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Molecular marker authenticating method for related gene of cucumber fruit length

A technology of molecular markers and identification methods, applied in the field of plant biology, to achieve the effects of easy mastery, reduced waste of manpower and material resources, and high 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

Previous studies on cucumber fruit length mostly focused on cytology, the relationship between endogenous hormones and fruit length, environmental factors (temperature, light, moisture, CO 2 , mineral nutrition) on cucumber fruit development, etc., but there are few reports on the genetic analysis and molecular markers of cucumber fruit length.

Method used

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  • Molecular marker authenticating method for related gene of cucumber fruit length
  • Molecular marker authenticating method for related gene of cucumber fruit length

Examples

Experimental program
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Effect test

Embodiment 1

[0019] A molecular marker identification method for cucumber fruit length-related genes, the method comprising the following steps:

[0020] Use the DNA of a cucumber variety or strain with unknown fruit length as a template, and use CSWCT28 as a primer to carry out PCR amplification. The amplified product is detected by 3% agarose gel electrophoresis or 7.5% denatured polyacrylamide gel for the amplified product. Gel electrophoresis detection, if the amplified band pattern is consistent with the known short fruit length cucumber Dongnong 803 band pattern, then the cucumber variety or line with unknown fruit length is a short fruit length variety or line, and the PCR amplification The added program is: pre-denaturation at 94°C for 5 minutes, denaturation at 94°C for 30 seconds, annealing at 47°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 CSWCT28 described is:

[0021] Primer 1: 5'-GAATTCAAAAAGCATTT...

Embodiment 2

[0024] In the molecular marker identification method of cucumber fruit length-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, 5U / μl TaqDNA polymerase: 0.2 μl, 60ng / μl template DNA: 1 μl, ddH20: 10.7 μl.

Embodiment 3

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

[0027] Using Dongnong 804, a new cucumber hybrid variety cultivated by the cucumber research group of the College of Horticulture, Northeast Agricultural University, as a material; extract its single plant DNA as a template, and use a cucumber fruit length molecular marker CSWCT28 provided by the present invention as a primer for PCR amplification. The results showed that the PCR amplification band pattern of Dongnong 804 single plant was completely consistent with the short fruit length control Dongnong 803 amplification band pattern, and the coincidence rate of PCR detection results and field identification was 100%, which proved that CSWCT28 is a practical Molecular markers for cucumber fruit length detection.

[0028]

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Abstract

The invention relates to a molecular marker authenticating method for the related gene of cucumber fruit length, which comprises the following steps of: taking the deoxyribose nucleic acid (DNA) of cucumber variety or strain with unknown fruit length as a template, taking CSWCT28 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 short fruit length, the cucumber variety or strain with the unknown fruit length is the variety or strain with short fruit length , 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 47 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 fruit length.

Description

Technical field: [0001] The invention relates to a molecular marker identification method of cucumber fruit length-related genes, belonging to the field of plant biotechnology. Background technique: [0002] The improvement of cucumber quality traits is an important goal of breeding research, and fruit length is one of the important quality traits of cucumber, which directly affects economic benefits. With the rapid development of the commodity economy and the improvement of people's living conditions, the consumption forms of cucumber are becoming more and more diversified, such as fresh food, cold salad, fried food, soup food, kimchi, salted, candied, dried, canned, etc. People have different requirements for the length of cucumbers for different purposes. For example, pickling processing requires short-fruited varieties, while salt-slicing processing requires long-fruited varieties; cucumber fruits with a uniform length can meet the requirements of mechanized picking and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 秦智伟周秀艳武涛王成辛明
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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