Nucleic acid, and method and kit used for identifying fungal strain pathogenic type

A kit and pathogenic technology, applied in the field of crop disease prevention and control, can solve the problems of cotton minor disease farmland yield, impact, cotton severe disease farmland yield, etc.

Active Publication Date: 2013-08-21
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the pathogenic types of different Verticillium dahliae strains are very different. For example, some strains with high pathogenicity will cause serious diseases of cotton after infecting cotton and cause a decline in farmland yield, while some strains with low pathogenicity The strains infected cotton only cause mild diseases of cotton and have little impact on farmland yield

Method used

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  • Nucleic acid, and method and kit used for identifying fungal strain pathogenic type
  • Nucleic acid, and method and kit used for identifying fungal strain pathogenic type
  • Nucleic acid, and method and kit used for identifying fungal strain pathogenic type

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] 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).

[0033] 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.

[0034] 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

[0036] This example is used to illustrate the detection results of the pathogenic types of the above 80 different Verticillium dahliae strains.

[0037] 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.

[0038] 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

[0044] 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.

[0045] In this example, the full genome sequences of the above two different Verticillium dahliae strains were obtained. Through sequence comparison, it was identified that the full genome sequence of VDG1 does not contain the sequence shown in SEQ ID NO: 1, nor does it contain SEQ ID NO: 1. The sequence shown in ID NO: 2; and the whole genome sequence of VDG2 contains the sequence shown in SEQ ID NO: 1 and also contains the sequence shown in SEQ ID NO: 2.

[0046] 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, ...

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Abstract

The invention provides a nucleic acid with a sequence represented by SEQ ID NO:1 or SEQ ID NO:2, or comprising a specific sequence of a sequence represented by the SEQ ID NO:1 or a specific sequence of a sequence represented by the SEQ ID NO:2. The invention also provides a nucleic acid complementary to the above nucleic acid. The invention also provides a method for identifying a fungal strain pathogenic type. The fungal strain is verticillium dahliae. The method comprises the steps that: (1) a total genomic DNA test sample comprising verticillium dahliae is prepared; (2) whether a first molecular marker exists in the test sample is detected, wherein the first molecular marker is the nucleic acid; and (3) verticillium dahliae strain pathogenic type is determined according to the detection result of 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 (Verticillium dahliae) is a plant pathogenic fungus, which can cause diseases such as Verticillium wilt in plants (especially cotton). However, the pathogenic types of different Verticillium dahliae strains are very different. For example, some strains with high pathogenicity will cause serious diseases of cotton after infecting cotton and cause a decline in farmland yield, while some strains with low pathogenicity The strains infected cotton only caused mild diseases of cotton and had little impact on farmland yield. In addition, the pathogenic type of Verticillium dahliae also shows a certain degree of latency, that is, the highly pathogenic strains infected in the current year may not cause disease, but will cause serious ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68C12Q1/04
Inventor 戴小枫李蕾陈捷胤陈相永柳少燕王金龙肖红利
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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