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Development and application of SSR (simple sequence repeats) marker for identifying prematurity of upland cotton

An early-maturing and auxiliary identification technology, which is applied in the determination/inspection of microorganisms, DNA/RNA fragments, DNA preparation, etc., can solve the problems of no early-maturing cotton breeding work, and no reliable molecular markers for identifying cotton early-maturity

Active Publication Date: 2017-08-29
INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, no reliable molecular markers have been obtained to identify or assist in the identification of cotton precociousness, and no relevant early-maturing cotton breeding work has been carried out through MAS in production.

Method used

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  • Development and application of SSR (simple sequence repeats) marker for identifying prematurity of upland cotton
  • Development and application of SSR (simple sequence repeats) marker for identifying prematurity of upland cotton
  • Development and application of SSR (simple sequence repeats) marker for identifying prematurity of upland cotton

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1. Acquisition of cotton precociousness-related SSR markers and related primer design and detection methods

[0053] The inventor's team located a chromosomal segment that is very closely related to precocity, located on the D3 chromosome, and a very prominent QTL site (qFT-D3-3) was detected in five environments from 2011 to 2015. And the contribution rate is about 20%, so there are genes that are very closely related to precocity at this locus; based on the key chromosomal segments ( figure 1 ), further screened primers in this segment, searched for polymorphic SSR markers between the parental materials—cotton variety 'Zhong36' and cotton line 'G2005', and finally obtained cotton precocity-related SSR markers, denoted as D03_1348 .

[0054] 1. Synthesis of primers related to cotton prematurity-related SSR marker D03_1348

[0055] The primer pairs synthesized in Suzhou Jinweizhi Biotechnology Co., Ltd. for the identification of precocity are as follows:

[0...

Embodiment 2

[0066] Embodiment 2, the precocity identification of genetic segregation population

[0067] The present invention firstly uses the recombinant inbred line group comprising 192 strains as material to carry out experiments, and the hybrid parents of the group are cotton variety 'Zhong 36' and cotton line 'G2005'. Among them, 'Zhong 36' is an early-maturing variety, and 'G2005' is a late-maturing line, both of which are upland cotton materials. The florescence (FT) of population was investigated in 2013-2015, and identified population and parents by primer pair and corresponding PCR, polyacrylamide gel electrophoresis detection method provided in Example 1. Combining the phenotype data of the population with the genotype, based on the homozygous genotypes of 'Zhong 36' and 'G2005' (the band type detected by polyacrylamide gel electrophoresis is consistent with that of 'Zhong 36' as the CC genotype, Those consistent with 'G2005' are recorded as GG genotype), and 192 lines were g...

Embodiment 3

[0072] Embodiment 3, the mark verification of precocious natural population

[0073] The present invention takes 88 precocious materials as objects, and verifies D03_1348. The names and sources of materials are shown in Table 2, and some materials are recorded in the literature "Han Yongliang, Lu Zhengying, Li Shiyun, Yang Yufeng, Cui Hongyin. Gray correlation analysis of early-maturing cotton varieties. Anhui Agricultural Sciences, 2015, 43(8): 37-38" . Using D03_1348 as the marker and 'Zhong 36' and 'G2005' as references, 88 early-maturing materials were genotyped. Refer to Examples 1 and 2 for specific operations and result determination.

[0074] The identification results showed that 70 of the 88 precocious materials had the same genotype as 'Zhong 36', 12 had the same genotype as 'G2005', 5 were heterozygous, and 1 had no amplified product ( image 3 ). It can be seen that about 80% of the early-maturing materials obtained the same genotype band as 'Zhong 36', which...

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Abstract

The invention discloses a development and application of an SSR (simple sequence repeats) marker for identifying prematurity of upland cotton. The invention provides a primer pair for identifying or assisting identification on prematurity of upland cotton. A primer pair of a following DNA (deoxyribonucleic acid) fragment is obtained through amplification, and the DNA fragment is obtained through PCR (polymerase chain reaction) amplification upon primer pairs of sequence 1 and sequence 2 in a sequence table by taking cotton genome DNA as a template. According to the development, a molecular marker within a target section is developed for a QTL (quantitative trait loci) site (qFT-D3-3) positioned on D3 chromosome of upland cotton, an SSR marker which is tightly interlinked with the target QTL is obtained, the correlativity with prematurity of cotton is marked through multi-group verification, then the SSR marker can be applied to selective application of prematurity of cotton, and significant resources for auxiliary selective breeding of molecular markers for premature cotton varieties are provided.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to the development and application of an SSR marker for identifying the early maturity of upland cotton. Background technique [0002] Cotton is a very important economic crop in the world and plays an important role in the national economy and social development. However, due to our country's basic national conditions of more people and less land, the contradiction between grain and cotton competition for land is very prominent, and the area of ​​cotton planting is continuously decreasing, while the traditional method of planting cotton once a year limits the development of cotton production. Therefore, there is an urgent need for early-maturing, high-yield, high-quality new cotton varieties suitable for double-cropping or multi-cropping planting. Among the four cotton cultivars, the annual production of upland cotton (Gossypium hirsutum) accounts for more than 95% of the total cotton p...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C12N15/10
CPCC12Q1/689C12Q2600/13
Inventor 喻树迅贾晓昀宿俊吉宋美珍魏恒玲庞朝友范术丽王寒涛马启峰
Owner INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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