Molecular Characterization of Grapevine yellow speckle viroid 1 Detected in the Aegean Region Vineyards of Turkey

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Proceedings of the 18th Congress of ICVG, Ankara, TURKEY | 7-11 September 2015
Dear Colleagues
It will a great pleasure for me to invite you to 18th Congress of the International
Council for the Study of Virus and Virus-like Diseases of the Grapevine (ICVG) which
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ICVG in Turkey. Its venue will be Sheraton Hotel in Ankara. We feel honored to host
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Hoping to seeing you in Ankara,
Prof. Dr. Filiz ERTUNÇ
Chair,18th ICVG Organizing Committee
Editor: Prof.Dr. Filiz ERTUNÇ
Address : Ankara University Faculty of Agriculture, Department of Plant Protection
06110, Ankara / TURKEY
Phone Number : +90 312 596 11 20
Fax : +90 312 318 70 29
E-Mail : fertunc@gmail.com, info@icvg2015.org

4 ICVG 2015 Abstracts
Proceedings of the 18th Congress of ICVG, Ankara, TURKEY | 7-11 September 2015
Committees & Invited Speakers
ICGV Steering Committee
Giovanni P. Martelli (Italy)
Mark Fuchs (USA)
Johan T. Burger (Republic of South Africa)
Fiona Constable (Australia)
Nicola Fiore (Chile)
Deborah A. Golino (USA)
Nuredin Habili (Austuralia)
Raymond Johnson (Canada)
Nikos Katis (Greece)
Olivier Lemaire (France)
Michael Maixner (Germany)
Gustavo Nolasco (Portugal)
Pasquale Saldarelli (Italy)
Edna Tanne (Israel)
Honorary Committee Members
Guiseppe Belli (Italy)
Antoine Caudwell (France)
Oscar A. De Sequeira (Portugal)
Hans M. Kassemeyer (Germany)
Gawrie Kriel (Rebuplic of South Africa)
Ali Rezaian (Australia)
Iannis Rumbos (Greece)
Guenther Stellmach (Germany)
Daniel Teliz (Mexico)
Bernard Walter (France)
18th ICVG Invited Speakers Members
Assunta Bertaccini
Giovanni P. Martelli
18th ICVG Organizing Committee
Filiz Ertunç
Özer Elibüyük
Didem Canik Orel
<D÷PXU7UNPHQ
Meetings
1. Changins (Switzerland) 17-20 August 1964
2. Davis (California) 7-11 September 1965
3. Bernkastel-Kues (W. Germany) September 1967
4. Colmar (France) 16-18 June 1970
5. Salice Terme (Italy) 16-19 September 1973
6. Cordoba and Madrid (Spain) 12-17 September 1973
7. Niagara Falls (Canada) 7-12 September 1980
8. Bari (Italy) 2-7 September 1984
9. Kiryat Anavim (Israel) 6-11 September 1987
10. Volos (Greece) 3-7 September 1990
11. Montreux (Switzerland) 5-10 September 1993
12. Lisbon (Portugal) 28 September - 2 October 1997
13. Adelaide (South Australia) 12-17 March 2000
14. Locorotondo (Italy) 12-17 September 2003
15. Stellenbosch (South Africa) 3-7 April 2006
16. Dijon (France) 31 Aug - 4 Sep 2009
17. Davis (California) 7-14 October 2012
18. Ankara (Turkey) 7-11 September 2015
18th ICVG Conference Correspondent
Organizing Secretariat :
Bilkon Tourism and Travel Agency
Address : Cinnah Cad. Gelibolu Sk. 3/11 Kavaklidere
06680 Ankara Turkey
Phone Number : +90 312 466 14 66
Fax Number : +90 312 466 14 68
E-Mail : kongre@bilkonturizm.com.tr
Web : www.bilkonturizm.com.tr

