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目的:采用PCR-DGGE的方法对安徽、江苏、四川、广西等10个产地蝉花的真菌多样性进行分析,以期获得蝉花优势寄生真菌。方法:通过提取样品总DNA,采用真菌通用引物NS1、Fung扩增18S rDNA,将PCR产物DGGE电泳后,对16个差异性条带进行了切胶回收、克隆测序以及BLAST数据库比对验证;对不同产地蝉花的电泳图谱进行了相似度以及多样性指数分析。结果:广西玉林产地蝉花的条带数最多为16条,H=2.679;四川北川、四川绵阳、广东河源3个产地的蝉花真菌多样性相似度较高。结论:克隆测序结果表明共有条带3和9代表的真菌为不同产地蝉花的优势寄生真菌蛹虫草Cordyceps militaris;除了3种未获得纯培养的节担菌纲和真核细胞外,其余的为Eurotium athecium、Magicicada cassini、Myrothecium roridum、Aspergillus penicillioides等菌。
Objective: The PCR-DGGE method was used to analyze the fungal diversity of 10 cicadas in Anhui, Jiangsu, Sichuan and Guangxi so as to obtain the dominant parasitic fungus. Methods: The total DNA of samples was extracted. The common fungal primers NS1 and Fung were used to amplify 18S rDNA. The PCR products were electrophoresed on DGGE, and 16 different bands were excised, cloned and sequenced, and compared with BLAST database. Similarities and diversity indices were analyzed on the electrophoresis patterns of the cicadas from different places. Results: The maximum number of bands in the cicadae flower of Yulin in Guangxi was 16 with H = 2.679. The similarities of the cicadae fungi diversity were found in three producing areas, Beichuan in Sichuan, Mianyang in Sichuan and Heyuan in Guangdong. Conclusion: The results of cloning and sequencing showed that the fungi represented by common bands 3 and 9 were Cordyceps militaris, the dominant parasitic fungus of Cordyceps from different origins. Except for 3 Cordyceps militaris strains which were not cultured in pure culture, the others were Eurotium athecium, Magicicada cassini, Myrothecium roridum and Aspergillus penicillioides.