTGTC

[0115] TACTCCAAAAATATCAAAGATACAGTCTCAGAAGACCAAAGGGCAATT

[0116] GAGACTTTTCAACAAAGGGTAATATCCGGAAACCTCCTCGGATTCCATT

[0117] GCCCAGCTATCTGTCACTTTATTGTGAAGATAGTGGAAAAGGAAGGTGG

[0118] CTCCTACAAATGCCATCATTGCGATAAAGGAAAGGCCATCGTTGAAGAT

[0119] GCCTCTGCCGACAGTGGTCCCAAAGATGGACCCCCACCCACGAGGAGC

[0120] ATCGTGGAAAAAGAAGACGTTCCAACCACGTCTTCAAAGCAAGTGGAT

[0121] TGATGTGATATCTCCACTGACGTAAGGGATGACGCACAATCCCACTATC

[0122] CTTCGCAAGACCCTTCCTCTATATAAGGAAGTTCATTTCATTTGGAGAG

[0123] AACACGGGGGACTCTAGAGGATCCCCGGGGATCTACCATGAGCCCAGA

[0124] ACGACGCCCGGCCGACATCCGCCGTGCCACCGAGGCGGACATGCCGGC

[0125] GGTCTGCACCATCGTCAACCACTACATCGAGACAAGCACGGTCAACTT

[0126] CCGTACCGAGCCGCAGGAACCGCAGGAGTGGACGGACGACCTCGTCC

[0127] GTCTGCGGGAGCGCTATCCCTGGCTCGTCGCCGAGGTGGACGGCGAGG

[0128] TCGCCGGCATCGCCTACGCGGGCCCCTGGAAGGCACGCAACGCCTACG

[0129] ACTGGACGGCCGAGTCGACCGTGTACGTCTCCCCCCGCCACCAGCGGA

[0130] CGGGACTGGGCTCCACGCTCTACACCCACCTGCTGAAGTCCCTGGAGG

[0131] CACAGGGCTTCAAGAGCGTGGTCGCTGTCATCGGGCTGCCCAACGACC

[0132] CGAGCGTGCGCATGCACGAGGCGCTCGGATATGCCCCCCGCGGCATGC

[0133] TGCGGGCGGCCGGCTTCAAGCACGGGAACTGGCATGACGTGGGTTTCT

[0134] GGCAGCTGGACTTCAGCCTGCCGGTACCGCCCCGTCCGGTCCTGCCCG

[0135] TCACCGAGATCTGATGACCCGAATTTCCCCGATCGTTCAAACATTTGGC

[0136] AATAAAGTTTCTTAAGATTGAATCCTGTTGCCGGTCTTGCGATGATTATC

[0137] ATATAATTTCTGTTGAATTACGTTAAGCATGTAATAATTAACATGTAATGC

[0138] ATGACGTTATTTATGAGATGGGTTTTTATGATTAGAGTCCCGCAATTATAC

[0139] ATTTAATACGCGATAGAAAACAAAATATAGCGCGCAAACTAGGATAAAT

[0140] TATCGCGCGCGGTGTCATCTATGTTACTAGATCGGGAATTCGTAATCATG

[0141] GTCATAGCTGTTTCCTGTGTGAAATTGTTATCCGCTCACAATTCCACACA

[0142] ACATACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAG

[0143] TGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTC

[0144] GGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGG

[0145] GAGAGGCGGTTTGCGTATTGGCTAGAGCAGCTTGCCAACATGGTGGAG

[0146] CACGACACTCTCGTCTACTCCAAGAATATCAAAGATACAGTCTCAGAAG

[0147] ACCAAAGGGCTATTGAGACTTTTCAACAAAGGGTAATATCGGGAAACC

[0148] TCCTCGGATTCCATTGCCCAGCTATCTGTCACTTCATCAAAAGGACAGT

[0149] AGAAAAGGAAGGTGGCACCTACAAATGCCATCATTGCGATAAAGGAAA

[0150] GGCTATCGTTCAAGATGCCTCTGCCGACAGTGGTCCCAAAGATGGACC

[0151] CCCACCCACGAGGAGCATCGTGGAAAAAGAAGACGTTCCAACCACGT

[0152] CTTCAAAGCAAGTGGATTGATGTGATAACATGGTGGAGCACGACACTC

[0153] TCGTCTACTCCAAGAATATCAAAGATACAGTCTCAGAAGACCAAAGGG

[0154] CTATTGAGACTTTTCAACAAAGGGTAATATCGGGAAACCTCCTCGGATT

[0155] CCATTGCCCAGCTATCTGTCACTTCATCAAAAGGACAGTAGAAAAGGA

