Fig. 4: Supplementation with GR24^5DS and ectopic expression of CCaMK restore ethylene-suppressed root colonization.

A Percent root length colonization, of wild-type L. japonicus, with R. irregularis at 4 wpi and treatment with the indicated chemicals at indicated concentrations. B Percent root length colonization of L. japonicus hairy roots transformed with empty vector (EV), pUbi:CCaMK, with R. irregularis at 5 wpi treated with solvent (water) or ACC. A, B The experiments were repeated twice with similar results. C RT-qPCR-based expression of AM marker genes in absence of the fungus in L. japonicus hairy roots carrying the indicated transgenes. Expression of SbtM1, BCP1 and RAM1 was normalized to the housekeeping gene Ubiquitin. Statistics: (A) Individual data-points and mean ± SE (N = 8) are shown. Kruskal-Wallis test (Kruskal-Wallis H statistic = 19.80 total, 12.34 arbuscules, 0.9343 vesicles) with Dunn’s posthoc comparison was used to assess significant differences between treatments for each AMF structure. B, C Two-way ANOVA with Tukey’s multiple comparisons test was used to assess significant differences between treatments and genotypes. B Individual data-points and mean ± SE (N = 8) are shown. For total: interaction F (1, 28) = 85.81 (P < 0.0001), treatment F (1, 28) = 131.1 (P < 0.0001), genotype F (1, 28) = 0.4159 (P = 0.5242); for arbuscules: interaction F (1, 28) = 71.81 (P < 0.0001), treatment F (1, 28) = 116.0 (P < 0.0001), genotype F (1, 28) = 1.254 (P = 0.2723); for vesicles: interaction F (1, 28) = 42.46 (P < 0.0001), treatment F (1, 28) = 89.84 (P < 0.0001), genotype F (1, 28) = 2.263 (P = 0.1437). Letters indicate significant differences. C Individual data-points and mean ± SE (N = 3) are shown. For SbtM1: interaction F (1, 8) = 37.06 (P = 0.0003), treatment F (1, 8) = 32.72 (P = 0.0004), genotype F (1, 8) = 1592 (P < 0.0001); for BCP1: interaction F (1, 8) = 10.47 (P = 0.0119), treatment F (1, 8) = 19.61 (P = 0.0022), genotype F (1, 8) = 798.3 (P < 0.0001); for RAM1: interaction F (1, 8) = 1.618 (P = 0.2391), treatment F (1, 8) = 2.957 (P = 0.1238), genotype F (1, 8) = 454.7 (P < 0.0001). Asterisks indicate significance level: * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001 **** p < 0.0001.