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Wheat powdery mildew resistance gene and application thereof

A powdery mildew and genetic technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of single disease-resistant germplasm resources, affecting the production potential of new varieties and genetic diversity, etc.

Inactive Publication Date: 2019-12-31
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, in the breeding practice of major crops such as wheat, the available disease-resistant germplasm resources are still too single, which affects the yield potential and genetic diversity of new varieties

Method used

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  • Wheat powdery mildew resistance gene and application thereof
  • Wheat powdery mildew resistance gene and application thereof
  • Wheat powdery mildew resistance gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1, the acquisition of TaJAZ1 protein and its coding gene

[0044] Total RNA of common wheat cultivar KN199 was extracted and reverse transcribed into cDNA. After a large number of sequence analysis, expression analysis and functional verification, a DNA coding sequence was found from the cDNA, as shown in sequence 2 of the sequence listing, and its encoded protein was shown in sequence 1 of the sequence listing.

[0045] The protein shown in Sequence 1 of the Sequence Listing is named TaJAZ1 protein. The gene encoding TaJAZ1 protein is named TaJAZ1 gene, and its open reading frame is shown in sequence 2 of the sequence listing.

Embodiment 2

[0046] Example 2, TaJAZ1 protein subcellular localization

[0047] 1. Replace the fragment between the two EcoRV restriction sites of the pQBV3 vector (also known as the entry vector) with the DNA molecule shown in sequence 2 of the sequence table to obtain a recombinant vector. Cloning the target fragment (the DNA molecule shown in Sequence 2 of the Sequence Listing) from the recombinant vector to the target vector pGWB5 by "gateway cloning technology" (gateway cloning technology, Invitrogen) to obtain the expression vector 35S:TaJAZ1-GFP (sequencing verification) .

[0048] 2. Introduce the expression vector 35S:TaJAZ1-GFP obtained in step 1 into Agrobacterium strain GV3101 (refer to literature: Liu, J., Zhang, T., Jia, J. and Sun, J. The Wheat Mediator Subunit TaMED25 Interactswith the Transcription Factor TaEIL1to Negatively Regulate Disease Resistance against Powdery Mildew.Plant physiology, 2016, 170, 1799-1816.), to obtain recombinant bacteria.

[0049] 3. Take the re...

Embodiment 3

[0051] Embodiment 3, TaJAZ1 gene expression pattern analysis

[0052] 1. The wheat cultivar KN199 was hydrocultured in a light incubator for one week, treated with 10 mM methyl jasmonate, and samples were taken at different time points (0h, 1h, 12h and 24h) after the treatment.

[0053] 2. Extract the samples obtained at each time point in step 1, extract total RNA, and reverse transcribe it into cDNA. Using the cDNA as a template, the relative expression of the TaJAZ1 gene was identified by real-time fluorescent quantitative PCR amplification using a primer pair consisting of primer F1: 5'-GACACGCCGAAGCCAAAGAC-3' and primer R1: 5'-GGCAAAGGAGGTGAAACACG-3' (taking TaGAPDH as As an internal reference, a primer pair consisting of primer F2: 5'-TTAGACTTGCGAAGCCAGCA-3' and primer R2: 5'-AAATGCCCTTGAGGTTTCCC-3' was used to identify the internal reference).

[0054] The result is as figure 2 shown. The results showed that the expression of TaJAZ1 was induced by jasmonic acid.

...

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Abstract

The invention discloses a wheat powdery mildew resistance gene and an application thereof. A wheat TaJAZ1 gene from an A genome is identified from a common wheat cultivar KN199 by the inventors, overexpression of TaJAZ1 missing a jas domain is proved to be capable of leading to wheat resistance enhancement through the stable transgenic technology, and positive regulation of wheat resistance by TaJAZ1 is indicated. The invention proves for the first time that TaJAZ1 is a key regulatory gene for wheat resistance.

Description

technical field [0001] The invention relates to a wheat powdery mildew resistance gene and its application. Background technique [0002] Wheat is an important food crop worldwide. About 35% to 40% of the world's population uses wheat as their main food. Since the 1980s, with the promotion of dwarf and semi-dwarf varieties and the improvement of water and fertilizer conditions, the occurrence and damage of wheat diseases have become increasingly serious in my country. Although chemical control of wheat diseases can achieve certain results, it is a waste of manpower, material and financial resources, especially causing environmental pollution. Therefore, it is the most economical and effective way to control wheat diseases by breeding and breeding disease-resistant varieties. Studies have shown that using gene editing technology to mutate the three TaMLO alleles in hexaploid wheat can endow wheat with heritable broad-spectrum resistance to powdery mildew. Ethylene negativ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8282
Inventor 孙加强荆叶醒刘杰
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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