(SI Appendix, Fig. S8C), confirming the distinct impact of cyp79b2/ b3 mutations on Trp derivatives

(SI Appendix, Fig. S8C), confirming the distinct impact of cyp79b2/ b3 mutations on Trp derivatives in roots of plants used in our BRDT Storage & Stability experiments. We tested the extent to which the various branches of Trp metabolism could contribute for the maintenance of fungal homeostasis in roots plus the BFO-mediated plant growth promotion applying a set of mutants that, as outlined by the literature, needs to be defective within the accumulation of camalexin [pad3 (53), cyp71a27 (25), and cyp71a12/a13 (54)], ICAs [cyp71a12/a13 (54)], IGs [myb34/51/122 (55)], and some of their hydrolysis solutions [pen2 (56) and pyk10/bglu21 (57)] (SI Appendix, Fig. S10A and Dataset S2). By repopulating these mutants and WT plants together with the BFO SynCom inside the gnotobiotic FlowPot technique, we observed that none of your tested mutants phenocopied plant development (SI Appendix, Fig. S10 B and C) and fungal load (SI Appendix, Fig. S10 D ) phenotypes observed within the context on the cyp79b2/b3 mutant. To validate deficiency of tested lines in the accumulation of particular4 of 11 j PNAS doi.org/10.1073/pnas.-0.metabolites, we analyzed their accumulation in roots of those mutants inoculated using the fungal pathogen Plectosphaerella cucumerina, a species that is definitely widespread within a. thaliana roots (three) and present in our fungal SynCom. This evaluation proved lack of camalexin in roots of pad3 and cyp71a12/a13 lines as well as IG deficiency in myb34/51/122 mutant (SI Appendix, Fig. S11); on the other hand, it did not confirm partial ICA deficiency observed earlier in infected leaves of cyp71a12/a13 plants (58). Strikingly, we also found a cyp79b2/b3-like reduction in no cost IAA levels in roots of myb34/51/122 plants, which indicated that within a. thaliana roots significant amounts of this hormone is usually derived from IAOx by means of IGs, as already postulated (59). Collectively, our metabolic analysis, combined with benefits on fungal load (SI Appendix, Fig. S10 D ) and plant development promotion (SI Appendix, Fig. S10 B and C), JAK2 manufacturer excluded person contributions of IAA, IGs, and camalexin but not of ICAs to fungal overgrowth in cyp79b2/b3 plants.Dysbiotic Phenotype in the cyp79b2/b3 Mutant Is Retained in the Reproductive Stage. To test the robustness from the dysbiotic phe-notype (i.e., improved fungal load and altered plant development)Wolinska et al. Tryptophan metabolism and bacterial commensals avoid fungal dysbiosis in Arabidopsis rootsA20 bacteria/plant/ref ratioBacterial loadB6 fungi/plant/ref ratioFungal loadC150 oomycetes/plant/ref ratioOomycetes loadP = 0.1 rar -301 bri1 ::BRI1 three b 35S 9b2/ 7 cyp four p a ds depy33 wr k 33/40 y wr k two hub x ape 1 hub 5 /cerk1 k1 lyk r fls2 /ce efr/ /bkk1 1 1 bak1/bkk bak WT1 1 rar -30 bri1 ::BRI three b 35S 9b2/ 7 cyp four pad s depy33 w r k 33/40 y wr k two hub x a p e1 hub 1 5 /cerk rk1 lyk fls2 /ce efr/ /bkk1 1 1 bak1/bkk bak WT1 rar -301 bri1 ::BRI 3 b 35S 9b2/ 7 cyp 4 pad s depy33 wrk 33/40 y wr k two hubx ape1 hu b rk1 5 lyk ls2/ce cerk1 / f efr/ /bkk1 1 1 bak1/bkk bak WTD1.2 Mean Relative FWBacteria P = 0.4028, R2 = -0.E1.Fungi P = 0.005374, R2 = 0.FOomycetes P = 0.3435, R2 = -0.0.Imply Relative FW1.0.0.0.0.0.0 0.0 0.five 1.0 Imply B load (log)0.0.0 0.0 0.5 Mean F load (log) 1.0 -1 0 1 two Imply O load (log)-0.WT bak1/bkk1 bak1/bkk1/cerk1 efr/fls2/cerk1 lyk5 hub1 apex hub2 wrky33 wrky33/40 deps pad4 cyp79b2/b3 35S::BRI1 rarFig. three. Fungal load in roots explains BFO-mediated plant growth phenotypes. (A ) Bacterial (A), fungal (B), and oomycetes (C) load in plant root samples, calculated depending on qPCR information r