Molecular marker linked with gummy stem blight resistance gene Gsb-2 and application thereof

A molecular marker, gsb-2 technology, applied in the field of agricultural biology, can solve the problems of individual plants that are difficult to screen for disease resistance genes, unsuitable disease conditions, affecting selection efficiency, etc., so as to improve the selection accuracy, shorten the breeding cycle, select targeted effect

Inactive Publication Date: 2011-08-03
XINJIANG AGRI SCI ACAD CANTALOUPE RES CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the prior art, when breeding disease-resistant breeding materials through traditional methods, artificial inoculation at the seedling stage is used for identification, and screening is carried out according to the phenotypic resistance characteristics of the plants. Traditional methods sometimes affect the selection efficiency due to insufficient inoculation or unsuitable disease conditions. It is difficult to accurately and quickly screen out individual plants with disease-resistant genes

Method used

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  • Molecular marker linked with gummy stem blight resistance gene Gsb-2 and application thereof
  • Molecular marker linked with gummy stem blight resistance gene Gsb-2 and application thereof
  • Molecular marker linked with gummy stem blight resistance gene Gsb-2 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: ISSR molecular marker ISSR-57 linked to muskmelon blight resistance gene Gsb-2 560 the acquisition

[0046] 1) F 1 , F 1 F 2 . By planting parents, F 1 Generation and F 2 generation groups. It is artificially inoculated and the incidence of each individual plant is counted, F 2 The population statistics of generation segregation showed that in the population of 134 strains, 83 strains were disease-resistant and 36 strains were susceptible, which conformed to the inheritance pattern of 3:1 single dominant gene.

[0047] 2) Inoculation of creeping blight and disease classification: the method of spraying inoculation with conidia suspension and 5-level disease classification (Zhang et al., 1997), spraying the spore suspension with a micro-sprayer, the concentration of the spore suspension is 5 × 105 pcs mL -1 , Spray until the leaves of the plants start to drip. After inoculation, use a small shed to keep moisture, and the relative humidity is above 90...

Embodiment 2

[0056] Example 2: Amplification detection of the parents by primer ISSR-57

[0057] Using the parental PI 157082 and the genomic DNA of Baipicrisp as templates, ISSR-PCR amplification was carried out with primer ISSR-57, and the ISSR-PCR reaction system was: 10×buffer (without Mg 2+ ) 2.0μL; dNTP (2mmol L -1 ) 2.0 μL; MgCl 2 (25mmol·L -1 ) 1.5μL; Primer (10mmol L -1 ) 1.0 μL; template DNA (30ng·μL -1 ) 1.0 μL; Taq polymerase (5U·μL -1 ) 0.2 μL; ddH 2 O 12.3 μL, a total of 20 μL. The PCR amplification reaction program is: pre-denaturation at 94°C for 5 minutes, then denaturation at 94°C for 30 seconds, annealing at 55°C for 45 seconds, extension at 72°C for 2 minutes, and 35 cycles of extension at 72°C for 10 minutes, and storage at 4°C. The entire reaction procedure is about 2.5h.

[0058] The amplified product was mixed with 1.5 μL bromophenol blue (0.25%), and the sample was spotted on a medium containing 0.5 tg·mL -1 In 1.5% agarose gel of EB, with 1×TAE as the ele...

Embodiment 3

[0059] Example 3: Primer ISSR-57 to F 1 amplification detection

[0060] Take Hybrid F 1 Genomic DNA of Genomic DNA was used as a template, ISSR-PCR amplification and electrophoresis detection were carried out with primer ISSR-57, the amplification and detection method was: ISSR-PCR reaction system was: 10×buffer (excluding Mg 2+ ) 2.0μL; dNTP (2mmol L -1 ) 2.0 μL; MgCl 2 (25mmol·L -1 ) 1.5μL; Primer (10mmol L -1 ) 1.0 μL; template DNA (30ng·μL -1 ) 1.0 μL; Taq polymerase (5U·μL -1 ) 0.2 μL; ddH 2 O 12.3 μL, a total of 20 μL. The PCR amplification reaction program was as follows: pre-denaturation at 94°C for 5 min, followed by denaturation at 94°C for 30 s, annealing at 55°C for 45 s, extension at 72°C for 2 min, and 35 cycles, further extension at 72°C for 10 min, and storage at 4°C. The amplified product was mixed with 1.5 μL bromophenol blue (0.25%), and the sample was spotted on a medium containing 0.5 μg·mL -1 In 1.5% agarose gel of EB, with 1×TAE as the electro...

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Abstract

The invention discloses a method for obtaining a molecular marker linked with gummy stem blight resistance gene Gsb-2 and application of the molecular marker. An F2 group is constructed by PI 157082 carrying the gummy stem blight resistance gene Gsb-2 and a local susceptible variety of white skin crisp, gummy stem blight conidiospore suspension is artificially inoculated in a seedling period, and the molecular marker ISSR-57560 linked with the Gsb-2 is obtained through a simple sequence repeat (ISSR) technology. The molecular marker ISSR-57560 is used for detecting genotypes of melon varieties or strains taking PI 157082 as a parent, and can judge that whether the varieties or strains resist gummy stem blight. The invention overcomes the defects of high difficulty and longer period in breeding stable gummy stem blight resistant strains in the prior art, and provides a marker-assisted selection technology for gummy stem blight resistant breeding so as to reduce cost, improve selection efficiency, and quicken the progress of culturing good stem blight resistant melon strains.

Description

field of invention [0001] The invention relates to the field of agricultural biotechnology. Specifically, the present invention relates to the technical field of a molecular marker linked to the muskmelon blight resistance gene Gsb-2, its obtaining method and application. Background technique [0002] Melon is an important economic crop. The planting area and output of melon in my country rank first in the world. Melon blight is one of the most serious global fungal diseases. Its occurrence often leads to devastating consequences. In addition to harming melons, the fungus also infects other cucurbit crops such as cucumbers, watermelons, pumpkins and zucchini, resulting in different degrees of yield reduction. Chemical control of vine blight is not only inefficient, but also easy to cause environmental pollution, and cultivating disease-resistant varieties is one of the safest, most effective and cost-saving measures to control the damage of melon vine blight. [0003] Acc...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 张永兵伊鸿平张龑张学军李寐华吴海波王登明
Owner XINJIANG AGRI SCI ACAD CANTALOUPE RES CENT
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