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Method for identifying authenticity of cucurbita pepo variety

A technology of zucchini and locus, which is applied in the field of identifying the authenticity of zucchini varieties and can solve the problems of lack of DNA molecular detection methods and the like

Active Publication Date: 2020-11-24
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no DNA molecular detection method for the identification of zucchini varieties in my country

Method used

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  • Method for identifying authenticity of cucurbita pepo variety
  • Method for identifying authenticity of cucurbita pepo variety
  • Method for identifying authenticity of cucurbita pepo variety

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1. Acquisition of primer combinations for identifying the authenticity of zucchini varieties

[0039] 1. Discovery of 48 SNP sites

[0040] The present invention obtains 48 SNP sites based on the resequencing data of 30 zucchini representative resources. These 30 zucchini represent rich types of resources, covering mosaic zucchini (8 parts), seedless zucchini (7 parts), green-skinned zucchini (8 parts) and long-vine zucchini (7 parts), basically including the main ecology of zucchini type, with high genetic diversity in terms of agronomic traits, reflecting germplasm representation as much as possible.

[0041] Specifically, the screening criteria for SNP loci are as follows: evenly selected positions throughout the genome, good polymorphism, low heterozygosity, MAF>0.3, good PCA clustering effect, high discrimination, and conservative 50bp sequences on both wings (no InDel , no SSR, no other SNP).

[0042] For the basic information of the 48 SNP sites, see c...

Embodiment 2

[0057] The validity check of the primer combination that embodiment 2, embodiment 1 develop

[0058] Randomly select 265 zucchini varieties for testing to check the effectiveness of the primer combinations developed in Example 1.

[0059] The basic information of the 265 tested zucchini varieties is shown in Table 3. The 265 tested zucchini varieties are all common fine varieties or some imported varieties.

[0060] Table 3. Names and sources of 265 tested zucchini varieties

[0061]

[0062]

[0063]

[0064]

[0065] 1. Obtaining the genomic DNA of the tested zucchini varieties

[0066] Genomic DNA of 265 leaves (true leaves of 30 seeds mixed) of 265 tested zucchini varieties were extracted by CTAB method to obtain genomic DNA of tested zucchini varieties.

[0067] The quality and concentration of the genomic DNA of the tested zucchini varieties must meet the requirements of PCR, and the standard is: agarose electrophoresis shows a single DNA band without obvio...

Embodiment 3

[0079] Embodiment 3, establishing the method for detecting which variety the zucchini to be tested belongs to among the 265 zucchini varieties in embodiment 2

[0080] One, establish the method for detecting which kind the zucchini to be tested belongs to among the 265 zucchini varieties in embodiment 2

[0081]1. Obtaining the genomic DNA of the zucchini to be tested

[0082] According to the method of step 1 in Example 2, "leaves of the tested zucchini variety" was replaced with "leaves of the zucchini to be tested", and other steps were kept unchanged to obtain the genomic DNA of the zucchini to be tested.

[0083] 2. Using the genomic DNA of the zucchini to be tested as a template, 48 primer sets were used for PCR amplification to obtain PCR amplification products. In each PCR reaction system, the concentration ratio of the primers containing "F1" in the name, the primers containing "F2" in the name and the primer containing "R" in the name is 2:2:5.

[0084] The reactio...

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Abstract

The invention discloses a method for identifying authenticity of a cucurbita pepo variety. The method provided by the invention can be used for carrying out early identification on the cucurbita pepovariety in a seed or seedling stage, guarantees the authenticity of the cucurbita pepo variety, practically protects the rights and interests of producers and breeders, and provides a technical support for cucurbita pepo germplasm resources and novel variety protection. The method provided by the invention can be used for identifying an unknown cucurbita pepo variety, and also can be used for identifying the authenticity of a known variety. The method provided by the invention has the advantages of high throughput, accuracy, low cost, simplicity in operation, saving of manpower and material resources and the like, and has extremely wide application prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for identifying the authenticity of zucchini varieties, which can be applied to biotechnology extension services and provides technical support for zucchini germplasm resources and variety protection. Background technique [0002] Zucchini, also known as American squash, is an important cultivated vegetable of the Cucurbitaceae family. It is rich in nutrients and contains an anti-interferon inducer, which can improve human immunity. Zucchini has the advantages of short growth period, high yield, storage and transportation, and suitable for intercropping and intercropping. It has become the melon vegetable with the cultivation area second only to cucumber. The planting area of ​​zucchini in my country exceeds 5 million mu, and the daily output of zucchini in Shouguang, Shandong is 75,000-100,000 kilograms, which has broad application prospects and huge economic po...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6895C12Q2600/156C12Q2531/113C12Q2537/161C12Q2563/107
Inventor 温常龙李海真张建杨静静罗江张晓飞李向晶赵泓
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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