Fig. 2

CaWRKY70 overexpression induces susceptibility in chickpea under Foc1 infection. a Diagram of gene construct used for CaWRKY70 overexpression in resistant chickpea. b qRT-PCR determination of CaWRKY70 mRNA level in control transgenic (VC) and overexpressing chickpea. Error bars indicate ±SD of three independent biological samples. Data was normalized to CaGAPDH. Fold change values relative to control vector transformed plants. c Development of disease symptom in control transgenic and CaWRKY70 overexpressing chickpea. Disease phenotype of transgenic plants were photographed under control treatment (0 dpi) and Foc1infection at 3, 7 and 12 dpi, respectively. d Assessment of disease intensity index in control transgenic and CaWRKY70 overexpressing chickpea under control treatment (0 dpi) and Foc1infection. Chickpea plants grown in the soil-rite mixture were inoculated with Foc1. e Effect of Foc1 infection on the control transgenic and CaWRKY70 overexpressing chickpea based on incidence of dead or wilted plants. In d and e, each data point represents mean values of three pots with five plants per pot. Error bars indicate ±SD of three independent biological samples. f Electrolyte leakage in control transgenic (VC) and CaWRKY70 overexpressing chickpea roots under control treatment (0 dpi) and Foc1 infection. Each time point represents ±SD of three biological replicates. *P ≤ 0.05 and **P ≤ 0.01 indicate values show significant differences between control transgenic (VC) and CaWRKY70 overexpressing chickpea as determined by Student’s t test. g Quantitation of Foc1 5.8S rDNA at root tissues of control transgenic (VC) and CaWRKY70 overexpressing chickpea plants by real-time PCR. Chickpea DNA amount was normalized by CaGAPDH expression. Each bar represents mean ± SD of three independent biological replicates and fold change is relative to control treatment (0 dpi). **P ≤ 0.01 indicate mean values significantly different from control vector transformed plant determined by Student’s t test. h Relative water content percent (RWC%) in control transgenic and CaWRKY70 overexpressing chickpea under control treatment and Foc1 infection. Asterisks (*) indicate significant difference at ***P ≤ 0.001 by one-way ANOVA followed by multiple comparison of means using tukey’s post-hoc test. i Disease symptoms in chickpea leaves for Foc1 varied from yellowing to browning or cell death at different infection time-points. j Chlorophyll A and (k) Chlorophyll B content in control transgenic and CaWRKY70 overexpressing chickpea leaves at different infection time-points. In j and k, error bars represent ±SD (n = 3). *P ≤ 0.05 indicate mean values are significantly different from control vector transformed plants as determined by Student’s t test