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      Catharanthus roseus (Apocynaceae) naturally infected with diverse phytoplasmas in Costa Rica Translated title: Catharanthus roseus (Apocynaceae) naturalmente infectada con diversos fitoplasmas en Costa Rica

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          Abstract

          Abstract Phytoplasmas (class Mollicutes) are causal agents of plant diseases with an economic impact on crops or threatening local biodiversity. A survey was conducted from 2012 to 2016 on infected Catharanthus roseus plants that exhibited symptoms reminiscent of phytoplasma infection throughout Costa Rica. A total of 73 plants were collected exhibiting symptoms such as virescence, phyllody, axillary proliferation, little leaf, leaf malformation, chlorosis, or yellowing. All samples were tested by nested PCR using phytoplasma universal and specific primer pairs. Phytoplasma infection was detected in 52 (71.2 %) of the plants collected. Phytoplasmas of six subgroups belonging to 16Sr groups I, III, IX, XIII and XV were identified based on sequencing and in silico RFLP analyses. ‘Candidatus Phytoplasma asteris’ (16SrI) was the predominant group among the positive samples (n = 30) showing variety of symptoms and wide distribution from sea level to ca. 1 400 m.a.s.l. in six of the seven Costa Rican provinces. Group 16SrIII was the second most abundant (14 samples); and the remaining three groups were seldom found in C. roseus (8 samples). Moreover, group 16SrXIII phytoplasma was detected for the first time in the country. To the best of our knowledge, this is the first report of natural infection of C. roseus with phytoplasma subgroups 16SrI-B, 16SrI-P, 16SrIII-F, 16SrIX-F, 16SrXIII-A, and 16SrXV-B in Costa Rica and Central America.

          Translated abstract

          Resumen Los fitoplasmas (clase Mollicutes) son agentes causales de enfermedades de plantas que provocan pérdidas económicas o amenazan la biodiversidad local. Una recolecta de plantas de Catharanthus roseus que mostraban síntomas de posible infección con fitoplasmas se realizó en diferentes lugares de Costa Rica desde 2012 a 2016. Un total de 73 plantas fueron recolectadas con síntomas tales como viriscencia, filodia, brotación axilar múltiple, reducción foliar, deformación foliar, clorosis, y amarillamiento. Todas las muestras fueron evaluadas mediante PCR anidado usando los pares de imprimadores universales y específicos para fitoplasmas. Infección por fitoplasmas se detectó en 52 (71.2 %) de las muestras. Fitoplasmas de seis subgrupos dentro de los grupos 16Sr I, III, IX, XIII y XV fueron identificados basados en secuenciación del ADN y análisis de polimorfismos de restricción (RFLP) in silico. El grupo predominante encontrado en las muestras positivas (n = 30) fue el 16SrI (‘CandidatusPhytoplasma asteris’), éste mostró variedad de síntomas y amplia distribución desde el nivel del mar hasta casi los 1 400 m.s.n.m. en seis de las siete provincias de Costa Rica. El grupo 16SrIII fue el segundo más abundante (14 muestras); y los restantes tres grupos se encontraron en pocas muestras de C. roseus (8 muestras). Además, fitoplasmas del grupo 16SrXIII se detectaron por primera vez en el país. De acuerdo a nuestro conocimiento, este es el primer informe de infección natural de C. roseus con fitoplasmas de los subgrupos 16SrI-B, 16SrI-P, 16SrIII-F, 16SrIX-F, 16SrXIII-A y 16SrXV-B en Costa Rica y Centroamérica.

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          Phytoplasmas: bacteria that manipulate plants and insects.

