Gray leaf spot resistance-related protein ZmWAK-RLK and encoding gene and applications thereof

A kind of gray spot disease and coding technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the economic loss of corn production and other problems

Pending Publication Date: 2019-05-03
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Gray spot disease can cause a 20-60% yield reduction in cases of disease, and up to 100% in severe cases, causing severe economic losses to maize production

Method used

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  • Gray leaf spot resistance-related protein ZmWAK-RLK and encoding gene and applications thereof
  • Gray leaf spot resistance-related protein ZmWAK-RLK and encoding gene and applications thereof
  • Gray leaf spot resistance-related protein ZmWAK-RLK and encoding gene and applications thereof

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Experimental program
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Effect test

Embodiment 1

[0059] Embodiment 1, the discovery of ZmWAK-RLK protein and its coding gene

[0060] Maize inbred line Y32 (as the donor parent) and maize inbred line Q11 (as the recurrent parent) were used to construct primary mapping populations and fine mapping populations. Mapping qRgls1 to maize chromosome 8 between IDP2 and M2, the physical location is about 120kb.

[0061] The Y32BAC library of the disease-resistant parent was screened by PCR using molecular markers located in the finely mapped QTL-qRgls1 region. Perform BAC clone fingerprint analysis to construct BAC contigs covering the entire gene segment. The clone that covers the least amount of gene region is selected for sequencing. Through sequence alignment and expression analysis, a new gene was found, encoding the protein shown in Sequence 1 of the Sequence Listing.

[0062] The protein shown in Sequence 1 of the sequence listing is named ZmWAK-RLK protein. The gene encoding ZmWAK-RLK protein was named ZmWAK-RLK gene. T...

Embodiment 2

[0063] Embodiment 2, verify the function of the fragment of 7.2kb

[0064] 1. Obtaining of transgenic plants

[0065] 1. A fragment of about 7.2kb in maize inbred line Y32 (this fragment is shown in sequence 4 of the sequence table; in sequence 4, the 1st-2103rd nucleotide is a promoter, and the 2104-4316th nucleotide (identical to sequence 3 in the sequence listing) was inserted into the BamH I restriction site of the pCAMBIA3301 vector to obtain a recombinant plasmid.

[0066] 2. Introduce the recombinant plasmid obtained in step 1 into Agrobacterium EHA105 to obtain recombinant Agrobacterium.

[0067] 3. Take the recombinant Agrobacterium obtained in step 2, and use the Agrobacterium-mediated method to genetically transform the immature embryos of the maize inbred line B73-329 to obtain T0 generation plants.

[0068] 4. The T0 generation plants are self-crossed, the grains are harvested, and the grains are cultivated into plants, which are T1 generation plants.

[0069] ...

Embodiment 3

[0104] Embodiment 3, verify the function of open reading frame

[0105] 1. Obtaining of transgenic plants

[0106] 1. Insert the double-stranded DNA molecule shown in nucleotides 87-2084 of Sequence 2 in the sequence listing into the XcmI restriction site of pBCXUN vector to obtain a recombinant plasmid.

[0107] 2. Introduce the recombinant plasmid obtained in step 1 into Agrobacterium EHA105 to obtain recombinant Agrobacterium.

[0108] 3. Take the recombinant Agrobacterium obtained in step 2, and use the Agrobacterium-mediated method to genetically transform the immature embryos of the maize inbred line B73-329 to obtain T0 generation plants.

[0109] 4. The T0 generation plants are self-crossed, the grains are harvested, and the grains are cultivated into plants, which are T1 generation plants.

[0110] 5. The T1 generation plants were identified by PCR, and the transgenic plants were screened. The transgenic plants selected from the T1 generation plants are T1 transgen...

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Abstract

The invention discloses a gray leaf spot resistance-related protein ZmWAK-RLK and an encoding gene and applications thereof. The protein provided by the present invention is obtained from maize inbredline Y32 and designated as ZmWAK-RLK protein which is as shown in SEQ ID NO:1 of a sequence list. Nucleic acid molecules encoding the ZmWAK-RLK protein also belongs to the protection scope of the invention. The present invention also protects a method of preparing a transgenic plant, and the method comprises the steps of introducing the nucleic acid molecules into a starting plant to obtain the transgenic plant with enhanced resistance to gray leaf spot. The present invention also protects a plant breeding method, and the method comprises the steps of: increasing the content and / or activity of the ZmWAK-RLK protein in the target plant so as to improve the gray leaf spot resistance of the target plant. The protein has great application value for the anti-gray leaf spot breeding of maize.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to gray spot disease resistance-related protein ZmWAK-RLK, its coding gene and application. Background technique [0002] Maize gray spot (GLS) is a disease of maize leaves that affects the yield and quality of maize. First discovered in Alexandria County, Illinois, USA in the 1920s, gray spot has gradually developed into a serious global leaf disease. Gray spot is widespread in major corn-growing areas of the United States, Asia, Europe, and Africa. In the case of the disease, gray spot can cause a 20-60% yield reduction, which can reach 100% in severe cases, causing serious economic losses to maize production. [0003] Maize gray spot is a fungal disease. It is generally believed that its pathogenic bacteria mainly include Cercospora zeae-maydis (Czm, Cercospora zeae-maydis) and Cercospora zeina (Cz, Cercospora zeina). In 2013, Liu et al. extensively collected disease s...

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
CPCC07K14/415C12N15/8282A01H5/10
Inventor 徐明良钟涛番兴明朱芒张艳徐凌刘丽
Owner CHINA AGRI UNIV
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