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Molecular marker for assisted selective breeding of upland cotton with greensickness resistant traits

A Verticillium wilt resistance and assisted selection technology, applied in the field of cotton genetics and breeding, can solve the problems of no field breeding groups, etc., and achieve the effects of improving efficiency and selection accuracy, improving efficiency and accuracy, and reducing workload

Inactive Publication Date: 2011-06-01
INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chinese Science Series C, 2009, 39: 849-861), or when verifying the assisted selection effect of molecular markers, it is only limited to parental segregation populations, and it is not applied to the entire field breeding population (Wang Furong et al., Upland Cotton Varieties Molecular markers of Verticillium wilt resistance traits and their assisted selection effects

Method used

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  • Molecular marker for assisted selective breeding of upland cotton with greensickness resistant traits
  • Molecular marker for assisted selective breeding of upland cotton with greensickness resistant traits
  • Molecular marker for assisted selective breeding of upland cotton with greensickness resistant traits

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1. Get 96 upland cotton lines (such as figure 2 Shown), with Changkang cotton and TM-1 as contrast respectively, described material is planted in Verticillium wilt disease nursery, single-row area, repeats twice;

[0026] 2. Inoculate Verticillium dahliae of the Anyang strain with moderate pathogenicity before sowing, and when the incidence of Verticillium dahliae peaks in June, July, and August, the rate of susceptible control strains reaches 80% or the disease index reaches about 50 , investigate the incidence of each test material, adopt the 5-level system to grade cotton Verticillium wilt (Table 1), and calculate the disease index and relative disease index, and finally calculate the average according to the material as the average disease index. The formulas for calculating the condition index and the relative condition index are as follows:

[0027] Disease index DI = [(number of first-level diseased plants × 1 + number of second-level diseased plants × 2 + numb...

Embodiment 2

[0048] 1. Take different upland cotton lines and plant them in the field (such as figure 2 ).

[0049] 2. Extracting its total DNA from the tender green leaves on the field material plants.

[0050] 3. Use the disease nursery to identify the Verticillium wilt resistance of the upland cotton strains, see Example 1 for the specific identification method.

[0051] 4. Utilize single SSR primer sequence of the present invention to analyze, concrete steps are as follows:

[0052] 4.1 SSR primer labeling analysis:

[0053] PCR amplification reaction system is 10 μL: 25ngDNA, 1μM upstream primer, 1μM downstream primer, 1×buffer (67mmol / L Tris-HCl, pH8.8, 16mmol / L (NH 4 ) 2 SO 4 ), 0.2mmol / L dNTPs, 2.5mmol / L MgCl 2 ,, TaqDNA polymerase 0.5U.

[0054] PCR reaction program: denaturation at 95°C for 3min; denaturation at 94°C for 45s, annealing at 60°C for 45s, extension at 72°C for 1min, a total of 30 cycles; extension at 72°C for 3min. Store at 10°C. PCR amplification was carr...

Embodiment 3

[0068] 1. Take different upland cotton strains (such as figure 2 ) planted in the field.

[0069] 2. Extracting its total DNA from the tender green leaves on the field material plants.

[0070] 3. Use the disease nursery to identify the Verticillium wilt resistance of the upland cotton strains, see Example 1 for the specific identification method.

[0071] 4. comprehensively utilize three of the present invention SSR primer sequence For analysis, the specific steps are as follows:

[0072] 4.1 SSR primer labeling analysis:

[0073] PCR amplification reaction system is 10 μL: 25ngDNA, 1μM upstream primer, 1μM downstream primer, 1×buffer (67mmol / L Tris-HCl, pH8.8, 16mmol / L (NH 4 ) 2 SO 4 ), 0.2mmol / L dNTPs, 2.5mmol / L MgCl 2 ,, TaqDNA polymerase 0.5U.

[0074] PCR reaction program: denaturation at 95°C for 3min; denaturation at 94°C for 45s, annealing at 60°C for 45s, extension at 72°C for 1min, a total of 30 cycles; extension at 72°C for 3min. Store at 10°C. PCR ampl...

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Abstract

The invention relates to a molecular marker for assisted selective breeding of upland cotton with greensickness resistant traits, belonging to the technical field of genetic breeding of cotton. Three SSR (Simple Sequence Repeat) markers for assisted selective breeding of the upland cotton resisting greensickness can be obtained by constructing a molecular marker genetic linkage map in upland cotton seeds to carry out QTL (Quantitative Trait Locus) positioning on the greensickness resistant traits and selecting remarkable SSR markers entering a linkage group section and the SSR makers linked with disease-resistant QTLs to screen different upland cotton plant systems. The three markers comprise marker primers and a primer combination and are suitable for greensickness-resistant molecular marker-assisted selective breeding of the upland cotton. The plant systems resisting the greensickness can be obtained by using the obtained molecular marker to carry out greensickness-resistant molecular marker-assisted selection on different upland cotton plant systems.

Description

technical field [0001] The invention belongs to the technical field of cotton genetics and breeding, and relates to molecular markers used for assisted selection breeding of upland cotton resistance to Verticillium wilt, in particular to molecular marker positioning, molecular marker assisted selection effect evaluation and breeding application of upland cotton resistance to Verticillium wilt. Background technique [0002] Using molecular marker technology to screen molecular markers that are not affected by environmental factors and are closely linked to the resistance genes of the existing Verticillium wilt germplasm in my country can be applied to assisted selection breeding for disease resistance, thereby accelerating the resistance to Verticillium wilt The breeding process. [0003] A common method for screening molecular markers linked to quantitative trait genes is QTL mapping. QTL mapping is carried out by molecular markers for genetic linkage analysis, detection of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 王红梅赵云雷陈伟李运海龚海燕桑晓慧
Owner INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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