Ellneskog-Staam Pernilla, Taketa Shin, Salomon Björn, Anamthawat-Jónsson Kesara, von Bothmer Roland
Department of Crop Science, Swedish University of Agricultural Sciences, Alnarp, Sweden.
Hereditas. 2006 Dec;143(2006):103-12. doi: 10.1111/j.2006.0018-0661.01953.x.
Wood barley, Hordelymus europaeus, was compared with other Triticeae species by Southern and fluorescence in situ hybridisation using total genomic DNA and repetitive sequences as probes. On Southern blots, the total genomic probe from H. europaeus hybridised strongly to DNA of its own species and to Leymus and Psathyrostachys, indicating the presence of Ns genome in H. europaeus. Furthermore, the total genomic probe from P. fragilis hybridised to DNA of H. europaeus as much as to all of the Psathyrostachys and Leymus species examined. Ns genome-specific DNA sequences isolated from L. mollis (pLmIs1, pLmIs44 and pLmIs53) hybridised essentially to H. europaeus and all of the species of Leymus and Psathyrostachys. Chromosomal localization of these clones on H. europaeus confirmed the presence of Ns genome-specific DNA on all chromosomes, indiscriminately. Under moderate hybridisation stringency the Ns genome-specific probes, together with repetitive sequences pTa71 and pAesKB7, produced species-specific RFLP banding profiles on Southern blots. A phenetic tree based on these profiles revealed a distinct Ns species cluster within the Triticeae, represented by Leymus and Psathyrostachys species. Hordelymus europaeus belonged to this Ns cluster. Chromosomal mapping of the 18S-25S and the 5S ribosomal genes, together with the repetitive sequence pLrTaiI, corroborated that H. europaeus was most probably related to Leymus, especially the European/Eurasian members of sect. Leymus. In an attempt to identify the genome of H. europaeus, different approaches were employed; the results clearly showed that wood barley had the Ns basic genome and nothing else.
利用总基因组DNA和重复序列作为探针,通过Southern杂交和荧光原位杂交,将欧洲大麦草(Hordelymus europaeus)与其他小麦族物种进行了比较。在Southern杂交印迹上,欧洲大麦草的总基因组探针与其自身物种以及赖草属(Leymus)和新麦草属(Psathyrostachys)的DNA强烈杂交,表明欧洲大麦草中存在Ns基因组。此外,脆弱新麦草(P. fragilis)的总基因组探针与欧洲大麦草DNA的杂交程度与所有检测的新麦草属和赖草属物种相同。从软穗赖草(L. mollis)中分离出的Ns基因组特异性DNA序列(pLmIs1、pLmIs44和pLmIs53)基本上与欧洲大麦草以及所有赖草属和新麦草属物种杂交。这些克隆在欧洲大麦草染色体上的定位证实了所有染色体上均存在Ns基因组特异性DNA,无差别。在中等杂交严谨度下,Ns基因组特异性探针与重复序列pTa71和pAesKB7一起在Southern杂交印迹上产生了物种特异性的RFLP条带图谱。基于这些图谱构建的聚类树显示,在小麦族中存在一个以赖草属和新麦草属物种为代表的独特的Ns物种簇。欧洲大麦草属于这个Ns簇。18S - 25S和5S核糖体基因以及重复序列pLrTaiI的染色体定位证实,欧洲大麦草很可能与赖草属相关,特别是赖草属sect. Leymus的欧洲/欧亚成员。为了鉴定欧洲大麦草的基因组,采用了不同的方法;结果清楚地表明,欧洲大麦草具有Ns基本基因组,别无其他。