紫花苜蓿丛枝病植原体的分子检测及鉴定  

Molecular detection and identification of a phytoplasma associated with lucerne witches'-broom

作　　者：刘涛[1] 蔡红[1] 赵丹[1] 陈海如[1] 

机构地区：[1]云南农业大学云南省植物病理重点实验室,昆明650201

出　　处：《植物病理学报》2007年第4期362-367,共6页Acta Phytopathologica Sinica

基　　金：国家自然科学基金资助项目(30260064);云南省自然科学基金资助项目(2000C0014Q)

摘　　要：利用植原体16S rRNA基因通用引物对云南昆明发生的苜蓿丛枝病感病植株总DNA进行巢式PCR扩增,得到1.2kb的特异片段,从分子水平证实了苜蓿丛枝病的病原是植原体。从PCR产物的RFLP酶切图谱可看出,该植原体株系的酶切图谱与马里兰翠菊黄化植原体(AY1)相同。对扩增片段进行克隆及序列测定后,利用最小进化法做Bootstrap验证的系统进化树,表明苜蓿丛枝病植原体为Candidatus Phytoplasma asteris成员之一,与植原体16SrI-B亚组成员关系密切。Using phytoplasma universal primers for 16S rRNA gene to detect phytoplasma associated with lucerne witches＇ -broom （LuWB） by nested-PCR. DNA fragment of 1.2 kb was amplified from the total DNA of diseased lucerne samples. The RFLP profile of the 1.2 kb fragment of the LuWB phytoplasma swain is as the same as that of Maryland aster yellows （ AY1 ）. After cloning and nucleotide sequencing of the amplified fragment, a phylogenetic tree based on 16S rDNA sequences was constructed by the minimum evolution method and delineated that lucerne witches ＇-broom phytoplasma is a member of Candidatus Phytoplasma asteris, most closely related to the phytoplasma subgroup 16SrI-B.

关 键 词：巢式PCR 紫花苜蓿 丛枝病 植原体 系统进化 

分 类 号：S432.42[农业科学—植物病理学]