European Journal of Plant Pathology

, Volume 139, Issue 1, pp 185–193 | Cite as

Incidence of Bois Noir phytoplasma in different viticulture regions of Spain and Stolbur isolates distribution in plants and vectors

  • Jordi Sabaté
  • Amparo Laviña
  • Assumpció Batlle


The incidence of Bois Noir (BN) disease in grapevine plots, the population of the vector Hyalesthes obsoletus, and the distribution of stolbur phytoplasma isolates were studied over a 4 year period in five regions of Spain. BN incidence in affected plots ranged from 1 to 75 %. A study of the H. obsoletus population indicated that individuals of this insect vector were identified in most of the sampled plots with low populations, from 0.25–5 individuals per aspiration. The population peaks of H. obsoletus were reached at different dates between June 6th and July 24th, depending on the sampled zone and the year. The percentage of H. obsoletus individuals carrying the phytoplasma showed an average of 55 %. In Aragon, this percentage rose to 76 %. The strain tuf-b of stolbur phytoplasma was the most prevalent type in grapevine plants and H. obsoletus, except in grapevine plants from La Rioja where the strain tuf-a was detected in most plants. A study of the vmp1 gene revealed the presence of four different isolates in grapevine plants and H. obsoletus.


Stolbur Hyalesthes Phytoplasma strains Bois Noir Grapevine 



We would like to thank J. Legorburu from Neiker (Vitoria-Gasteiz), J. Fortanete and A. Sorolla from Centro de Protección Vegetal (Zaragoza), J.J. Perez de Obanos and L. Caminero from EVENA (Navarra), J.L. Perez Marin from CIDA-La Rioja, and J. Reyes and G. Barrios from DAAM- Servei Sanitat Vegetal (Catalonia) for their help in conducting the surveys.

This work was funded by grant RTA11-067-C4 of the Programa Sectorial de I + D, M.A.P.A., Spain.


