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      Detection and identification of the phytoplasma associated with China ixeris ( Ixeridium chinense) fasciation

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          Phytoplasmas are always associated with symptoms in host plants such as stunting of stems, witches’-broom, yellowing of leaves, formation of sterile-deformed flowers, virescence and phyllody. Recently also symptom of fasciation was reported associated with phytoplasma presence. In the present work, China ixeris fasciation was observed associated with phytoplasmas in Guanzhong Area, Shaanxi, China.


          Phytoplasma-like bodies were observed under transmission electron microscope in stem tissues of symptomatic samples. The 16S rRNA operon and tuf genes from phytoplasmas were amplified by PCR assays. Phylogenetic trees were calculated respectively based on sequences data of these two genes. The pattern of restriction fragment length polymorphism (RFLP) was generated via digesting the PCR products of 16S rRNA gene with eight restriction enzymes.


          The presence of phytoplasma in China ixeris exhibiting fasciation symptom was confirmed by the results of TEM observation and PCR testing. Based on sequence data, phylogeny analysis and actual restriction fragment length polymorphism (RFLP) analysis, the associated phytoplasma was classified as related to 16SrI-C subgroup. This was the first record of phytoplasmas in China ixeris.

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          The online version of this article (doi:10.1186/1999-3110-54-52) contains supplementary material, which is available to authorized users.

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          Most cited references 20

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          'Candidatus Phytoplasma', a taxon for the wall-less, non-helical prokaryotes that colonize plant phloem and insects.

          The trivial name 'phytoplasma' has been adopted to collectively name wall-less, non-helical prokaryotes that colonize plant phloem and insects, which were formerly known as mycoplasma-like organisms. Although phytoplasmas have not yet been cultivated in vitro, phylogenetic analyses based on various conserved genes have shown that they represent a distinct, monophyletic clade within the class Mollicutes. It is proposed here to accommodate phytoplasmas within the novel genus 'Candidatus (Ca.) Phytoplasma'. Given the diversity within 'Ca. Phytoplasma', several subtaxa are needed to accommodate organisms that share 97.5% similar to those of other 'Ca. Phytoplasma' species, are characterized by distinctive biological, phytopathological and genetic properties. These include 'Ca. Phytoplasma pyri' (associated with pear decline), 'Ca. Phytoplasma prunorum' (associated with European stone fruit yellows), 'Ca. Phytoplasma spartii' (associated with spartium witches'-broom), 'Ca. Phytoplasma rhamni' (associated with buckthorn witches'-broom), 'Ca. Phytoplasma allocasuarinae' (associated with allocasuarina yellows), 'Ca. Phytoplasma ulmi' (associated with elm yellows) and an additional taxon for the stolbur phytoplasma. Conversely, some organisms, despite their 16S rRNA gene sequence being <97.5% similar to that of any other 'Ca. Phytoplasma' species, are not presently described as Candidatus species, due to their poor overall characterization.
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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.

                Author and article information

                Bot Stud
                Bot Stud
                Botanical Studies
                Springer Berlin Heidelberg (Berlin/Heidelberg )
                31 October 2013
                31 October 2013
                December 2013
                : 54
                [1 ]GRID grid.144022.1, ISNI 0000000417604150, State Key Laboratory of Crop Stress Biology for Arid Areas/Key Laboratory of Integrated Pest Management on Crop in Northwestern Loess Plateau, Ministry of Agriculture/Key Laboratory of Plant Protection Resources and Pest Management, Ministry of Education, College of Plant Protection, , Northwest A&F University, ; Yangling, 712100 Shaanxi Province P. R. China
                [2 ]GRID grid.144022.1, ISNI 0000000417604150, College of Forestry, , Northwest A&F University, ; Yangling, Shaanxi 712100 P. R. China
                © Li et al.; licensee Springer. 2013

                This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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