8 ICVG 2015 Abstracts
Proceedings of the 18th Congress of ICVG, Ankara, TURKEY | 7-11 September 2015
Session 10: Phytoplasmas
OP 45 - (Invited Lecture) Phytoplasma diseases in grapevine a threat to worldwide viticulture 112
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OP 46 - Characterization of vmp1 gene of grapevine stolbur isolates from Bosnia and Herzegovina 119
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showing yellows symptoms
121
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OP 48 - ‘Bois Noir’ phytoplasma disease in grapevine in Azerbaijan 124
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OP 49 - Epidemiology of aster yellows phytoplasma: alternate host plants and the vector Mgenia
fuscovaria (Hemiptera: Cicadellidae) in South Africa
126
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OP 50 - The genetic variability of AY in South African vineyards and its spatial and temporal distribution
in individual vines
128
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OP 51 - Grapevine phytoplasma Infections in Turkey 130
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Session 11: Phytoplasmas - II
OP 52 - Results from the ‘Epidemiological studies on reservoir hosts and potential vectors of grapevine
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132
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OP 53 - Molecular epidemiology of ‘Candidatus Phytoplasma solani’ by multilocus sequence analsis 135
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‘Candidatus Phytoplasma solani’
137
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OP 55 - Incidence and distribution of aster yellows disease of grapevine in the Olifants River wine
producing area of South Africa
139
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OP 56 - Successful elimination of Grapevine Rupestris stem pitting-associated virus and its gradual
Re-infection in the vineyard but not in the greenhouse
141
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POSTER PRESENTATIONS
Session 1: Nepoviruses
PP 01 - Molecular characterization of Grapevine fan leaf virus from non Vitis hosts 149
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PP 02 - Grapevine virus diseases testing in the seedlings introduced to Ukraine 151
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10 ICVG 2015 Abstracts
Proceedings of the 18th Congress of ICVG, Ankara, TURKEY | 7-11 September 2015
PP 21 - First report of Grapevine rupestris stem pitting associated virus (GRSPaV) in Turkey 186
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PP 22 - Grapevine rupestris stem pitting-associated virus improves tolerance to water stress and miRNAs
are involved in these virus-plant-drought interactions.
188
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PP 23 - Improved RT-PCR detection and prevalence of GVA, GVB and GRSPaV in Greek vineyards 190
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PP 24 - Molecular detection of GVA and GVB variants in Portuguese grapevine varieties 192
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Session 5-6: New viruses, disaeses of unknown etiology and viroids
PP 25 - Characterization of a novel reovirus species in Cabernet grapevine in California 194
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PP 26 - First Report of Australian grapevine viroid (AGVd) in the Aegean Region Vineyards of Turkey 196
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PP 27 - Molecular Characterization of Grapevine yellow speckle viroid 1 Detected in the Aegean Region
Vineyards of Turkey
198
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PP 28 - Survey of Grapevine Viruses in the East and Southeast Regions of Turkey 200
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PP 29 - Survey on a new emergent grapevine disease and Grapevine Pinot gris virus (GPGV) in Veneto,
Northeast Italy.
201
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PP 30 - Investigation of newly-emerging grapevine viruses in The Central Anatolia region of Turkey 203
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PP 31 - Grapevine yellow speckle viroid-3 (GYSVd-3), a tentative viroid species in Turkish vineyards 204
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PP 32 - Comparison of molecular hybridization, real time PCR and classical PCR techniques for diagnosis
of GYSVD-1 (Grapevine yellow speckle viroid-1)
206
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PP 33 - Symptom alterations in Australian grapevine viroid chimeras 208
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PP 34 - Hop stunt viroid, a good candidate for internal control in detection of viroids and viruses in
grapevine
210
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PP 35 - Spread of GPGV-associated disease in two vineyards in Trentino (Italy) 212
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PP 36 - Molecular variability of grapevine virus D isolates from naturally infected vineyards in Tunisia 214
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PP 37 - Preliminary data on the transmission of Grapevine Pinot Gris Virus by Colomerus vitis 217
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POSTER PRESENTATIONS

