Phloem Limited Pathogens on Grapevine in the Republic of Macedonia

Phytoplasmic changes as well as virus diseases in one of the most promising crops in the Republic of Macedonia, the grapevine, are among the least studied and researched pathogenic changes in the country. In contrast, in other European countries, these changes have been investigated very intensively with results in publications relevant to the sciences and practice. During the period dedicated to the research in this PhD thesis (2008-2013), the situation was constantly monitored from the aspect of symptomatology. The collected material was analyzed in the Laboratory of plant and environment protection, and typification of the present pathogens was carried out by using modern molecular methods of PCR / RFLP, by studying seven phytoplasmic gene loci: 16S rRNA, tuf, vmp1 gene (stol - 1H0), stamp gene, trxA-truB gene, rplS - csdB gene, cbiQ - glyA gene for the detection of the type of the present phytoplasmas. By applying serological ELISA technique and RT-PCR molecular diagnostics, we determining the presence of most distributed phloemic viruses in grapevine - Grapevine leafroll associated virus GLRaV (-1, -2, -3, -7). In addition to grapevine as a cultivated plant and a very important crop in the country, we also monitored the surrounding vegetation in the vineyards from which symptomatic material was collected for analysis in order to determine the transition of phytoplasmas from the weed vegetation to the grapevine. The study covered a total of 766 isolates of grapevine phytoplasmas, 40 isolates of phytoplasmas from the surrounding vegetation and 382 isolates of grapevine viruses. During the investigated period, the presence of stolbur phytoplasmas in the grapevine and the weed vegetation was established (Bois noir, 16SrXII - A subgroup - stolbur) - Type I and II (type a & type b) (VKI & VKII). In relation to grapevine viruses, we monitored the presence of the leafroll virus in grapevine (Grapevine leafroll associated virus) with groups -1 and -3 of this virus (GLRaV-1, GLRaV-3). Key words: phytoplasmas, Mollicutes, Grapevine leafroll associated virus,PCR/RFLP, ELIS