A nested PCR assay was employed to detect the presence of phytoplasmas in 127 blueberry plants exhibiting typical or a portion of blueberry stunt (BBS) syndrome collected in 2010 and 2011, from 11 commercial farms predominantly located in two counties in New Jersey, USA. Ninety plants exhibiting typical stunt syndrome tested positive for phytoplasma infection. Restriction fragment length polymorphism (RFLP) analysis indicated that two distinct phytoplasmas were associated with BBS-diseased plants. About 95% of phytoplasmas detected were very closely related to BBS phytoplasma strains BBS3-AR (subgroup 16SrI-E) and BBS1-MI (unidentified) identified previously, and 4.4% of phytoplasmas detected belonged to the pigeon pea witches'-broom phytoplasma group (16SrIX). Sequence and phylogenetic analysis of cloned 16S rDNA further indicated the subgroup 16SrI-E related phytoplasmas represented a variant of 16SrI-E reference strain BBS3-AR, while the 16SrIX related phytoplasmas were closely related to juniper witches'-broom (JunWB) phytoplasma (16SrIX-E), representing a 16SrIX-E variant. Ribosomal protein (rp) and secY gene-based phylogenies revealed that BBS3-AR and BBS-NJ 16SrI-E strains belonged to a closely related lineage, while BBS-NJ 16SrIX-E strains and JunWB strains represented two distinct lineages. Single nucleotide polymorphisms (SNPs) analyses of rp and secY gene sequences further revealed that no specific rp gene SNPs and only two specific secY gene SNPS were present between BBS-NJ 16SrI-E strains and BBS3-AR. In contrast, BBS-NJ 16SrIX-E strains/clones had 15 consensus rp SNPs and 28 consensus secY SNPs that separated them from JunWB strains/clones. For the first time, two distinct phytoplasmas that cause BBS-disease in the U.S. was revealed.

译文

采用巢式PCR测定法检测了127株蓝莓植物中植原体的存在,这些植物表现出典型的或部分的蓝莓特技 (BBS) 综合征,2010年和2011年从主要位于美国新泽西州两个县的11个商业农场收集。90种表现出典型特技综合症的植物对植原体感染呈阳性反应。限制性片段长度多态性 (RFLP) 分析表明,两个不同的植原体与患BBS的植物有关。检测到的约95% 的植物原质与BBS3-AR (16SrI-E亚组) 和先前鉴定的BBS1-MI (未鉴定) 的BBS植物原质非常密切相关,检测到的4.4% 的植物原质属于木豆巫婆-扫帚植物原质组 (16srix)。克隆的16S rDNA的序列和系统发育分析进一步表明,与16SrI-E相关的植物质亚群代表了16SrI-E参考菌株BBS3-AR的变体,而与16srix相关的植物质与杜松巫婆扫帚 (JunWB) 植物质密切相关 (16SrIX-E),代表16SrIX-E变体。核糖体蛋白 (rp) 和基于secY基因的系统发育表明,BBS3-AR和BBS-NJ 16SrI-E菌株属于密切相关的谱系,而BBS-NJ 16SrIX-E菌株和JunWB菌株代表两个不同的谱系。rp和secY基因序列的单核苷酸多态性 (snp) 分析进一步表明,在BBS-NJ 16SrI-E菌株和BBS3-AR之间不存在特定的rp基因snp和仅两个特定的secY基因snp。相反,BBS-NJ 16SrIX-E菌株/克隆具有15个共有的rp snp和28个共有的secY snp,将它们与JunWB菌株/克隆分开。首次揭示了在美国引起BBS疾病的两种不同的植原体。

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