  1. Ahrens, U., & Seemüller, E. (1992). Detection of DNA of plant pathogenic mycoplasma like organisms by a polymerase chain reaction that amplifies a sequence of the 16SrRNA gene. Phytopathology, 82, 828–832.CrossRefGoogle Scholar
  2. Batlle, A., Martinez, M. A., & Laviña, A. (2000). Occurrence, distribution and epidemiology of grapevine yellows in Spain. European Journal of Plant Pathology, 106, 811–816.CrossRefGoogle Scholar
  3. Batlle, A., Altabella, N., Sabaté, J., & Laviña, A. (2008). Study of the transmission of Stolbur Phytoplasma to different crop species, by Macrosteles quadripunctulatus (Kirschbaum). Annals of Applied Biology, 152, 235–242.CrossRefGoogle Scholar
  4. 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 Environmental Microbiology, 75, 2951–2957.PubMedCentralPubMedCrossRefGoogle Scholar
  5. Cvrković, A., Jović, J., Mitrović, M., Krstić, O., & Toševski, I. (2013). Experimental and molecular evidence of Reptalus panzeri as a natural vector of bois noir. Plant Pathology. doi: 10.1111/ppa.12080.Google Scholar
  6. Deng, S., & Hiruki, C. (1991). Amplification of 16S rRNA genes from culturable and non-culturable mollicutes. Journal of Microbiological Methods, 14, 53–61.CrossRefGoogle Scholar
  7. García-Chapa, M., Sabaté, J., Laviña, A., & Batlle, A. (2005). Role of Cacopsylla pyri in the epidemiology of pear decline in Spain. European Journal of Plant Pathology, 111, 9–17.CrossRefGoogle Scholar
  8. Langer, M., & Maixner, M. (2004). Molecular characterization of grapevine yellows associated phytoplasmas of the stolbur-group based on RFLP-analysis of non-ribosomal DNA. Vitis, 43(4), 191–199.Google Scholar
  9. Laviña, A., Batlle, A., Larrue, J., Daire, X., Clair, D., & Boudon-Padieu, E. (1995). First report of grapevine bois noir phytoplasma in Spain. Plant Disease, 79, 1075.Google Scholar
  10. Lee, I. M., Gundersen, D. E., Davis, R. E., & Bartoszyk, I. M. (1998). Revised classification scheme of phytoplasmas based on RFLP analyses of 16S rRNA and ribosomal protein gene sequences. International Journal of Systematic Bacteriology, 48, 1153–1169.CrossRefGoogle Scholar
  11. Lessio, M., Tedeschi, R., & Alma, A. (2007). Population dynamics, host plants and infection rate with stolbur phytoplasma of Hyalesthes obsoletus Signoret in North-Western Italy. Journal of Plant Pathology, 89(1), 97–102.Google Scholar
  12. Lorenz, K. H., Schneider, B., Ahrens, U., & Seemüller, E. (1995). Detection of the apple proliferation and pear decline phytoplasmas by PCR amplification of ribosomal and nonribosomal DNA. Phytopathology, 85, 771–776.CrossRefGoogle Scholar
  13. Maixner, M. (1994). Research note: transmission of German grapevine yellows (Vergilbungskrankheit) by the planthopper Hyalesthes obsoletus (Auchenorrhyncha: Cixiidae). Vitis, 33, 103–104.Google Scholar
  14. Maixner, M., Ahrens, U., & Seemüller, E. (1995). Detection of the german grapevine yellows (vergilbungskrankheit) MLO in grapevine, alternative hosts and a vector by a specific PCR procedure. European Journal Plant Pathology, 101, 241–250.CrossRefGoogle Scholar
  15. Maixner, M., Johannesen, J., & Seitz, A. (2009). Aspects of the interaction of stolbur phytoplasma, vectors and host plants in the two epidemic systems of Bois Noir. Proceedings of 16th Meeting of ICVG. France: Dijon.Google Scholar
  16. Mori, N., Pavan, F., Bondavalli, R., Reggiani, N., Paltrinieri, S., & Bertaccini, A. (2007). Factors affecting the spread of “bois noir” disease in grapevine cv. Lambrusco in northern Italy. Bulletin of Insectology, 60, 295–296.Google Scholar
  17. Murolo, S., Marcone, C., Prota, V., Garau, R., Foissac, X., & Romanazzi, G. (2010). Genetic variabilty of the stolbur phytoplasma vmp1 gene in grapevines, bindweeds and vegetables. Journal of Applied Microbiology, 109(6), 2049–2059.PubMedCrossRefGoogle Scholar
  18. Pacifico, D., Alma, A., Bagnoli, B., Foissac, X., Pasquini, G., Tessitori, M., et al. (2009). Characterization of Bois noir isolates by restriction fragment length polymorphism of a stolbur-specific putative membrane protein gene. Phytopathology, 99, 711–715.PubMedCrossRefGoogle Scholar
  19. Palermo, S., Elekes, M., Botti, S., Ember, I., Alma, A., Orosz, A., et al. (2004). Presence of stolbur phytoplasma in Cixiidae in Hungarian vineyards. Vitis, 43, 201–203.Google Scholar
  20. Pinzauti, F., Trivellone, V., & Bagnoli, B. (2008). Ability of Reptalus quinquecostatus (Hemiptera: Cixiidae) to inoculate stolbur phytoplasma to artificial feeding medium. Annals of Applied Biology, 153, 299–305.CrossRefGoogle Scholar
  21. Quaglino, F., Zhao, Y., Casati, P., Bulgari, D., Bianco, P.A., Wei, al. (2013). ‘Candidatus Phytoplasma solani’, a novel taxon associated with stolbur and bois noir related diseases of plants. International Journal Systematic Evolutionary Microbiology. PMID:23334879.Google Scholar
  22. Sabaté, J., Laviña, A. & Batlle, A. (2006). Molecular characterization of stolbur phytoplasma isolates in grapevines and insect vectors. 15th ICGV Symposium, Stellenbosch (South Africa). Extended Abstracts pp-157–158.Google Scholar
  23. Sabaté, J., Laviña, A., Legorburu, J., Fortanete, J., de Pérez Obanos, J. J., Pérez Marín, J. L., et al. (2007). Incidence of Bois Noir phytoplasma in different wine-growing regions of Spain and its relation to Hyalesthes obsoletus. Bulletin of Insectology, 60(2), 367–368.Google Scholar
  24. Sforza, R., Clair, D., Daire, X., Larrue, J., & Boudon-Padieu, E. (1998). The role of Hyalesthes obsoletus (Hemiptera:Cixiidae) in the occurrence of bois noir of grapevines in France. Journal of Phytopathology, 146, 549–556.CrossRefGoogle Scholar
  25. Smart, C. D., Schneider, B., Blomquist, C. L., Guerra, L. J., Harrison, N. A., Ahrens, U., et al. (1996). Phytoplasma-specific PCR primers based on sequences of the 16S-23S rRNA spacer region. Applied Environmental Microbiology, 62, 1988–1993.Google Scholar
  26. Trivellone, V., Pinzauti, F., & Bagnoli, B. (2005). Reptalus quinquecostatus (Dufour) (Auchenorrhyncha Cixiidae) as a possible vector of Stolbur-phytoplasma in a vineyard in Tuscany. Redia, 88, 103–108.Google Scholar
  27. Weber, A., & Maixner, M. (1998). Survey of populations of the planthopper Hyalesthes obsoletus Sign. (Auchenorrhyncha, Cixiidae) for infection with the phytoplasma causing grapevine yellows in Germany. Journal of Applied Entomology, 122, 375–381.CrossRefGoogle Scholar

Copyright information

© KNPV 2014

Authors and Affiliations

  • Jordi Sabaté
    • 1
  • Amparo Laviña
    • 1
  • Assumpció Batlle
    • 1
  1. 1.Institut de Recerca i Tecnología AgroalimentàriesCabrilsSpain

Personalised recommendations