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DNA fingerprint spectrum of eggplant varieties as well as establishment method and application of spectrum

A DNA fingerprinting and fingerprinting technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of eggplant seeds being easily mixed, time-consuming and labor-intensive, and difficult to identify, so as to protect legitimate rights and interests and achieve results. Accurate and easy to promote the application effect

Active Publication Date: 2018-03-06
SHANGHAI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in the process of hybrid seed production, eggplant seeds are easily mixed due to artificial castration and pollination, and the plant morphology of some varieties is very close, so it is difficult to identify
If judged by the phenotypic characteristics of eggplant, the identification period is long and time-consuming and labor-intensive; DNA molecular markers have been widely used in the quality detection of crop seeds due to their high individual specificity and environmental stability.

Method used

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  • DNA fingerprint spectrum of eggplant varieties as well as establishment method and application of spectrum
  • DNA fingerprint spectrum of eggplant varieties as well as establishment method and application of spectrum
  • DNA fingerprint spectrum of eggplant varieties as well as establishment method and application of spectrum

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Experimental program
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Effect test

Embodiment 1

[0050] 1. Materials

[0051] The eggplant hybrid variety "Huqie No. 5" and its parents "Fu-2" and "Jiangqie" were provided by the Institute of Horticulture, Shanghai Academy of Agricultural Sciences.

[0052] 2. Reagents

[0053] The extracted DNA and PCR reagents used in the experiment were purchased from Shanghai Sangon Bioengineering Co., Ltd.

[0054] 3. Method

[0055] 3.1 DNA extraction and detection

[0056] DNA extraction steps using CTAB micro method:

[0057] (1) Add 1 to 2 pieces of fresh leaves the size of the cap of the centrifuge tube into a 2ml centrifuge tube;

[0058] (2) Add 200ul CTAB buffer solution and grind for 5min with a grinder;

[0059] (3) After grinding, add 500ul CTAB buffer;

[0060] (4) Water bath at 65°C for 1 hour;

[0061] (5) Add 700 μl of chloroform / isoamyl alcohol with a volume ratio of 24:1, and mix up and down for 15 minutes to ensure that the sample and chloroform are fully mixed;

[0062] (6) Centrifuge at 12000rpm for 10min;

[...

Embodiment 2

[0120] The DNA fingerprint spectrum that the present invention establishes is used for the identification of the hybrid variety purity of "Huqie No. 5":

[0121] Randomly select 120 germinated hybrids and 2 parents each, extract DNA with micro-CTAB method, use the extracted DNA as a template, amplify with primer SSR116, and analyze the amplified products by electrophoresis on 6% PAGE. If the DNA fingerprint of a single plant detected by SSR116 marker analysis is 1111111, it is the true hybrid of "Huqie No. 5", and if the DNA fingerprint is 1011111, it is the female parent "Jiangqi" inbred, which is a false hybrid . Then divide the number of false hybrids by the total number of samples with bands, and you can get the purity of the eggplant hybrids produced in the seed production base. It is time-saving and convenient to investigate the purity of larger field planting, and can provide timely and reliable information for the sale of eggplant hybrid seeds. Purity Information. If...

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Abstract

The invention discloses a method for establishing a DNA fingerprint spectrum of eggplant varieties. The method comprises the following steps: extracting eggplant genome DNA; performing SSR primer amplification and screening; performing PCR (Polymerase Chain Reaction) amplification and electrophoresis detection of the product; establishing DNA digital fingerprinting, namely the fingerprint spectrum; and establishing digital two-dimensional codes. The invention further provides a DNA fingerprint spectrum established according to the method and application of the spectrum in purity identificationof seeds of a hybrid eggplant 'Hu Eggplant V'. The method for identifying the seeds by utilizing the DNA fingerprint spectrum of the variety and the digital two-dimensional codes is a molecular marker method by taking polymorphism of DNA as basis, and multiple defects and difficulties in morphological identification of the seed purity as well as isozyme and seed protein electrophoresis identification can be overcome and solved. Specifically, the DNA fingerprint spectrum and digital two-dimensional codes of the facility cultivation variety 'Hu Eggplant V'and parents 'Fu-2' and 'Jiang Eggplant'are established by utilizing the SSR technology, and a scientific basis can be provided for identification of the variety and the purity detection.

Description

technical field [0001] The invention belongs to the technical field of eggplant variety purity identification, and relates to an eggplant variety DNA fingerprint and its establishment method and application. Background technique [0002] Eggplant (Solanum melongena L.) is a cultivated species in the genus Solanum of Solanaceae. It is one of the important vegetable crops, and its hybrids are widely cultivated at home and abroad. As the key to seed quality, the purity of hybrids is closely related to the interests of breeding units and farmers. However, in the process of hybrid seed production, eggplant seeds are easily mixed due to artificial castration and pollination, and the plant morphology of some varieties is very close, so it is difficult to identify them. If judged by the phenotypic characteristics of eggplant, the identification cycle is long and time-consuming; while DNA molecular markers have been widely used in the quality detection of crop seeds because of their...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6858
CPCC12Q1/6858C12Q1/6895C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 吴雪霞查丁石朱宗文张爱冬李贤
Owner SHANGHAI ACAD OF AGRI SCI
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