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禾本科psbA-trnH基因的序列分析及其在斑点野生稻鉴定中的应用
Analysis of psbA-trnH Gene Sequences of Gramineae and Its Application in Identification of Oryza punctata
【摘要】 通过对稻属(Oryza L.)及其近缘种等禾本科(Gramineae)植物叶绿体psbA-trnH基因进行扩增和测序,分析其系统进化关系及序列差异,并设计鉴定斑点野生稻(O.punctata)的特异分子标记。序列分析的结果表明,栽培稻材料间psbA-trnH基因序列没有或很少存在碱基差异,野生稻相对变异丰富,其中靠近3′端下游序列碱基变异较为丰富,5′端相对保守。根据斑点野生稻psbA-trnH基因序列的特异位点设计的引物,扩增的目的片段ptBD118长118bp,退火温度在62℃时特异性最佳。在条码基因差异位点分析的基础上,开发的特异分子标记用于物种的快速鉴定,为珍贵物种的口岸鉴定提供了检测方法。
【Abstract】 In this paper,after amplifying and sequencing psbA-trnH genes of Oryza L.and its relatives in Gramineae,we analyzed the phylogenetics and differences of these sequences and designed the specific identification primers for Oryza punctata.We found that there are no or a few variations in cultivar rice,but many variations in wild rice variants,especially in the 3′ terminal sequences.According to the specific primers we amplified the target fragment ptBD118 with length of 118 bp in annealing temperature of 62℃,the result of which showed that the work flow could distinguish Oryza punctata from other Gramineae and is appropriate to identification of Oryza punctata.The successful trial for inspection and identification of species showed that discovering specific molecular markers to identify species with high efficiency and quickly based on the analysis of sequence difference of barcode genes.
【Key words】 psbA-trnH gene Oryza punctata Molecular marker Species identification;
- 【文献出处】 生物技术通报 ,Biotechnology Bulletin , 编辑部邮箱 ,2011年02期
- 【分类号】S511.9
- 【被引频次】12
- 【下载频次】227