稻蝗属特异性SCAR分子标记的鉴定

Identification of Oxya-Specific SCAR Molecular Marker

【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650bp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(550bp),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。

【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 developed,an expected specific band（550 bp） was obtained by using SCAR primers in seven species of Oxya examined but absent in Pseudoxya diminuta and other Acridoidea species.【Conclusion】 Oxya-specific SCAR molecular marker was identified,and SCAR primers can be used for rapid identification of Oxya species.