• Title/Summary/Keyword: virtual RFLP

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Molecular Detection of Phytoplasmas of the 16SrI and 16SrXXXII Groups in Elaeocarpus sylvestris Trees with Decline Disease in Jeju Island, South Korea

  • Geon-Woo, Lee;Sang-Sub, Han
    • The Plant Pathology Journal
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    • v.39 no.1
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    • pp.149-157
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    • 2023
  • 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).

Phytoplasma Infection could Affect Chemical Composition of Artemisia sieberi

  • Hemmati, Chamran;Nikooei, Mehrnoosh
    • The Plant Pathology Journal
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    • v.35 no.3
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    • pp.274-279
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    • 2019
  • Artemisia sieberi showing symptoms resembling those caused by phytoplasma were observed in Geno Mountain, Hormozgan Province, Iran, and were examined for phytoplasma presence by PCR assays. In addition, the essential oils hydrodistilled from the aerial parts of phytoplasma-infected and healthy plants have been analyzed and compared by GC and GC/MS. Phylogenetic and virtual RFLP analysis of the 16S rRNA gene sequences revealed that the phytoplasma associated with A. sieberi witches' broom (AsWB) was a strain of 'Candidatus Phytoplasma aurantifolia'. The presence of the disease, however, induced a further enrichment (from 4.9 to 45.2%, a relative increase of 90%) of the entire monoterpene class as compared to the abundance in healthy samples. Conversely, a matching decrease in monoterpenoid (from 48.7 to 2%, a relative decrease of 90.2%) was observed in the infected plants. Besides the first report of phytoplasma infection of A. sieberi, the changes of its essential oils are reported.