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Allele of succinate dehydrogenase B subunit gene sdHB of Corynespora cassiicola and use thereof

A technology of succinate dehydrogenase and Corynebacterium multiprime, which is applied in the directions of application, genetic engineering, plant genetic improvement, etc., can solve the problems of short duration of effect and high variability of Corynebacterium multiprime.

Active Publication Date: 2017-11-24
INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Corynespora multiprimum has highly variable characteristics, so the use of resistant varieties for disease control has the problem of short duration

Method used

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  • Allele of succinate dehydrogenase B subunit gene sdHB of Corynespora cassiicola and use thereof
  • Allele of succinate dehydrogenase B subunit gene sdHB of Corynespora cassiicola and use thereof
  • Allele of succinate dehydrogenase B subunit gene sdHB of Corynespora cassiicola and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1, the acquisition of mutation site

[0029] 1. Determination of sensitivity level of field strains of Corynespora polybasicum to fluopyram

[0030] Tested strains: 1 sensitive strain HG14102524-4 (a field strain of Corynespora multiprimum that has not used any succinate dehydrogenase inhibitor fungicides); 2 suspected resistant strains HG15050729-5 and HG15050825-6. Among them, HG14102524-4 was collected from Hebei Province, and other strains were collected from Liaoning Province, all of which were isolated and purified by single spore.

[0031] Test drug: fluopyram technical product (purity: 96%), kindly provided by Shenyang Research Institute of Chemical Industry.

[0032] Test medium: yeast peptone acetate medium (YBA): yeast powder 10g, peptone 10g, sodium acetate 20g, agar 15g, distilled water to 1L;

[0033] Potato dextrose agar medium (PDA): 200 g of potatoes, 20 g of glucose, 15 g of agar powder, and distilled water to a volume of 1 L. The above me...

Embodiment 2

[0056] Example 2, Verification of SdhB-V280 in making Corynespora multiprimum resistant to fluopyram

[0057] (1), materials and reagents

[0058] Tested strains: Corynespora cassiicola, numbers: HG15050729-5 (SdhB-V280), HG14102524-4 (SdhB-I280).

[0059] In the following examples, the CTAB method was used for DNA extraction.

[0060] Reagents: 2×CTAB extract (Coolaber Science&Technology), chloroform:isoamyl alcohol (24:1) (Coolaber Science&Technology), absolute ethanol (Beijing Chemical Plant), ddH 2 o

[0061] Instruments: Christ ALPHA 1-4 LSC freeze dryer (Martin Christ, Germany), pipette (Eppendorf, Germany), HQ-60 vortex mixer (Northern Tongzheng, Beijing), XMTD-700 water bath (Han Kang Electronics, Jiangsu), 3K15 high-speed centrifuge (Sigma, Germany), Spectrophotometers spectrophotometer (NanoDrop company, the United States)

[0062] In the following examples, PCR amplification

[0063] Reagents: Xerox DNA polymerase (Beijing Biomed Biotechnology Co., Ltd.), TaKaR...

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Abstract

The invention discloses an allele of a succinate dehydrogenase B subunit gene sdHB of Corynespora cassiicola and a use thereof. The invention provides a protein. The protein is obtained through mutation of a succinate dehydrogenase B subunit amino acid I at 280th site in the amino acid sequence into V, wherein other amino acid residues are not changed. Novel unreported allele mutation is found in the Corynespora cassiicola group collected in the Liaoning province and can cause high resistance of the strain to fluopyram. The mutation has the important guiding significance for detection of SDHIs resistance in the field and the later drug resistance management.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an allele and application of the succinate dehydrogenase B subunit gene sdhB of Corynespora polymata. Background technique [0002] Leaf spot caused by Corynespora cassiicola (Berk. & M.A. Curtis) C.T.Wei is a worldwide disease. The fungus is distributed in 95 countries and regions around the world, and can infect more than 530 kinds of plants, including economic crops such as soybean, cotton and rubber, vegetable crops such as cucumber, tomato and cowpea, and other plants. Since 2005, the research group has been working in my country's Liaoning, Hebei, Inner Mongolia Autonomous Region, Heilongjiang, Shandong, Shanxi, Henan, Gansu, Tianjin, Beijing, Hunan, Hubei, Guangdong, Guangxi, Chongqing, Sichuan, Yunnan, Hainan, Zhejiang , 21 provinces and cities in Fujian and Xinjiang have found diseases caused by the infection of Corynebacterium multiprime, and the hosts are mainl...

Claims

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Application Information

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IPC IPC(8): C12N9/02C12N15/53C12N15/80
CPCC12N9/001C12N15/80C12Y103/99001
Inventor 李宝聚朱发娣石延霞谢学文柴阿丽
Owner INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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