Method for constructing lichi core collection by using EST-SSR (Expresses Sequence Tag-Simple Sequence Repeat) molecular marker

A core germplasm and construction method technology, applied in DNA preparation, recombinant DNA technology, etc., can solve the problems of lack of litchi core germplasm and molecular fingerprint construction methods, achieve stable results, clear amplification bands, and improve utilization efficiency Effect

Inactive Publication Date: 2011-09-07
POMOLOGY RES INST GUANGDONG ACADEMY OF AGRI SCI
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Problems solved by technology

[0007] At this stage, there is a lack of methods for constr

Method used

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  • Method for constructing lichi core collection by using EST-SSR (Expresses Sequence Tag-Simple Sequence Repeat) molecular marker
  • Method for constructing lichi core collection by using EST-SSR (Expresses Sequence Tag-Simple Sequence Repeat) molecular marker
  • Method for constructing lichi core collection by using EST-SSR (Expresses Sequence Tag-Simple Sequence Repeat) molecular marker

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Embodiment Construction

[0022] Below in conjunction with embodiment, further illustrate the present invention.

[0023] 1 Materials and methods

[0024] 1.1 Plant material :

[0025] The 96 litchi varieties used in this experiment were all taken from the National Litchi Resource Garden of the Institute of Pomology, Guangdong Academy of Agricultural Sciences. When sampling, the young leaves with strong growth and no leaves of diseases and insect pests were selected, put into fresh-keeping bags, and then refrigerated. After taking it out, take it back to the laboratory quickly, and store it in an ultra-low temperature freezer at -80°C for later use.

[0026] 1.2 Method

[0027] 1.2.1 Extraction of DNA

[0028] Genomic DNA was extracted by the improved SDS method (Chen Liang et al., 1997) (Chen Liang, Chen Daming, Gao Qikang et al. Purification and Identification of Tea Plant Genomic DNA, Tea Science, 1997, 17(2):172-176). Measure its ultraviolet absorption spectrum at 230nm, 260nm, and 280n...

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Abstract

The invention discloses a method for constructing lichi core collection, which comprises the following steps: selecting tender lichi leaves, and extracting genome DNAs according to varieties to obtain genome DNA samples of lichi; searching EST-SSR (Expresses Sequence Tag-Simple Sequence Repeat) of the lichi according to an EST sequence of the lichi, and designing an EST-SSR primer; carrying out PCR (Polymerase Chain Reaction) by using the EST-SSR primer to amplify the genome DNAs; carrying out electrophoretic analysis on PCR products to obtain existence data of alleles of the PCR products; and analyzing the obtained data to construct the core collection. In the method disclosed by the invention, the processes of EST-SSR-marked primer formation and experiment operation are simple and are easy to operate, an amplification band is clear, results are stable, and the method is suitable for analyzing genetic diversity among lichi resources, the core collection constructed on the basis almost represents all the genetic diversity of the original germplasm and has better representativeness, and the method has realistic significances for lichi germplasm management, new germplasm collection and germplasm reproduction and renewal and has great importance for promoting the exploration in the utilization of the lichi germplasm resources, so that the utilization efficiency of the lichi germplasm resources is improved.

Description

technical field [0001] The invention relates to a method for constructing litchi core germplasm. Background technique [0002] litchi( Litchi chinensis Sonn.) for Sapindaceae ( Sapindaceae ) Litchi ( Litchi ) plant, native to southern my country, is an important tropical and subtropical fruit, and one of the oldest cultivated fruit trees in the world. It is distributed in the tropical and subtropical regions of the eastern and western hemispheres. Litchi in my country is mainly distributed at 18° to 24° north latitude The main cultivation provinces are Guangdong, Guangxi, Fujian, Hainan, Taiwan, and Yunnan in the tropical and subtropical regions of 30′. In addition, there are also a small amount of cultivation in some special microclimate areas of Sichuan, Guizhou, and Zhejiang (Zhang Zhanwei et al., 1997). Wu Shuxian et al. (1996) conducted statistics on litchi germplasm resources in various places and believed that 222 litchi germplasms have been found in my country, ...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 向旭欧良喜孙清明袁沛元邱燕萍陈洁珍李志强蔡长河
Owner POMOLOGY RES INST GUANGDONG ACADEMY OF AGRI SCI
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