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Molecular marker authenticating method for related gene of cucumber propamocarb residue

A technology of molecular markers and identification methods, applied in the field of plant biology, can solve the problems of low pesticide residue content, achieve simple operation, high efficiency, and improve breeding efficiency

Inactive Publication Date: 2012-07-18
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that the pesticide residues in cucumbers are closely related to the ecological types. The South China type cucumbers have the highest amount of pesticide residues, followed by the North China type. , Qin Zhiwei, Zhou Xiuyan. Screening of cucumber germplasm resources with low pesticide residues. Journal of Northeast Agricultural University, 2010, 41(7): 32-36), however, the research on genes related to cucumber pesticide residues has not been reported

Method used

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  • Molecular marker authenticating method for related gene of cucumber propamocarb residue
  • Molecular marker authenticating method for related gene of cucumber propamocarb residue
  • Molecular marker authenticating method for related gene of cucumber propamocarb residue

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Embodiment 1

[0020] A molecular marker identification method for cucumber propamocarb residue-related genes, the method comprising the steps of:

[0021] Use the DNA of cucumber varieties or strains with unknown propamocarb residues as templates, and use CSWCT14 as primers to perform PCR amplification, and perform 3% agarose gel electrophoresis on the amplified products or carry out 7.5% denaturation polymerization on the amplified products. Acrylamide gel electrophoresis detection, if the amplified band pattern is consistent with the known low propamocarb residual cucumber Dongnong 803 band pattern, then the cucumber variety or line with unknown propamocarb residue is low propamocarb For residual varieties or strains, the PCR amplification procedure is: pre-denaturation at 94°C for 5 minutes, denaturation at 94°C for 30 seconds, annealing at 57°C for 1 minute, 35 cycles, extension at 72°C for 1 minute, extension at 72°C for 10 minutes, and finally storage at 4°C. The CSWCT14 is:

[0022]...

Embodiment 2

[0025] In the molecular marker identification method of cucumber propamocarb residue-related genes, in the PCR amplification, the PCR reaction system of every 20 μl is as follows: 10×buffer: 2 μl, 2mM dNTP: 2 μl, 10 pmol / μl of Primer 1: 0.8 μl, 10 pmol / μl Primer 2: 0.8 μl, 25 mM MgCl 2 : 2.5 μl, 5 U / μl TaqDNA polymerase: 0.2 μl, 60 ng / μl template DNA: 1 μl, ddH2O: 10.7 μl.

Embodiment 3

[0027] Application of molecular marker identification method for cucumber propamocarb residue-related genes in assisting selection of pollution-free cucumber materials.

[0028] Using the new cucumber hybrid varieties Dongnong 804 and Dongnong 805 bred by the cucumber research group of the College of Horticulture of Northeast Agricultural University as materials; extract the DNA of a single plant as a template, and use a cucumber propamocarb residue-related molecular marker provided by the present invention CSWCT14 was used as a primer for PCR amplification. The results showed that the PCR amplification band patterns of Dongnong 804 and Dongnong 805 were completely consistent with the low propamocarb residue control Dongnong 803, and the PCR detection results were consistent with the experimental results. The coincidence rate of laboratory identification results was 100%, which proved that CSWCT14 is a practical molecular marker for the detection of propamocarb residues in cucu...

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Abstract

The invention relates to a molecular marker authenticating method for the related gene of cucumber propamocarb residue. The method of the invention comprises the following steps of: taking the deoxyribose nucleic acid (DNA) of cucumber variety or strain with unknown propamocarb residue as a template, taking CSWCT14 as a primer to carry out polymerase chain reaction (PCR) amplification, and carrying out 3% agarose gel electrophoresis detection on an amplified product or carrying out 7.5% denaturing polyacrylamide gel electrophoresis detection on the amplified product, wherein if the amplified banding pattern is consistent with the banding pattern of cucumber Dongnong 803 with known low-propamocarb residue, the cucumber variety or strain with the unknown propamocarb residue is the variety or strain with low propamocarb residue, and the PCR amplification has the procedures: previously denaturating for 5min under 94 DEG C, denaturating for 30s under 94 DEG C, annealing for 1min under 57 DEG C, carrying out 35 cycles, extending for 1min under 72 DEG C, extending for 10min under 72 DEG C, and finally storing under 4 DEG C. The method is used for authenticating the related gene of the cucumber propamocarb residue.

Description

Technical field: [0001] The invention relates to a molecular marker identification method of a cucumber propamocarb residue-related gene, which belongs to the field of plant biotechnology. Background technique: [0002] my country is the country with the largest production area and the highest total output of cucumbers in the world, and they are planted all over the country. Cucumber is one of the vegetables with the most pests and diseases, and is often harmed by various diseases during the growth process. It has a large demand for pesticides, and there are many types of demand. It is the main vegetable with excessive pesticide residues. One of the varieties. With the industrialization of cucumber production in my country, the requirements for varieties are becoming more and more stringent. Not only high yield, disease resistance, and high quality are required, but the fruit shape, taste, and low pesticide residues of cucumbers have also become important planting indicator...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 秦智伟武涛周秀艳马佰慧辛明
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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