|
| Status |
Public on Oct 05, 2012 |
| Title |
GRSPaV-free_berries_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
berries from grapevine plants non-infected (e.g. control) by Grapevine rupestris stem pitting-associated virus (GRSPaV), replicate 2
|
| Organism |
Vitis vinifera |
| Characteristics |
cultivar: Bosco
treatment: GRSPaV-free
tissue: berries
|
| Treatment protocol |
Grapevine rupestris stem pitting-associated virus (GRSPaV) infection. All the plants of ‘Bosco’ derived by vegetative propagation from a single mother plant originally infected by GRSPaV and further subjected to sanitation. The in vitro thermotherapy and meristem tip culture sanitation techniques successfully eliminated most of the viruses present in the mother vine, but they did not provide satisfactory results in the eradication of GRSPaV, leaving some plants still infected by the virus. The sanitary status of all plants of ‘Bosco’ was tested in 2010 by multiplex RT-PCR (Gambino and Gribaudo, 2006), in order to detect the most important 9 viruses that commonly affect grapevine in Italy and in other viticultural countries: Arabis mosaic virus (ArMV), Grapevine fanleaf virus (GFLV), Grapevine virus B (GVB), Grapevine fleck virus (GFkV), GLRaV-1,-2,-3, GVA and GRSPaV. The multiplex RT-PCR analyses were repeated in 2011 to confirm the sanitary status of plants selected for the trial.
|
| Growth protocol |
The study was carried out in a vineyard planted in 2002 in Albenga (Liguria), North-West Italy, where a row was established with the white grape cultivar Bosco (V. vinifera L.). Vines were grafted onto 1103 P rootstock, spaced 2.10 m between rows and 1.00 m between plants for a total plant density of about 4700 vines/hectare. Vines were vertically trained and one-cane pruned (Guyot), leaving 12 buds per vine.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Samples from each organ were arbitrary pooled in 3 independent biological replicates and total RNA was extracted according to the method described by Gambino et al. (2008) [PMID: 16240119].
|
| Label |
Cy3
|
| Label protocol |
cDNA synthesis and labeling reactions were performed according to the NimbleGen Arrays User’s Guide (V 3.2).
|
| |
|
| Hybridization protocol |
Hybridization and washing procedures were performed according to the NimbleGen Arrays User’s Guide (V 3.2).
|
| Scan protocol |
Each microarray was scanned using a Axon GenePix 4400 A at 532 nm (Cy-3 absorption peak) and GenePix Pro7 software (Molecular Devices, Sunnyvale, CA, USA) according to the manufacturers' instructions.
|
| Data processing |
Images were analyzed using NimbleScan v2.5 software (Roche), background correction and standard RMA normalization were selected.
|
| |
|
| Submission date |
Mar 20, 2012 |
| Last update date |
Oct 05, 2012 |
| Contact name |
Marianna Fasoli |
| E-mail(s) |
marianna.fasoli@univr.it
|
| Phone |
+39 045 842 5625
|
| Organization name |
University of Verona
|
| Department |
Biotechnology
|
| Lab |
Molecular Viticulture
|
| Street address |
Via della Pieve 70
|
| City |
San Pietro in Cariano |
| State/province |
Verona |
| ZIP/Postal code |
37029 |
| Country |
Italy |
| |
|
| Platform ID |
GPL13936 |
| Series (1) |
| GSE36632 |
Co-evolution between Grapevine rupestris stem pitting-associated virus and Vitis vinifera L. induces a decrease in defence responses and physiological performance associated with an increase in photosynthesis-related gene transcription |
|
