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Method for assisting identification of 100-seed weight related loci of soybeand special primer thereof

An auxiliary identification and 100-grain weight technology, applied in the fields of botanical equipment and methods, applications, and plant genetic improvement, can solve the problems of low selection efficiency and long breeding cycle, shorten the breeding cycle, improve the breeding efficiency and soybean yield. horizontal effect

Inactive Publication Date: 2013-11-27
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • 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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  • Method for assisting identification of 100-seed weight related loci of soybeand special primer thereof
  • Method for assisting identification of 100-seed weight related loci of soybeand special primer thereof
  • Method for assisting identification of 100-seed weight related loci of soybeand special primer thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1. Acquisition of soybean 100-kernel weight-related locus (high 100-kernel weight-related locus) hsw1

[0032] 1. Primer synthesis

[0033] The sequence of the GNE070 primer pair was obtained from the SOYBASE database; the upstream forward primer sequence was: 5'-TGCTTTCCTCTATTGGTTGT-3'; the downstream reverse primer sequence was: 5'-TCTTTTCTCTGATCCATCACC-3', which was synthesized by Beijing Osaibo Biological Company.

[0034] 2. Parental genome amplification detection

[0035] 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, and its grain volume is relatively large (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).

[0036] Genomic DNA from parental leaves was extracted by CTAB method, and the amplification e...

Embodiment 2

[0042] Example 2, Correlation verification between the amplification product of the GNE070a primer pair and the grain weight traits

[0043] The experimental samples were 48 soybean plant materials (homozygous varieties or lines).

[0044] The genomic DNA of each soybean plant leaf was extracted by CTAB method, and the PCR amplification experiment was carried out using the genomic DNA as a template and GNE070a primer pair. PCR reaction system (20μl): 10×Buffer 2μl (containing Mg 2+ ), dNTP (10mM) 0.4μl, upstream forward primer (5μM) and downstream reverse primer (5μM) each 2μl, genomic DNA (40ng / μl) 1μl, Taq enzyme (5U / μl) 0.5μl, the rest is ddH 2 O, add a drop of mineral oil to cover. PCR reaction program: 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.

[0045] PCR amplification products were analyzed by 9% polyacrylamide gel electrophoresis, stained with rapid silver staining and observed under ultraviolet light. The PCR ampli...

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Abstract

The invention discloses a method for assisting identification of 100-seed weight related loci of soybean and a special primer thereof. The method for assisting identification of 100-seed weight related loci of soybean comprises the following steps: with the genomic DNAs (deoxyribonucleic acids) of to-be-detected soybean as a template, carrying out PCR (polymerase chain reaction) amplification by using a GNE070a primer pair consisting of DNAs shown in a sequence 1 and a sequence 2, detecting amplification products, and amplifying a to-be-detected soybean (with a specific DNA fragment) in the amplification products to be a candidate soybean with a high 100-seed weight trait, wherein the size of the specific DNA fragment is 275-285bp. By using the method disclosed by the invention, the problems of low breeding efficiency and the like since a phenotypic selection is interfered easily by environmental factors in conventional breeding are solved, the breeding period can be shortened through carrying out early-generation selection and identification on 100-seed weight related loci of soybean by using the special primer, thereby rapidly screening out high-yield soybean material with large seeds. The method and special primer disclosed by the invention can be applied to the high-yield breeding of soybean so as to improve the breeding efficiency and yield level of soybean.

Description

technical field [0001] The invention relates to a method for assisting identification of soybean hundred-grain weight-related sites and special primers 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 of traits showing ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11A01H1/02
Inventor 朱保葛任海红周国安李素坤杨瑞潘毅
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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