小麦AFLP片段序列多态性分析和AFLP-SCAR标记的研究  

Sequence Polymorphism of Wheat AFLP Fragments and Conversion of AFLP Marker to SCAR Marker

作　　者：王立新[1] 常利芳[1] 黄岚[2] 王晓维[3] 赵昌平[1] 

机构地区：[1]北京市农林科学院/北京杂交小麦工程技术研究中心,北京100097 [2]华中农业大学生命科学技术学院,湖北武汉430070 [3]内蒙古农业大学生物工程学院,内蒙古呼和浩特010018

出　　处：《麦类作物学报》2007年第6期943-951,共9页Journal of Triticeae Crops

基　　金：北京市自然科学基金项目(5053026);北京市农业育种基础研究创新平台项目(YZPT01-06)

摘　　要：为了解相同长度的AFLP片段中DNA序列的多态性,对70条小麦AFLP片段进行回收、克隆以及测序分析,比较了不同品种间相同长度的AFLP片段、一条AFLP片段的不同单克隆以及品种间非多态性AFLP片段的DNA序列。对70个片段构成的98对同长片段进行DNA序列比对,结果表明：（1）只有3对片段的序列100%相同,其他95对片段之间DNA序列差异为1%-70%;（2）不同小麦品种间同长的AFLP片段中普遍存在DNA序列完全不同、部分不同和具有SNP的现象;（3）一条回收的AFLP带纹的不同单克隆之间也存在DNA序列完全不同、部分不同和具有SNP的现象;（4）品种间有些非多态性AFLP片段的DNA序列也存在多态性。揭示了小麦三套基因组间AFLP等位基因的多态性,以及不同AFLP位点同长片段的DNA序列多态性,对AFLP标记的使用提出了一些不同看法,并提出了利用AFLP片段的序列多态性设计SCAR引物的方法。Amplified fragment length polymorphism（AFLP） is an efficient technique to identify DNA fragment length polymorphism,which was widely used for genetic diversity analysis,construction of genetic maps and genes mapping in fungi,animals,plants and insect.But some works could not get expectant result while amplified fragment length polymorphism included sequences polymorphism,single nucleotide polymorphisms（SNPs） and insertions and deletions（indel）.To understand DNA sequence polymorphism of wheat AFLP fragments,we compared 30 pair fragments in which every pair fragments were the same size but from different cultivars,and other 15 pair fragments in which every pair fragments from the same band.The sequences of every pair fragments were not all identical.The difference of each pair fragments were 1%～70%.Even non-polymorphic bands between cultivars contained multiple fragments with sequence difference.In this study we present sequence differences of 95 pair fragments with the same size from 64 wheat AFLP bands and they should locate different loci or alleles among chromosome A,B and D.That is the reason of some AFLP markers could not be mapped and some wheat varieties could not be clustered in the right group by AFLP markers.This paper also describes the method for the conversion of AFLP markers to sequence characterized amplified region（SCAR） markers based on sequence polymorphisms of AFLP fragments for single locus and SNP assays.

关 键 词：小麦 AFLP标记 多态性 AFLP-SCAR标记 

分 类 号：S512.1[农业科学—作物学] S330