黑麦基因组特异DNA片段的分离与SCAR标记的建立

Specific DNA Band Isolation and SCAR Marker Construction of Rye Genome

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。

One primer,Hll,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 OPH11679. According to OPHl1679 specific PCR primers H11-F and H11-R were designed and the other Tritlceae species and the species carrying rye chromatin were examined and a band of 643 bp（ named as pScH643） was only amplified from the species carrying rye chromatin,indicating that it was rye specific. pScH643 was amplified from a set of China spring-Imperial addition lines with H11-F and Hll-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, pScH613 could be used as the SCAR marker to detect materials carrying rye chromatin .