丁香卷叶病植原体的分子鉴定  

Molecular detection and identification of a phytoplasma associated with lilac leaf roll

作　　者：李正男[1] 张磊[2] 吴云锋[1] 

机构地区：[1]旱区作物逆境生物学国家重点实验室农业部西北黄土高原作物有害生物综合治理重点实验室西北农林科技大学植保学院,杨凌712100 [2]西北农林科技大学林学院,杨凌712100

出　　处：《植物病理学报》2012年第2期131-138,共8页Acta Phytopathologica Sinica

基　　金：国家自然科学基金(30970133);高等学校博士学科点基金(20100204110004);高等学校学科创新引智计划(B07049)

摘　　要：通过透射电子显微镜,在表现卷叶、褪绿症状的丁香(Syringa oblata)样品的叶脉韧皮部筛管细胞内观察到大量植原体粒子。应用植原体16S rRNA基因通用引物对P1/P7和R16F2n/R16R2对表症丁香植株总DNA进行巢式PCR扩增,得到了约1.2 kb的目标片段,通过对扩增片段进行测序、系统发育分析和同源性分析,结果表明,该片段长度为1 246 bp,在系统发育进化树上与翠菊黄化组(Candidatus Phytoplasma asteris)成员是聚集在一起的,与该组成员同源性均在98%以上。用16Sr RNAⅠ组和Ⅴ组特异引物确定了该病害非混合侵染所致,相似性系数和RFLP分析表明该植原体属于16SrⅠ-B亚组。这是国内关于翠菊黄化组植原体在丁香上感染的首次报道。In those of Syringa oblata plants showing symptoms of leaf roll, chlorotic, a great quantity of structures resembling phytoplasmas were observed in small pieces of phloem sieve elements by Electron microscopy. Nested PCR using a combination of phytoplasma-specific universal primer pairs （P1/P7- R16F2n/ R16R2） amplified 16S rDNA with the expected size （1.2 kb） from all samples of symptomatic S. oblata plants. On the basis of sequencing, phylogenetic analysis and nucleotide alignments to PCR products, a 1 246 bp fragment was obtained that clustered together with the members of ＇ Candidatus Phytoplasma asteris＇ and shared a 98% similarity. Group specific primer pairs R16 （ I ） F1/R16 （ I ） R1 and R16 （ V ） F1/R16 （ V ） proved that this disease was caused by unmixed infection. Similarity coefficient and RFLP analysis indicated that it was belonged to phytoplasmas members of 16Sr I-B. This is the first time to report group ‘Candidatus Phytoplasma asteris＇ infecting S. oblata plants in China.

关 键 词：植原体 16S RRNA基因 PCR 相似性系数 RFLP 

分 类 号：S432[农业科学—植物病理学]