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A pair of special primers for assisted evaluation of oil content locus of soybean kernel and method thereof

A technology of oil content and grain, which is applied in a pair of special primers for assisting in the identification of soybean grain oil content sites and its field, can solve the problems of long breeding cycle and low selection efficiency, so as to increase the oil content of soybean grain and improve breeding efficiency Effect

Inactive Publication Date: 2012-06-06
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Conventional breeding methods based on phenotypic selection have disadvantages such as low selection efficiency and long breeding cycle. It is urgent to inject modern molecular technology, supplemented by high-efficiency genotype-directed selection, in order to quickly and efficiently breed excellent new soybean varieties.

Method used

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  • A pair of special primers for assisted evaluation of oil content locus of soybean kernel and method thereof
  • A pair of special primers for assisted evaluation of oil content locus of soybean kernel and method thereof
  • A pair of special primers for assisted evaluation of oil content locus of soybean kernel and method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Obtaining of soybean grain oil content-related loci (high oil content-related loci) oil6

[0021] 1. Primer synthesis

[0022] The sequence of the GNE165 primer pair is obtained from the SOYBASE database (the sequence of the upstream and downstream primers selected from the soybean EST sequence contains a SSR characteristic sequence consisting of 2-3 base tandem repeats); the upstream primer is 5'-CCCACTGTCTCACT CACACT- 3'; the downstream primer is 5'-TGCACAACTTCACATTGTCT-3'; the characteristic sequence of SSR is contained between the upstream and downstream primers. Primer synthesis was entrusted to Beijing Aoke Biological Company.

[0023] 2. Parental genome amplification detection

[0024] The parents used to create soybean recombinant inbred line (recombinant inbred line, RIL) population were Zhonghuang 4 (National Soybean Germplasm Bank) and Zhongpin 661 (National Soybean Germplasm Bank). Zhonghuang 4 is the female parent, its grain oil content is low ...

Embodiment 2

[0031] Example 2 Correlation verification between the amplified product of the GNE165 primer pair and the oil content traits of soybean seeds

[0032] 48 soybean resource materials (homozygous varieties or lines) were collected from different soybean ecological regions in my country.

[0033] The genomic DNA of each soybean plant leaf was extracted by CTAB method, and the amplification experiment was carried out with the GNE165 primer pair. The PCR reaction system is: 10×Buffer 2μl (containing Mg 2+ ), dNTP 0.4μl (10mM), upstream primer and downstream primer 2μl (5μM), genomic DNA 1μl (40ng / μl), Taq enzyme 0.5μl (5U / μl), the reaction volume is 20μl, add a drop of mineral oil to cover . The PCR reaction program was: 94°C for 5min; 35 cycles of 94°C for 60s, 53.5°C for 60s, and 72°C for 60s; 72°C for 10min.

[0034] PCR amplification products were analyzed by 9% polyacrylamide gel electrophoresis, stained by rapid silver staining and observed and photographed under ultraviole...

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Abstract

The invention discloses a pair of special primers for assisted evaluation of oil content locus of soybean kernel and a method thereof. The method for assisted screening of soybean with high kernel oil content property provided by the invention comprises the following steps: the gene group DNA of soybean to be tested serves as a template, a GNE 165 primer pair performs PCR amplification, and an amplification product is tested; a soybean to be tested, which contains a DNA fragment between 275-285bp in the amplification product, is a candidate soybean with high kernel oil content property ; the GNE 165 primer pair is composed of ribotide represented by a sequence 1 and a sequence 2 in a sequence table; the high kernel oil content property is the property that kernel oil content is 21.5 or more than 21.5% (percentage by weight).The method of the invention overcomes the problem that breeding efficiency is low and the like because the phenotypic selection is easy to be disturbed by environmental factors in normal breeding; the method of the invention can use the special primer marked by the GNE 165 to carry out early generation choose for the relevant locus of high oil content property of soybean kernels so as to shorten breeding cycle, and then, soybean materials with higher kernel oil content can be quickly screened. The invention can be used for soybean high-oil breeding for improving breeding efficiency and soybean kernel oil content.

Description

technical field [0001] The invention relates to a pair of special primers and a method thereof for assisting in identifying oil content sites in soybean grains. Background technique [0002] Conventional breeding methods based on phenotypic selection have disadvantages such as low selection efficiency and long breeding cycle. It is urgent to inject modern molecular technology, supplemented by high-efficiency genotype-directed selection, in order to quickly and efficiently breed excellent new soybean varieties. With the rapid development of molecular biology and genomics, the application of molecular marker technology is more extensive. PCR-based molecular markers such as SSR are favored by breeders because of their high polymorphism rate, relative stability, simple and fast detection method, and easy operation. Since molecular marker-assisted selection is not easily affected by environmental factors and the interference of traits showing recessiveness, etc., the target trai...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11A01H1/04
Inventor 朱保葛田苗苗任海红陈修文张利明徐民新赵丽
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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