[0156] AGGTGGCACCTACAAATGCCATCATTGCGATAAAGGAAAGGCTATCGTT

[0157] CAAGATGCCTCTGCCGACAGTGGTCCCAAAGATGGACCCCCACCCACG

[0158] AGGAGCATCGTGGAAAAAGAAGACGTTCCAACCACGTCTTCAAAGCA

[0159] AGTGGATTGATGTGATATCTCCACTGACGTAAGGGATGACGCACAATCC

[0160] CACTATCCTTCGCAAGACCTTCCTCTATATAAGGAAGTTCATTTCATTTG

[0161] GAGAGGACACGCTGAAATCACCAGTCTCTCTCTACAAATCTATCTCTCT

[0162] CGAGCTTTCGCAGATCCCGGGGGGCAATGAGATATGAAAAAGCCTGAA

[0163] CTCACCGCGACGTCTGTCGAGAAGTTTCTGATCGAAAAGTTCGACAGC

[0164] GTCTCCGACCTGATGCAGCTCTCGGAGGGCGAAGAATCTCGTGCTTTC

[0165] AGCTTCGATGTAGGAGGGCGTGGATATGTCCTGCGGGTAAATAGCTGCG

[0166] CCGATGGTTTCTACAAAGATCGTTATGTTTATCGGCACTTTGCATCGGCC

[0167] GCGCTCCCGATTCCGGAAGTGCTTGACATTGGGGAGTTTAGCGAGAGC

[0168] CTGACCTATTGCATCTCCCGCCGTGCACAGGGTGTCACGTTGCAAGACC

[0169] TGCCTGAAACCGAACTGCCCGCTGTTCTACAACCGGTCGCGGAGGCTA

[0170] TGGATGCGATCGCTGCGGCCGATCTTAGCCAGACGAGCGGGTTCGGCC

[0171] CATTCGGACCGCAAGGAATCGGTCAATACACTACATGGCGTGATTTCAT

[0172] ATGCGCGATTGCTGATCCCCATGTGTATCACTGGCAAACTGTGATGGAC

[0173] GACACCGTCAGTGCGTCCGTCGCGCAGGCTCTCGATGAGCTGATGCTTT

[0174] GGGCCGAGGACTGCCCCGAAGTCCGGCACCTCGTGCACGCGGATTTCG

[0175] GCTCCAACAATGTCCTGACGGACAATGGCCGCATAACAGCGGTCATTG

[0176] ACTGGAGCGAGGCGATGTTCGGGGATTCCCAATACGAGGTCGCCAACA

[0177] TCTTCTTCTGGAGGCCGTGGTTGGCTTGTATGGAGCAGCAGACGCGCTA

[0178] CTTCGAGCGGAGGCATCCGGAGCTTGCAGGATCGCCACGACTCCGGGC

[0179] GTATATGCTCCGCATTGGTCTTGACCAACTCTATCAGAGCTTGGTTGACG

[0180] GCAATTTCGATGATGCAGCTTGGGCGCAGGGTCGATGCGACGCAATCGT

[0181] CCGATCCGGAGCCGGGACTGTCGGGCGTACACAAATCGCCCGCAGAAG

[0182] CGCGGCCGTCTGGACCGATGGCTGTGTAGAAGTACTCGCCGATAGTGG

[0183] AAACCGACGCCCCAGCACTCGTCCGAGGGCAAAGAAATAGAGTAGAT

[0184] GCCGACCGGGATCTGTCGATCGACAAGCTCGAGTTTCTCCATAATAATG

[0185] TGTGAGTAGTTCCCAGATAAGGGAATTAGGGTTCCTATAGGGTTTCGCT

[0186] CATGTGTTGAGCATATAAGAAACCCTTAGTATGTATTTGTATTTGTAAAAT

[0187] ACTTCTATCAATAAAATTTCTAATTCCTAAAACCAAAATCCAGTACTAAA

[0188] ATCCAGATCCCCCGAATTAATTCGGCGTTAATTCAGTACATTAAAAACGT

[0189] CCGCAATGTGTTATTAAGTTGTCTAAGCGTCAATTTGTTTACACCACAAT

[0190] ATATCCTGCCACCAGCCAGCCAACAGCTCCCCGACCGGCAGCTCGGCA

[0191] CAAAATCACCACTCGATACAGGCAGCCCATCAGTCCGGGACGGCGTCA

[0192] GCGGGAGAGCCGTTGTAAGGCGGCAGACTTTGCTCATGTTACCGATGCT

[0193] ATTCGGAAGAACGGCAACTAAGCTGCCGGGTTTGAAACACGGATGATC

[0194] TCGCGGAGGGTAGCATGTTGATTGTAACGATGACAGAGCGTTGCTGCCT

[0195] GTGATCACCGCGGTTTCAAAATCGGCTCCGTCGATACTATGTTATACGCC

[0196] AACTTTGAAAACAACTTTGAAAAAGCTGTTTTCTGGTATTTAAGGTTTT

[0197] AGAATGCAAGGAACAGTGAATTGGAGTTCGTCTTGTTATAATTAGCTTC

