PCR/LDR molecular marker and method for identifying high-temperature-resistant TT1 genotype of rice

A molecular marker, high temperature-resistant technology, used in biochemical equipment and methods, microbial assay/inspection, recombinant DNA technology, etc., can solve problems such as unfavorable high-throughput analysis, cumbersome detection process, etc. easy effect

Inactive Publication Date: 2020-07-31
SHANGHAI ACAD OF AGRI SCI
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Problems solved by technology

Based on this, the present invention designs a molecular marker based on PCR/LDR technology in view of the relatively cumbersome detection process of the traditional molecular marker technology based on PCR and electrophoresis technology, which is not conducive to the high-throughput analysis of man

Method used

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  • PCR/LDR molecular marker and method for identifying high-temperature-resistant TT1 genotype of rice
  • PCR/LDR molecular marker and method for identifying high-temperature-resistant TT1 genotype of rice
  • PCR/LDR molecular marker and method for identifying high-temperature-resistant TT1 genotype of rice

Examples

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Effect test

Embodiment 1

[0041] 1. Rice material

[0042] Shenfan 14 is a restorer line of the three-line hybrid japonica rice Huayou 14, WYJ CG14 It is a near isogenic line carrying the high temperature resistant allele OgTT1, donated to Academician Lin Hongxuan of the Institute of Plant Physiology and Ecology, Chinese Academy of Sciences. Shenfan 14 and WYJ identified by sequencing CG14 The SNP2 loci of the TT1 gene are A and G respectively, indicating that Shen Fan 14 and WYJ CG14 carrying OsTT1 and OgTT1 alleles ( figure 1 ), the F1 used comes from Shenfan 14 and WYJ CG14 hybridization.

[0043] 2. Rice genomic DNA extraction:

[0044] Shen Fan 14, WYJ CG14 Take about 50 mg of leaves from each of its hybrid F1 generation rice plants, cut them into pieces with scissors, put them into a 2ml centrifuge tube, add 600 μl of 1.5×CTAB solution (1.5% CTAB, 75mM Tris-HCl, 15mM EDTA, 1.05M NaCl, pH8. 0) and a steel ball with a diameter of 5 mm, vibrate and grind at a frequency of 70 Hz for 90 s on a ...

Embodiment 2

[0062] Rice samples used in Example 2 are Shen Fan 14 and WYJ CG14 The 24 strains of the F2 population were crossed, and other detection steps and methods were the same as in Example 1. The test results are shown in Table 1.

[0063]

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Abstract

The invention relates to a PCR/LDR molecular marker and method for identifying a high-temperature-resistant TT1 genotype of rice, and belongs to the technical field of rice breeding. The molecular marker comprises a pair of PCR primers and three LDR probes, the forward primer sequence of the PCR primer pair is shown as SEQ ID NO. 1, and the reverse primer sequence is shown as SEQ ID NO. 2. The LDRprobes comprise a fluorescence labeling probe Probe-FAM of which the sequence is shown as SEQ ID NO.3, a non-high-temperature-resistant variety allele specific probe Probe-OsTT1 of which the sequenceis shown as SEQ ID NO.4, and a high-temperature-resistant allele specific probe Probe-OgTT1 of which the sequence is shown as SEQ ID NO.5. The method disclosed by the invention not only can accurately and quickly identify rice varieties and the genotype of the high-temperature-resistant gene TT1 in each strain in a breeding population, but also can realize high-throughput detection of more samples. Compared with a conventional molecular marker group, the molecular marker has the advantages of simplicity, convenience, quickness and high efficiency. The method provided by the invention can improve the selection efficiency of the high-temperature-resistant OgTT1 homozygous genotype rice strain, and accelerates the breeding process of the high-temperature-resistant rice variety.

Description

technical field [0001] The invention relates to the technical field of rice breeding, in particular to a PCR / LDR molecular marker and method for identifying rice high temperature resistant gene TT1 genotype. 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. However, global warming caused by greenhouse gas emissions after global industrialization has increasingly become an important unfavorable factor hindering food security. In my country, high temperature heat damage has become one of the most important factors affecting rice production, and the middle and lower reaches of the Yangtze River, as the main rice producing area, have also become the hardest hit areas by high temperature heat damage. [0003] Under the background of continuous global warming and frequent occurre...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/686C12N15/11
CPCC12Q1/6895C12Q1/686C12Q2600/13C12Q2600/156C12Q2531/137
Inventor 曹黎明储黄伟程灿涂荣剑牛付安周继华孙滨
Owner SHANGHAI ACAD OF AGRI SCI
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