Method for detecting resistance of cotton plants to greensickness by molecular marker-assisted selection

A verticillium wilt and plant technology, applied in the field of agricultural biology, can solve the problems of large differences between years, poor disease resistance of bred varieties, inaccurate selection of progeny materials, etc., and achieve the effect of rapid detection

Active Publication Date: 2014-02-26
INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After the 1970s, the occurrence of Fusarium wilt of cotton gradually increased, and breeding for resistance to Fusarium wilt began. However, the breeding technology basically used natural disease-susceptible test fields for disease resistance identification. The identification results of hybrid offspring materials are inaccurate, and the selection or elimination of materials is relatively blind
After the 1990s, the occurrence of Verticillium wilt became more and more serious, and there was an urgent need to breed varieties resistant to Verticillium wilt. However, methods similar to those for res

Method used

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  • Method for detecting resistance of cotton plants to greensickness by molecular marker-assisted selection
  • Method for detecting resistance of cotton plants to greensickness by molecular marker-assisted selection
  • Method for detecting resistance of cotton plants to greensickness by molecular marker-assisted selection

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Experimental program
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Effect test

Embodiment 1

[0037] 1) Collect 31 kinds of cotton seed materials, according to the artificial seedbed disease in the literature (Zhang Xinghua, Cotton blight resistance, Verticillium wilt research progress and resistance identification method, Jiangxi Agricultural Journal, 2008,20(3):43-49) The results were listed in Table 1. Among them, 3 copies of disease-resistant germplasm were represented as "R" in the column of Verticillium wilt-resistant materials in Table 1; 28 copies of susceptible germplasm , which is indicated as "S" in the column of Verticillium dahliae-sensitive materials in Table 1.

[0038] (2) According to the method in the "Molecular Cloning Experiment Guide", isolate the genomic DNA of each germplasm.

[0039] Using the genomic DNA of each germplasm as a template, the forward primer is shown in SEQ ID NO: 1 (5′-AAACAATAATAAACGAGTTGAATTA-3′) and the reverse primer is shown in SEQ ID NO: 2 (5′-TTGTTTCATAATTTTAAAGTATGTA- 3′) for PCR amplification, the denaturation temperatu...

Embodiment 2

[0048] In this example, the cotton plants to be tested are the F2 generation seeds of Zhongzhimian 2 (GK44) selection line and Xin 59-4. The resistance to Verticillium wilt of the selected line Zhongzhimian 2 (GK44) and the F2 generation seeds of Xin 59-4 was not known in advance.

[0049] The tissue of the cotton plant that died of Verticillium dahliae was crushed, soaked in water of equal weight for 48 hours, and filtered to obtain the soaking solution containing the pathogenic bacteria of Verticillium dahliae.

[0050] Soak the F2 generation seeds of Zhongzhimian No. 2 (GK44) and Xin 59-4 in the above-mentioned soaking solution containing Verticillium dahliae pathogenic bacteria for 24 hours to obtain soaked seeds, wherein, relative to each part by weight seeds, the consumption of soaking liquid is 5 parts by weight.

[0051] The tissue of the cotton plant that died of Verticillium wilt was pulverized and mixed into the soil. The amount of the tissue of the cotton plant th...

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Abstract

The invention provides a method for detecting the resistance of cotton plants to greensickness by molecular marker-assisted selection. The method comprises the following steps: (1), soaking seeds of cotton plants to be detected in a soaking solution; (2), planting the soaked seeds in a disease garden; (3), selecting a cotton plant capable of surviving in the disease garden as a preliminary screened cotton plant; and (4), respectively carrying out PCR (Polymerase Chain Reaction) amplification on genome DNA (Desoxvribose Nucleic Acid) of the cotton plant by a first primer pair and a second primer pair to obtain a first amplification product and a second amplification product, wherein the first primer pair is shown as SEQ ID NO:1 and SEQ ID NO:2, the second primer pair is shown as SEQ ID NO:3 and SEQ ID NO:4, and if the first amplification product has nucleic acid shown as SEQ ID NO:5 and the second amplification product has nucleic acid shown as SEQ ID NO:6, the cotton plant to be detected is a greensickness-resistant plant. According to the method, the resistance to greensickness is predicted, the cotton plants are screened to discard plants which do not show symptoms but are susceptible in a disease garden screening process, the waste of manpower and material resources is reduced, and the breeding efficiency is increased.

Description

technical field [0001] The invention relates to the field of agricultural biotechnology, in particular to a method for detecting the resistance of cotton plants to Verticillium wilt. Background technique [0002] Cotton verticillium wilt is a major plant disease worldwide, causing 10-15% yield loss to cotton production all the year round, and more than 60% of the country's cotton planting area in serious years, causing a heavy blow to cotton production. The pathogenic bacteria spread along with the seeds, broken branches and leaves, and the soil. The sclerotia can survive in the soil for many years, and chemical drugs, cultivation and other methods are ineffective against it. It is called the "cancer" of cotton by the phytopathological circle, and it has become a long-standing unsolved problem in the world. sexual conundrum. Breeding disease-resistant varieties is recognized as the most economical and effective means to solve cotton Verticillium wilt. The reason why it has ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 戴小枫陈捷胤李蕾马雪峰桂月晶孔志强包郁明
Owner INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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