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Application of OsMYB63 gene in improving disease resistance of rice

An osmyb63, gene technology, applied in the application field of rice MYB transcription factor OsMYB63

Active Publication Date: 2021-01-19
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many plant-specific transcription factors, as disease resistance-related genes, play an important and unique role in regulating plant responses to biotic stress, such as WRKY, NAC, MYB, bZIP, and ERF, among which MYB transcription factors participate in rice resistance. disease rarely reported

Method used

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  • Application of OsMYB63 gene in improving disease resistance of rice
  • Application of OsMYB63 gene in improving disease resistance of rice
  • Application of OsMYB63 gene in improving disease resistance of rice

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1: Obtaining of OsMYB63 gene overexpression material

[0028] (1) Construction of OsMYB63 gene overexpression vector and knockout vector

[0029] This example is a general description about the construction of pU1301-OsMYB63 vector.

[0030]Using the cDNA of the rice variety Zhonghua 11 (or ZH11, a well-known and public rice verification land model variety from the Institute of Crop Science, Chinese Academy of Agricultural Sciences, whose genetic background is relatively clear) as a template, the primer MYB63F (5'-TTACGAACGATAGCC was designed GGTACC ATGGGGAAGGGCCGGGCACCGTGCTGC-3') and MYB63R (5'-ATGATCTTTGTAATC GGATCC CACGTAGTAATCCATGGGGTTACTCGT-3') was amplified by high-fidelity DNA polymerase PCR to obtain the full-length CDS fragment of the OsMYB63 gene (the 128th to 268th deoxyribonucleotides in the sequence shown in SEQ ID NO: 1, the 375th to 504th Deoxyribonucleotides, the 609th to 1453rd deoxyribonucleotides. Electrophoresis reclaims the PCR produc...

Embodiment 2

[0043] Example 2: Correlation analysis and functional verification of OsMYB63 transgenic material

[0044] (1) Overexpression of OsMYB63 gene and phenotypic identification of osmyb63 mutant materials inoculated with Xanthomonas blight at booting stage

[0045] In China, Hubei, and Wuhan, the overexpression material of the OsMYB63 gene, the osmyb63 mutant (obtained in Example 1) and the wild-type control were inoculated with bacterial blight in the summer field. The results show that the OsMYB63 gene overexpression rice line of the present invention is inoculated with bacterial blight (bacterial strain number PXO347, provided by the International Rice Research Institute, Philippines) at the booting stage, and wild type Zhonghua 11 (wild type, in this field) The conventional concept is that the non-transgenic variety is called wild type, the same below), the disease length of the positive single plant of OsMYB63 gene overexpression rice line 15 and line 31 was significantly shor...

Embodiment 3

[0048] Example 3: Analysis of OsMYB63 regulating plant cell wall

[0049] (1) Overexpression of OsMYB63 gene and detection of cellulose content in osmyb63 mutant materials

[0050] Flag leaves at the booting stage of OsMYB63 gene overexpression material, osmyb63 mutant and wild type were taken, greened at 105°C for 15 minutes, dried at 60°C to constant weight, and then pulverized through a 60-mesh sieve. Take by weighing 0.1000 gram sample and determine the relative content of cell wall cellulose, each sample is done 3 technical repetitions, main steps are as follows:

[0051] 1) Weigh 0.1000 g of powder, add an appropriate amount of single distilled water and mix into a homogenized state. Transfer to a 15mL centrifuge tube. Centrifuge at 4000 rpm for 5 minutes. Discard the supernatant and save the residue.

[0052] 2) Add 10 mL of dimethyl sulfoxide: ultrapure water (9:1, v / v), shake at room temperature overnight. Centrifuge at 4000 rpm for 5 minutes. The supernatant wa...

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Abstract

The invention belongs to the technical field of plant genetic engineering, and particularly relates to application of an OsMYB63 gene in improving disease resistance of rice. Through research of the OsMYB63 gene overexpression, it is found that the OsMYB63 gene can improve the resistance of rice to bacterial leaf blight, and the resistance of rice to bacterial leaf blight is reduced by knocking out the OsMYB63 gene, that is, the resistance of rice to bacterial leaf blight can be improved by overexpression of the OsMYB63 gene. The OsMYB63 gene of the invention can be applied to disease-resistant breeding of rice.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and specifically relates to the application of rice MYB transcription factor OsMYB63. The gene is the rice OsMYB63 gene, and the gene can be used to enhance the cultivation of rice varieties resistant to bacterial blight. Background technique [0002] Rice is one of the most important food crops in the world. Its yield has been threatened by various pathogens for a long time in the past, and rice bacterial blight is one of the most serious pathogens. my country is an important rice producing and consuming country in the world, and the yield and quality of rice seriously affect the sustainable development of my country's economy and society. Bacterial blight is a disease of rice vascular bundles. The pathogen enters the rice body through the wound or water hole of the leaf and reproduces. It often causes the rice leaf to form long and dead spots, eventually leading to the death o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82C07K14/415A01H5/00A01H6/46
CPCC07K14/415C12N15/8281
Inventor 袁猛谢文亚
Owner HUAZHONG AGRI UNIV
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