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Primer and method for fast differentiation of walnut varieties

A variety and walnut technology, applied in the field of primers for quickly distinguishing walnut varieties, can solve the problems of poor judgment of primers for distinguishing varieties, lack of timeliness, not intuitive enough, etc., to achieve early identification, wide versatility, and convenient operation. Effect

Inactive Publication Date: 2012-07-11
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This patent uses a lot of primers, and the amount of operation in the quality identification is very large, and it is not time-sensitive. It is not easy to judge the primers to distinguish varieties through the clustering tree, and it is not intuitive enough.

Method used

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  • Primer and method for fast differentiation of walnut varieties
  • Primer and method for fast differentiation of walnut varieties
  • Primer and method for fast differentiation of walnut varieties

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Embodiment Construction

[0036] The present invention will be further described below in conjunction with the accompanying drawings.

[0037] 1. Extraction of DNA

[0038] The walnut samples of the present invention are common important walnut varieties in the market, and these 20 walnut varieties include: Daifeng, N8-19, Luhe No. 1, S11-1, YZ 1-6, Heart-shaped walnut, Fenghui, N1 -2, Luguang, N10-15, Zhonglin 1, Luguo 2, Yuanfeng, Italy, S13-7, N12-6, S11-8, Liaoning 2, S1-19, Daixiang, the above varieties Mainly refer to "Chinese Fruit Tree Records Walnut Volume", edited by Xi Rongting and others, published by China Forestry Press, and some varieties are newly bred varieties by Shandong Fruit Tree Institute in recent years.

[0039] Using young walnut leaves as materials, on the basis of the traditional DNA extraction method-CTAB method, adding phenols and antioxidants (β-mercaptoethanol) to effectively remove protein and phenolic substances, using this improved CTAB method to extract walnut genomi...

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Abstract

The invention discloses a primer and a method for fast differentiation of walnut varieties, belonging to the field of molecular markers of molecular biology. The method comprises the following steps of: designing RAPD (random amplified polymorphic DNA (deoxyribonucleic acid)) random primers against a walnut gene sequence, screening, then getting two random primers, extracting the DNA of the walnut varieties, further utilizing the primers to amplify the DNA of the unknown walnut varieties one by one, differentiating out the varieties with unique specific spectrum bands, and establishing a tree-shaped identification diagram. The primers disclosed by the invention are obtained by strict screening, the stability of the RAPD reaction is improved in a targeted manner, 20 varieties of the walnuts can be easily differentiated only through the two primers, and the operation is convenient, fast and accurate. According to the method disclosed by the invention, 20 walnut varieties can be fast and accurately differentiated under any situations, the tree-shaped identification diagram is more intuitive than a cluster tree, and the primers which can differentiate any two varieties can be found out according to the tree-shaped identification diagram.

Description

technical field [0001] The invention relates to the field of molecular biology molecular markers, in particular to a primer and a method for quickly distinguishing walnut varieties. Background technique [0002] Walnut (Juglans regia L.) is one of the four largest dried fruits in the world. The distribution of walnuts is vast. Except for the severe cold regions in the north, there are no or very few walnuts in Taiwan, Guangdong, Jiangxi and southern Fujian, there are certain cultivation areas in other places. Due to the continuous development and expansion of the walnut industry in recent years, the promotion and deepening of walnut clone varieties, and the destruction of the resource environment, walnut varieties tend to develop towards a single aspect, resource confusion often occurs in resource conservation, germplasm The importance of resource research has attracted more and more attention. Therefore, the establishment of a fast and reliable identification technology s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 韩键张美勇徐颖相昆上官凌飞王晨
Owner NANJING AGRICULTURAL UNIVERSITY