黑麦基因组特异DNA片段的分离与SCAR标记的建立

以2个栽培黑麦、2个野生黑麦和4个普通小麦为材料,从200条10碱基RAPD随机引物中筛选出1条引物H11。H11在小麦中有1条低拷贝扩增,而在黑麦中却有极高拷贝的扩增。对H11在黑麦中的高拷贝片段进行克隆、测序,得其全长679 bp,记作OPH11679。根据OPH11679设计特异PCR引物H11-F和H11-R,对小麦族其它物种和含黑麦染色质的物种进行验证,结果发现仅含黑麦染色质的物种能扩增出长为643 bp的片段(命名为pScH643),这表明该片段为黑麦所特有。用H11-F和H11-R对1套中国春-Imperial黑麦附加系等进行扩增,结果显示pScH643片段分布在黑麦整套染色体上,这一结果在小麦-黑麦异源材料的分析中得到进一步验证。即表明pScH643片段可作为SCAR标记用于含黑麦染色质材料的检测。

Specific DNA Band Isolation and SCAR Marker Construction of Rye Genome

One primer,H11,was screened from 200 ten-base RAPD random primers with two domesticated Secale cereale L.materials,two wild S.cereale L.materials and four Triticum aestivum L.materials.H11 had one lowly amplifiable copy in T.aestivum L.and a extremely highly amplifiable copy in S.cereale L..The copy in S.cereale L.was cloned and sequenced to have a full length of 679 bp and recorded as OPH11_(679).According to OPH11_(679) specific PCR primers H11-F and H11-R were designed and the other Triticeae species and the species carrying rye chromatin were examined and a band of 643 bp(named as pScH_(643)) was only amplified from the species carrying rye chromatin,indicating that it was rye specific.pScH_(643) was amplified from a set of China spring-Imperial addition lines with H11-F and H11-R and revealed to distribute in the whole chromosome set of rye;this result was far verified in the analysis of wheat-rye materials,that is to say,pScH_(643) could be used as the SCAR marker to detect materials carrying rye chromatin.