稻蝗属特异性SCAR分子标记的鉴定.pdfVIP

中国农业科学 2010,43(23):4834-4839 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2010.23.008 稻蝗属特异性SCAR分子标记的鉴定 张建珍，李大琪，李 涛，马恩波，郭亚平 （山西大学应用生物学研究所，太原 030006 ） 摘要：【目的】研究稻蝗属特异性 DNA 分子标记，为稻蝗属物种的分类鉴定提供快速有效的分子检测方法。 【方法】基于稻蝗属物种及其近缘属种的大量 RAPD-PCR 结果，筛选出稻蝗属物种特异性的 RAPD 条带，对该特异 RAPD条带进行克隆、测序。基于所测序列设计特异引物，以稻蝗属不同物种和蝗总科其它物种基因组DNA为模板 进行 PCR 扩增。【结果】随机引物 S823 可在稻蝗属不同物种扩增出约 650 bp 的 RAPD 条带，经对该条带的克隆、 测序，发现在 3个受试的稻蝗属物种中序列同源度达92.3%—96.6%，序列 G+C含量大于15%, 并富含大量的A、T 重复区；基于已知序列设计的特异引物对稻蝗属7个物种可以扩增出目的条带 (550 bp)，而对赤胫伪稻蝗及蝗总 科其它物种均无扩增条带。【结论】鉴定了稻蝗属特异的 RAPD 条带，基于该条带序列设计的特异引物可用于稻蝗 属物种的快速分子鉴定。 关键词：稻蝗属；RAPD-PCR；SCAR分子标记 Identification of Oxya-Specific SCAR Molecular Marker ZHANG Jian-zhen, LI Da-qi, LI Tao, MA En-bo, GUO Ya-ping (Research Institute of Applied Biology, Shanxi University, Taiyuan 030006) Abstract: 【Objective 】 Oxya-specific molecular marker was studied in order to provide a rapid and effective molecular identification method. 【Method 】 Oxya-specific RAPD band was screened out from lots of RAPD-PCR results, the specific band was cloned and sequenced, a pair of primer was designed based on homologue region after sequence alignment. PCR was performed by using designed specific primers, genomic DNA from different Oxya species and other Acridoidea species were used as template. 【Result 】 A clear and reproductive band about 650 bp amplified from random primer S823 was present in different Oxya species. Sequence analysis showed high nucleotide homology (92.3%-96.6%) among three Oxya species, each sequence determined consisted of 651-667 nucleotides, with G+C content 15% and repetitive A or T regions. Sequence characterized amplified region (SCAR) primers for specific PCR were develope