First Report on ‘ Candidatus Phytoplasma asteris’ (16SrI-B subgroup) Strain Associated with Pineapple Shoot Proliferation and Witches’ Broom Symptoms in Tripura, India
2019; American Phytopathological Society; Volume: 103; Issue: 11 Linguagem: Inglês
10.1094/pdis-05-19-0900-pdn
ISSN1943-7692
AutoresSurabhi Mitra, Prasenjit Debnath, Amar Bahadur, Sukhen Das, Amit Yadav, G. P. Rao,
Tópico(s)Cocoa and Sweet Potato Agronomy
ResumoHomePlant DiseaseVol. 103, No. 11First Report on 'Candidatus Phytoplasma asteris' (16SrI-B subgroup) Strain Associated with Pineapple Shoot Proliferation and Witches' Broom Symptoms in Tripura, India PreviousNext DISEASE NOTES OPENOpen Access licenseFirst Report on 'Candidatus Phytoplasma asteris' (16SrI-B subgroup) Strain Associated with Pineapple Shoot Proliferation and Witches' Broom Symptoms in Tripura, IndiaSurabhi Mitra, Prasenjit Debnath, Amar Bahadur, Sukhen Chandra Das, Amit Yadav, and G. P. RaoSurabhi MitraDivision of Plant Pathology, Indian Agricultural Research Institute, New Delhi, 110012, India, Prasenjit DebnathCollege of Agriculture, Tripura, Lembucherra, West Tripura, 799210, India, Amar BahadurCollege of Agriculture, Tripura, Lembucherra, West Tripura, 799210, India, Sukhen Chandra DasCollege of Agriculture, Tripura, Lembucherra, West Tripura, 799210, India, Amit Yadavhttp://orcid.org/0000-0002-4882-9075National Centre for Microbial Resource, National Centre for Cell Science, Sai Trinity Complex, Sus Road, Pashan, Pune 411 021, India, and G. P. Rao†Corresponding author: G. P. Rao; E-mail Address: gprao_gor@rediffmail.comhttp://orcid.org/0000-0003-1057-7826Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi, 110012, IndiaAffiliationsAuthors and Affiliations Surabhi Mitra1 Prasenjit Debnath2 Amar Bahadur2 Sukhen Chandra Das2 Amit Yadav3 G. P. Rao1 † 1Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi, 110012, India 2College of Agriculture, Tripura, Lembucherra, West Tripura, 799210, India 3National Centre for Microbial Resource, National Centre for Cell Science, Sai Trinity Complex, Sus Road, Pashan, Pune 411 021, India Published Online:13 Sep 2019https://doi.org/10.1094/PDIS-05-19-0900-PDNAboutSectionsSupplemental ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat Pineapple (Ananas comosus [L.] Merr., family Bromeliaceae) is a perennial crop grown for its delicious fruit having pronounced flavor and fiber. India ranked fifth position in pineapple production in the world. During a recent survey of pineapple fields of queen cultivar at Boromura, West Tripura, India, shoot proliferation and witches' broom (PiSP&WB) symptoms were observed in June 2018 with an incidence of 10 to 15%. Molecular tests were carried out to detect and identify the possible association of phytoplasmas with PiSP&WB symptoms. Three each of symptomatic and asymptomatic pineapple samples were collected, and DNA was extracted from the leaves, which was used as a template for direct and nested PCR assays by phytoplasma universal primer pairs (P1/P7; R16F2n/R16R2) and secA gene primer pair (SecAfor1/SecArev3; SecAfor2/SecArev3) using previously described conditions (Hodgetts et al. 2008; Schneider et al. 1995). The ∼1.2 kb and ∼480 bp amplified products of 16S rRNA and secA genes of phytoplasma DNA were detected, respectively, in all the three symptomatic samples, but not from any asymptomatic samples. Both the amplified fragment products of PiSP&WB strain of 16S rRNA and secA genes were purified using the WizardR SV Gel and PCR Clean-up System (Promega, Madison, WI) and sequenced in both directions at ABA Biotech, India. The sequences of PCR products were assembled using DNA Base version 4. The 16S rRNA and secA gene sequences were aligned with phytoplasma group/subgroup representatives available in GenBank using