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Pea cold resistance correlated SSR primer compositions and application thereof

A primer combination and cold-resistant technology, applied in pea cold-resistant related SSR primer combination, identification of cold-resistant pea, cold-resistant or non-cold-resistant pea fingerprint field, can solve the problems of subjective bias, easy to be affected by growing season and environment, and heavy workload. , to achieve the effect of low cost, good application prospect and easy operation

Active Publication Date: 2017-05-24
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to: aim at a series of problems such as the identification of the cold-resistant characteristics of pea varieties can only be carried out in the field, the period of phenotypic identification is long, the workload is heavy and cumbersome, the growing season and environment are easily affected, and there are subjective deviations, etc. A set of SSR primers related to cold tolerance of pea and identification of cold tolerance of pea varieties using these primers

Method used

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  • Pea cold resistance correlated SSR primer compositions and application thereof
  • Pea cold resistance correlated SSR primer compositions and application thereof
  • Pea cold resistance correlated SSR primer compositions and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Screening of SSR primers related to pea cold resistance traits

[0040] In this example, 30 representative germplasm resources of pea were selected, and after 3 years and 3 fields of cold-resistant phenotype identification, including 13 cold-resistant resources and 17 cold-resistant resources (see Table 1).

[0041] According to the results of pea genome-wide association analysis, selection is significantly related to cold-resistant traits of pea, and there is a significant difference between cold-resistant and non-cold-resistant pea resources, with high genetic consistency and good repeatability within the resources, and at the same time, it meets the requirements of stable PCR amplification and electrophoresis detection of amplified products 35 pairs of SSR primers with clear bands were used as the primer set for cold tolerance identification of pea germplasm resources (see Table 2).

[0042] Table 2 35 pairs of cold tolerance-related SSR primers

[0043] ...

Embodiment 2

[0046] Example 2 Construction of characteristic fingerprint of cold-resistant pea and establishment of identification method for cold-resistant pea

[0047] 1. Extraction of pea total DNA

[0048] The improved CTAB method was used to extract the DNA of the test resources: take about 2g of fresh young leaves, grind them into powder in liquid nitrogen, put them into a 2.0ml centrifuge tube, and store them in a freezer for later use; To the CTAB extract, add β-mercaptoethanol in proportion to 1% of the solution volume, and mix thoroughly; add 800 μL of preheated CTAB extract to each sample, vortex for 1-2 min, and place in a 65°C water bath for 1 h (every 15 min Mix up and down once); after heating in the water bath, add 800 μL of chloroform / isoamyl alcohol (24:1) solution to the centrifuge tube, mix for 15-20 minutes, and then centrifuge at 10,000 rpm for 15 minutes in a high-speed centrifuge; absorb about 600 μL Transfer the supernatant to a new centrifuge tube (absorb an appr...

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Abstract

The invention provides pea cold resistance correlated SSR primer compositions and application thereof and belongs to the technical field of crop molecule assisted breeding. 35 pairs of SSR primer compositions correlated with pea cold resistance are provided, nucleotide sequences of the SSR primer compositions are respectively shown as SEQ ID NO.70, cold-resistant or non-cold-resistant peas are identified by utilizing a PCR amplification technique and non-denaturing polyacrylamide gel electrophoresis, the pea cold resistance characteristic identification work can be completed within short time, and the pea cold resistance correlated SSR primer compositions have the advantages of being efficient, accurate and convenient to operate, making identification results not prone to environment and season influence and the like. The method discloses the heritable variation and genetic relationship of cold-resistant or non-cold-resistant pea varieties at DNA level, a technical support is provided for reasonable utilization of excellent varieties in the pea variety breeding process, the pea cold resistance correlated SSR primer compositions can be applied to cold resistance identification and molecular assisted breeding of pea genetic resources or varieties in the agricultural field and have a good application prospect.

Description

technical field [0001] The invention relates to the technical field of crop molecular assisted breeding, in particular to a pea cold-resistant related SSR primer combination, a cold-resistant or non-cold-resistant pea fingerprint based on the SSR primer combination, and a method for identifying cold-resistant pea. Background technique [0002] Pea (Pisum sativum L.) is one of the main edible beans in my country. It has a long history of cultivation and is rich in nutrition. , can improve soil fertility and increase crop yield. Pea has strong adaptability and wide geographical distribution. It is widely planted in more than 90 countries around the world. As the fourth largest edible bean crop in the world, its cultivation area and output are second only to soybeans, kidney beans and chickpeas. Statistics show that whether it is dry peas or green peas, China ranks among the top in the world in terms of planting area and total output, which has established China's important pos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 宗绪晓刘荣杨涛方俐张红岩张晓艳郝俊杰
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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