198 ICVG 2015 Abstracts
Proceedings of the 18th Congress of ICVG, Ankara, TURKEY | 7-11 September 2015
PP 27 - Molecular characterization of Grapevine yellow speckle viroid 1 detected
in the Aegean Region vineyards of Turkey
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INTRODUCTION
*UDSHYLQH\HOORZVSHFNOHYLURLG1 (GYSVd-1) belonging to the genus $SVFDYLURLG (3RVSLYLURLGDH) is one of the causal
agents of Grapevine yellow speckle disease (Koltunow and Rezaian, 1988; Koltunow et al., 1989). Grapevine yellow
speckle disease is appeared in the worldwide vineyards extensively and it causes vein banding, yellow speckle on leaves
and stunting in grapevine plants (Randless, 2003). GYSVd-1 is one of the two viroids (GYSVd-1 and 2) which can induce
symptoms in the grapevine and also widespread in the main vineyards of our country and region (Gazel and Önelge, 2003;
Gökçek, 2007; Çopul, 2012).
MATERIALS AND METHODS
Plant Sample Collection and Reverse Transcriptase PCR Analyses
In the present study, forty nine grapevine leaf samples were collected from different vineyards from Aegean region.
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GGC GTA ACG C 3’ and reverse 5’ GGA CGC GAA CGT GAA TAG G 3’ (Koltunow and Rezaian, 1988)]. Also, RT-PCR
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ACC TCG GAA GGC CGC C-3’ and reverse 5’-TTG GAT CCT AAC CAC AGG AAC CAC A-3’).
Molecular Cloning and Sequence Data Analyses
For each grapevine cultivar, a GYSV-1 isolate from positive plant samples were selected randomly to perform molecular
characterization. The complete genome of selected isolates (GYSVd-1-TR) were cloned and sequenced. RNA genomes
of different GYSVd-1 Turkish isolates were analyzed by computer programs such as Blastn, Vector NTI and CLC Main
Workbench. The sequences of Turkish isolates were aligned with other world isolates. Also, following the cloning and
sequencing of complete genomes of GYSVd-3 positive isolates, the obtained results were then compared with the nucleotide
sequences characterized by Jiang et al. (2012) (GenBank accession no. DQ371469 and DQ371470) using Vector NTI
software. Finally, phylogenetic tree that showed identity of GYSVd-1 and GYSVd-3 Turkish isolates with world isolates was
constructed using CLC Main Workbench computer software.
RESULTS AND DISCUSSION
RT-PCR Results and Positive Isolates for GYSVd-1 and 3
In consequence of RT-PCR analyses, twenty one of 49 plant samples (42,86%) belonging to Sultani Çekirdeksizi,
2UDøVD(UJLQdHNLUGHNVL]L(PLU&ULPVRQ6HHGOHVVDQG&DOPHULDFXOWLYDUVZHUHIRXQGSRVLWLYHIRU*<6YG,Q573&5
DQDO\VHVSHUIRUPHG WR GHWHUPLQH *<69G WZHQW\ ¿YH RI  SODQW VDPSOHV  EHORQJLQJ WRGLIIHUHQWJUDSHYLQH
FXOWLYDUV(PLU(UJLQdHNLUGHNVL]L$XWXPQ5R\DO+D¿]$OL2UD'DQXWD&DOPHULDZHUHIRXQGSRVLWLYH
Results of Phylogenetic Analyses
As a result of the alignments, the identity among different GYSVd-1 Turkish isolates varied between 79-96% (Table 1).
Phylogenetic analyses revealed that GYSVd-1 Turkish isolates showed the highest identity with the isolates from Germany
(GenBank accession no. X87906), USA (KF137564), Italy (EU682453), China (DQ371471), Australia (X06904), Hungary
(GQ995473), India (AB742223), Iran (KF916046), Canada (AF462163), Thailand (AY639607) and Japan (AB028466)
(Figure 1). Also, the identity among GYSVd-3 Turkish isolates and GYSVd-3 isolates characterized by Jiang et al. (2012)
(GenBank accession no. DQ371469 and DQ371470) varied between 80-91% (Table 2).

ICVG 2015 Abstracts 199
Proceedings of the 18th Congress of ICVG, Ankara, TURKEY | 7-11 September 2015
GYSVd-1 Crimson S. Emir Ergin C. Isa Ora Sultani C.
Calmeria 87 89 93 96 84 92
Crimson S. 84 87 90 82 91
Emir 89 91 79 87
Ergin C. 96 84 93
øVD 87 95
Ora 85
GYSVd-3 DQ371469 and DQ371470
Emir 91
Ergin C. 90
Autumn R. 89
+D¿]$OL 89
Ora 86
Danuta 85
Calmeria 80
Figure 1. Phylogenetic tree that showed identity of GYSVd-Turkish isolates with GYSVd-1 world isolates.
ACKNOWLEDGMENTS
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2$OVRZH WKDQN RXU FROOHDJXHVIURP (JH 8QLYHUVLW\<]QF<ÕO8QLYHUVLW\DQG 0DQLVD 9LWLFXOWXUH 5HVHDUFK
Station who provided insight and expertise that greatly assisted the research.
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dRSXO6(JHE|OJHVLED÷ DODQODUÕQGDYLURLGOHULQ EHOLUOHQPHVL]HULQH oDOÕúPDODU(JHhQLYHUVLWHVL)HQ%LOLPOHUL(QVWLWV%LWNL.RUXPD
$QDELOLP'DOÕ<NVHN/LVDQV7H]Lø]PLU7UNL\HVXQSXEOLVKHG
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Table 1. Similarity table of
molecularly characterized
GYSVd-1 Turkish isolates
selected from different grapevine
cultivars (%).
Table 2. Similarity table of molecularly characterized GYSVd-3 Turkish
isolate and the isolates characterized by Jiang et al. (2012) (GenBank
accession no. DQ371469 and DQ371470) (%).