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Wheat seedling stage antifreeze-related CAPS marker and method for identifying wheat seedling stage freezing tolerance

A freezing resistance and labeling technology, which can be used in biochemical equipment and methods, recombinant DNA technology, and microbial assay/inspection, etc., and can solve problems such as time-consuming and long-term

Active Publication Date: 2018-11-30
ANHUI AGRICULTURAL UNIVERSITY
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AI Technical Summary

Problems solved by technology

The former requires the construction of a mapping population, which takes a long time, and the identified loci / molecular markers are limited to the differences between the two parents

Method used

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  • Wheat seedling stage antifreeze-related CAPS marker and method for identifying wheat seedling stage freezing tolerance
  • Wheat seedling stage antifreeze-related CAPS marker and method for identifying wheat seedling stage freezing tolerance
  • Wheat seedling stage antifreeze-related CAPS marker and method for identifying wheat seedling stage freezing tolerance

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Embodiment Construction

[0021] 1. Identification of frost resistance phenotype in wheat seedling stage

[0022] From December 2015 to February 2016, there were continuous low temperature weather in Huaibei, Anhui. The present invention carried out seedling freezing injury (Freezing tolerance, FT) identification on 403 wheat varieties planted in Huaibei Wupu Farm in three periods: December 17, 2015, January 19, 2016, and February 16, respectively. Recorded as 15FT, 16FT-1, 16FT-2. Calculate the average value of the freezing damage phenotype in the three periods, and record it as FT-M. The level records of freezing damage in wheat fields refer to the agricultural industry standards of the People's Republic of China and are divided into 5 levels:

[0023]

[0024] 2. Extraction of wheat genomic DNA by SDS-Tris saturated phenol method

[0025] (1) Take 2-3 wheat seeds and place them in a 2ml sterilized centrifuge tube, and use MP high-throughput tissue grinder to grind them into powder.

[0026] (...

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Abstract

The invention discloses a wheat seedling stage antifreeze-related CAPS marker and a method for identifying the wheat seedling stage freezing tolerance, which relates to the technical field of wheat molecular breeding. The CAPS marker is named as CAPS-Ta-2-4, and a nucleotide sequence of the CAPS marker is SEQ ID NO. 1. The method for identifying the wheat seedling freezing tolerance by using the CAPS marker comprises the following steps: step 1, extracting a to-be-detected wheat genome DNA; step 2, performing the PCR amplification for the wheat genome DNA obtained in step 1 by utilizing the sequences as shown by SEQ ID NO. 2 and SEQ ID NO. 3, and obtaining an amplification product; step 3, performing enzyme digestion for the amplification product by utilizing BtsaI incision enzyme, and obtaining an enzyme digestion product; and step 4, when the enzyme digestion product is 185bp and 70bp, naming a genotype of the corresponding amplification product as CAPS-Ta-2-4-A, wherein the corresponding wheat seedling stage freezing tolerance is poor; and the CAPS marker developed in the invention can provide a molecular marker for genetically improving the freeze tolerance of the wheat.

Description

technical field [0001] The invention relates to the technical field of wheat molecular breeding, in particular to a CAPS marker related to frost resistance at seedling stage of wheat and a method for using the marker to identify frost resistance at seedling stage of wheat. Background technique [0002] About 40% of the world's population uses wheat as a staple food, and China is the world's largest wheat producer and consumer. Therefore, high and stable yields have always been an important goal of wheat breeding (He Zhonghu et al. 2018). However, wheat suffers from low temperature and freezing damage (<-5°C) at the seedling stage, which seriously affects the high and stable yield of wheat. Breeding wheat varieties with strong frost resistance is an effective way to solve this problem. [0003] Freezing tolerance (FT) traits in wheat are jointly controlled by multiple genes. The reported frost resistance-related genes in wheat are mainly located on chromosomes 1B, 2B, 1D...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 张海萍卢杰常成张璨姜昊董连生邵辉司红起马传喜
Owner ANHUI AGRICULTURAL UNIVERSITY
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