云南农业大学学报（自然科学）2018，Vol.33Issue(5)：826-835,10.DOI:10.12101/j.issn.1004-390X(n).201712004
云南蔓草虫豆丛枝植原体16S rDNA和secY基因序列分析
Analysis of the Sequences of 16S rDNA and secY of Cajanus scarabaeoides Witches'-broom Phytoplasma in Yuanmou, Yunnan Province
摘要
Abstract
[Purpose] To determine the pathogen of Cajanus scarabaeoides witches'-broom disease samples which were collected in Yuanmou of Yunnan Province.[Method] Near-full-length phytoplasmal 16S rDNA gene sequences and secY gene sequences were amplified via direct-PCR and nested-PCR using phytoplasma universal primer pairs from DNAs extracted from C. scarabaeoides symptomatic plants. The amplicons were cloned and sequenced. DNA fragments about 1.8 kb of 16S rDNA and 1.7 kb of secY were obtained.[Result]Classification of 16S rDNA by using computersimulated restriction fragment length polymorphism (RFLP) and phylogenetic analysis in iPhyClassifier indicated that the pathogen of C. scarabaeoides witches'-broom is a member of phytoplasma subgroup A, group 16SrII (similarity coefficient 1.00), which related to Candidatus Phytoplasma australasiae. From the homology comparison and the phylogenetic tree based on secY gene sequences showed that genetic relationship of C. scarabaeoides witches '-broom phytoplasma strain (CaSWBYNym) was closely related with PnWB from Yunnan and Taiwan Province and confirmed that CaSWB-Ynym belongs to the member of subgroup A, group 16SrII. The classification result based on secY gene analysis coincided with those of results based on 16SrDNA gene.[Conclusion]To our best knowledge, it is the first report of a 16SrII group phytoplasma associated with C. scarabaeoides witches'-broom disease in Yunnan Province. And it is the first study to determine the phytoplasma pathogen of C. scarabaeoides witches'-broom disease in Yunnan Province and identify taxonomic status using molecular biological techniques. These results will provide a theoretical basis for early diagnosis and detection of C. scarabaeoides witches'-broom disease.关键词
蔓草虫豆丛枝病/植原体/16S rDNA/secY基因/序列分析Key words
Cajanus scarabaeoides witches'-broom/phytoplasma/16S rDNA/secY/sequence analysis分类
农业科技引用本文复制引用
王柱华,刘俊男,杨子祥,赵宇,蔡红,陈海如..云南蔓草虫豆丛枝植原体16S rDNA和secY基因序列分析[J].云南农业大学学报（自然科学）,2018,33(5):826-835,10.基金项目
云南省高校科技创新团队支持计划(云科教[2014]22号) （云科教[2014]22号）
国家自然科学基金项目(31060239). （31060239）
