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Identification method for hybrid rice variety hybrid source superior 69 based on InDel (insertion-deletion length polymorphism) marker

An identification method and technology of hybrid rice, which is applied in the field of identification of hybrid rice variety Jiaoyuanyou 69, can solve the problems of not being able to obtain test results on the same day, expensive SNP chips, cumbersome data processing, etc., and achieve good application prospects and high accuracy , the effect of the simple method

Active Publication Date: 2015-07-22
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The method of identifying whether a specific variety is a specific rice variety is limited to a method of SNP chips, but SNP chips are expensive, the analysis cycle is long, and subsequent data processing is cumbersome, and the test results cannot be obtained on the same day

Method used

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  • Identification method for hybrid rice variety hybrid source superior 69 based on InDel (insertion-deletion length polymorphism) marker
  • Identification method for hybrid rice variety hybrid source superior 69 based on InDel (insertion-deletion length polymorphism) marker
  • Identification method for hybrid rice variety hybrid source superior 69 based on InDel (insertion-deletion length polymorphism) marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1. Map-based cloning of rice functional genes

[0028] Using the primer pair used in the invention and a mutant positioning population constructed based on a japonica-indica hybrid to carry out map-based cloning of rice genes.

[0029] The amplification system used is: 20μL, including 30-50ng template DNA, 0.25 units of DNA polymerase (Sheneng Group), 2.0μL of 10×Buffer (containing Mg2+), 2.0mM dNTPs 2μL, 1.0μL of two Methyl sulfoxide (DMSO);

[0030] The amplification program used was: 95°C pre-denaturation for 5 minutes; 95°C denaturation for 30s, 56°C renaturation for 30s, 72°C extension for 30s, 32 cycles; finally 72°C for 10min extension.

[0031] Electrophoresis detection: add an appropriate amount of loading buffer to the amplified product described in step 2, under 22V / cM constant electric field intensity, after 100 minutes of 10% denaturing polyacrylamide gel electrophoresis, silver staining develops; contains rice When the DNA sample is combined with the afo...

Embodiment 2

[0032] Example 2: Identification of germplasm resources of japonica rice varieties and indica rice varieties

[0033] The following primer pairs of the invention were used to identify the rice variety Jiaoyuanyou 69.

[0034]

[0035]

[0036] InDel molecular markers were used to identify rice varieties Nipponbare, Jiaoyuan 5A, JP69 and Jiaoyuanyou 69.

[0037] The amplification system used is: 20μL, including 30-50ng template DNA, 0.25 units of DNA polymerase (Shenergy Group), 2.0μL of 10×Buffer (containing Mg 2+ ), 2.0mM dNTPs 2μL, 1.0μL dimethyl sulfoxide (DMSO);

[0038] The amplification program used was: 95°C pre-denaturation for 5 minutes; 95°C denaturation for 30s, 56°C renaturation for 30s, 72°C extension for 30s, 32 cycles; finally 72°C for 10min extension.

[0039] Electrophoresis detection: add an appropriate amount of loading buffer to the amplified product described in step 2, under 22V / cM constant electric field intensity, after 100 minutes of 10% denaturing polyacrylam...

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Abstract

The invention belongs to the technical field of creature identification, and particularly relates to an identification method for a hybrid rice variety hybrid source superior 69 based on an InDel (insertion-deletion length polymorphism) marker. In the method, the whole genome DNA sequence of a japonica rice variety Nipponbare and the whole genome DNA sequence of an indica type rice variety 93-11 are compared to obtain 14 pairs of specific DNA primers designed for an insertion / deletion differential fragment; the hybrid source superior 69 and parent hybrid sources 5A and JP69 of the hybrid source superior 69 are subjected to DNA extraction, DNA fragment amplification and electrophoretic separation, and the electrophoretogram is analyzed for identifying the hybrid source superior 69; to be specific, a finger-print spectrum obtained through polymerase chain reaction and gel vertical slab electrophoresis is analyzed by virtue of the combination of the 14 pairs of InDel primers, and further, whether the variety is the hybrid rice variety hybrid source superior 69 or not is determined according to the electrophoretic band type of 14 InDel sites. According to the invention, the sample amount required to be detected is small, the method is simple, convenient and quick, and the identification result is accurate, so that the method can be applied to the variety identification of the hybrid source superior 69 on the market.

Description

Technical field [0001] The invention belongs to the technical field of biological identification, and relates to a method for identifying hybrid rice variety Jiaoyuanyou 69 based on InDel markers, that is, a method for identifying hybrid rice variety Jiaoyuanyou 69 using rice DNA insertion or deletion molecular markers. Background technique [0002] Jiaoyuanyou 69 is a large-spike indica-japonica hybrid super high-yield variety, which was bred by the School of Life Science and Technology, Shanghai Jiao Tong University. It has important agricultural and commercial value. This variety is characterized by large panicle shape, dense grain, good grain filling, plant height about 129cm, compact plant shape, fertilizer tolerance, strong resistance, long flag leaves, suitable maturity, high quality rice, and total grains per panicle About 291.4 grains, the number of solid grains is about 258.2, the seed setting rate is about 88.6%, the 1,000-grain weight is about 25 grams, and the whole ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156C12Q2600/16
Inventor 张大兵吕阳袁政罗治靖陈明姣
Owner SHANGHAI JIAO TONG UNIV
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