Method for rapidly identifying broccoli variety Tailv1 hybrid

An identification method, the technology of broccoli, applied in the direction of microbial measurement/inspection, biochemical equipment and methods, etc., to achieve the effects of high accuracy, reduced identification cost, and reliable results

Active Publication Date: 2014-08-20
台州市农业科学研究院
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with SSR, ISSR is more versatile among different species and easier to develop primers. Compared with RAPD, it has higher stability. It is a simple and low-cost detection method. Currently, researchers have used ISSR The hybrid purity identification of rice, cucumber and other crops has been carried out, but there is no report on the application of ISSR to the genetic purity identification of broccoli hybrids

Method used

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  • Method for rapidly identifying broccoli variety Tailv1 hybrid
  • Method for rapidly identifying broccoli variety Tailv1 hybrid
  • Method for rapidly identifying broccoli variety Tailv1 hybrid

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Parents, hybrids and false hybrids of 'Tailv No. 1' are used as materials to prove the feasibility of the method of the present invention and its special primers for identifying true and false hybrids of broccoli 'Tailu No. 1'.

[0034] (1) Extraction of genomic DNA from seedlings of 'Tailu No. 1' artificially mixed with pseudo-hybrids. Among them, broccoli 'Tailu No. 1' has one male parent and one female parent, 14 hybrid seeds of 'Tailu No. 1', and 8 other random mixed seeds. The individual plants are numbered sequentially, and the broccoli is quickly extracted by CTAB simple extraction method. Genomic DNA of seedlings, the specific steps are as follows:

[0035] ①Take 0.2g of young leaves at the seedling stage, cut them into small pieces, put them into 2mL centrifuge tubes, and add 1 stainless steel ball (diameter about 0.5cm) to each centrifuge tube, cover the centrifuge tubes and put them in liquid Stand still in the nitrogen foam box for 10 minutes, pour out the ...

Embodiment 2

[0046] The 'Tai Lv No. 1' hybrid produced in the broccoli breeding base of Taizhou Academy of Agricultural Sciences in 2013 and randomly mixed with a small amount of false hybrids was used as the sample to be tested, numbered randomly, and blue and white were identified by the conventional field planting identification method and the method of the present invention. The purity of the hybrid of vegetable 'Tailu 1' was compared, and the identification results of the two methods were compared, and the feasibility, reliability and timeliness of the method of the present invention were further investigated.

[0047] A total of 210 samples to be tested were numbered for individual plants, and conventional field planting identification and method identification of the present invention were carried out.

[0048] (1) Field planting identification method: on August 3, 2013, 210 samples to be tested were sown in sequence according to the numbering order, and transplanted in the field on ...

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Abstract

The invention provides a method for rapidly, accurately and efficiently identifying the molecules of a broccoli variety Tailv1 hybrid. The method comprises the following steps: extracting genomic DNA from a sample to be detected, carrying out PCR amplification on the genomic DNA by a PCR primer of 5'-TCTCTCTCTCTCTCTCN-3', carrying out gel electrophoresis imaging analysis on the obtained PCR amplification products, and judging the sample to be detected as a Tailv1 hybrid single plant if about 800bp and about 600bp brands are simultaneously obtained through amplification; and calculating according to a formula of genetic purity of hybrid = number of hybrid plants/number of sample plants to be detected *100% to obtain the genetic purity of the sample to be detected. The accuracy of the identification of the purity of the broccoli variety Tailv1 hybrid by an ISSR primer TNISSR15 used in the invention is high, has a 99% consistency with a field identification result, and is reliable; the identification can be completed within 10d, the tedious program of field planting identification is avoided, the labor force and the land resource are saved, and the identification cost is reduced; and the method can determine the genetic purity of seeds in the same year of seed production, is convenient for seed graduation and purchase, and improves the identification efficiency.

Description

(1) Technical field [0001] The invention relates to a rapid identification method for a broccoli variety 'Tailu No. 1' hybrid. (2) Background technology [0002] Broccoli (Brassica oleracea var. italica), also known as green cauliflower, broccoli, etc., is a variety of Brassica oleracea Brassica species. Its edible part is the fat and tender flower stem with flower buds, which is rich in nutrition, unique in flavor, and has anti-cancer and anti-oxidation effects, and is deeply loved by consumers. In recent years, the cultivated area of ​​broccoli in my country has increased year by year, and the demand for dominant broccoli varieties has also continued to increase. The use of heterosis to produce broccoli F 1 Hybrids have become a trend. However, in the process of hybrid seed production, the female parent self-crosses, the mutual pollination of people in the operation process, and the pollination caused by birds, bees, etc. will all form false hybrids. In addition, broccol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6895C12Q2600/156
Inventor 张志仙何道根朱长志檀国印
Owner 台州市农业科学研究院
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