Bois noir is one of the most important diseases caused by ‘Candidatus Phytoplasma solani’ affecting grapevine in the Euro-Mediterranean area. ‘Ca. P. solani’ characterization plays a significant role in its diagnosis, control and understanding of its epidemiology. In the present study, two genes encoding membrane proteins, namely vmp1 and stamp, were used to assess the genetic variability of 18 ‘Ca. P. solani’ grapevine isolates from the Herzegovina region (B&H), previously characterised as tuf-b type. Nested-PCR/RFLP and sequence analyses of vmp1 gene showed the presence of three profiles V17, V14 and V4 where V17 was highly prevalent. Phylogenetic analysis of stamp gene revealed the presence of four different genotypes, where Rqg50 was the most frequent. Finally, the combined results of tuf, vmp1 and stamp genes, reported three previously identified genotypes from Serbia, Austria and Montenegro, and two newly described ones (DB1 and DB2). Although performed only on ‘Ca. P. solani’ isolates from infected grapevines, this study provides useful molecular information to get better insight on BN epidemiology in B&H.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
Bosnia and Herzegovina
Polymerase chain reaction
Restriction fragment length polymorphisms
Aryan, A., Brader, G., Mörtel, J., Pastar, M., & Riedle-Bauer, M. (2014). An abundant ‘Candidatus Phytoplasma solani’ tuf b strain is associated with grapevine, stinging nettle and Hyalesthes obsoletus. European Journal of Plant Pathology, 140, 213–227.
Atanasova, B., Jakovljević, M., Spasov, D., Jović, J., Mitrović, M., Toševski, I., & Cvrković, T. (2015). The molecular epidemiology of bois noir grapevine yellows caused by ‘Candidatus Phytoplasma solani’ in the Republic of Macedonia. European Journal of Plant Pathology. doi:10.1007/s10658-015-0649-0.
Balech, B., Vicario, S., Donvito, G., Monaco, A., Notarangelo, P., & Pesole, G. (2015). MSA-PAD: DNA multiple sequence alignment framework based on PFAM accessed domain information. Bioinformatics, 31(15), 2571–2573.
Cimerman, A., Pacifico, D., Salar, P., Marzachi, C., & Foissac, X. (2009). Striking diversity of vmp1, a variable gene encoding a putative membrane protein of the stolbur phytoplasma. Applied and Environmental Microbiology, 75, 2951–2957.
Cvrković, T., Jović, J., Mitrović, M., Krstić, O., & Toševski, I. (2014). Experimental and molecular evidence of Reptalus panzeri as a natural vector of bois noir. Plant Pathology, 63, 42–53.
Delić, D., Seljak, G., Martini, M., Ermacora, P., Carraro, L., Myrta, A., & Đurić, G. (2007). Surveys for grapevine yellows phytoplasmas in Bosnia and Herzegovina. Bulletin of Insectology, 60, 369–370.
Delić, D., Contaldo, N., Paltrinieri, S., Lolić, B., Ðurić, Z., Hrnčić, S., & Bertaccini, A. (2011a). Grapevine yellows in Bosnia and Herzegovina: surveys to identify phytoplasmas in grapevine, weeds and insect vectors. Bulletin of Insectology, 64, 245–246.
Delić, D., Lolić, B., & Karačić, A. (2011b). Screening for phytoplasma presence in West Herzegovina vineyards. Phytopathogenic Mollicutes, 1, 87–90.
Delić, D., Đurić, Z., Jović, J., Lolić, B., Toševski, I., & Karačić, A. (2013). “Bois noir” phytoplasma and Auchenorryncha species in Bosnia and Herzegovina vineyards. In E. Torres, A. Lavina, & A. Batlle (Eds.), Third European Bois Noir Workshop (pp. 19–21). Barcelona.
Ewing, B., & Green, P. (1998). Base-calling of automated sequencer traces using phred. II. Error probabilities. Genome Research, 8, 186–194.
Ewing, B., Hillier, L., Wendl, M. C., & Green, P. (1998). Base-calling of automated sequencer traces using phred. I. Accuracy assessment. Genome Research, 8(3), 175–185.
