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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。
A large number of phytoplasma particles were observed in the ventral phloem phloem cell of the Syringa oblata sample showing leafy, chlorotic symptoms by transmission electron microscopy. The total DNA of Clove plants was amplified by nested PCR using phytoplasma 16S rRNA gene universal primers, and a target fragment of about 1.2 kb was obtained. The amplified fragment was sequenced and phylogenetic analysis And homology analysis showed that the fragment was 1 246 bp in length and was clustered with members of Candidatus Phytoplasma asteris in the phylogenetic tree, More than 98%. The 16Sr RNAⅠ and Ⅴ specific primers were used to determine the non-mixed infection of this disease. The similarity coefficient and RFLP analysis indicated that this phytoplasma belonged to 16SrⅠ-B subgroup. This is the first report on Ascochyta spp. Infection on clove in China.