[0198] TTGGGGTATCTTTAAATACTGTAGAAAAGAGGAAGGAAATAATAAATGG

[0199] CTAAAATGAGAATATCACCGGAATTGAAAAAACTGATCGAAAAATACC

[0200] GCTGCGTAAAAGATACGGAAGGAATGTCTCCTGCTAAGGTATATAAGCT

[0201] GGTGGGAGAAAATGAAAACCTATATTTAAAAATGACGGACAGCCGGTAT

[0202] AAAGGGACCACCTATGATGTGGAACGGGAAAAGGACATGATGCTATGG

[0203] CTGGAAGGAAAGCTGCCTGTTCCAAAGGTCCTGCACTTTGAACGGCAT

[0204] GATGGCTGGAGCAATCTGCTCATGAGTGAGGCCGATGGCGTCCTTTGCT

[0205] CGGAAGAGTATGAAGATGAACAAAGCCCTGAAAAGATTATCGAGCTGT

[0206] ATGCGGAGTGCATCAGGCTCTTTCACTCCATCGACATATCGGATTGTCCC

[0207] TATACGAATAGCTTAGACAGCCGCTTAGCCGAATTGGATTACTTACTGAA

[0208] TAACGATCTGGCCGATGTGGATTGCGAAAACTGGGAAGAAGACACTCC

[0209] ATTTAAAGATCCGCGCGAGCTGTATGATTTTTTAAAGACGGAAAAGCCC

[0210] GAAGAGGAACTTGTCTTTTCCCACGGCGACCTGGGAGACAGCAACATC

[0211] TTTGTGAAAGATGGCAAAGTAAGTGGCTTTATTGATCTTGGGAGAAGCG

[0212] GCAGGGCGGACAAGTGGTATGACATTGCCTTCTGCGTCCGGTCGATCA

[0213] GGGAGGATATCGGGGAAGAACAGTATGTCGAGCTATTTTTTGACTTACT

[0214] GGGGATCAAGCCTGATTGGGAGAAAATAAAATATTATATTTTACTGGATG

[0215] AATTGTTTTAGTACCTAGAATGCATGACCAAAATCCCTTAACGTGAGTTT

[0216] TCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCT

[0217] TGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAAC

[0218] CACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCT

[0219] TTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTC

[0220] CTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCAC

[0221] CGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAG

[0222] TGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCG

[0223] GATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCC

[0224] CAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGA

[0225] GCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGT

[0226] ATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTT

[0227] CCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACC

[0228] TCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCT

[0229] ATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGC

[0230] TGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGA

[0231] TAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGA

[0232] ACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCT

[0233] GATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATAT

[0234] GGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGTAT

[0235] ACACTCCGCTATCGCTACGTGACTGGGTCATGGCTGCGCCCCGACACCC

[0236] GCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATC

