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Method and kit for authenticating variety of castor seed by means of dimolecular markers

A technology for castor seeds and authenticity, which is applied in the directions of biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of few castor genotypes and low genome diversity, achieve accurate results and improve identification accuracy. , good repeatability

Active Publication Date: 2012-04-25
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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Problems solved by technology

However, the common problem encountered by these molecular markers is that the genome diversity is low. On the one hand, there are fewer genotypes in castor, and on the other hand, it is related to the defects of various molecular markers

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  • Method and kit for authenticating variety of castor seed by means of dimolecular markers
  • Method and kit for authenticating variety of castor seed by means of dimolecular markers
  • Method and kit for authenticating variety of castor seed by means of dimolecular markers

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Embodiment Construction

[0036] The embodiments of the present utility model are described in further detail below in conjunction with the accompanying drawings; it should be noted that: the present embodiments are descriptive, not restrictive, and cannot be used to limit the protection scope of the present utility model.

[0037] The main principle of the present invention is: by screening 20 AP-PCR primers and 84 pairs of RMAPD primers, the best primer combination with clear bands and high polymorphism is obtained and synthesized by a relevant biotechnology company. Different parts of the genome of each castor variety were marked by AP-PCR and RMAPD, and then combined analysis was performed to obtain the map of known varieties, and then the authenticity and purity of unknown varieties were identified.

[0038] The castor varieties described in this example are all common commercial varieties at home and abroad, numbered 1 to 31, and number 10 is the variety to be tested.

[0039] A method for identify...

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Abstract

The invention relates to a method and kit for authenticating the variety of a castor seed by means of dimolecular markers. The method comprises the following steps: (1) extracting the genome DNA (deoxyribonucleic acid) of the castor seed; (2) using the genome DNA obtained in the step (1) as the template to carry out AP-PCR (arbitrarily primed polymerase chain reaction) amplification, wherein the primer is shown in sequence 1; (3) carrying out gel electrophoresis on the AP-PCR amplification product obtained in the step (2); (4) using the genome DNA obtained in the step (1) as the template to carry out RMAPD (random microsatellite amplification polymorphic DNA) amplification, wherein the primers are shown in sequence 2 and sequence 3; (5) carrying out polyacrylamide gel electrophoresis on the RMAPD amplification product obtained in the step (4); and (6) comparing the bands of the unknown variety with a data base of known varieties to judge the variety of the castor seed. By using the self-developed kit and the laboratory conventional PCR (polymerase chain reaction) technology and electrophoretic technology, the whole process from sampling to detection can be finished in about 7 hours, and the method can be implemented throughout the whole year. Besides, the invention has the advantages of high simplicity and practicability, high speed, high sensitivity and accurate result.

Description

technical field [0001] The invention belongs to the field of oil crop seed screening and identification, in particular to a method and a kit for identifying the authenticity of castor seed varieties by using bimolecular markers. Background technique [0002] Castor (Ricinus communis L.) is a dicotyledonous annual or perennial herb belonging to the genus Ricinus in the Euphorbiaceae family. It is one of the top ten oil crops in the world and has significant planting and industrial value. Castor is planted all over the world, among which countries such as India, China and Brazil have the most extensive planting areas. There are many varieties of castor. As of November 2011, there are 1,430 castor varieties in my country's castor planting resource database. With the advancement of breeding technology, new castor varieties are constantly emerging. The variety of varieties brings certain difficulties to the identification of varieties. At present, the commonly used methods for ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 王昌禄何东叶锋王玉荣陈勉华李风娟李贞景杨朝晖李伟
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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