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Method for inspecting purity of two-line hybrid rice seeds by utilizing SSR (Simple Sequence Repeat) method

A technology for purity inspection and hybrid rice, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., and can solve problems such as not being able to meet the needs of seed production and management.

Inactive Publication Date: 2011-10-19
HEFEI FENGLE SEED +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, this method cannot well meet the needs of seed production and management.

Method used

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  • Method for inspecting purity of two-line hybrid rice seeds by utilizing SSR (Simple Sequence Repeat) method
  • Method for inspecting purity of two-line hybrid rice seeds by utilizing SSR (Simple Sequence Repeat) method
  • Method for inspecting purity of two-line hybrid rice seeds by utilizing SSR (Simple Sequence Repeat) method

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Embodiment Construction

[0028] A method for testing the purity of Fengliangyou No. 1 or Fengliangyouxiang No. 1 two-line hybrid rice seeds by SSR method, which is characterized in that the specific steps are:

[0029] 1) Extraction of sample DNA: After soaking the seeds for 24 hours, take the sprouts 4-5 days after germination, cut the sprouts about 1 cm, and put them into 96-well PCR tubes, and add 40 μL 0.30mol / L NaOH to each tube The solution was boiled in boiling water for 45 sec, then 40 μL of 0.17 mol / L Tris-HCI (pH 3) was added, boiled in boiling water for 2 minutes, then cooled on ice, and stored at 4°C for later use.

[0030] 2) PCR amplification

[0031] ①Reaction system

[0032] The reaction system consists of sample DNA extract, buffer, dNTP, primer, ddH 2 O and Taq enzyme composition. The volume ratio is 1:0.8:1:0.5:6.5:0.2. Among them, the primer sequences used for the purity test of Fengliangyou No. 1 are: P1F: 5'-CTTACAGAGAAACGGCATCG-3', P1R: 5'-GCTGGTTTGTTTCAGGTTCG-3'. The primer sequence...

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Abstract

The invention discloses a method for inspecting purity of two-line hybrid rice seeds by utilizing an SSR (Simple Sequence Repeat) method, in particular a method for inspecting the purity of Fengliangyou No. 1 rice or Fengliangyouxiang No. 1 rice seeds by utilizing SSR molecular markers. The method comprises the following steps of: performing amplification and agarose electrophoretic separation on genome DNA (Deoxyribonucleic Acid) of the two-line hybrid rice varieties Fengliangyou No. 1 and parents thereof or Fengliangyouxiang No. 1 and parents thereof respectively by utilizing primers P1 and P2 to find out a characteristic spectrum band of difference between a first-filial generation and the parents; inspecting the samples of Fengliangyou No. 1 or the Fengliangyouxiang No. 1 by calculating the purity of the inspected seeds; and comparing and contrasting the inspected purity with purity tested by field planting to confirm that the results of the two are highly consistent. The method is suitable for inspecting the authenticity and variety purity of the Fengliangyou No. 1 or Fengliangyouxiang No. 1 two-line hybrid rice and parents thereof.

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 rice seeds, in particular to a method for inspecting the purity of Fengliangyou No. 1 or Fengliangyouxiang No. 1 two-line hybrid rice seeds by using SSR markers. Background technique [0002] my country's first two-line hybrid rice technology has been widely used in a large area after years of research. However, since the female parent of the two-line hybrid rice is a light- and temperature-sensitive genic male sterile line, it is easily affected by the external environment. According to research, when the ambient temperature is lower than 23.5 ℃, the fertility of the sterile line begins to transform and can automatically Cross-bearing, affect the purity of hybrid seeds. In order to ensure the safety of Fengliangyou No. 1 or Fengliangyouxiang No. 1 two-line hybrid rice seed, the purity testing must be strengthened. [0003] Cu...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 周桂林陈庆全王兆贤蒋继武高前宝朱先飞汪华春程国旺
Owner HEFEI FENGLE SEED
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