中国农业科学Issue(19)：3887-3898,12.DOI:10.3864/j.issn.0578-1752.2015.19.011
苹果部分种质资源分子身份证的构建
Establishment of Molecular ID for Some Apple Germplasm Resources
高源 1刘凤之 1王昆 1王大江 1龚欣 1刘立军1
作者信息
- 1. 中国农业科学院果树研究所/农业部园艺作物种质资源利用重点实验室，辽宁兴城 125100
- 折叠
摘要
Abstract
Objective] A total of 131 apple germplasms including landraces, bred cultivars and related species selected from the National Repository of Pear and Apple Germplasm Resources in Xingcheng, China were studied with tailed primer M13 microsatellite markers (TP-M13-SSR). An analysis was made to establish the molecular ID of 131 apple germplasms.[Method]Based on genetic fingerprints, germplasms were distinguished with selected SSR markers, alleles that were amplified by each marker were coded, then was combined as a molecular ID.[Result]Two accessions selected from a total of 131 accessions were used for optimization of the first PCR detecting conditions and SSR primer screening. 16 pairs of TP-M13-SSR primers with high stability and good repeatability were used to establish the fingerprints of 131 accessions ofMalus Mill.. By using 16 selected SSR markers, 326 polymorphic sites were detected with a mean value of 20.3. The amplification of CH05d04 for all germplasms obtained 49 alleles with the highest expected heterozygosity 0.878, and the amplification of CH01f07a for all germplasms obtained 48 alleles. The polymorphism information content was calculated by PopGen32 with 0.7558 on average. 16 SSR markers could distinguish 11 accessions at least, 71 accessions at most, and 49 accessions on average. The identified rate was 8.09%-52.21%. The identified rate of CH01f07a was the highest, and the identified rate of BGT23b was the lowest. Based on the PIC of amplification and identified rate, each parir of two SSR primers was combined together to identify all the accessions. The combination of CH04h02 and CH01f07a could distinguish 120 accessions at most. More SSR primers were combined together. Finally three SSR primers could distinguish all the accessions. All the alleles of the three core SSR markers were sequenced from small to large, and the assignment was from number 01. A character string was constituted by combining all the codes of the three primers for every accession. By using barcode technology molecular ID can be transferred into a barcode ID that can be quickly scanned by machine.[Conclusion]Based on the PIC of amplification and identified rate, core SSR primers can be screened out to distinguish all the landraces, bred cultivars, and related species. And by constructing fingerprints, every apple germplasm obtains its differentiable molecular ID that can be recognized by the machine. The purpose was to distinguish the most apple germplasms by using the least and the most specific primers.关键词
苹果/种质资源/TP-M13-SSR/分子身份证Key words
apple(Malus Mill.)/germplasm resources/TP-M13-SSR/molecular ID引用本文复制引用
高源,刘凤之,王昆,王大江,龚欣,刘立军..苹果部分种质资源分子身份证的构建[J].中国农业科学,2015,(19):3887-3898,12.