[0237] CGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAG

[0238] GTTTTCACCGTCATCACCGAAACGCGCGAGGCAGGGTGCCTTGATGTG

[0239] GGCGCCGGCGGTCGAGTGGCGACGGCGCGGCTTGTCCGCGCCCTGGTA

[0240] GATTGCCTGGCCGTAGGCCAGCCATTTTTGAGCGGCCAGCGGCCGCGAT

[0241] AGGCCGACGCGAAGCGGCGGGGCGTAGGGAGCGCAGCGACCGAAGG

[0242] GTAGGCGCTTTTTGCAGCTCTTCGGCTGTGCGCTGGCCAGACAGTTATG

[0243] CACAGGCCAGGCGGGTTTTAAGAGTTTTAATAAGTTTTAAAGAGTTTTA

[0244] GGCGGAAAAATCGCCTTTTTTCTCTTTTATATCAGTCACTTACATGTGTG

[0245] ACCGGTTCCCAATGTACGGCTTTGGGTTCCCAATGTACGGGTTCCGGTT

[0246] CCCAATGTACGGCTTTGGGTTCCCAATGTACGTGCTATCCACAGGAAAG

[0247] AGACCTTTTCGACCTTTTTCCCCTGCTAGGGCAATTTGCCCTAGCATCTG

[0248] CTCCGTACATTAGGAACCGGCGGATGCTTCGCCCTCGATCAGGTTGCGG

[0249] TAGCGCATGACTAGGATCGGGCCAGCCTGCCCCGCCTCCTCCTTCAAAT

[0250] CGTACTCCGGCAGGTCATTTGACCCGATCAGCTTGCGCACGGTGAAAC

[0251] AGAACTTCTTGAACTCTCCGGCGCTGCCACTGCGTTCGTAGATCGTCTT

[0252] GAACAACCATCTGGCTTCTGCCTTGCCTGCGGCGCGGCGTGCCAGGCG

[0253] GTAGAGAAAACGGCCGATGCCGGGATCGATCAAAAAGTAATCGGGGTG

[0254] AACCGTCAGCACGTCCGGGTTCTTGCCTTCTGTGATCTCGCGGTACATC

[0255] CAATCAGCTAGCTCGATCTCGATGTACTCCGGCCGCCCGGTTTCGCTCT

[0256] TTACGATCTTGTAGCGGCTAATCAAGGCTTCACCCTCGGATACCGTCAC

[0257] CAGGCGGCCGTTCTTGGCCTTCTTCGTACGCTGCATGGCAACGTGCGTG

[0258] GTTTCTACCAGGTCGTCTTTCTGCTTTCCGC

[0259] TTCGGTTAGATGGGAAACCGCCATCAGTACCAGGTCGTAATCCCACACA

[0260] TTCGGTTAGATGGGAAACCGCCATCAGTACCAGGTCGTAATCCCACACA

[0261] TTCGGTTAGATGGGAAACCGCCATCAGTACCAGGTCGTAATCCCACACA

[0262] TTCGGTTAGATGGGAAACCGCCATCAGTACCAGGTCGTAATCCCACACA

[0263] TTCGGTTAGATGGGAAACCGCCATCAGTACCAGGTCGTAATCCCACACA

[0264] TTCGGTTAGATGGGAAACCGCCATCAGTACCAGGTCGTAATCCCACACA

[0265] TTCGGTTAGATGGGAAACCGCCATCAGTACCAGGTCGTAATCCCACACA

[0266] TTCGGTTAGATGGGAAACCGCCATCAGTACCAGGTCGTAATCCCACACA

[0267] TTCGGTTAGATGGGAAACCGCCATCAGTACCAGGTCGTAATCCCACACA

[0268] TTCGGTTAGATGGGAAACCGCCATCAGTACCAGGTCGTAATCCCACACA

[0269] TTCGGTTAGATGGGAAACCGCCATCAGTACCAGGTCGTAATCCCACACA

[0270] TTCGGTTAGATGGGAAACCGCCATCAGTACCAGGTCGTAATCCCACACA

[0271] ​​​​​​​​​​​​​CAGGGCCGGCAGACAACCCAGCCGCTTACGCCTGGCCAACCGCCCGTT

[0272] CCTCCACACATGGGGCATTCCACGGCGTCGGTGCCTGGTTGTTCTTGAT

[0273] TTTCCATGCCGCCTCCTTTAGCCGCTAAAATTCATCTACTCATTTATTCAT

[0274] TTGCTCATTTACTCTGGTAGCTGCGCGATGTATTCAGATAGCAGCTCGGT

[0275] AATGGTCTTGCCTTGGCGTACCGCGTACATCTTCAGCTTGGTGTGATCC

[0276] TCCGCCGGCAACTGAAAGTTGACCCGCTTCATGGCTGGCGTGTCTGCC

[0277] AGGCTGGCCAACGTTGCAGCCTTGCTGCTGCGTGCGCTCGGACGGCCG

[0278] GCACTTAGCGTGTTTGTGCTTTTGCTCATTTTCTCTTTACCTCATTAACTC

