First detection of stolbur phytoplasma in grapevines (Vitis vinifera, cv Chardonnay) affected with grapevine yellows in the Ukraine

B. Milkus¹*, D. Clair², S. Idir², N. Habili³ and E. Boudon-Padieu²

¹ Laboratory of Virology and Microbiology, Nursery Complex, Close Corporation "Odessa Brandy Company", Meljnickaja st.13, Odessa, 65005, The Ukraine
² INRA, UMR Plante Microbe Environnement, Biologie et écologie des phytoplasmes, BP 86510, 21065 Dijon Cedex, France
³ Waite Diagnostics, School of Agriculture & Wine, Waite Campus, The University of Adelaide, PMB1, Glen Osmond, South Australia 5064

*milkus@milkus.intes.odessa.ua

Accepted for publication 25/10/04

During summer 2003, symptoms typical of a grapevine yellows (GY) disease were seen in grapevine plots planted in 2000 with cv Chardonnay in the Ovidiopolskij region (Odessa area) of the Ukraine (Fig. 1). Ten to 20 % of the vines showed typical GY symptoms i.e. rolling and discolouration of leaves and uneven lignification of shoots. Important GYs in Europe are Flavescence dorée (FD) and Bois noir (BN). FD, transmitted by the vine leafhopper Scaphoideus titanus, is a quarantine disease in the EU and Ukraine, and is epidemic in certain areas of France, Italy and Spain. BN is found in Asia Minor and is associated with the endemic stolbur phytoplasma and transmitted by a polyphagous planthopper, Hyalesthes obsoletus.

Figure 1: Chardonnay with symptoms of grapevine yellows in summer 2003 in Odessa.

DNA extracted from leaf samples taken from 5 diseased grapevines was tested by PCR using standard generic primers (P1/P7 and R16F2n/R16R2) that amplify phytoplasma rDNA. Positive amplification was obtained from all 5 samples (not shown). Cuttings were taken in December 2003 of lignified canes from 15 grapevines that had shown symptoms during the summer. DNA extracted from phloem scrapings was amplified using a bi-specific PCR procedure for the detection of FD and BN phytoplasmas (Clair et al., 2003). Seven cuttings out of the 15 tested were positive for a stolbur phytoplasma (Fig. 2). The other 8 cuttings tested negative, as did the healthy grapevine control.

Figure 2: Agarose gel electrophoresis of amplimers obtained with DNA from different origins using the (FD + BN) specific multiplex-nested PCR procedure. Lane 1, 1kb ladder (BRL); lane 2-16, winter phloem scrapings from 15 Chardonnay plants showing Yellows symptoms; lane 17, FD-infected grapevine from France; lane 18, stolbur-infected periwinkle (Catharanthus roseus); lane 19, healthy grapevine; lane 20, double (FD + stolbur) infected periwinkle. The size of FD and stolbur amplimers are 1160 bp (FD9) and 720 bp (Stol11), respectively.

Earlier reports have described the presence of GY symptoms on cv Regina in Romania (Rafaila & Costache, 1968) and on cv Chardonnay in Moldavia (Marinesku et al., 1990). At that time the phytoplasma associated with this disease could not be detected in grapevine and the etiology and vector of BN were unknown, although H. obsoletus had been described in the South of Ukraine (Zirka, 1967). Since no direct control methods are available for BN, affected plants will be mapped over the next years to check for recovery. Further surveys will include monitoring of GYs, to confirm the role of H. obsoletus and to screen for S. titanus, a harmful pest actively spreading in Europe. This is the first record of stolbur phytoplasma infecting grapevines in the Ukraine.

Clair D, Larrue J, Aubert G, Gillet J, Cloquemin G, Boudon-Padieu E, 2003. A multiplex nested-PCR assay for sensitive and simultaneous detection and direct identification of phytoplasma in the Elm yellows group and Stolbur group and its use in survey of grapevine yellows in France. Vitis 42, 151-157.

Marinesku VG, Kalashyan YA, Verderevskaya TD, 1991. Grapevine yellows in Moldavian SSR. In: Proceedings of the 10th meeting of ICVG, 1990, Volos, Greece: ORES Publishing, 218.

Rafaila C, Costache M, 1968. Ingalbenirea aurie (Flavescence dorée), o boala noua a Vitei de Vie in Romania. Analele Institutului de Cercetari pentru Protectia Plantelor 6, 151-156.

Zirka TZ, 1967. Peredatcha virusu stolburu percu ta baklazanov scheplenjam ta cicadkoj Hyalesthes obsoletus Sign. Microbilologitchni zurnal 29, 334-338.

The British Society for Plant Pathology