摘要
以非洲黑麦、小麦-非洲黑麦双二倍体、安岳排灯麦等为材料筛选100条ISSR引物。其中,引物UBC815可在非洲黑麦中扩增出1条长561bp的特异性片段(命名为pSaUBC815561),而小麦对照均未扩出该片段。引物UBC815同样能在黑麦属的瓦维洛夫黑麦(Secale vavilovii Grossh.)、森林黑麦(Secale sylvestre Host.)等5个种扩增出pSaUBC815561。根据pSaUBC815561设计特异PCR引物U815-F、U815-R,对小麦族多物种进行扩增,表明pSaUBC815561为黑麦属特有。进而利用一套中国春-Imperial黑麦二体附加系及小麦-黑麦异源材料进行扩增,结果显示,pSaUBC815561分布在黑麦整套染色体上,并且所有后代材料都能扩增出pSaUBC815561,表明pSaUBC815561可作为特异性标记用来检测小麦背景中的黑麦染色质。
Genome in situ hybridization （GISH） analysis of wheat-Secale africanum amphiploid revealed that the S. africanum genome displayed significant divergence to the Secale cereale genome. It is thus valuable to deploy genes from S. africanum. We performed the PCR analysis on S. africanum, wheat-S, aflicanum amphiploid, T. eastivum cv. Anyuepaideng and other genetic stocks by 100 ISSR primers. A specific segment of 561 bp, named pSaUBC815 561, was obtained from S. africanum using primer UBC815. This segment was not amplified from the control wheat lines. Primer UBC815 also amplified fragments from wild species of genus Secale, including S. vavilovii, S. sylvestre, and other cultivated ryes. Based on the sequence of pSaUBC815 561, a pair of special primers U815-F and U815-R was designed and was used to amplify the DNA of wheat related species in Triticeae aimed at validating the specificity of pSaUBC815 561. PCR analysis indicated that this specific DNA fragment was amplified not only from a set of Chinese Spring wheat-Imperial rye chromosome addition lines but also from certain wheat-rye introgression lines. Therefore, pSaUBC815 561 can be used as a specific marker for detection of chromosomes of Secale genome in wheat.
出处
《遗传》
CAS
CSCD
北大核心
2008年第8期1056-1062,共7页
Hereditas（Beijing)
基金
国家自然科学基金(编号:30671288和30671136)
教育部新世纪优秀人才资助项目(编号:NCET-06-0810)资助~~
关键词
非洲黑麦
ISSR分子标记
特异PCR
Secale africanum
ISSR （Inter-simple sequence repeat） marker
specific PCR
