Maize Molecular Breeding Core SNP Site Combination and Its Application

A maize and site technology, which is applied to the combination of core SNP sites in maize molecular breeding and its application field, can solve the problems of application limitation, site fixation and high detection cost, and achieve the effect of improving breeding efficiency

Active Publication Date: 2022-05-03
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the high cost of detection of a single sample of the chip and the fixed position, the application in molecular marker-assisted selection breeding is limited to a certain extent.

Method used

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  • Maize Molecular Breeding Core SNP Site Combination and Its Application
  • Maize Molecular Breeding Core SNP Site Combination and Its Application
  • Maize Molecular Breeding Core SNP Site Combination and Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0365] Example 1, Screening of Molecular Breeding Core SNP Site Combinations and Design of KASP Primers

[0366] 1. Screening of core SNP loci in molecular breeding

[0367] 1. Preliminary screening of 740 SNP sites

[0368] From the Maize6H90K chip developed by the Maize Research Center of Beijing Academy of Agriculture and Forestry Sciences, 740 SNP sites were selected according to the comprehensive indicators such as site typing effect, chromosome uniform distribution, PIC value, and MAF value, and 740 sets of KASP primers were designed and synthesized. A primer corresponds to a SNP site.

[0369] 2. Re-screen 498 pairs of SNP markers with ideal typing effect

[0370] The genomic DNA of 157 maize materials to be identified was extracted and diluted to 200ng / μl (RNAase enzyme treatment) respectively, and prepared into two 96-well mother liquid plates, and the remaining sample wells were used as reference NTC.

[0371] A total of 740 pairs of primers were synthesized to pe...

Embodiment 2

[0705] Example 2, Molecular Breeding Core SNP Sites and Application of KASP Primers

[0706] 1. The application of molecular breeding core SNP loci and their KASP primers in detecting the genetic similarity of inbred lines and detecting the heterozygosity of hybrids

[0707] 1. Detection of genetic similarity of inbred lines

[0708] Genomic DNA was extracted from the 124 corn inbred lines shown in Table 1 as templates, respectively amplified with the KASP primers shown in Table 3, and the genotypes of 300 core SNP loci of the 124 corn inbred lines were detected.

[0709] The brief workflow of the KASP screening primer test is as follows: Dilute the DNA from the mother plate to 20ng / μl as the Master plate; use the Replikator hole plate duplicator of LGC Company to select the appropriate mode to divide the liquid into the 1536-well Working plate, and Import the liquid separation data into the Kraken system; use the drying oven to dry the DNA in the Working plate; use the Merdi...

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Abstract

The invention discloses the core SNP site combination of corn molecular breeding and its application. The invention provides the application of 300 SNP sites in at least one of the following (1)-(4): (1) detecting the genetic similarity of corn inbred lines; (2) detecting the heterozygosity of corn hybrids; (3) ) Molecular marker-assisted background selection for maize backcross breeding; (4) maize molecular breeding; the present invention screens and determines a set of SNP site combinations suitable for maize molecular breeding from 740 pairs of SNP primers. Therefore, based on the high-throughput KASP detection platform of samples, the present invention screens the combination of core SNP sites suitable for molecular breeding of maize, which can significantly improve the breeding efficiency.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a combination of core SNP sites of corn molecular breeding and its application. Background technique [0002] At present, the breeding of new corn varieties in my country still largely relies on conventional breeding techniques, which have long breeding cycles, low efficiency, high chance, and time-consuming and laborious. The rapid development of genomics and DNA molecular marker technology has given birth to crop molecular breeding. Molecular marker technologies mainly include RAPD, AFLP, SSR, SNP, etc. Among them, SSR technology is widely used in crop genetics and breeding in my country because of its advantages of high polymorphism and simple operation. However, because SSR marker technology cannot meet the needs of large-scale, high-throughput, and automated detection, international seed companies such as Pioneer and Monsanto have adopted SNP marker technology as t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156C12Q2600/13
Inventor 宋伟赵久然苏爱国王帅帅李春辉张如养王凤格田红丽王蕊
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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