Fig. 3

Highly divergent GhRVD1 between Hap. B and Hap. E. a An illustration of a 26 base point mutation in the CDS, and 22 nonsynonymous mutations in amino acid sequences among GhRVD1_R and GhRVD1_S; R and S are shown in green and black words, respectively. Mutation positions are marked on the top and bottom. b Phenotype of GhRVD1_R and GhRVD1_S overexpression cotton lines. c Transcript detection of both GhRVD1 genotypes using semi-quantitative PCR with universal primers; GhHiston3 was used as an internal control. d Phenotypes with a significant reduction of plant height and internode distances. Different letters indicate significant difference at α = 0.05 level via one-way ANOVA analysis. e Trypan blue staining to detect dead cells (top), and DAB staining to detect accumulation of ROS species (bottom) in 10-week-old sample leaves. f SA content of 10-week-old sample leaves via HPLC. g Expression of SAR markers (e.g., NPR1 and PR1) and HR markers (e.g., HIN1 and HSR203J) in GhRVD1 overexpression lines from leaves collected from 10-week-old plants. h Plant wilt phenotype, leaves wilt phenotype, and stem browning symptoms were photographed at 3 weeks post-inoculation. i DI was the mean value of three independent experiments and was measured at 21 dpi