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A method for testing the purity and consistency of conventional rice varieties

A technology of species purity and consistency, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of large workload, low efficiency and high cost, and achieve the effect of overcoming high cost

Inactive Publication Date: 2019-12-31
INST OF QUALITY STANDARD & DETECTION TECH YUNNAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Based on the technical problems existing in the background technology, the main purpose of the present invention is to overcome the problems of heavy workload, high cost and low efficiency of the single plant determination method using SSR markers in the current variety purity and consistency identification, and provide a fast, simple and convenient method. Method for Accurately Testing the Purity and Consistency of Conventional Rice Varieties

Method used

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  • A method for testing the purity and consistency of conventional rice varieties
  • A method for testing the purity and consistency of conventional rice varieties

Examples

Experimental program
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Effect test

Embodiment 1

[0023] The plants of the conventional rice variety Chujing 27 were planted in 10 rows×10 columns, and each individual plant was positioned and numbered according to the rows and columns; , width are 1cm leaf tissue, after mixing, DNA is extracted according to the conventional CTAB method, and it is recorded as DNAa; the length and width of the first leaf middle section of each rice plant in column b are cut from the leaf tissue that is 1 cm in length and width, and after mixing The DNA was extracted according to the conventional CTAB method, which was recorded as DNAb, and the 12 SSR markers in group A published in the agricultural industry standard "SSR Marker Method for Identification of Rice Varieties" (NY / T 1433-2014) were used to amplify the mixed extracted DNA , and the amplified products were electrophoresed and stained. Specific amplification primers are as follows:

[0024]

[0025]

[0026] After analyzing the SSR amplification products, it was found that the ...

Embodiment 2

[0031] The plants of the conventional rice variety Xiujing 12 were planted in 10 rows×10 columns, and each individual plant was positioned and numbered according to the rows and columns; , width are 1cm leaf tissue, after mixing, DNA is extracted according to the conventional CTAB method, and it is recorded as DNAa; the length and width of the first leaf middle section of each rice plant in column b are cut from the leaf tissue that is 1 cm in length and width, and after mixing The DNA was extracted according to the conventional CTAB method, which was recorded as DNAb, and the 12 SSR markers in group A published in the agricultural industry standard "SSR Marker Method for Identification of Rice Varieties" (NY / T 1433-2014) were used to amplify the mixed extracted DNA , and the amplified products were electrophoresed and stained.

[0032] After analyzing the SSR amplification products, it was found that primers RM583 and RM19 amplified heterotypic bands in row 5 (R5) and column ...

Embodiment 3

[0037] Plant the plants of the conventional rice variety Yunjingyou No. 1 in 10 rows×10 columns, and position and number each individual plant according to the rows and columns; when the rice grows to the booting stage, cut the middle part of the first leaf of each rice plant in row a The length and width are 1cm leaf tissue, after mixing, DNA is extracted according to the conventional CTAB method, and it is recorded as DNAa; the length and width of the first leaf middle section of each rice plant in column b are cut from the leaf tissue of 1 cm in length, After mixing, the DNA was extracted according to the conventional CTAB method, which was recorded as DNAb, and the 12 SSR markers of group A published in the agricultural industry standard "SSR Marker Method for Identification of Rice Varieties" (NY / T 1433-2014) were used to carry out the DNA extraction. Amplify, and electrophoresis and stain the amplified product.

[0038] After analyzing the SSR amplification products, it ...

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Abstract

The invention discloses a method for detecting the purity and consistency of conventional rice varieties, comprising the following steps: planting rice of the same variety in the form of an array, wherein any rice plant is marked as R a C b When rice grows to booting stage, get the leaf tissue of every rice plant in the same row, get the leaf tissue of every rice plant in the same column again, extract DNA according to routine CTAB method after mixing; Carry out SSR-PCR amplification to obtain amplified Amplify the product, then perform electrophoresis separation and staining; according to the band type of the amplified product, amplify two or more pairs of SSR primers to obtain the first heterotypic band a line and number b The intersection point of the column was determined as a suspected heterotypic strain; then the true heterotype was determined according to the size of the heterotypic band on the same primer in the mixed sample or according to the SSR fingerprint of the individual plant corresponding to the intersection point. Compared with the rice variety consistency test based on phenotypic traits, the invention is simpler and faster, reduces workload, improves efficiency and reduces cost.

Description

technical field [0001] The invention relates to the technical field of variety detection, in particular to a method for detecting the purity and consistency of conventional rice varieties. Background technique [0002] Variety purity is an important index to measure the quality of a variety. It refers to the degree of typical consistency of varieties in terms of characteristics, that is, the percentage of the number of plants with typical characteristics of this variety in the number of plants of the crop sample for inspection. It is consistent with the out-of-shape rate expression in the consistency determination in the variety DUS test. With the rapid development of molecular marker technology research, DNA detection methods have been widely used in the identification of variety purity. The International Union for the Protection of New Varieties of Plants (UPOV) recommended SSR simple sequence repeat markers (SSR markers) and single nucleotide polymorphism markers (SNP ma...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895
Inventor 刘艳芳张建华黄清梅杨晓洪王江民李彦刚张惠张鹏管俊娇
Owner INST OF QUALITY STANDARD & DETECTION TECH YUNNAN ACAD OF AGRI SCI
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