Inspection method for purity of melon seeds

A technology for purity inspection and seeds, which is applied in the field of purity inspection of the seeds of Melon Jinlu No. 3 by using SSR molecular markers, can solve the problems of high cost, unsuitable for the development of modern seed industry, incomplete emasculation of the female parent, etc.

Inactive Publication Date: 2014-03-12
HEFEI FENGLE SEED
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Problems solved by technology

During the process of hybrid seed production, the female parent needs to be detasselled artificially, and the male parent pollen is given to complete the hybridization process. Incomplete or missing emasculation of the female parent will greatly affect the purity of the seed production.
The traditional hybrid purity detection method is planting identification, which depends on the phenotypic differences of varieties, and is identified through the comparison of the appea

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  • Inspection method for purity of melon seeds

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[0030] A method for seed purity inspection of Muskmelon Jinlu No. 3 is performed according to the following steps:

[0031] 1) DNA extraction:

[0032] Put melon Jinlu No. 3 seeds in a nutrient bowl to germinate and germinate. When the seedlings have 2-3 true leaves, take 3cm 2 Place the left and right leaves in a small mortar, add 700ul of 4×CTAB extraction buffer to grind them, transfer them to a 1.5ml centrifuge tube, and bath them in a water bath at 65°C for half an hour;

[0033] Add the same volume of phenol-chloroform-isoamyl alcohol mixture as the extraction buffer, shake it by hand for 10 minutes, centrifuge at 12000 rpm for 10 minutes, aspirate 300ul of the supernatant in another centrifuge tube, add 700ul of frozen absolute ethanol, -20℃ Deposit DNA in the freezer for 60 minutes;

[0034] Centrifuge at 12000rpm for 10 minutes, discard the supernatant; add 400ul of 75% ethanol, let stand for 4 minutes to remove the salt, and discard the ethanol;

[0035] The centrifuge tube i...

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Abstract

The invention discloses an inspection method for the purity of melon seeds, especially an inspection method for the purity of Jinlu No. 3 melon seeds by using SSR molecular markers. The method utilizes a pair of SSR primers, i.e., FLWSSRL and FLWSSRR, for amplification and agarose gel electrophoresis separation of male and female parents of a hybrid melon variety, i.e., the Jinlu No. 3, and the genome DNA of a first-filial generation; characteristic bands of difference between the first-filial generation and the male and female parents are found out; inspection is carried out on seed samples of the first-filial generation of the Jinlu No. 3 through calculation of the purity of to-be-inspected seeds; and inspected purity is compared with purity obtained in field planting identification, and results show that the inspected purity is highly identical to field planting identification purity. The method is applicable to inspection of authenticity of the Jinlu No. 3 and parents thereof and inspection of seed purity.

Description

technical field [0001] The invention belongs to the technical field of seed quality inspection, and relates to a method for inspecting the purity of hybrid melon seeds, in particular to a method for inspecting the purity of melon Jinlu No. 3 seeds by using SSR molecular markers. Background technique [0002] Jinlu No. 3 is a new high-yield and high-quality hybrid melon variety bred by Hefei Fengle Seed Industry Co., Ltd. (see the "Registration Form for Disclosure of Genetic Resources Sources" for details), and has excellent field agronomic characteristics. During the process of hybrid seed production, the female parent needs to be artificially detasselled, and the male parent pollen is given to complete the hybridization process. Incomplete or missing emasculation of the female parent will greatly affect the purity of the seed production. The traditional hybrid purity detection method is planting identification, which depends on the phenotypic differences of varieties, and i...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6895C12Q2600/156
Inventor 周贤达王兆贤陈会中王浩波吴义兵周桂林林茂朱先飞徐丽媛王丽梅
Owner HEFEI FENGLE SEED
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