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Sample GSM3259678 Query DataSets for GSM3259678
Status Public on Jul 26, 2019
Title B. cinerea-treated_T2_rep3
Sample type RNA
 
Source name berry pericarp B. cinerea-injected T2 (10 days after injection), replicate 3
Organism Vitis vinifera
Characteristics cultivar: Muller-Thurgau
tissue: Berry pericarps
type of treatment: B. cinerea-infected
Treatment protocol Among the sterilized berries, 200 berries were frozen, divided into three different biological replicates (a pool of 65 berries each) and sampled as time point 0 (T0) of the experiment. Botrytis cinerea B05.10 was cultured and inoculated into 400 berries (1x105 conidia per ml). The remaining 400 berries were water-injected and used as control. Berries were incubated into sterile 24-well plates under controlled conditions (15°C/15h dark; 18°C/9h light) to induced noble rot. Three biological replicates (about 50 berries each) of infected/control berries were collected and frozen 5 days after the injection when inoculated berries were at the initial stage of noble rot development (time point 1, T1). Moreover, infected/control berries were sampled 10 days after the injection when the inoculated berries were at the pourri plain stage of noble rot (time point 2, T2) characterized by a complete and homogeneous chocolate hue.
Growth protocol Vitis vinifera cv. Muller-Thurgau bunches were harvested in August 2017 in Custoza (Italy) and dehydrated into the withering setting of the “La Prendina” farm located in Monzambano (Mantova, Italy). Muller-Thurgau grapes were collected with a soluble solids content of 18.25 ± 0.05 °Brix and placed into plateaux (about 20 kg each) in a ventilated withering facility (14-15°C, 53-60% of relative humidity). Brix degree and weight loss of berry bunches where monitored weekly during withering. After one month, berry Brix degree was about 24 and the percentage of weight loss was ~30% of the initial weigh. One thousand Muller-Thurgau berries (with pedicels) were sampled, surface sterilized for 5 minutes in 70% Ethanol and dried in sterility.
Extracted molecule total RNA
Extraction protocol Infected/uninfected berry pericarps were ground into liquid nitrogen and total RNA was isolated from 200 mg of powders using the Spectrum™ Plant Total RNA Kit (Sigma-Aldrich, St. Louis, MO, USA). The RNA quantity, integrity and purity were controlled using a Nanodrop 2000 instrument (Thermo Scientific, Wilmington, DE, USA) and a Bioanalyzer Chip RNA 7500 serie II (Agilent).
Label Cy3
Label protocol Cyanine-3 (Cy3) labeled cRNA was prepared from 0.2 ug RNA using the Low Input Quick Amp Labeling kit (Agilent Technologies) according to the manufacturer's instructions, followed by RNAeasy column purification (QIAGEN, Valencia, CA). Dye incorporation and cRNA yield were checked with the NanoDrop 2000 instrument (Thermo Scientific, Wilmington, DE, USA).
 
Hybridization protocol 1.65 ug of Cy3-labelled cRNA (specific activity >9.0 pmol Cy3/ug cRNA) was fragmented at 60°C for 30 minutes in a reaction volume of 55 ul containing 25x Agilent fragmentation buffer and 10x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 55 ul of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent-048771 4x44K Grape all custom microarray chip (GPL22427) for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then dried immediately by brief centrifugation.
Scan protocol Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner using one color scan setting for 4x44k array slides (Scan Area 61x21.6 mm, Scan resolution 5um, Dye channel is set to Green and Green PMT is set to XDR Hi100%, XDR Lo 10%).
Description Gene expression 10 days after the injection (Time 2) in B. cinerea-injected berry pericarps, replicate 3
Data processing The scanned images were analyzed with Feature Extraction Software 10.5.1 (Agilent) using default parameters (protocol GE1-105_Dec08 and Grid:048771_D_F_20130410) to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded.
 
Submission date Jul 06, 2018
Last update date Jul 26, 2019
Contact name Teresa Colombo
Organization name Consiglio Nazionale delle Ricerche (CNR)
Department Institute of Molecular Biology and Pathology (IBPM)
Street address p.le A. Moro 7
City Rome
ZIP/Postal code 00185
Country Italy
 
Platform ID GPL22427
Series (2)
GSE116740 Whole-transcriptome microarray analysis of grapevine-Botrytis cinerea interaction during latent infection of berries (“noble rot”)
GSE116741 Whole-transcriptome analysis of grapevine-Botrytis cinerea interaction during latent infection of berries

Data table header descriptions
ID_REF
VALUE T-mev data normalized by percentile shifts at the 75th percentile using TMeV v4.8 (http://mev.tm4.org)

Data table
ID_REF VALUE
CUST_100_PI428914343 13000.20216
CUST_10000_PI428914343 373.7332256
CUST_10002_PI428914343 1174.462179
CUST_10003_PI428914343 70.97011295
CUST_10004_PI428914343 462.5314309
CUST_10005_PI428914343 264.9551704
CUST_10006_PI428914343 618.7838855
CUST_10007_PI428914343 121.4015722
CUST_10008_PI428914343 71.72067884
CUST_10009_PI428914343 810.8811504
CUST_10011_PI428914343 14.41043452
CUST_10013_PI428914343 101.4608373
CUST_10014_PI428914343 45.44768085
CUST_10016_PI428914343 822.0535364
CUST_10017_PI428914343 122.3672093
CUST_10018_PI428914343 329.0035209
CUST_10020_PI428914343 68.57216441
CUST_10027_PI428914343 74.3196762
CUST_10031_PI428914343 82.70812947
CUST_10040_PI428914343 1495.228233

Total number of rows: 24085

Table truncated, full table size 812 Kbytes.




Supplementary file Size Download File type/resource
GSM3259678_T2.T3.txt.gz 1.7 Mb (ftp)(http) TXT
Processed data included within Sample table

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