Large-Scale Phenomics Identifies Primary and Fine-Tuning Roles for CRKs in Responses Related to Oxidative Stress
An age-matched collection of T-DNA insertion lines in CRK genes was analyzed for developmental and stress-related phenotypes. (A) Analysis of developmental phenotypes of crk mutant lines: senescence, germination (endosperm rupture), epidermal cell segmentation, bolting, flowering, and root length. (B) Analysis of abiotic stresses phenotypes of crk lines: germination of crk lines on medium containing NaCl, cell death (measured by electrolyte leakage) in response to Xanthine-Xanthine Oxidase (X+XO), ultraviolet light (UV-AB), ozone (O3), or light stress. (C) Analysis of photosynthesis responses upon treatment with DCMU or methyl viologen (MV). (D) Pathogen phenotypes. ROS production in response to treatment with the bacterial elicitor flagellin (flg22). Stomatal aperture ratio in response to flg22 and chitin treatments and crk susceptibility to the hemibiotrophic bacterial pathogen Pseudomonas syringae pv. tomato DC3000 (Pto infection) or the biotrophic fungal pathogens Golovinomyces orontii (Go) (virulent on Arabidopsis) or Blumeria graminis f.sp. hordei (Bgh, a barley pathogen, non-pathogenic on Arabidopsis). (E) Analysis of stomatal parameters: fresh weight (for determination of water loss), density, length, aperture, stomatal aperture in response to ABA treatment, steady state stomatal conductance, stomatal closure in response to elevated CO2, O3, and darkness. Experiments were made comparable by bootstrap sampling to n = 15 followed by averaging over bootstrap estimates. Red and blue indicate statistically significant increase or decrease in response compared to Col-0 wild type, respectively, while white indicates a response that is similar to wild type Col-0. The intensity of color is proportional to the Benjamini-Hochberg false discovery rate (FDR) adjusted Z statistic which takes the estimated means and their variation into account. As a rough guideline, |Z|>1.67 corresponds to a FDR<10% (shown with light hue), and |Z|<2.6 to a strong FDR<1% (intense color). White: non-significant response; grey: not measured. A corresponding plot displaying the adjusted Z statistics without thresholding is shown in S25 Fig.