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Molecular marker identification method of cucumber branchiness related gene

A technology of molecular marker and identification method, applied in the field of plant biology, can solve the problems of redundant labor of cucumber, increased cost of cucumber cultivation, infection of plant wounds, etc., and achieves the effects of easy control, improved breeding efficiency and low cost.

Inactive Publication Date: 2012-07-04
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many branches and strong branch growth will bring great inconvenience to the cultivation of cucumbers, and the growers must often interrupt the field, which will not only bring a lot of redundant labor to the cultivation of cucumbers, but also the wounds of the interrupted plants will be contagious. Diseases will increase the cost of cucumber cultivation invisibly

Method used

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  • Molecular marker identification method of cucumber branchiness related gene
  • Molecular marker identification method of cucumber branchiness related gene
  • Molecular marker identification method of cucumber branchiness related gene

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Experimental program
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Embodiment 1

[0019] A molecular marker identification method for cucumber branching-related genes, the method comprising the steps of:

[0020] Use the DNA of a cucumber variety or line with unknown branching as a template, and use CSWCT17 as a primer to perform PCR amplification, and perform 3% agarose gel electrophoresis detection on the amplified product or perform 7.5% denatured polyacrylamide on the amplified product. Gel electrophoresis detection, if the amplified band type is consistent with the known weakly branched cucumber Dongnong 803 band type, then the cucumber variety or line with unknown branching is a weakly branched variety or line, and the The program of PCR amplification is: pre-denaturation at 94°C for 5 minutes, denaturation at 94°C for 30 seconds, annealing at 56°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 CSWCT17 described is:

[0021] Primer 1: 5'-TTGAATTATGGGTTCATTTTT-3',

[0022] Pri...

Embodiment 2

[0024] In the molecular marker identification method of cucumber branching-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, Primer 1 of 10 pmol / μl: 0.8 μl , 10pmol / μl Primer 2: 0.8μl, 25mM 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

[0026] Application of molecular marker identification method of cucumber branching related genes in assisted selection of cucumber breeding materials.

[0027] 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 were used as materials; the DNA of a single plant was extracted as a template, and a cucumber branching molecular marker CSWCT17 provided by the present invention was used as a primer PCR amplification was carried out, and the results showed that the PCR amplification band patterns of Dongnong 804 and Dongnong 805 individual plants were completely consistent with the weak branching control Dongnong 803 amplification band pattern, and the PCR detection results were consistent with field branching identification. The coincidence rate was 100%, which proved that CSWCT17 is a practical molecular marker for cucumber branching detection.

[0028]

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Abstract

The invention relates to a molecular marker identification method of cucumber branchiness related gene. The method comprises the steps of: carrying out PCR (polymerase chain reaction) amplification with DNA (deoxyribonucleic acid) of a cucumber variety or line with unknown branchiness and with CSWCT17 as a primer, carrying out 3% agarose gel electrophoresis detection on the amplification product or carrying out 7.5% denaturalized polyacrylamide gel electrophoresis detection on the amplification product, and judging the cucumber variety or line with unknown branchiness to be a variety or line with weak branchiness if the amplified band form is consistent with the band form of cucumber Dongnong 803 with weak branchiness, wherein the PCR amplification process comprises the steps of: pre-denaturizing at 94 DEG C for 5 minutes, denaturalizing at 94 DEG C for 30 seconds, annealing at 67 DEG C for 1 minute, circulating 35 times, drawing at 72 DEG C for 1 minute, drawing at 72 DEG C for 10 minutes, and finally storing at 4 DEG C. The molecular marker identification method provided by the invention is used for identifying the cucumber branchiness related gene.

Description

Technical field: [0001] The invention relates to a molecular marker identification method of cucumber branching-related genes, belonging to the field of plant biotechnology. Background technique: [0002] The growth of branches is one of the important characters of cucumber plant type. Branches can grow on the main branch of cucumber, and branches can also regenerate on the branches. The number of branches is related to the variety and cultivation conditions. Generally, there are more middle-late maturing varieties with strong growth vigor than early-maturing varieties. The length of the stem and the number of branches and other characteristics and habits are often the basis for the adjustment of cucumber plants, while the thickness of the stem and the length of the internodes are one of the signs for diagnosing the strength of the plant and the level of yield. The plants are weak and the bristles are underdeveloped, so it is difficult to obtain high yields; the stems are ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 秦智伟周秀艳武涛李开盛辛明
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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