Glutathione (GSH) and indole glucosinolates (IGs) exert key functions in the immune system of the model plant Arabidopsis (). Appropriate GSH levels are important for execution of both pre- and postinvasive disease resistance mechanisms to invasive pathogens, whereas an intact PENETRATION2 (PEN2)-pathway for IG metabolism is essential for preinvasive resistance in this species. Earlier indirect evidence suggested that the latter pathway involves conjugation of GSH with unstable products of IG metabolism and further processing of the resulting adducts to biologically active molecules. Here we describe the identification of Glutathione--Transferase class-tau member 13 (GSTU13) as an indispensable component of the PEN2 immune pathway for IG metabolism. mutant plants are defective in the pathogen-triggered biosynthesis of end products of the PEN2 pathway, including 4-O-β-d-glucosyl-indol-3-yl formamide, indole-3-ylmethyl amine, and raphanusamic acid. In line with this metabolic defect, lack of functional GSTU13 results in enhanced disease susceptibility toward several fungal pathogens including ,, and Seedlings of plants fail also to deposit the (1,3)-β-glucan cell wall polymer, callose, after recognition of the bacterial flg22 epitope. We show that GSTU13 mediates specifically the role of GSH in IG metabolism without noticeable impact on other immune functions of this tripeptide. We postulate that GSTU13 connects GSH with the pathogen-triggered PEN2 pathway for IG metabolism to deliver metabolites that may have numerous functions in the innate immune system of Arabidopsis.

Summary Effective defense of Arabidopsis against filamentous pathogens requires two mechanisms, both of which involve biosynthesis of tryptophan (Trp)‐derived metabolites. Extracellular resistance involves products of PEN2‐dependent metabolism of indole glucosinolates (IGs). Restriction of further fungal growth requires PAD3‐dependent camalexin and other, as yet uncharacterized, indolics. This study focuses on the function of CYP71A12 monooxygenase in pathogen‐triggered Trp metabolism, including the biosynthesis of indole‐3‐carboxylic acid (ICA). Moreover, to investigate the contribution of CYP71A12 and its products to Arabidopsis immunity, we analyzed infection phenotypes of multiple mutant lines combining pen2 with pad3, cyp71A12, cyp71A13 or cyp82C2. Metabolite profiling of cyp71A12 lines revealed a reduction in ICA accumulation. Additionally, analysis of mutant plants showed that low amounts of ICA can form during an immune response by CYP71B6/AAO1‐dependent metabolism of indole acetonitrile, but not via IG hydrolysis. Infection assays with Plectosphaerella cucumerina and Colletotrichum tropicale, two pathogens with different lifestyles, revealed cyp71A12‐, cyp71A13‐ and cyp82C2‐associated defects associated with Arabidopsis immunity. Our results indicate that CYP71A12, but not CYP71A13, is the major enzyme responsible for the accumulation of ICA in Arabidopsis in response to pathogen ingression. We also show that both enzymes are key players in the resistance of Arabidopsis against selected filamentous pathogens after they invade.

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Four species of flowering plants comprising Arctic populations of Cerastium alpinum and Poa arctica var. vivipara and indigenous Antarctic species Colobanthus quitensis and Deschampsia antarctica were investigated. Plants derived from natural origins were grown in an experimental greenhouse in Poland (53°47 0 N and 20°30 0 E latitude). Plants for experiment were collected during spring of 2010. Soluble carbohydrates in the intact shoots of C. alpinum and C. quitensis, polar plants of the family Caryophyllaceae, and D. antarctica and P. arctica var. vivipara, representatives of the family Poaceae, were analyzed by gas chromatography, and their involvement in the plants' response to chilling stress was examined. Plant tissues of the examined families growing in a greenhouse conditions (18-20°C, short day 10/14 h light/darkness) differed in the content and composition of soluble carbohydrates. In addition to common monosaccharides, myo-inositol and sucrose, Caryophyllaceae plants contained raffinose family oligosaccharides (RFOs), D-pinitol and mono-galactosyl pinitols. RFOs and D-pinitol were not detected in plants of the family Poaceae which contain 1-kestose, a specific tri-saccharide. The accumulation of significant quantities of sucrose in all investigated plants, RFOs in Caryophyllaceae plants and 1-kestose in Poaceae plants in response to chilling stress (4°C for 48 h with a long day photoperiod, 20/4 h) indicates that those compounds participate in the stress response. The common sugar accumulating in cold stress response and probably most important for chilling tolerance of four investigated plants species seems to be sucrose. On the other hand, the accumulation of above-mentioned carbohydrates during chilling stress can be a return to sugars metabolism, occurring in natural environmental conditions. No changes in D-pinitol concentrations were observed in the tissues of C. alpinum and C. quitensis plants subjected to both low and elevated temperatures, which probably rules out the protective effects of D-pinitol in response to cold stress.

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