Detection and Identification of the Coconut Lethal Yellowing Phytoplasma in Weeds Growing in Coconut Farms in Côte D’Ivoire
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Canadian Journal of Plant Pathology ISSN: 0706-0661 (Print) 1715-2992 (Online) Journal homepage: http://www.tandfonline.com/loi/tcjp20 Detection and identification of the coconut lethal yellowing phytoplasma in weeds growing in coconut farms in Côte d’Ivoire Y. Arocha Rosete, H. ATTA Diallo, J.L. Konan Konan, A.E.P. Kouamé, K. Séka, K.D. Kra, M.N. Toualy, K.E. Kwadjo, W.A.M.P. Daramcoum, N.I. Beugré, B.W.M. Ouattara, C.G. Kouadjo Zaka, K. Allou, N.D. Fursy-Rodelec, O.N. Doudjo- Ouattara, N. Yankey, S. Dery, A. Maharaj, M. Saleh, R. Summerbell, N. Contaldo, S. Paltrinieri, A. Bertaccini & J. Scott To cite this article: Y. Arocha Rosete, H. ATTA Diallo, J.L. Konan Konan, A.E.P. Kouamé, K. Séka, K.D. Kra, M.N. Toualy, K.E. Kwadjo, W.A.M.P. Daramcoum, N.I. Beugré, B.W.M. Ouattara, C.G. Kouadjo Zaka, K. Allou, N.D. Fursy-Rodelec, O.N. Doudjo-Ouattara, N. Yankey, S. Dery, A. Maharaj, M. Saleh, R. Summerbell, N. Contaldo, S. Paltrinieri, A. Bertaccini & J. Scott (2016) Detection and identification of the coconut lethal yellowing phytoplasma in weeds growing in coconut farms in Côte d’Ivoire, Canadian Journal of Plant Pathology, 38:2, 164-173, DOI: 10.1080/07060661.2016.1191044 To link to this article: http://dx.doi.org/10.1080/07060661.2016.1191044 Accepted author version posted online: 19 Submit your article to this journal May 2016. Published online: 20 Jun 2016. Article views: 97 View related articles View Crossmark data Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalInformation?journalCode=tcjp20 Download by: [University of Toronto Libraries] Date: 14 February 2017, At: 08:27 Can. J. Plant Pathol., 2016 Vol. 38, No. 2, 164–173, http://dx.doi.org/10.1080/07060661.2016.1191044 Bacteria and phytoplasmas/Bactèries et phytoplasmes Detection and identification of the coconut lethal yellowing phytoplasma in weeds growing in coconut farms in Côte d’Ivoire Y. AROCHA ROSETE1, H. ATTA DIALLO2, J.L. KONAN KONAN3, A.E.P. KOUAMÉ2, K. SÉKA2, K.D. KRA2, M.N. TOUALY2, K.E. KWADJO2, W.A.M.P. DARAMCOUM3, N.I. BEUGRÉ3, B.W.M. OUATTARA2, C.G. KOUADJO ZAKA3, K. ALLOU3, N.D. FURSY-RODELEC2, O.N. DOUDJO-OUATTARA4, N. YANKEY5, S. DERY5, A. MAHARAJ1, M. SALEH1, R. SUMMERBELL1, N. CONTALDO6, S. PALTRINIERI6, A. BERTACCINI6 AND J. SCOTT1,7 1Sporometrics, 219 Dufferin Street, Suite 20C, Toronto, ON M6K 3J1, Canada 2Université Nangui Abrogoua, 02 BP 801, Abidjan 02, Abidjan, Côte d’Ivoire 3Station de Recherche ‘Marc Delorme’, Centre National de Recherche Agronomique (CNRA), Abidjan, 07 BP 13, Côte d’Ivoire 4Swiss Center of Scientific Research, Abidjan, Côte d’Ivoire 5Council for Scientific Research Program–Oil Palm Research Institute–Coconut Research Program (CSIR-OPRI), Off Well Road Box 254, Sekondi, Western Region, Ghana 6Department of Agricultural Sciences, Alma Mater Studiorum, University of Bologna viale Fanin 42, Bologna, 40127, Italy 7Department of Agricultural Sciences, Plant Pathology, Alma Mater Studiorum, University of Bologna, viale G. Fanin 42, 40127 Bologna, Italy (Accepted 15 May 2016) Abstract: Coconut farms located in the southern coast of Grand-Lahou in Côte d’Ivoire are severely affected by a lethal yellowing disease (CILY) associated with the group 16SrXXII-B, ‘Candidatus Phytoplasma palmicola’-related strains. Given the high prevalence of weed species on most of the farms, plants growing within and in the periphery of five selected coconut farms were assessed for the presence of the CILY phytoplasma to identify potential alternative hosts. A total of 396 plant samples belonging to 84 plant species and 35 botanical families were collected. Total DNA was extracted and tested by nested PCR with primers targeting the 16S rRNA and the translocation protein (secA) phytoplasma genes, and sequenced. Twenty samples from six plant species and five botanical families yielded PCR amplicons of the expected size, and both the secA and 16S rDNA sequences showed over 99% similarity with that of the Côte d’Ivoire lethal yellowing phytoplasma previously identified from coconut palms grown in Grand-Lahou coconut farms. Plant species from the families Poaceae (Paspalum vaginatum, Pennisetum pedicillatum), Verbenaceae (Stachytarpheta indica), Plantaginaceae (Scoparia dulcis), Phyllanthaceae (Phyllantus muellerianus) and Cyperacea (Diplacrum capitatum) were positive for the presence of the CILY phytoplasma, suggesting they may have epidemiological implications for disease spread in coconut farms in Grand-Lahou. Keywords: 16S rRNA, 16SrXXII phytoplasma, alternative hosts, ‘Candidatus Phytoplasma palmicola’-related strains, coconut, Côte d’Ivoire lethal yellowing, secA Résumé: Les exploitations agricoles de la côte sud de Grand-Lahou, en