Multilocus Genotyping Reveals New Molecular Markers for Differentiating Distinct Genetic Lineages among “Candidatus Phytoplasma Solani” Strains Associated with Grapevine Bois Noir

Pathogens

MariaR. Silletti¹  Emilija Arsov²  Sasa Mitrev²  Fabio Quaglino³  PieroA. Bianco³  Alessandro Passera³  Paola Casati³  Abdelhameed Moussa³  Sergio Murolo⁴  Gianfranco Romanazzi⁴  Roberto Pierro⁵  Alberto Materazzi⁵  Alessandra Panattoni⁵  Andrea Luvisi⁶  Nicola Mori⁷  RobertE. Davis⁸  Yan Zhao⁸  Wei Wei⁸ 

[1] Centro di Ricerca e Sperimentazione in Agricoltura Basile Caramia, 70010 Locorotondo, Italy;Department for Plant and Environment Protection, Faculty of Agriculture, Goce Delcev University, Štip 2000, North Macedonia;Department of Agricultural and Environmental Sciences, Production, Landscape, Agroenergy, University of Milan, 20133 Milano, Italy;Department of Agricultural, Food and Environmental Sciences, Marche Polytechnic University, 60131 Ancona, Italy;Department of Agriculture, Food and Environment, University of Pisa, 56124 Pisa, Italy;Department of Biological and Environmental Sciences and Technologies, University of Salento, 73100 Lecce, Italy;Department of Biotechnology, University of Verona, 37134 Verona, Italy;Molecular Plant Pathology Laboratory, USDA-Agriculture Research Service, Beltsville, MD 20705, USA;

关键词: elongation Factor-Tu;    hemolysin;    restriction fragment length polymorphism (RFLP);    Vitis vinifera L.;   

DOI  :  10.3390/pathogens9110970

来源: DOAJ

【 摘 要 】

Grapevine Bois noir (BN) is associated with infection by “Candidatus Phytoplasma solani” (CaPsol). In this study, an array of CaPsol strains was identified from 142 symptomatic grapevines in vineyards of northern, central, and southern Italy and North Macedonia. Molecular typing of the CaPsol strains was carried out by analysis of genes encoding 16S rRNA and translation elongation factor EF-Tu, as well as eight other previously uncharacterized genomic fragments. Strains of tuf-type a and b were found to be differentially distributed in the examined geographic regions in correlation with the prevalence of nettle and bindweed. Two sequence variants were identified in each of the four genomic segments harboring hlyC, cbiQ-glyA, trxA-truB-rsuA, and rplS-tyrS-csdB, respectively. Fifteen CaPsol lineages were identified based on distinct combinations of sequence variations within these genetic loci. Each CaPsol lineage exhibited a unique collective restriction fragment length polymorphism (RFLP) pattern and differed from each other in geographic distribution, probably in relation to the diverse ecological complexity of vineyards and their surroundings. This RFLP-based typing method could be a useful tool for investigating the ecology of CaPsol and the epidemiology of its associated diseases. Phylogenetic analyses highlighted that the sequence variants of the gene hlyC, which encodes a hemolysin III-like protein, separated into two clusters consistent with the separation of two distinct lineages on the basis of tufB gene sequences. Alignments of deduced full protein sequences of elongation factor-Tu (tufB gene) and hemolysin III-like protein (hlyC gene) revealed the presence of critical amino acid substitutions distinguishing CaPsol strains of tuf-type a and b. Findings from the present study provide new insights into the genetic diversity and ecology of CaPsol populations in vineyards.

【 授权许可】

Unknown