摘要
以植原体通用引物P1/P7和R16F2n/R16R2对云南省峨山县发现的山黄麻丛枝病感病植株,总DNA进行巢式PCR扩增、克隆和测序,获得1 246bp原体16S rDNA序列.经iPhyClassifier植原体在线分类鉴定工具分析,显示该植原体与翠菊黄化植原体(Candidatus Phytoplasma asteris)相关,与该组参考株系(M30790)的序列相似性为99.6%,属16SrI组成员,16S rDNA的F2nR2片段的RFLP模型与16Sr I-B亚组的参考模型(AP006628)最为相似,相似系数F为0.96,推测是一个16SrI组新亚组.以MEGA软件和MUSCLE在线工具进行多序列比对和系统进化树构建,显示山黄麻丛枝植原体(SHMWB-ES)与16Sr I组成员聚为一支,在亚组构建中显示16Sr I组与16Sr I-B亚组的菌株BSP-Br1(EU423898)、16Sr I-X亚组的TLWBYN01(MN513329)、BG01(LT594117)及BG02(LT594118)菌株序列同源性都在99%以上,与前两者更高达为99.76%和99.84%.
For the Trema witches’-broom disease found in Eshan county,Yunnan province,the total DNA of affected plants were amplified by using nested-PCR with universal primers P1/P7 and R16F2n/R16R2,then the amplified fragments were cloned and sequenced.Finally,the 16S rDNA sequence(1 246 bp)of Trema witches’-broom phytoplasma was obtained.The analysis by iPhyClassifier which was used for online classification and identification of phytoplasmas reveals that the phytoplasma was related to‘Candidatus Phytoplasma asteris’;the 16S rDNA sequence shares 99.6% similarity with that of the‘Candidatus Phytoplasma asteris’ reference strain(GenBank accession:M30790).And the phytoplasma under study was a member of 16Sr group I.The virtual RFLP pattern derived from the 16S rDNA F2nR2 fragment was most similar to the reference pattern of the 16Sr group I,subgroup B(GenBank accession:AP006628),with a similarity coefficient of 0.96.This strain may represent a new subgroup within the 16Sr group I.The multi-sequence alignment and phylogenetic tree construction that were demonstrated using MEGA software and MUSCLE online tool showed that Trema witches’-broom phytoplasma(SHMWB-ES)and members of 16SrI group were clustered into one branch,and it was more closely related to 16Sr I-B strain BSP-Br1(EU423898)and 16Sr I-X strains TLWBYN01(MN513329),BG01(LT594117),BG02(LT594118)among the 16SrI subgroup,the homology with the former two strains was the highest,99.76% and 99.84%respectively,and the rest were almost above 99%.Trema was a new host of phytoplasma.
作者
万琼莲
苏帆
许杏萍
蔡红
王连春
WAN Qionglian;SU Fan;XU Xingping;CAI Hong;WANG Lianchun(School of Chemical Biology and Environment,Yuxi Normal University,Yuxi,Yunnan 653100;Key Laboratory of Agro-Biodiversity and Pest Management of Education Ministry of China,Yunnan Agricultural University,Kunming,Yunnan 650201)
出处
《玉溪师范学院学报》
2020年第3期46-52,共7页
Journal of Yuxi Normal University
基金
云南省教育厅科学研究基金项目资助性项目(2017ZZX176)
云南省教育厅科学研究基金项目指导性项目(2017ZDX136)
关键词
植原体
山黄麻
丛枝
分子检测
16S
rDNA
Phytoplasma
Trema Lour.
witches’-broom
molecular identification
16S rDNA
