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Addition of 0.01–0.2 mM methyl salicylate (MeSA) to the culture media, the antioxidative capacities of maca fresh calli were increased and were 1.19–1.88 times of the control. But MeSA inhibited maca cell growth and this negative effect was dose-dependent. The dry weights of maca calli were 0.60–0.95 times of the control when addition of 0.01–0.2 mM MeSA to the culture media. Elicited by 0.05 mM MeSA, the total phenolic contents in maca calli increased and were 1.25–1.37 times of those in the control in the period from the 10th to the 20th culture day. H₂O₂ and malondialdehyde (MDA) contents increased as well. At the same time, the activities of antioxidative enzymes had corresponding changes. In conclusions, the antioxidative capacity of maca calli could be enhanced by addition of MeSA to the culture media. Considering the increase of antioxidative capacity and inhibition of maca cell growth, 0.05 mM MeSA was optimal concentration.
Calcium (Ca2+)/calmodulin (CaM), a core component of calcium messenger system, is a multiple second messenger that is involved in multiple stress tolerance including heat tolerance in plants, hydrogen sulfide (H2S) is fast emerging similar functions, but Ca2+ and H2S crosstalk in the acquisition of thermotolerance is not completely understood. In this article, Ca2+ and CaM activated the activity of cysteine desulfhydrase (L-DES), followed by inducing endogenous H2S accumulation in tobacco suspension cultured cells, while treatment with Ca2+ chelator ethylene glycol-bis(b-aminoethylether)- N,N,N0,N0-tetraacetic acid and CaM antagonists chlorpromazine lowered L-DES activity and H2S accumulation. Interestingly, Ca2+- and CaM-activated L-DES activity and endogenous H2S accumulation were eliminated by H2S synthesis inhibitors DL-propargylglycine (PAG), aminooxy acetic acid (AOA), potassium pyruvate (PP) and hydroxylamine (HA), and the H2S scavenger hypotaurine (HT), indicating that Ca2+ and CaM regulated endogenous H2S generation via activating L-DES activity in tobacco cells. Furthermore, H2S donor NaHS, Ca2+ and CaM treatment alone significantly improved the thermotolerance of tobacco cells, and heat tolerance induced by Ca2+ and CaM was enhanced by exogenous NaHS, while weakened by PAG, AOA, PP, HA or HT. All above-mentioned results suggest that endogenous H2S generated by L-DES was involved, at least partly, in the thermotolerance induced by Ca2+ and CaM in tobacco suspension cell cultures.
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