{"title":"Molecular Detection of Phytoplasmas of the 16SrⅠ and 16SrXXXⅡ Groups in Elaeocarpus sylvestris Trees with Decline Disease in Jeju Island, South Korea.","authors":"Geon-Woo Lee, Sang-Sub Han","doi":"10.5423/PPJ.OA.07.2022.0091","DOIUrl":null,"url":null,"abstract":"<p><p>Phytoplasmas were discovered in diseased Elaeocarpus sylvestris trees growing on Jeju Island that showed symptoms of yellowing and darkening in the leaves. Leaf samples from 14 symptomatic plants in Jeju-si and Seogwipo-si were collected and phytoplasma 16S rRNA was successfully amplified by nested polymerase chain reaction using universal primers. The sequence analysis detected two phytoplasmas, which showed 99.5% identity to 'Candidatus Phytoplasma asteris' and 'Ca. P. malaysianum' affiliated to 16SrI and 16SrXXXII groups, respectively. Through polymerase chain reaction-restriction fragment length polymorphism (RFLP) analyses using the AfaI (RsaI) restriction enzyme, the presence of two phytoplasmas strains as well as cases of mixed infection of these strains was detected. In a virtual RFLP analysis with 17 restriction enzymes, the 16S rRNA sequence of the 'Ca. P. asteris' strain was found to match the pattern of the 16SrI-B subgroup. In addition, the phytoplasmas in the mixed-infection cases could be distinguished using specific primer sets. In conclusion, this study confirmed mixed infection of two phytoplasmas in one E. sylvestris plant, and also the presence of two phytoplasmas (of the 16SrⅠ and 16SrXXXⅡ groups) in Jeju Island (Republic of Korea).</p>","PeriodicalId":20173,"journal":{"name":"Plant Pathology Journal","volume":"39 1","pages":"149-157"},"PeriodicalIF":1.8000,"publicationDate":"2023-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/45/54/ppj-oa-07-2022-0091.PMC9929163.pdf","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Plant Pathology Journal","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.5423/PPJ.OA.07.2022.0091","RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"PLANT SCIENCES","Score":null,"Total":0}
引用次数: 0
Abstract
Phytoplasmas were discovered in diseased Elaeocarpus sylvestris trees growing on Jeju Island that showed symptoms of yellowing and darkening in the leaves. Leaf samples from 14 symptomatic plants in Jeju-si and Seogwipo-si were collected and phytoplasma 16S rRNA was successfully amplified by nested polymerase chain reaction using universal primers. The sequence analysis detected two phytoplasmas, which showed 99.5% identity to 'Candidatus Phytoplasma asteris' and 'Ca. P. malaysianum' affiliated to 16SrI and 16SrXXXII groups, respectively. Through polymerase chain reaction-restriction fragment length polymorphism (RFLP) analyses using the AfaI (RsaI) restriction enzyme, the presence of two phytoplasmas strains as well as cases of mixed infection of these strains was detected. In a virtual RFLP analysis with 17 restriction enzymes, the 16S rRNA sequence of the 'Ca. P. asteris' strain was found to match the pattern of the 16SrI-B subgroup. In addition, the phytoplasmas in the mixed-infection cases could be distinguished using specific primer sets. In conclusion, this study confirmed mixed infection of two phytoplasmas in one E. sylvestris plant, and also the presence of two phytoplasmas (of the 16SrⅠ and 16SrXXXⅡ groups) in Jeju Island (Republic of Korea).
在济州岛生长的病树中发现了植物原体,叶子变黄变黑。收集济州和西归浦地区14株病状植物的叶片样本,利用通用引物巢式聚合酶链反应成功扩增出植物原体16S rRNA。序列分析结果显示,2个植物原体分别属于16SrI和16SrXXXII类群的‘Candidatus Phytoplasma asteris’和‘Ca. P. malaysium’,同源性达99.5%。采用AfaI (RsaI)限制性内切酶进行聚合酶链反应-限制性片段长度多态性(RFLP)分析,检测到两株植物原体菌株的存在以及两株植物原体混合感染的病例。通过17种限制性内切酶的虚拟RFLP分析,发现‘Ca. P. asteris’菌株的16S rRNA序列与16SrI-B亚群的模式相匹配。此外,混合感染病例中的植物原体可以通过特异性引物进行区分。结果表明,在韩国济州岛,一株西尔维斯特植物中存在两种植物原体的混合侵染,并且存在两种植物原体(16SrⅠ和16SrXXXⅡ群)。