Nucleic acid, method for identifying pathotype of fungus strain and kit

A nucleic acid and bacterial strain technology, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve problems such as great differences in pathogenic manifestations, impact on cotton farmland yield, and decline in cotton farmland yield

Active Publication Date: 2013-08-14
INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, the pathogenicity of different pathogenic types of Verticillium dahliae strains varies greatly. For example, some highly pathogenic strains can cause serious diseases of cotton after infecting cotton and cause a decline in farmland yield, while some low-pathogenic strains After the pathogenic strains infect cotton, they only cause mild diseases of cotton and have little impact on farmland yield

Method used

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  • Nucleic acid, method for identifying pathotype of fungus strain and kit
  • Nucleic acid, method for identifying pathotype of fungus strain and kit
  • Nucleic acid, method for identifying pathotype of fungus strain and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] This example is used to illustrate the preparation of samples to be tested. Specifically, the preparation of genomic DNA from 80 different Verticillium dahliae strains (numbered VDG1 to VDG80).

[0030] Cut the stem of the cotton plant about 10-35cm above the ground and cut it into 1mm 3 Small pieces of about the size are inoculated in PDA medium (containing 200 g / L of potato, 20 g / L of glucose, and 20 g / L of agar) containing antibiotics (ampicillin, rifampicin and streptomycin, all at a concentration of 100 μg / ml) Above, cultivated at 25°C for 7 days, and identified the colonies of Verticillium dahliae according to the standard colony morphology of Verticillium dahliae. The identified colonies of Verticillium dahliae were taken as samples of Verticillium dahliae strains.

[0031] According to the above method, obtain 80 different Verticillium dahliae bacterial strains from 80 different cotton plants, and the above-mentioned 80 different Verticillium dahliae bacterial...

Embodiment 2

[0033] This example is used to illustrate the detection results of the pathogenic types of the above 80 different Verticillium dahliae strains.

[0034] Refer to literature (Zhu Heqin, Feng Zili, Li Zhifang, Zhao Lihong, Shi Yongqiang. Identification of Verticillium wilt resistance of cotton varieties (lines) by quantitative dipping in vermiculite sandy bottomless paper bowl. China Cotton, 2010, 37(12) : 15-17) in the vermiculite sandy soil bottomless paper pot quantitatively dipped in bacteria liquid method, 80 different Verticillium dahliae bacterial strains described in Example 1 were carried out the pathogenic type identification of cotton seedling stage. The level of the pathogenic type was characterized by the disease index of the 80 different Verticillium dahliae strains after inoculating healthy cotton plants.

[0035] Specifically, Verticillium dahliae was cultured in Chapic medium (2g / L NaNO 3 , K of 1g / L 2 HPO 4 , 0.5g / L KCl, 0.5g / L MgSO 4 , 0.01g / L FeSO 4 , th...

Embodiment 3

[0041] The whole genome sequence of Verticillium dahliae strains (coded as VDG1 and VDG2) was determined according to the method specified in the instruction manual (Solexa technology) of Genome Analyzer of Illumina Company.

[0042] In this example, the whole genome sequences of the above two different Verticillium dahliae strains were obtained. Through sequence comparison, it was identified that the whole genome sequence of VDG1 contained the sequence shown in SEQ ID NO: 1, while the whole genome sequence of VDG2 The sequence does not contain the sequence shown in SEQ ID NO:1.

[0043] By the same method as above, 61 of the above 80 different Verticillium dahliae strains (respectively VDG3, VDG63, VDG61, VDG18, VDG35, VDG32, VDG73, VDG31, VDG33, VDG26, VDG55, VDG23, VDG57, VDG36, VDG11, VDG67, VDG47, VDG5, VDG29, VDG59, VDG49, VDG44, VDG21, VDG48, VDG8, VDG50, VDG25, VDG43, VDG37, VDG46, VDG16, VDG4, VDG4, VDG38, VDG2 VDG45, VDG12, VDG76, VDG27, VDG10, VDG41, VDG64, VDG17, ...

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Abstract

The invention provides a nucleic acid. The nucleic acid has the sequences shown in SEQ ID NO:1 or has specific sequences in the sequences shown in SEQ ID NO:1. The invention also provides a nucleic acid complementary with the above nucleic acid. The invention also provides a method for identifying the pathotype of a fungus strain. The fungus strain is verticillium dahliae. The method comprises the following steps of: (1) preparing a sample to be detected of complete genome DNA containing verticillium dahliae; (2) detecting whether a first molecular marker exists in the sample to be detected, wherein the first molecular marker is the above nucleic acid; and (3) judging the pathotype of verticillium dahliae according to the detection result in the step (2). The invention also provides a kit.

Description

technical field [0001] The invention relates to the field of crop disease prevention and control, in particular to a nucleic acid, a method for identifying pathogenic types of fungal strains and a kit. Background technique [0002] Verticillium dahliae is a phytopathogenic fungus that can cause Verticillium wilt in plants, especially cotton. However, the pathogenicity of different pathogenic types of Verticillium dahliae strains varies greatly. For example, some highly pathogenic strains can cause serious diseases of cotton after infecting cotton and cause a decline in farmland yield, while some low-pathogenic strains The pathogenic strains only cause mild cotton diseases and have little impact on the yield of the farmland after infecting cotton. In addition, the pathogenic type of Verticillium dahliae also showed a certain degree of latency, that is, the highly pathogenic strains may not cause disease in the year of infection, but cause serious disease on a large scale in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68C12Q1/04
Inventor 戴小枫陈相永李蕾柳少燕王金龙陈捷胤田李
Owner INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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