Identification and characterization of new 'Candidatus Phytoplasma solani' strains associated with bois noir disease in Vitis vinifera L. cultivars showing a range of symptoms severity in Georgia, the Caucasus region

Evidence from a preliminary survey highlighted that ‘Candidatus Phytoplasma solani’, the etiological agent of bois noir (BN) disease of grapevine, infects grapevine varieties in Georgia, a country of the South Caucasus. In this study, field surveys were carried out to investigate the BN symptom severity in international and Georgian native varieties. ‘Ca. P. solani’ was detected and identified by polymerase chain reaction-based amplification and restriction fragment length polymorphism analysis of 16S ribosomal DNA, and further characterized by multiple gene typing analysis (vmp1 and stamp genes). Obtained data highlighted that the majority of Georgian grapevine varieties showed moderate and mild symptoms, whereas international cultivars exhibited severe symptoms. Molecular characterization of ‘Ca. P. solani’ from grapevine revealed the presence of 11 distinct phytoplasma types. Only one type (VmGe12/StGe7) was identical to a strain previously reported in periwinkle from Lebanon; the other ‘Ca. P. solani’ types are described here for the first time. Phylogenetic analyses of vmp1 and stamp gene concatenated nucleotide sequences showed that ‘Ca. P. solani’ strains in Georgia are associated mainly with the bindweed-related BN host system. Moreover, the fact that ‘Ca. P. solani’ strains are distributed in grapevine cultivars showing a range of symptom intensity suggests a different susceptibility of such local cultivars to BN.