[0279] AAATGAGTTTTGATTTAATTTCAGCGGCCAGCGCCTGGACCTCGCGGGC

[0280] AGCGTCGCCCTCGGGTTCTGATTCAAGAACGGTTGTGCCGGCGGCGGC

[0281] AGTGCCTGGGTAGCTCACGCGCTGCGTGATACGGGACTCAAGAATGGG

[0282] CAGCTCGTACCCGGCCAGCGCCTCGGCAACCTCACCGCCGATGCGCGT

[0283] GCCTTTGATCGCCCGCGACACGACAAAGGCCGCTTGTAGCCTTCCATCC

[0284] GTGACCTCAATGCGCTGCTTAACCAGCTCCACCAGGTCGGCGGTGGCC

[0285] CATATGTCGTAAGGGCTTGGCTGCACCGGAATCAGCACGAAGTCGGCT

[0286] GCCTTGATCGCGGACACAGCCAAGTCCGCCGCCTGGGGCGCTCCGTCG

[0287] ATCACTACGAAGTCGCGCCGGCCGATGGCCTTCACGTCGCGGTCAATC

[0288] GTCGGGCGGTCGATGCCGACAACGGTTAGCGGTTGATCTTCCCGCACG

[0289] GCCGCCCAATCGCGGGCACTGCCCTGGGGATCGGAATCGACTAACAGA

[0290] ACATCGGCCCCGGCGAGTTGCAGGGCGCGGGCTAGATGGGTTGCGATG

[0291] GTCGTCTTGCCTGACCCGCCTTTCTGGTTAAGTACAGCGATAACCTTCA

[0292] TGCGTTCCCCTTGCGTATTTGTTTATTTACTCATCGCATCATATACGCAGC

[0293] GACCGCATGACGCAAGCTGTTTTACTCAAATACACATCACCTTTTTAGA

[0294] CGGCGGCGCTCGGTTTCTTCAGCGGCCAAGCTGGCCGGCCAGGCCGCC

[0295] AGCTTGGCATCAGACAAACCGGCCAGGATTTCATGCAGCCGCACGGTT

[0296] GAGACGTGCGCGGGCGGCTCGAACACGTACCCGGCCGCGATCATCTCC

[0297] GCCTCGATCTCTTCGGTAATGAAAAACGGTTCGTCCTGGCCGTCCTG

[0298] 3) Gene editing of HOVUSG0811000 was performed on the barley receptor C1 to construct the HOVUSG0811000 gene-edited line (named HOVUSG0811000-Crisp#).

[0299] First, dual-guide RNAs (sgRNAs) targeting the gene need to be designed and synthesized. Target 1: GTAGGTTAGAACCGATGCCGAGG; Target 2: GTCCCGAAGCACCCTGAAGCCGG; then, the 19 nucleotides N in the forward and reverse primers are replaced with the target sequence and its reverse complementary sequence. Next, PCR is performed based on the PCR fragment sequence, using specific reaction mixtures and cycling conditions to construct the recombinant vector. Then, a Golden Gate reaction is set up to ligate the purified PCR fragment and vector under specific conditions. After ligation, the reaction mixture is transformed into competent *E. coli* cells, and positive clones are screened on LB agar plates containing kanamycin and induced to differentiate into complete plants. Finally, the edited plants are identified by PCR and sequencing, confirming that HAVUSG0811000 has been edited. Figure 2 ).

[0300] 4) VIGS was performed on HOVUSG0811000 in the barley receptor C1 to construct the HOVUSG0811000 gene-silenced VIGS line (named HOVUSG0811000-VIGS).

