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High-efficiency breeding kit for self-fertilizing progenies of 'Hui' based on ssr markers

A self-flowering, molecular marker technology, applied in the direction of determination/inspection of microorganisms, DNA/RNA fragments, biochemical equipment and methods, etc.

Active Publication Date: 2020-11-24
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Research work related to apple SSR has been carried out, but there is no research on the association analysis of apple self-fruiting traits using SSR markers

Method used

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  • High-efficiency breeding kit for self-fertilizing progenies of 'Hui' based on ssr markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Analysis of SSR loci of self-flowering offspring of 'Hui'

[0027] 1. Extraction of genomic DNA from 'Hui' self-bred offspring

[0028] 1) Take about 0.1 g of apple leaves and grind them into powder with liquid nitrogen, then add 700 μl of DNA extraction buffer, and bathe in water at 65° C. for 30 minutes. Or use scissors to cut an appropriate amount of tissue samples into a 2ml centrifuge tube, add a steel ball to the centrifuge tube, and then add 700 μl of preheated DNA extraction buffer. After being violently shaken by a tissue breaker (25 Hz, 1 min), the leaves are finally powdered. Then put it into a water bath at 65°C for 30 minutes, take out the sample and shake it two to three times during this period, so that the sample is fully in contact with the extraction buffer.

[0029] 2) Add 700 μl of chloroform:isoamyl alcohol (24:1), mix well, fully extract some impurities, and then let it stand at room temperature for 5 minutes.

[0030] 3) Centrifuge a...

Embodiment 2

[0043] Example 2: Analysis of SSR loci of self-fertilizing offspring of 'Hui'×'Changfu 2'

[0044] 1. Extraction of genomic DNA from hybrid offspring of 'Hui'×'Changfu 2'

[0045] 1) Take about 0.1 g of apple leaves and grind them into powder with liquid nitrogen, then add 700 μl of DNA extraction buffer, and bathe in water at 65° C. for 30 minutes. Or use scissors to cut an appropriate amount of tissue samples into a 2ml centrifuge tube, add a steel ball to the centrifuge tube, and then add 700 μl of preheated DNA extraction buffer. After being violently shaken by a tissue breaker (25 Hz, 1 min), the leaves are finally powdered. Then put it into a water bath at 65°C for 30 minutes, take out the sample and shake it two to three times during this period, so that the sample is fully in contact with the extraction buffer.

[0046] 2) Add 700 μl of chloroform:isoamyl alcohol (24:1), mix well, fully extract some impurities, and then let it stand at room temperature for 5 minutes....

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PUM

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Abstract

The invention relates to a high-efficiency breeding kit for a 'Hui' self-fruitful offspring based on an SSR marker, and discloses an SSR molecular marker Ch04d02, which is related to the 'Hui' self-fruitfulness. 114bp:126bp genetic typing is manifested in the 'Hui' self-fruitful offspring. The invention further provides a primer for identifying the above SSR molecular marker, through the molecularmarker and the primer, an efficient molecular marker-assisted selection breeding system can be established, which can efficiently screen the apple germplasm with self-fruitfulness in the 'Hui' offspring, so as to create self-fruitful apple varieties.

Description

technical field [0001] The invention relates to the technical field of biological breeding, in particular to the development and application of an SSR molecular marker located on the No. 12 apple chromosome and related to the self-flowering trait of 'Hui' apple. Background technique [0002] Apple is the largest fruit in my country. It has high nutritional value and is deeply loved by consumers. However, due to the existence of self-incompatibility of apples, it is necessary to configure different S genes in the actual production process. Varieties are pollinated varieties, and the yield can be guaranteed only by pollination by visiting insects or artificial pollination during the flowering period, which not only increases the production cost, but also increases the difficulty of management. And cultivating apple varieties with self-fertilizing ability is one of the effective means to realize the light and simplified cultivation of fruit trees. Now there are few self-fertile ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11C12Q1/686
CPCC12Q1/686C12Q1/6895C12Q2600/13C12Q2565/125C12Q2563/107
Inventor 李天忠刘春生刘志姜峰李洋于杰
Owner CHINA AGRI UNIV