偃麦草E染色体组特异RAPD和SCAR标记的建立  

Establishment of E-Genome-Specific RAPD and SCAR Markers for Thinopyrum spp.

作　　者：尤明山[1] 李保云[1] 唐朝晖[1] 刘守斌[1] 宋健民 毛善锋[1] 刘广田[1] 

机构地区：[1]中国农业大学作物学院,北京100094

出　　处：《中国农业大学学报》2002年第5期1-6,共6页Journal of China Agricultural University

基　　金：国家自然科学基金重点资助项目 (39930 110 );北京市自然科学基金重大资助项目 (6 990 0 0 1)

摘　　要：用 10 0条 10碱基随机引物 ,以普通小麦中国春、中间偃麦草为材料进行 RAPD分析 ,筛选到一个偃麦草染色体组特异引物 OPF0 3,并从中间偃麦草中克隆了该引物的特异 DNA片段 OPF0 312 91。将该片断与比萨偃麦草中的 OPF0 312 96(Gen Bank序号 U4 35 16 )比较 ,同源性为 88%。根据 OPF0 312 91的序列 ,设计了 2对SCAR引物 ,利用 OPF0 3和引物 P3、P4对普通小麦、普通小麦 -中间偃麦草的部分双二倍体、长穗偃麦草、中间偃麦草、小麦 -二倍体长穗偃麦草代换系、附加系共 6类材料进行了 RAPD和 SCAR分析 ,发现 RAPD标记OPF0 312 91没有出现在含 Ee染色体组的材料中 ,而标记 SCAR982 则出现于所有含 E染色体组的材料中。说明 ,根据 Eb 染色体组特异 RAPD标记转化的 SCAR标记 ,可以同时检测小麦背景下的 EeA total of 100 decamer oligonucleotides were used as primers to perform random amplified polymorphic DNA (RAPD) analysis on Triticum aestivum var. Chinese Spring and Thinopyrum intermedium. Out of these primers, one could amplify a specific DNA fragment in the accession SZ of Th. intermedium. This fragment, OPF03 1291 , was cloned and sequenced. Sequence data searching in GenBank showed that this fragment had a homologous degree of 88% to a RAPD marker OPF03 1296 (GenBank accession U43516) in Th. bessarabicum. Based on the sequence of OPF03 1291 , two pairs SCAR primers were designed. With OPF03 and primers P3 and P4, RAPD and SCAR analyses were performed on T.aestivum, T.aestivum Th.intermedium partial amphidiploid, Th.elongatum, Th.intermedium, wheat Th.elongatum substitution and addition lines. The results of PCR amplification showed that the RAPD marker was not appeared in materials with genome E e, whereas SCAR marker was appeared in all E genome possessed materials. The SCAR marker derived from the E b genome specific RAPD fragment could be used in the meantime to detect the chromosome of E e genome under common wheat background.

关 键 词：偃麦草 E染色体组 特异RAPD SCAR标记 

分 类 号：S512.1[农业科学—作物学]