[0301] Virus-Induced Gene Silencing (VIGS) system was applied. In this study, the function of HOVUSG0811000 in C1 was identified by using Barley Stripe Mosaic Virus (BSMV)-mediated VIGS technology. First, HOVUSG0811000 was amplified and inserted into the enzyme-digested BSMV-γ to construct the recombinant virus vector BSMV:HOVUSG0811000. The efficiency of virus infection and gene silencing was verified by observing the leaf bleaching phenomenon and using BSMV-mediated tomato carotenoid cleavage dioxygenase (TaPDS) as a control. Inoculation and quantitative analysis were performed according to the previous method. Six C1 seedlings at the three-leaf stage were inoculated with BSMV:HOVUSG0811000 and BSMV blank control. Real-time PCR was used to analyze the expression level of the target gene in the newly developed leaves after BSMV:HOVUSG0811000 infection, and to determine the silencing efficiency of HOVUSG0811000 gene Figure 3

[0302] Example 2. HOVUSG0811000 positively regulates the resistance of C1 to powdery mildew

[0303] Example 1. HOVUSG0811000-Crisp# and HOVUSG0811000-VIGS strains were highly susceptible to powdery mildew, and a large number of visible conidia were produced on the leaves compared with the wild type. However, the resistance of HOVUSG0811000-OE strain to powdery mildew was significantly enhanced. In addition, the microcolony index (MI) of each strain was calculated to evaluate the resistance of different transgenic strains to powdery mildew. The MI of HOVUSG0811000-Crisp# and HOVUSG0811000-VIGS was 31.2% and 29.1%, respectively, which was significantly higher than that of the wild type strain (16.7%). However, the colonization of conidia on the leaves of HOVUSG0811000-OE strain was significantly inhibited, with an MI of only 8.6% (Figs. 2A and 2B). Figure 4 Further, the leaves of different transgenic strains at 2 dpi were stained with NAB and NBT, and the DAB and DBT staining in HOVUSG0811000-OE strain was significantly deeper than that in the wild type, while the staining in HOVUSG0811000-Crisp# and HOVUSG0811000-VIGS strains was the shallowest.​Figure 4 C and 4D), indicating that H2O2 and superoxide anion free radicals (O 2-· ) significantly accumulated in HOVUSG0811000 overexpression lines, suggesting that the related defense responses were activated. In summary, the relevant experimental results of overexpression lines, knockout lines and VIGS lines strongly suggest that HOVUSG0811000 can positively regulate the resistance of highland barley to powdery mildew.

