Rapid method for identifying truth of maize variety

A technology for authenticity identification and variety, applied in the field of plant molecular biology, can solve the problems of cumbersome and time-consuming operation process, and achieve the effect of simple operation steps, stable quality and short duration.

Inactive Publication Date: 2009-07-01
BEIJING KINGS NOWER SEED S&T
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The use of SSR to identify the authenticity of corn varieties has been reported at home and abroad, such as Chinese patents 200310117180.3 and 20061006573.4, but the operation process of the prior art is cumbersome and time-consuming. To solve this problem, the inventor re-screened the primers and adopted an optimized TouchDown PCR program and DNA extraction method

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 Identify whether the female parent of the popular corn hybrid M is the inbred line N that has been applied for protection

[0019] The identification operation process is as follows:

[0020] (1) DNA extraction

[0021] Extraction of Genomic DNA from Sample M

[0022] The test materials are dry seeds, and only one dry seed is needed. Extract the embryos of dry seeds as follows: take 2 / 3 embryos of dry seeds and put them into a 0.5ml centrifuge tube, add 150μL 0.1mol / L NaOH boiling water for 5min, add 150μL 1×TE (pH 2.0) buffer after cooling to room temperature , and then placed at 4°C for later use;

[0023] Extraction of sample M pericarp DNA tissue:

[0024] ①Take 20 seeds of the hybrid species M, wash them with pure water, then wash them once with sterile water, soak them in 1.0% sodium hydroxide solution for about 20 minutes, take them out, and then rinse the seeds with pure water for 3 to 5 times; use tweezers carefully The pericarp of each seed was ...

Embodiment 2

[0037] Example 2 Identify whether the parental inbred line P being used by a company is the inbred line Q that has been applied for protection. The identification operation process is as follows:

[0038] (1) DNA extraction

[0039] Extraction of sample P, Q genomic DNA

[0040]The test materials are dry seeds, and only one dry seed is needed. Extract the embryos of dry seeds as follows: take 2 / 3 embryos of dry seeds and put them into a 0.5ml centrifuge tube, add 150μL 0.1mol / L NaOH boiling water for 5min, add 150μL 1×TE (pH 2.0) buffer after cooling to room temperature , mixed by pipetting and placed at 4°C for later use.

[0041] (2) Primer synthesis

[0042] 合成如下20对核心引物:bnlg1866,phi001,bnlg1017,umc1042,phi029,bnlg1108,bnlg2162,umc2188,bnlg1006,bnlg602,umc1023,phi031,umc1406,bnlg1792,bnlg1131,dupssr14,umc1636,bnlg1129,phi041,umc1291。 This example was synthesized by Beijing Aoke Biotechnology Co., Ltd., and each tube of primers is 4.0 OD.

[0043] (3) PCR amplification ...

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PUM

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Abstract

The invention relates to a rapid identification method for truth of corn varieties. In the method, an embryo of a single dry seed is adopted to extract DNA of a genome of a sample to be detected, and the method is time-saving and ensures that the extracted genome has high and steady quality; 20 pairs of SSR molecular marker core primers are used, have rich polymorphism and steady banding patterns, and are evenly distributed on 10 chromosomes; and the method also adopts an optimized TouchDown PCR amplification program. The rapid identification method for the truth of the corn varieties has low design cost for the primers, is simple in operation and time-saving, can conveniently and quickly identify the authenticity of the corn varieties, and is particularly suitable for material identification of a large amount of samples.

Description

technical field [0001] The invention belongs to the field of plant molecular biology, and relates to a method for quickly identifying the authenticity of corn varieties, in particular to a method for quickly identifying the authenticity of corn varieties by using SSR molecular markers. Background technique [0002] The protection of intellectual property rights of new plant varieties is a powerful measure to encourage breeders to keep cultivating new varieties (lines) or hybrids. It is very necessary to develop high-discrimination identification technology of corn materials for the authenticity identification of corn, as well as the registration of new varieties and the protection of property rights registration. [0003] The ideal variety identification technology should not only be accurate and reliable, but also simple, fast and economical, which is the prerequisite for the industrialization of variety identification. The traditional morphological identification methods ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 孔荣张红伟莫云谭振波李明源王晓娜刘玲霞
Owner BEIJING KINGS NOWER SEED S&T
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