          Superkingdom Prokaryota; Kingdom Monera; Domain Bacteria; Phylum Firmicutes (low-G+C, Gram-positive eubacteria); Class Mollicutes; Candidatus (Ca.) genus Phytoplasma. Ca. Phytoplasma comprises approximately 30 distinct clades based on 16S rRNA gene sequence analyses of approximately 200 phytoplasmas. Phytoplasmas are mostly dependent on insect transmission for their spread and survival. The phytoplasma life cycle involves replication in insects and plants. They infect the insect but are phloem-limited in plants. Members of Ca. Phytoplasma asteris (16SrI group phytoplasmas) are found in 80 monocot and dicot plant species in most parts of the world. Experimentally, they can be transmitted by approximately 30, frequently polyphagous insect species, to 200 diverse plant species. In plants, phytoplasmas induce symptoms that suggest interference with plant development. Typical symptoms include: witches' broom (clustering of branches) of developing tissues; phyllody (retrograde metamorphosis of the floral organs to the condition of leaves); virescence (green coloration of non-green flower parts); bolting (growth of elongated stalks); formation of bunchy fibrous secondary roots; reddening of leaves and stems; generalized yellowing, decline and stunting of plants; and phloem necrosis. Phytoplasmas can be pathogenic to some insect hosts, but generally do not negatively affect the fitness of their major insect vector(s). In fact, phytoplasmas can increase fecundity and survival of insect vectors, and may influence flight behaviour and plant host preference of their insect hosts. The most common practices are the spraying of various insecticides to control insect vectors, and removal of symptomatic plants. Phytoplasma-resistant cultivars are not available for the vast majority of affected crops.
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            Revised Classification Scheme of Phytoplasmas based on RFLP Analyses of 16S rRNA and Ribosomal Protein Gene Sequences

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              Construction of an interactive online phytoplasma classification tool, iPhyClassifier, and its application in analysis of the peach X-disease phytoplasma group (16SrIII)

              Phytoplasmas, the causal agents of numerous plant diseases, are insect-vector-transmitted, cell-wall-less bacteria descended from ancestral low-G+C-content Gram-positive bacteria in the Bacillus–Clostridium group. Despite their monophyletic origin, widely divergent phytoplasma lineages have evolved in adaptation to specific ecological niches. Classification and taxonomic assignment of phytoplasmas have been based primarily on molecular analysis of 16S rRNA gene sequences because of the inaccessibility of measurable phenotypic characters suitable for conventional microbial characterization. In the present study, an interactive online tool, iPhyClassifier, was developed to expand the efficacy and capacity of the current 16S rRNA gene sequence-based phytoplasma classification system. iPhyClassifier performs sequence similarity analysis, simulates laboratory restriction enzyme digestions and subsequent gel electrophoresis and generates virtual restriction fragment length polymorphism (RFLP) profiles. Based on calculated RFLP pattern similarity coefficients and overall sequence similarity scores, iPhyClassifier makes instant suggestions on tentative phytoplasma 16Sr group/subgroup classification status and ‘Candidatus Phytoplasma’ species assignment. Using iPhyClassifier, we revised and updated the classification of strains affiliated with the peach X-disease phytoplasma group. The online tool can be accessed at http://www.ba.ars.usda.gov/data/mppl/iPhyClassifier.html.
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                Author and article information

                Contributors
                Role: ND
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                Journal
                rbt
                Revista de Biología Tropical
                Rev. biol. trop
                Universidad de Costa Rica (San José, San José, Costa Rica )
                0034-7744
                0034-7744
                March 2019
                : 67
                : 1
                : 321-336
                Affiliations
                [4] orgnameUniversidad de Costa Rica Costa Rica federico.albertazzi@ 123456ucr.ac.cr
                [2] orgnameUSDA USA kristi.bottner@ 123456ars.usda.gov
                [3] orgnameUniversidad de Costa Rica Costa Rica mauricio.monteroastua@ 123456ucr.ac.cr
                [1] orgnameUniversidad de Costa Rica Costa Rica william.villalobos@ 123456ucr.ac.cr
                Article
                S0034-77442019000100321
                10.15517/rbt.v67i1.33972
                68842352-9db9-479c-a9e7-9211b832f081

                This work is licensed under a Creative Commons Attribution 3.0 International License.

                History
                : 08 August 2018
                : 10 December 2018
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 63, Pages: 16
                Product

                SciELO Costa Rica


                16SrXV,16SrXIII,16SrI,vincas,PCR anidado,16SrIII,16SrIX,periwinkle,nested PCR

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