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以黑木耳为试材,利用PCR技术扩增黑木耳18S rDNA序列并测序,再用生物软件Clustalx 2、MEGA 5.1对测序结果进行分析和RNA二级结构预测。根据18S rDNA序列多样性分析和预测的二级结构,研究来自中国东北的5个黑木耳栽培菌株的遗传多样性,以便为黑木耳栽培引种和新品种选育提供参考。结果表明:扩增的18S rDNA序列长度约为1 700bp,去除两端测序质量不好的区域后长度为1 695bp,GC含量为48.02%~48.32%,其核苷酸变异位点25个,信息位点10个,变异位点主要集中在88~107bp区域,N-J系统进化树中5个黑木耳聚为一支,毛木耳单独聚另一支,18S rDNA序列分支与传统的分类结果基本一致;18S rDNA区序列一、二级结构相结合可为黑木耳分类鉴定提供相关的分子结构信息。
Using black fungus as test material, the 18S rDNA sequence of black fungus was amplified by PCR and sequenced. The sequencing results were analyzed by biological software Clustalx 2 and MEGA 5.1, and RNA secondary structure was predicted. Based on the analysis of the 18S rDNA sequence diversity and the predicted secondary structure, the genetic diversity of five black fungus cultivars from Northeast China was studied in order to provide references for the introduction of black fungus and the breeding of new varieties. The results showed that the amplified 18S rDNA sequence was about 1 700 bp in length, with a length of 1 695 bp after removal of poorly sequenced regions at both ends, a GC content of 48.02% -48.32% and a nucleotide variation site of 25 There were 10 loci in these loci. The variation loci were mainly located in the region of 88 ~ 107bp. Five black fungus species in NJ phylogenetic tree were clustered together, and the other one was found in the tree species. The 18S rDNA sequence divergence was basically consistent with the traditional classification results. The combination of 18S rDNA sequence of primary and secondary structures can provide relevant molecular structure information for classification and identification of black fungus.