Fabre, A., Danet, J. L., & Foissac, X. (2011). The stolbur phytoplasma antigenic membrane protein gene stamp is submitted to diversifying positive selection. Gene, 472, 37–41.
Fialová, R., Válová, P., Balakishiyeva, G., Danet, J.-L., Săfářová, D., Foissac, X., & Navrátil, M. (2009). Genetic variability of stolbur phytoplasma in annual crop and wild plant species in South Moravia (Czech Republic). Journal of Plant Pathology, 91, 411–416.
Johannesen, J., Foissac, X., Kehrli, P., & Maixner, M. (2012). Impact of vector dispersal and host-plant fidelity on the dissemination of an emerging plant pathogen. PLoS ONE 7, e51809.
Kosovac, A., Radonjić, S., Hrnčić, S., Krstić, O., Toševski, I., & Jović, J. (2015). Molecular tracing of the transmission routes of bois noir in Mediterranean vineyards of Montenegro and experimental evidence for the epidemiological role of Vitex agnus-castus (Lamiaceae) and associated Hyalesthes obsoletus (Cixiidae). Plant Pathology. doi:10.1111/ppa.12409.
Kostadinovska, E., Quaglino, F., Mitrev, S., Casati, P., Bulgari, D., & Bianco, P. A. (2014). Multiple gene analyses identify distinct Bois noir phytoplasma genotypes in the Republic of Macedonia. Phytopathologia Mediterranea, 53, 300–310.
Kovačević, Z., Petrović, D., Herceg, N., Vego, D., & Arar, K. (2010). Adventive weed flora in vineyards of Bosnia andHerzegovina. IX. Alps-Adria Scientific Workshop. doi: 10.1556/Novenyterm.59.2010.Suppl.1.
Landi, L., Riolo, P., Murolo, S., Romanazzi, G., Nardi, S., & Isidoro, N. (2015). Genetic variability of stolbur phytoplasma in Hyalesthes obsoletus (Hemiptera: Cixiidae) and its main host plants in vineyard agroecosystems. Journal of Economic Entomology. doi:10.1093/jee/tov103.
Langer, M., & Maixner, M. (2004). Molecular characterisation of grapevine yellows associated phytoplasmas of the stolbur-group based on RFLP-analysis of non-ribosomal DNA. Vitis, 43, 191–199.
Murolo, S., & Romanazzi. (2015). In-vineyard population structure of ‘Candidatus Phytoplasma solani’ using multilocus sequence typing analysis. Infection, Genetics and Evolution. doi:10.1016/j.meegid.2015.01.028.
Plavec, J., Križanac, I., Budinščak, Ž., Škorić, D., & Šeruga Musić, M. (2015). A case study of FD and BN phytoplasma variability in Croatia: multigen sequence analyses approach. European Journal of Plant Pathology. doi:10.1007/s10658-015-0637-4.
Quaglino, F., Zhao, Y., Casati, P., Bulgari, D., Bianco, P. A., Wei, W., & Davis, R. E. (2013). ‘Candidatus Phytoplasma solani’, a novel taxon associated with stolbur and bois noir related diseases of plants. International Journal of Systematic and Evolutionary Microbiology, 63, 2879–2894.
Ronquist, F., Teslenko, M., Van der Mark, P., Ayres, D. L., Darling, A., Höhna, S., Larget, B., Liu, L., Suchard, M. A., & Huelsenbeck, J. P. (2012). MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Systematic Biology, 61, 539–542.
This work was supported by the B&H Ministry of Civil Affairs (project number 10-35-5-673/13-16). We are very thankful to Xavier Foissac for helpful suggestion in determination of vmp1 restriction types. Special thanks also the anonymous reviewers for the helpful comments with which we clarified and upgraded this paper.
About this article
Cite this article
Delić, D., Balech, B., Radulović, M. et al. Vmp1 and stamp genes variability of ‘Candidatus phytoplasma solani’ in Bosnian and Herzegovinian grapevine. Eur J Plant Pathol 145, 221–225 (2016). https://doi.org/10.1007/s10658-015-0828-z
- Bois noir disease
- Genetic variability
- Membrane protein
- Nested PCR