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TT1-dCAPS marker for detecting genotypes of high temperature resistant genes TT1 of rice and application

A high-temperature, genotypic technology that can be used in recombinant DNA technology, microbial determination/inspection, DNA/RNA fragments, etc.

Inactive Publication Date: 2019-02-15
SHANGHAI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

High temperature heat damage during the reproductive growth stage will cause abnormal pollen development, which will reduce the seed setting rate and lead to a decline in rice quality.

Method used

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  • TT1-dCAPS marker for detecting genotypes of high temperature resistant genes TT1 of rice and application
  • TT1-dCAPS marker for detecting genotypes of high temperature resistant genes TT1 of rice and application
  • TT1-dCAPS marker for detecting genotypes of high temperature resistant genes TT1 of rice and application

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Experimental program
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Effect test

Embodiment 1

[0028] 1. Rice material

[0029] Shen 9B is the maintainer line of the three-line hybrid japonica rice Huayou 14. The near-isogenic line NIL (CG14) was provided by researcher Lin Hongxuan of the Chinese Academy of Sciences. The F1 and F2 generation populations used in the present invention are derived from the hybridization of Shen 9B and NIL (CG14).

[0030] 2. Genomic DNA extraction from rice

[0031] Take about 50 mg of leaves from each rice variety, cut them into pieces with scissors, put them into a 2 mL centrifuge tube, add 600 μL of 1.5×CTAB solution (1.5% CTAB, 75 mM Tris-HCl, 15 mM EDTA, 1.05 M NaCl, pH8.0) and a grain of 5 mm in diameter The steel balls were oscillated and ground at a frequency of 70 Hz for 90 s on a fast grinder; the ground sample was incubated in a 56°C water bath for 20 minutes, 450 μL of chloroform was added, and after vigorous shaking, it was centrifuged at 12000 r / min for 10 minutes; 450 μL of the supernatant was transferred to another Add 900...

Embodiment 2

[0043] Using TT1-dCAPS markers to identify the TT1 genotypes of 48 individuals in the F2 population of Shen 9B and NIL (CG14) hybrids, the results showed that 12 individuals showed the same genotype as Shen 9B, and 13 individuals showed the same genotype as Shen 9B. The same genotype as NIL(CG14), while the other 23 individuals showed heterozygous genotypes (such as Figure 4 shown), conforming to a separation ratio of 1:2:1. This result further indicates that the TT1-dCAPS marker can accurately detect the genotype of TT1, and is suitable for marker-assisted selection to breed TT1-containing CG14 Genetic heat-tolerant rice varieties.

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Abstract

The invention provides a method for detecting high temperature resistant genes TT1 of rice and a dCAPS marker thereof. The primer sequence is: TT1-dcaps-F:5'-AGTCGAAAGCAAGCACAACAGGATTATC-3', and TT1-dcaps-R:5'-AGCAGAAACACCGATTCGGAATCAC-3'. The PCR product is digested by restriction endonuclease BsaBI, and detected by 8% polyacrylamide gel electrophoresis and silver staining. A 143 bp band is detected as a rice material containing a high temperature resistant TT1CG14 gene. The marker TT1-dCAPS is verified by using a high temperature sensitive three-line hybrid indica rice line retention line 9B, a near-isogenic line NIL (CG14) containing a high temperature resistant allele TT1CG14, and hybrid F1 and F2 generation populations between the two, and the results show that the marker can accurately identify different TT1WYJ, TT1CG14 and heterozygous genotypes of rice TT1 genes, and can be used for molecular marker-assisted breeding of the TT1 genes.

Description

technical field [0001] The invention relates to a dCAPS marker for detecting the TT1 genotype of rice high temperature resistant gene, which belongs to the technical field of rice breeding. Background technique [0002] Rice is an important grain crop in my country. The rice planting area is close to 30% of the total grain crop planting area in my country, and the total output is more than one-third of the total grain output in my country. China's rice planting area accounts for 18.5% of the world's rice planting area, second only to India, ranking second in the world. It is estimated that by 2050, the world's food production needs to increase by 38%, and by 2070, the world's food production needs to increase by 57%, in order to meet the food demand caused by the world's population growth. However, global warming caused by post-industrial greenhouse gas emissions has become a major obstacle to the realization of this goal. In my country, high temperature heat damage has be...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6895C12Q2600/13C12Q2600/156C12Q2531/113C12Q2521/301C12Q2565/125
Inventor 储黄伟曹黎明程灿涂荣剑周继华牛付安孙滨胡雪娇杨佳
Owner SHANGHAI ACAD OF AGRI SCI
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