[0304] Example 3. It was determined that HOVUSG0811000 regulates the accumulation of phenolic amine substances

[0305] The expression changes of HvTBTs genes in HOVUSG0811000 transgenic lines and WT lines after inoculation with powdery mildew for different times were determined by RT-qPCR experiments Figure 5 A and 5B). The results showed that the expression of HvTBT1 was significantly up-regulated in HOVUSG0811000-OE lines and significantly down-regulated in HOVUSG0811000-Crisp# lines after inoculation with powdery mildew compared with WT lines Figure 5 A). The expression of HvTBT2 was only significantly increased in the early stage of inoculation with powdery mildew in HOVUSG0811000-OE Figure 5 B). Further analysis of the enrichment of HOVUSG0811000 at the transcription start site (TSS) and coding region of HvTBT1 and HvTBT2 showed that HOVUSG0811000 was significantly enriched near the TSS region of HvTBT1 after Bgh inoculation, indicating that HOVUSG0811000 may specifically bind to the TSS region of HvTBT1. In addition, no significant enrichment of HOVUSG0811000 was detected at the TSS and coding region of HvTBT2 before and after Bgh inoculation, indicating that HvTBT2 is not a target gene of HOVUSG0811000 Figure 5 C and D). At the same time, the results of chromatin immunoprecipitation (ChIP) technology also showed that the level of histone H3K27me3 on the HvTBT1 gene was reduced in HOVUSG0811000-OE lines compared with WT, but significantly increased in HOVUSG0811000-Crisp# lines, especially after inoculation with powdery mildew Figure 5 E). The results of Dual luciferase assay also showed that HOVUSG0811000 can significantly activate the expression of HvTBT1 Figure 5 F and 5G). Based on these findings, it is suggested that HvTBT1 may act as a target gene directly regulated by HOVUSG0811000 to participate in the regulation of resistance of highland barley to powdery mildew.

[0306] In conclusion, the application finds a new gene HOVUSG0811000 in highland barley, the gene improves the resistance of highland barley to powdery mildew by regulating phenolic amine metabolism. The new gene, the recombinant carrier, the recombinant bacteria and the transgenic plant provided by the application have good application prospect.

Claims

1. A gene, characterized by: Its nucleotide sequence is shown in SEQ ID NO.

1.

2. A recombinant vector, characterized in that: It includes the nucleotide sequence shown in SEQ ID NO.

1.

3. The recombinant vector according to claim 2, characterized in that: The recombinant vector is a recombinant PC186 vector.

4. A recombinant bacterium, characterized in that: It comprises the recombinant vector as described in claim 2.

5. The recombinant bacteria according to claim 4, characterized in that: The recombinant bacteria is recombinant Agrobacterium.

6. A method for constructing a plant resistant to powdery mildew, characterized in that: The gene described in claim 1 can be transferred into a plant.

7. The construction method as described in claim 6, characterized in that: The method for transferring the substance into the plant is one of the following: Agrobacterium-mediated transformation, gene gun method, electroporation, PEG-mediated transformation, liposome method, and calcium phosphate-DNA coprecipitation method.

8. The construction method as described in claim 6 or 7, characterized in that, The plant in question is either tobacco or barley.

9. Use of the gene of claim 1, the recombinant vector of claim 2 or 3, and / or the recombinant bacteria of claim 4 or 5 in the preparation of plant varieties resistant to powdery mildew.

10. The use as described in claim 9, characterized in that, The plant in question is either tobacco or barley.

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