A method for identification of loquat varieties based on est-ssr markers

A variety, loquat technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as poor judgment of primers for distinguishing varieties, lack of timeliness, and lack of intuition, so as to achieve early identification, extensive The effect of versatility and easy operation

Inactive Publication Date: 2015-10-21
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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Problems solved by technology

The amount of operation in 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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  • A method for identification of loquat varieties based on est-ssr markers
  • A method for identification of loquat varieties based on est-ssr markers
  • A method for identification of loquat varieties based on est-ssr markers

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Embodiment

[0041] A method for identifying loquat varieties based on EST-SSR markers, the specific steps are as follows:

[0042] (1) Extract DNA:

[0043] The loquat samples in the examples of the present invention are all common and important loquat varieties. These 18 loquat varieties mainly refer to "Chinese Fruit Trees·Longan Loquat Volume" (edited by Qiu Wuling and Zhang Huizhi, published by China Forestry Publishing House), other parts The variety is a loquat germplasm newly collected in recent years by the Institute of Horticulture, Zhejiang Academy of Agricultural Sciences.

[0044] In this step, young loquat leaves are used as materials, and the sample DNA is extracted by the traditional CTAB method. The extracted loquat young leaf genomic DNA was detected by 0.8% agarose gel electrophoresis, and the concentration and purity of the DNA were detected by the Bio-Photometer nucleic acid detector produced by Eppendorf. The final concentration was diluted to 30-50ng / μL for PCR Amplificat...

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Abstract

The invention discloses a method for identifying loquat varieties based on an EST-SSR (expressed sequence tag-simple sequence repeat) marker, belonging to the field of molecular biology molecular markers. The method comprises the following steps: designing SSR primers specific to an EST sequence of loquat; screening to obtain five pairs of primers for variety identification; extracting DNA (deoxyribonucleic acid) of the loquat varieties; amplifying the DNA of the loquat varieties by using the primers one by one; distinguishing the obtained varieties having unique specific bands, and establishing a tree type identification chart. According to the method, the primers are obtained by strict screening, and 18 loquat varieties can be distinguished via five pairs of EST-SSR primers, so that convenience, rapidness and accuracy in operation are achieved. By adopting the method disclosed by the invention, 18 loquat varieties can be distinguished rapidly and accurately under any situation, the tree type identification chart is more intuitive than a cluster tree, and primers capable of distinguishing any two varieties can be found according to the tree type identification chart.

Description

Technical field [0001] The invention relates to the field of molecular markers for molecular biology, in particular to a method for identifying loquat varieties based on EST-SSR markers. Background technique [0002] Loquat (Eriobotrya japonica Lindl, 2n=2x=34) is a plant belonging to the genus Rosaceae, Maloideae, and is widely planted in the subtropical regions of my country and other Asian countries. Loquat originated in the southeast of my country (Lin and Hu, 2000). It has been cultivated in my country for more than two thousand years and has abundant germplasm resources. Loquat is a subtropical evergreen fruit tree. According to the characteristics of the fruit, loquats can be divided into red sand and white sand varieties (Fu et al., 2012); mature loquat fruits are mostly spherical or oval, soft and juicy, rich in glutamic acid , Phenolics (especially flavonoids), vitamins and carotenoids and other nutrients are important health fruits (Cao et al., 2009; Fu et al., 2011)....

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6895C12Q2600/156
Inventor 李晓颖陈俊伟徐红霞
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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