Department of Biotechnology, Faculty of Science, Taif University, Taif 21974, Saudi Arabia.
Department of Genetics, Faculty of Agriculture-Chatby, University of Alexandria, Alexandria, Egypt.
Biomolecules. 2012 Apr 13;2(2):228-39. doi: 10.3390/biom2020228.
orf256 is a wheat mitochondrial gene associated with cytoplasmic male sterility (CMS) that has different organization in various species. This study exploited the orf256 gene as a mitochondrial DNA marker to study the genetic fingerprint of Triticum and Aegilops species. PCR followed by sequencing of common parts of the orf256 gene were employed to determine the fingerprint and molecular evolution of Triticum and Aegilops species. Although many primer pairs were used, two pairs of orf256 specific primers (5:-94/C: 482, 5:253/C: 482), amplified DNA fragments of 576 bp and 230 bp respectively in all species were tested. A common 500 bp of nine species of Triticum and Aegilops were aligned and showed consistent results with that obtained from other similar chloroplast or nuclear genes. Base alignment showed that there were various numbers of base substitutions in all species compared to S. cereal (Sc) (the outgroup species). Phylogenetic relationship revealed similar locations and proximity on phylogenetic trees established using plastid and nuclear genes. The results of this study open a good route to use unknown function genes of mitochondria in studying the molecular relationships and evolution of wheat and complex plant genomes.
orf256 是一个与细胞质雄性不育(CMS)相关的小麦线粒体基因,在不同物种中有不同的组织。本研究利用 orf256 基因作为线粒体 DNA 标记,研究了小麦属和山羊草属物种的遗传指纹。采用 PCR 技术,对 orf256 基因的常见部分进行测序,以确定小麦属和山羊草属物种的指纹和分子进化。尽管使用了许多引物对,但测试了两对 orf256 特异性引物(5:-94/C:482, 5:253/C:482),它们分别在所有物种中扩增出 576 bp 和 230 bp 的 DNA 片段。对 9 种小麦属和山羊草属物种的共同 500 bp 进行了比对,并与从其他类似叶绿体或核基因获得的结果一致。碱基比对显示,与外群物种(Sc)相比,所有物种都有不同数量的碱基替换。系统发育关系显示,使用质体和核基因建立的系统发育树上的位置和接近度相似。本研究为利用线粒体未知功能基因研究小麦和复杂植物基因组的分子关系和进化开辟了良好的途径。