2.2. Inoculation of cotton bolls
Cotton bolls (28e30 dpa) on greenhouse-grown cotton plants (var. Coker 312) werewounded in the center of one of the locules to a depth of 5e10 mm with a 3 mm dia cork borer. A small aliquot (10 ml) of the A. flavus conidial suspension (104 conidia/ml) was pipetted into the hole. Control bolls received only PDB with no conidia. Bolls were harvested at 6, 24, 48 and 72 h after inoculation and were divided into several sub-samples for gene expression analysis. The sub-samples included pericarp and fiber-free seeds
from both inoculated (L1) and non-inoculated, adjacent (Adj)
locules. All the samples were quick-frozen in liquid nitrogen and
stored at 80 C for RNA isolation. Three bolls each from two
different plants (biological replicates) were used for each
treatment.