ClustalW software, and the consensus sequences were submitted to GenBank database (16Sr RNA gene: MK209105, MK328881; secA gene: MK751427, MK840445). BLAST analysis of 16S rRNA and secA gene sequences of PiSP&WB isolates revealed the highest sequence identity of 99.92 and 100%, respectively, with Catharanthus roseus aster yellows phytoplasma strain (accession no. CP035949) and sesame phyllody phytoplasma strain (KJ434315), members of 'Candidatus Phytoplasma asteris'. The phylogenetic trees were constructed using the 16S rRNA and secA gene sequences by the neighbor-joining method with MEGA 7.0 using 1,000 bootstrap replications (Kumar et al. 2016), which demonstrated the clustering of PiSP&WB isolates with members of 16SrI-B subgroups. The phytoplasma sequence corresponding to the R16F2n/R16R2 fragment of PiSP&WB strain was subjected to in silico restriction fragment length polymorphism (RFLP) comparison of 17 restriction enzymes using the iPhyClassifier tool (Zhao et al. 2009). The virtual RFLP pattern of PiSP&WB isolate was found identical (similarity coefficient 1.00) to the reference profile of a 16SrI-B strain of onion yellows phytoplasma (accession no. AP006628), confirming the association of a 16SrI-B subgroup phytoplasma strain with the PiSP&WB samples. Pineapple proliferation phytoplasma strain related to 16SrI-R and 16SrXII-A was reported earlier from Oceania (Davis et al. 2006) and 16SrX-B in Colombia (Satta et al. 2015). To the best of our knowledge, this is the first report of the association of 'Ca. P. asteris' (16SrI-B) with shoot proliferation and witches' broom symptoms of pineapple in the world. Because pineapple has been reported as a new host of 16SrI-B phytoplasma subgroup in India, which has already been reported to have a wide host range infecting various crops in India (Rao et al. 2017), future studies on epidemiology would be helpful in planning in the management of the disease.The author(s) declare no conflict of interest.References:Davis, R. I., et al. 2006. Aus.Pl. Path. 35:335. https://doi.org/10.1071/AP06029 Crossref, ISI, Google ScholarHodgetts, B. N., et al. 2008. Int. J. Syst. Evol. Microbiol. 58:1826. https://doi.org/10.1099/ijs.0.65668-0 Crossref, ISI, Google ScholarKumar, S., et al. 2016. Mol. Biol. Evol. 33:1870. https://doi.org/10.1093/molbev/msw054 Crossref, ISI, Google ScholarRao, G. P., et al. 2017. Phytopath.Moll. 7:1. https://doi.org/10.5958/2249-4677.2017.00001.9 Crossref, Google ScholarSatta, E., et al. 2015. Phytopath. Mollicutes. 5:S79. https://doi.org/10.5958/2249-4677.2015.00033.X Crossref, Google ScholarSchneider, B., et al. 1995. Mol. Diagn. Proced. Mycoplasmol. 1:369. https://doi.org/10.1016/B978-012583805-4/50040-6 Crossref, Google ScholarZhao, Y., et al. 2009. Int. J. Syst. Evol. Microbiol. 59:2582. https://doi.org/10.1099/ijs.0.010249-0 Crossref, ISI, Google ScholarThe author(s) declare no conflict of interest.Funding: Funding was provided by Department of Biotechnology, Ministry of Science and Technology.DetailsFiguresLiterature CitedRelated Vol. 103, No. 11 November 2019SubscribeISSN:0191-2917e-ISSN:1943-7692 DownloadCaptionAdvanced symptoms of bacterial blotch disease on mushroom caps (Osdaghi et al.). Photo credit: C. Bull. Powdery mildew caused by Golovinomyces neosalviae on Salvia fruticosa (Soylu et al.). Photo credit: S. Soylu. Metrics Article History Issue Date: 4 Nov 2019Published: 13 Sep 2019First Look: 9 Jul 2019Accepted: 3 Jul 2019 Pages: 2941-2941 Information© 2019 The American Phytopathological SocietyFundingDepartment of Biotechnology, Ministry of Science and TechnologyKeywordsAnanas comosusmolecular identification16S rRNA genesecA geneThe author(s) declare no conflict of interest.Cited byIdentification of 'Ca. 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