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BMC microbiology
Nov 25, 2017;17 (1): 221.

Classification of a new phytoplasmas subgroup 16SrII-W associated with Crotalaria witches' broom diseases in Oman based on multigene sequence analysis.

Ali Al-Subhi, Saskia A Hogenhout, Rashid A Al-Yahyai, Abdullah M Al-Sadi
Author Information
  1. Ali Al-Subhi: Department of Crop Sciences, College of Agricultural and Marine Sciences, Sultan Qaboos University, Al Khod 123, PO Box 34, Seeb, Oman.
  2. Saskia A Hogenhout: Department of Crop Genetics, John Innes Centre, Norwich, NR4 7UH, UK.
  3. Rashid A Al-Yahyai: Department of Crop Sciences, College of Agricultural and Marine Sciences, Sultan Qaboos University, Al Khod 123, PO Box 34, Seeb, Oman.
  4. Abdullah M Al-Sadi: Department of Crop Sciences, College of Agricultural and Marine Sciences, Sultan Qaboos University, Al Khod 123, PO Box 34, Seeb, Oman. alsadi@squ.edu.om. ORCID
PMID: 29178845 DOI: 10.1186/s12866-017-1130-3

Abstract

BACKGROUND: Crotalaria aegyptiaca, a low shrub is commonly observed in the sandy soils of wadis desert and is found throughout all regions in Oman. A survey for phytoplasma diseases was conducted. During a survey in a wild area in the northern regions of Oman in 2015, typical symptoms of phytoplasma infection were observed on C. aegyptiaca plants. The infected plants showed an excessive proliferation of their shoots and small leaves.
RESULTS: The presence of phytoplasma in the phloem tissue of symptomatic C. aegyptiaca leaf samples was confirmed by using Transmission Electron Microscopy (TEM). In addition the extracted DNA from symptomatic C. aegyptiaca leaf samples and Orosius sp. leafhoppers were tested by PCR using phytoplasma specific primers for the 16S rDNA, secA, tuf and imp, and SAP11 genes. The PCR amplifications from all samples yielded the expected products, but not from asymptomatic plant samples. Sequence similarity and phylogenetic tree analyses of four genes (16S rDNA, secA, tuf and imp) showed that Crotalaria witches' broom phytoplasmas from Oman is placed with the clade of Peanut WB (16SrII) close to Fava bean phyllody (16SrII-C), Cotton phyllody and phytoplasmas (16SrII-F), and Candidatus Phytoplasma aurantifolia' (16SrII-B). However, the Crotalaria's phytoplasma was in a separate sub-clade from all the other phytoplasmas belonging to Peanut WB group. The combination of specific primers for the SAP11 gene of 16SrII-A, -B, and -D subgroup pytoplasmas were tested against Crotalaria witches' broom phytoplasmas and no PCR product was amplified, which suggests that the SAP11 of Crotalaria phytoplasma is different from the SAP11 of the other phytoplasmas.
CONCLUSION: We propose to assign the Crotalaria witches' broom from Oman in a new lineage 16SrII-W subgroup depending on the sequences analysis of 16S rRNA, secA, imp, tuf, and SAP11 genes. To our knowledge, this is the first report of phytoplasmas of the 16SrII group infecting C. aegyptiaca worldwide.

Keywords

Crotalaria Phytoplasma phylogeny WBD

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Grants

  1. BBS/E/J/000C0623/Biotechnology and Biological Sciences Research Council
  2. SR/AGR/CROP/13/01 and SR/AGR/CROP/17/01/Sultan Qaboos University

MeSH Term

Crotalaria
DNA Primers
DNA, Bacterial
Genes, Bacterial
Microscopy, Electron, Transmission
Oman
Phylogeny
Phytoplasma
Plant Diseases
Polymerase Chain Reaction
RNA, Ribosomal, 16S
Sequence Analysis, DNA
Species Specificity

Chemicals

DNA Primers
DNA, Bacterial
RNA, Ribosomal, 16S
Journal Article
Article has an altmetric score of 1

Word Cloud

Created with Highcharts 10.0.0CrotalariaphytoplasmasphytoplasmaaegyptiacaOmanSAP11Csampleswitches'broomPCR16SsecAtufimpgenessubgroupobservedregionssurveydiseasesplantsshowedsymptomaticleafusingtestedspecificprimersrDNAPeanutWB16SrIIphyllodyPhytoplasmagroupnew16SrII-WanalysisBACKGROUND:lowshrubcommonlysandysoilswadisdesertfoundthroughoutconductedwildareanorthern2015typicalsymptomsinfectioninfectedexcessiveproliferationshootssmallleavesRESULTS:presencephloemtissueconfirmedTransmissionElectronMicroscopyTEMadditionextractedDNAOrosiusspleafhoppersamplificationsyieldedexpectedproductsasymptomaticplantSequencesimilarityphylogenetictreeanalysesfourplacedcladecloseFavabean16SrII-CCotton16SrII-FCandidatusaurantifolia'16SrII-BHoweverCrotalaria'sseparatesub-cladebelongingcombinationgene16SrII-A-B-DpytoplasmasproductamplifiedsuggestsdifferentCONCLUSION:proposeassignlineagedependingsequencesrRNAknowledgefirstreportinfectingworldwideClassificationassociatedbasedmultigenesequencephylogenyWBD

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