Côte d’Ivoire, où l’on cultive la noix de coco, sont gravement touchées par le jaunissement mortel (JM) associé au groupe 16SrXXII-B des souches apparentées à ‘Candidatus Phytoplasma palmicola’. Étant donnée la forte prévalence des mauvaises herbes sur la plupart des fermes, des plantes poussant dans cinq exploitations ciblées ou en périphérie de celles-ci ont été évaluées au regard du phytoplasme du JM afind’identifier de possibles hôtes alternatifs. En tout, 396 échantillons de plantes appartenant à 84 espèces et 35 familles botaniques ont été collectés. L’ADN total a été extrait et testé par PCR par amorces incluses ciblant l’ARNr 16S et les gènes du phytoplasme de la protéine de translocation (secA), puis séquencé. En tout, 20 échantillons provenant de 6 espèces Correspondence to: Y. Arocha Rosete. E-mail: [email protected] © 2016 The Canadian Phytopathological Society Detection and identification of a CILY phytoplasma 165 de plantes et de 5 familles botaniques ont fourni des amplicons de taille attendue découlant de la PCR, et les séquences de la secA ainsi que de l’ARNr 16S ont affiché une similitude de 99% avec celles du phytoplasme du jaunissement mortel de la Côte d’Ivoire préalablement identifié chez les cocotiers cultivés à Grand-Lahou. Des espèces de plantes de la famille des Poaceae (Paspalum vaginatum, Pennisetum pedicillatum), des Verbenaceae (Stachytarpheta indica), des Plantaginaceae (Scoparia dulcis), des Phyllanthaceae (Phyllantus muellerianus) et des Cyperacea (Diplacrum capitatum) se sont révélées positives quant à l’occurrence du phytoplasme du JM, ce qui suggère qu’elles peuvent avoir des implications épidémiologiques quant à la propagation de la maladie dans les exploitations de Grand-Lahou. Mots clés: ARNr 16S, cocotier, hôtes alternatifs, jaunissement mortel de la Côte d’, Ivoire, phytoplasme 16SrXXII, secA, souches apparentées à ‘Candidatus Phytoplasma palmicola’ fi Introduction conserved nature of the 16S rRNA gene makes it dif cult to discriminate among closely related strains (Duduk & Phytoplasmas are cell wall-less prokaryotic intracellular Bertaccini 2011; Marcone 2014). Therefore, assays based plant pathogens belonging to the class Mollicutes (Marcone on non-ribosomal target genes such as the secAgeneare 2014) which originated from Gram-positive bacteria. Lethal beingused(Dickinson&Hodgetts2013;Yankeyetal.2014; yellowing (LY)-like or Lethal Decline (LD)-like phyto- Valiunas et al. 2015) as a support to the 16S rRNA gene- plasma diseases have decimated millions of coconut groves based identification and classification system, and to distin- and affected livelihoods of thousands of smallholder farm guish among closely related phytoplasmas. communities that depend on coconut palm trees in Africa, Phytoplasmas possess a double life cycle that involves Central America and the Caribbean (Danyo 2011; Sullivan replication in phloem-feeding Hemipteran insects and plant &Harrison2013). Côte d’Ivoire lethal yellowing (CILY) hosts. The latter involve over 1000 plant species, including disease is rapidly expanding into new coconut-growing vil- crops of economic importance worldwide (Duduk & lages of Grand-Lahou, a coastal commune in southern Côte Bertaccini 2011). Phytoplasma vectors and alternative host d’Ivoire, and sub-prefecture of the Grand-Lahou Department plants play an active role in the development of epidemics in the Lagunes District. The disease has already destroyed (Lee et al. 2003; Weintraub & Beanland 2006). Alternative over 400 hectares of coconut groves and continues to expand host plants of phytoplasma-associated diseases have been from the most CILY-affected areas (Arocha-Rosete et al. identified worldwide, and include either cultivated species 2014). Moreover, the destruction of the coconut palms or weeds. Examples of phytoplasma diseases whose sec- makes lands previously covered more exposed and therefore ondary plant hosts have been identified include the alfalfa more prone to degradation, which adds serious environmen- witches’ broom phytoplasma in Iran (Esmailzadeh-Hosseini tal repercussions (Yankey et al. 2009). et al. 2011), Napier grass stunt phytoplasma in East Africa CILY symptoms resemble those of the Cape St. Paul Wilt (Arocha & Jones 2010), and 'bois noir' phytoplasma in Disease (CSPWD) that wreaked great havoc to the coconut grapevine in Italy (Mori et al. 2015). industry in Ghana during the last 30 years (Eziashi & Plant species found within surrounding crop fields Omamor 2010;Danyo2011). The CILY phytoplasma has have been shown to play a role in spreading phytoplasma become a phytosanitary risk for the Ivorian coconut industry diseases, as they may act as alternative reservoirs for the (Arocha-Rosete et al. 2014). Based on the analysis of the 16S phytoplasmas (Weintraub & Beanland 2006) or their rRNA gene, it was identified