Method for establishing cauliflower hybrid seed DNA (deoxyribonucleic acid) fingerprint spectrum and application thereof

A technology of DNA fingerprinting and method establishment, which is applied in the direction of biochemical equipment and methods, microbiological measurement/inspection, etc., can solve the problems that the quality of seeds cannot be supervised in advance, the information of seed purity cannot be provided in time, and the cycle is long, etc., to achieve protection Legitimate interest, readability and analysis effects

Inactive Publication Date: 2011-06-29
SHANGHAI ACAD OF AGRI SCI +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional field identification method uses the color, shape and other agronomic traits of seeds, seedlings, leaves, flowers, fruits, pollen and other tissues and organs as the observation indicators for identification. Although it is a relatively feasible method in production practice, field identification requires Additional land occupation, long period, high cost, not only limited by seasons, but also the performance of many traits is affected by cultivation measures and environmental factors, and the observation experience of the appraiser also restricts the ac

Method used

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  • Method for establishing cauliflower hybrid seed DNA (deoxyribonucleic acid) fingerprint spectrum and application thereof
  • Method for establishing cauliflower hybrid seed DNA (deoxyribonucleic acid) fingerprint spectrum and application thereof

Examples

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Embodiment 1

[0021] In this example, the SDS method was used to extract the genomic DNA of the cauliflower variety "Zaohua 60" and its parents, and its DNA fingerprint was constructed by RAPD and SRAP molecular markers for identification of seed purity. The specific operation is as follows:

[0022] 1. Selected test materials:

[0023] The materials are cauliflower hybrid "Zaohua 60" and its parents.

[0024] 2. Extraction and purification of cauliflower DNA:

[0025] The genomic DNA of each cauliflower material was extracted according to the following procedure:

[0026] Take 2g of fresh tissue, grind it into powder in liquid nitrogen, put it in a 50ml centrifuge tube, add 8ml of extraction solution (100mM Tris pH 8.0, 50mM EDTA, 500mM NaCl, 1.5% SDS), keep it at 65°C for 30 minutes, and mix it upside down from time to time Mix well, add 2.5ml 5M potassium acetate in ice bath for 10 minutes, add 0.5ml chloroform, mix upside down, centrifuge at 10000rmp for 8 minutes, take the supernata...

Embodiment 2

[0035] The DNA fingerprint of the present invention is used for the identification of " early flower 60 " seed purity:

[0036] Randomly select 50-80 individual plants from the "Zaohua 60" hybrid produced in the seed production base, extract the DNA of each individual plant sample by micro-DNA extraction method, use the extracted DNA as a template, and use the corresponding primers to carry out PCR amplification and electrophoretic analysis. For example, if the DNA fingerprint of a single plant detected by S 140 primer analysis is "001111111111", it is the true hybrid strain of "Zaohua 60", and the DNA fingerprint of "001110111111" or "110111111111" is the parental inbred strain of "Zaohua 60". That is, pseudo-hybrid strains. For another example, the DNA fingerprint of a single plant detected by M8-E18 primer analysis is "11111111111111111111", which is the true hybrid strain of "Zaohua 60", and the DNA fingerprint is "111111111111110011001" or "111011111101110110110", which is...

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Abstract

The invention discloses a method for establishing a cauliflower hybrid seed and a parent DNA (deoxyribonucleic acid) fingerprint spectrum and the application thereof. The establishment method comprises the following steps: 1) extracting and purifying cauliflower DNA; 2) using the obtained high-purity cauliflower DNA as a template for performing RAPD (random amplified polymorphic DNA) and SRAP (sequence-related amplified polymorphism) analysis; and 3) selecting selective polymorphism amplified bands to construct the DNA fingerprint spectrum of a tested material; and the cauliflower hybrid seed and parents thereof in the DNA fingerprint spectrum have specific DNA fingerprints which can be mutually differentiated. The DNA fingerprint spectrum is represented in the form of a diagram, thereby being relatively intuitive to look and being easy to understand; and the DNA fingerprint spectrum is converted to the digital representation form, thereby being convenient to be identified, read and analyzed by a computer. The method can be applied in identification of seed purity of the cauliflower hybrid seed, the result is accurate and reliable, and the detection is rapid.

Description

technical field [0001] The invention relates to cauliflower seed identification technology, in particular to a method for establishing DNA fingerprints of cauliflower hybrids and its application. Background technique [0002] At present, many varieties of cauliflower (Brassica oleracea L.var.botrytis L) popularized and applied in my country are the first generation of hybrids. The first generation of hybrids can synthesize many excellent characters of the parents, and the economic benefits are better. [0003] However, in the process of seed production, due to improper management measures and the defects of self-incompatibility itself, self-seeds, ie false hybrids, are often produced, resulting in a decrease in seed purity. Therefore, improving the purity of hybrids is an important topic in the field of heterosis utilization research. At present, there are roughly three methods for variety identification and seed purity analysis, namely field morphology, biochemical marker...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 薄天岳陈烨丽陈锦秀缪体云
Owner SHANGHAI ACAD OF AGRI SCI
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