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Fingerprint spectrum of cauliflower hybrid and construction method thereof

A fingerprint and construction method technology, which is applied in the field of cauliflower hybrid fingerprints and its construction, can solve the problems of severe cauliflower diseases and restrictions on large-scale cultivation of cauliflower, and achieve the effect of protecting interests and saving manpower and material resources

Inactive Publication Date: 2015-04-29
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to serious diseases on cauliflower, especially black rot and downy mildew, the large-scale planting of cauliflower is restricted.

Method used

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  • Fingerprint spectrum of cauliflower hybrid and construction method thereof
  • Fingerprint spectrum of cauliflower hybrid and construction method thereof
  • Fingerprint spectrum of cauliflower hybrid and construction method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The specific steps of the unique SSR primer screening in the present invention are as follows:

[0021] (1) Use the parents of the cauliflower hybrid "Shenxue 108 days" as the experimental material, the cytoplasmic male line AW-100 as the female parent, and the inbred line C-3 as the male parent, referring to the SSR analysis method of Uzunova and Ecke (1999) , optimize the SSR molecular labeling reaction system, the SSR molecular labeling reaction system is as follows: MgCl 2 , 2.5mmol·L -1 , dNTPs 0.2mmol L -1 , 100ng SSR primers, 0.1U Taq polymerase, 1μl 10×PCRBuffer, 35ng "Shenxue 108 days" DNA template;

[0022] (2) Using the parents of the cauliflower hybrid "Shenxue 108 days" as experimental materials, using the optimized SSR molecular marker reaction system, 26 pairs of polymorphic SSR marker primers were screened, including BRMS05, BRMS06, BRMS08, BRMS15 , BRMS16, BRMS25, BRMS30, BRMS36, BRMS42-2, BRMS54, BRMS71, BRMS296, BRMS324, Bn38A, Bn59A, ENA4, ENA7, E...

Embodiment 2

[0028] (1) Material

[0029] This example uses "Shen Xue 108 Days" and "COMS05×B1" as experimental materials.

[0030] (2) Experimental method

[0031] SSR analysis method: refer to Uzunova and Ecke (1999). The primers were BRMS05 primers and BRMS36 primers, which were synthesized by Shanghai Bioengineering Company.

[0032] SSR molecular labeling reaction system is as follows: MgCl 2 , 2.5mmol·L -1 , dNTPs 0.2mmol L -1 , SSR primer 100ng, Taq polymerase 0.1U, 10×PCRBuffer 1μl, DNA template 35ng.

[0033] PCR amplification program: pre-denaturation at 94°C for 2 min, 1 cycle; denaturation at 94°C for 1 min, annealing at 63°C for 1 min, extension at 72°C for 1 min, a total of 30 cycles; extension at 72°C for 10 min, 1 cycle.

[0034] Separation was carried out by vertical electrophoresis on 6% polyacrylamide gel, silver staining was performed on the electrophoresis gel plate, rinsed with water, dried, photographed and recorded.

[0035] (3) Drawing

[0036] At the migra...

Embodiment 3

[0038] (1) Material

[0039] In this example, "Shenxue 108 Days" and "COMS05×W3" are used as experimental materials.

[0040] (2) Experimental method

[0041] SSR analysis method: refer to Uzunova and Ecke (1999). The primers were BRMS05 primers and BRMS36 primers, which were synthesized by Shanghai Bioengineering Company.

[0042] SSR molecular labeling reaction system is as follows: MgCl 2 , 2.5mmol·L -1 , dNTPs 0.2mmol L -1 , SSR primer 100ng, Taq polymerase 0.1U, 10×PCRBuffer 1μl, DNA template 35ng.

[0043] PCR amplification program: pre-denaturation at 94°C for 2 min, 1 cycle; denaturation at 94°C for 1 min, annealing at 63°C for 1 min, extension at 72°C for 1 min, a total of 30 cycles; extension at 72°C for 10 min, 1 cycle.

[0044] Separation was carried out by vertical electrophoresis on 6% polyacrylamide gel, silver staining was performed on the electrophoresis gel plate, rinsed with water, dried, photographed and recorded.

[0045] (3) Drawing

[0046] At the ...

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Abstract

The invention belongs to the technical field of molecular organisms and particularly relates to a fingerprint spectrum of cauliflower hybrid and a construction method thereof. The cauliflower hybrid is temporarily named as 'shenxue 108 day', and is a new cauliflower variety bred by multi-year hybrid in Shanghai funong seed industry limited company. The fingerprint spectrum and the construction method have the advantages that SSR molecular markers are utilized for carrying out quantitative marking on the 'shenxue 108 day', and the fingerprint spectrum different from sibling species or the same series of cauliflower varieties is constructed; the applicability to identification, selling and truth-falseness discrimination of farmers for the shenxue 108 day of the cauliflower is achieved. The fingerprint spectrum and the construction method have the effects that the mastering is easy and the benefit for producers and seed buyers can be protected.

Description

technical field [0001] The invention relates to the field of molecular biology technology, in particular to a cauliflower hybrid fingerprint and a construction method thereof. Background technique [0002] my country is a country with a large population in the world, and needs a large amount of vegetables every day. Among them, cauliflower is rich in nutrition and delicious in flavor, and is one of the main vegetable types in my country. Cauliflower (commonly known as cauliflower) belongs to Brassicaceae Brassica Brassica species, native to the eastern coast of the Mediterranean Sea in Europe. In the middle of the 19th century, cauliflower was gradually widely cultivated in southern my country. At present, the annual cultivated area in the country has reached 4 million mu. However, due to serious diseases on cauliflower, especially black rot and downy mildew, the large-scale planting of cauliflower is restricted. Shanghai Rich Farm Seed Industry Co., Ltd. has been engaged...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q1/6895C12Q2600/13C12Q2600/156
Inventor 王新华刘杨高莉洁周腾夏胡继军
Owner SHANGHAI JIAO TONG UNIV
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