A pair of special primers for assisted evaluation of relevant locus of soybean seed weight and method thereof

An auxiliary identification and 100-grain weight technology, which is applied in the fields of botanical equipment and methods, biochemical equipment and methods, and applications, can solve the problems of rarely found trait gene molecular markers and map positioning, etc., to improve breeding efficiency and soybean. Yield level, effect of shortening breeding cycle

Inactive Publication Date: 2009-12-30
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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Problems solved by technology

Large grain size is one of the important target traits for improving soybean yield, but so

Method used

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  • A pair of special primers for assisted evaluation of relevant locus of soybean seed weight and method thereof
  • A pair of special primers for assisted evaluation of relevant locus of soybean seed weight and method thereof
  • A pair of special primers for assisted evaluation of relevant locus of soybean seed weight and method thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0022] Example 1. Obtainment of soybean 100-seed weight related sites (high-hundred-seed weight related sites) hsw3

[0023] 1. Primer synthesis

[0024] The sequence of the Satt126 primer pair was obtained from the SOYBASE database; the upstream forward primer sequence was: 5’-GCTTGGTAGCTGTAGGAA-3’; the downstream reverse primer sequence was: 5’-ATAAAACAAATTCG CTGATAT-3’, which was commissioned to be synthesized by Beijing Osaibo Biological Company.

[0025] 2. Parental genome amplification test

[0026] The parents used to create soybean recombinant inbred line (RIL) population are Zhonghuang 4 (National Soybean Germplasm Bank) and Zhongpin 661 (National Soybean Germplasm Bank). Zhonghuang 4 is the female parent and its grain volume is larger (22.0-26.5 g / 100 grains); Zhongpin 661 is the male parent and its grain volume is small (17.7-19.5 g / 100 grains).

[0027] The CTAB method was used to extract the genomic DNA of the parental leaves, and the Satt 126 primer pair was used for...

Example Embodiment

[0033] Example 2. Correlation verification between amplified product of Satt126 primer pair and grain weight traits

[0034] Collected 48 soybean resource materials (homozygous varieties or strains) from different soybean ecological regions in my country.

[0035] The genomic DNA of each soybean plant leaf was extracted by CTAB method, and PCR amplification experiment was carried out with Satt126 primer pair. The PCR reaction system is: 10×Buffer 2μl (including Mg 2+ ), dNTP 0.4μl (10mM), upstream forward primer and downstream reverse primer each 2μl (5μM), genomic DNA 1μl (40ng / μl), Taq enzyme 0.5μl (5U / μl), reaction volume is 20μl, add dropwise Cover with a drop of mineral oil. The PCR reaction program is: 94°C for 5min; 94°C for 60s, 53.5°C for 60s, 72°C for 60s, 35 cycles; 72°C for 10min.

[0036] The PCR products were analyzed by 9% polyacrylamide gel electrophoresis, stained with rapid silver staining, and observed and photographed under ultraviolet light. See the result fig...

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Abstract

The invention discloses a pair of special primers for assisted evaluation of soybean seed weight and a method thereof. The invention provides a method for assisted screening of soybean with seed weight property, comprising the following steps: the gene group DNA of soybean to be tested serves as a template, a Satt126 primer pair is used for carrying out PCR amplification, and an amplification product is tested; the soybean to be tested, which contains a DNA fragment between 165-175bp in the amplification product, is a candidate soybean with high seed weight property; the Satt126 primer pair is composed of ribotide represented by a sequence 1 and a sequence 2 in a sequence table; the high seed weight property is the property that kernel seed weight is 25.0 or more than 25.0 grams. The method of the invention overcomes the problem that breeding efficiency is low and the like because 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 Satt126 to carry out early generation choosing evaluation for the relevant locus of high seed weight of soybean so as to shorten breeding cycle, and then, high-yield soybean materials with bigger kernels can be quickly screened. The invention can be used for soybean high-yield breeding for improving breeding efficiency and soybean yield level.

Description

technical field [0001] The invention relates to a pair of special primers capable of rapidly identifying and selecting soybean 100-grain weight-related sites and a method thereof. 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 microsatellites or SSR (simple sequence repeat) are widely used 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...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11A01H1/04
Inventor 朱保葛田苗苗任海红陈修文徐民新张利明周国安
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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