Project description:Three grapevines cultivars (Merlot, Cabernet-Sauvignon and Ugni Blanc) were infected by E. lata. The expression profiles of the wood part near the infection point were determined for both infected and non infected plant for each cultivars with Nimblegen microarrays vitis. Three plants were used for biological replicates. Comparisons between infected and non infected conditions allow, for each cultivars, the identifcation of genes which the expression is modified by E. lata.
Project description:Cabernet Sauvignon in vitro plantlets were experimentally infected with the E. lata strain NE85-1 and compared to healthy controls.
Project description:25 ml of a modified basal salts medium (BSM; Hareland et al. 1975, J Bacteriol 121:272) supplemented with 0.05% casamino acids (Becton & Dickinson, Sparks, USA), 0.05 % yeast extract (Oxoid, Basingstoke, UK) and 0.4 % glucose (Fisher Scientific, Loughborough, UK) flasks were inoculated with 2 x 10^8 CFU and incubated at 30M-0C on an orbital shaker at 150 rpm. Growth was monitored spectrophotometrically. Burkholderia lata strains evaluated were: strain 383 (LMG22485) and strain 383-CMIT (a spontaneous mutant with increased tolerance to a commercially available blend of 3:1 methylisothiazolinone & chloromethylisothiazolinone preservatives). Sub-inhibitory concentrations of preservatives added at the time of inoculation were: 0.003% dimethylol dimethyl hydantoin (DDH, Lonza Ltd, Basal, Switzerland) or 0.01 % methylisothiazolinone/chloromethylisothiazolinone (MIT/CMIT, Romm & Haas, Coventry, UK). Only B. lata strain 383 was cultured with dimethylol dimethyl hydantoin. Strain 383 cultivated without preservatives was used as control condition.<br>Cultures were harvested at mid-exponential growth phase (optical density of 0.5, 2 x 10^8 CFU), promptly aliquoted into a microcentrifuge tube and immediately snap-cooled in liquid nitrogen before centrifuging at 20.000 x g at 4M-:C for 1 min. Pellets were immediately frozen at -80M-:C.<br>
