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【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】 The aim of this study was to study the molecular markers of rice genus Locust and to provide a rapid and effective molecular detection method for the classification and identification of the genus. 【Method】 Based on a large number of RAPD-PCR results of Ophioderma species and their related genera, RAPD bands were screened for species-specific RAPD. The specific RAPD bands were cloned and sequenced. Specific primers were designed based on the sequences tested, and genomic DNA from different species of Oxya species and other species of Acridoidea were used as templates for PCR amplification. 【Result】 A random amplified polymorphic DNA (RAPD) band of about 650bp was amplified from different species of Oxya. According to the cloning and sequencing of this band, we found that the sequence homology 92.3% -96.6%. The sequence G + C content was more than 15%, and was rich in A and T repeats. Based on the specific primers designed with known sequences, the target bands (550 bp ), But no amplification band was found in other species of Ch both wild rice and locust. 【Conclusion】 The RAPD bands specific to Oxya chrysanthum were identified. The specific primers designed based on this band sequence can be used for rapid molecular identification of Oxya spp. Species.