Melatonin confers thermotolerance and antioxidant capacity in Chinese cabbage.

Due to the damaging effect of high temperatures on plant development, global warming is predicted to increase agricultural risks. Chinese cabbage holds considerable importance as a leafy vegetable that is extensively consumed and cultivated worldwide. Its year-round production also encounters severe challenges in the face of high temperatures. In this study, melatonin (MT), a pivotal multifunctional signaling molecule that coordinates responses to diverse environmental stressors was used to mitigate the harmful effects of high temperatures on Chinese cabbage. Through the utilization of growth indices, cytological morphology, physiological and biochemical responses, and RNA-Seq analysis, alongside an examination of the influence of crucial enzymes in the endogenous MT synthesis pathway on the thermotolerance of Chinese cabbage, we revealed that MT pretreatment enhanced photosynthetic activity, maintained signaling pathways associated with endoplasmic reticulum protein processing, and preserved circadian rhythm in Chinese cabbage under high temperatures. Furthermore, pretreatment with MT resulted in increased levels of soluble sugar, vitamin C, proteins, and antioxidant enzyme activity, along with decreased levels of malondialdehyde, nitrate, flavonoids, and bitter glucosinolates, ultimately enhancing the capacity of the organism to mitigate oxidative stress. The knockdown of the tryptophan decarboxylase gene, which encodes a key enzyme responsible for MT biosynthesis, resulted in a significant decline in the ability of transgenic Chinese cabbage to alleviate oxidative damage under high temperatures, further indicating an important role of MT in establishing the thermotolerance. Taken together, these results provide a mechanism for MT to improve the antioxidant capacity of Chinese cabbage under high temperatures and suggest beneficial implications for the management of other plants subjected to global warming.

Role of melatonin in promoting plant growth by regulating carbon assimilation and ATP accumulation.

Melatonin (MT) is a phytohormone important in mediating diverse plant growth processes. In this study, we performed transcriptomic, qRT-PCR, physiological and biochemical analyses of Brassica rapa seedlings in order to understand how MT promotes plant growth. The results showed that exogenous MT increased the rate of cyclic electron flow around photosystem (PS) I, fluorescence quantum yield, and electron transport efficiency between PSII and PSI to promote the vegetative growth of B. rapa seedlings without affecting oxidative stress level, as compared to control. However, MT treatment significantly reduced photosynthetic rate (Pn), transpiration rate (Tr), and stomatal conductance (Gs) by 2.25-, 1.23- and 3.50-fold at 0.05 level, respectively. This occurred in parallel with the down-regulation of the genes for carbon fixation in photosynthetic organisms in a KEGG pathway enrichment. More accelerated plant growth despite the reduced photosynthesis rate and the enhanced electron transport rate suggested that NADPH and adenosine triphosphate (ATP) were preferentially diverted into other anabolic reactions than the Calvin cycle upon MT application. MT treatment increased ATP level and facilitated carbon assimilation into primary metabolism that led to a significant enhancement of soluble protein, sucrose, and fructose, but a significant decrease in glucose content. MT-induced carbon assimilation into primary metabolism was driven by up-regulation of the genes for glutathione metabolism, Krebs cycle, ribosome, and DNA replication in a KEGG pathway enrichment, as well as down-regulation of the genes for secondary metabolites. Our results provide an insight into MT-mediated metabolic adjustments triggered by coordinate changes in a wide range of gene expression profiles to help improve the plant functionality.

Effects of BrMYC2/3/4 on Plant Development, Glucosinolate Metabolism, and Sclerotinia sclerotiorum Resistance in Transgenic Arabidopsis thaliana.

MYC2/3/4, known as a basic helix-loop-helix (bHLH) transcription factor, directly activate the genes involved in diverse plant development and secondary metabolites biosynthesis. In this study, we identified and cloned five MYC paralogs (BrMYC2/3-1/3-2/4-1/4-2) from Chinese cabbage (Brassica rapa ssp. pekinensis). In-silico analyses for the physicochemical properties suggested that BrMYC2/3-1/3-2/4-2/4-3 are unstable hydrophobic and acidic proteins, while BrMYC4-1 is an unstable hydrophobic and basic protein. BrMYC2/3/4 belong to the bHLH superfamily and are closely related to AthMYC2/3/4 orthologs that mediate the regulation of various secondary metabolites. It was demonstrated that BrMYC2/3/4-GFP fusion protein localized in the nucleus and expression levels of five BrMYC2/3/4 homologous genes all elevated relative to control (Ctrl). When expressed in Arabidopsis under the control of 35S promoter, each of the BrMYC2/3-1/3-2/4-1/4-2 transgenes differentially influenced root and shoot elongation, vegetative phase change, flowering time, plant height and tiller number after flowering, and seed production. Despite the variation of phenotypes between the transgenic lines, all the lines except for BrMYC4-2 exhibited shorter seed length, less seed weight, higher accumulation of glucosinolates (GSs), and resistance to Sclerotinia sclerotiorum than Ctrl. Notably, BrMYC2 overexpression (OE) line significantly reduced the lengths of root and hypocotyl, seed length, and weight, along with faster bolting time and strikingly higher accumulation of total GSs. Accumulation of GSs at the highest levels in the BrMYC2 OE line conferred the highest resistance to S. sclerotiorum. Unlike BrMYC3 OE and BrMYC4 OE , BrMYC2 OE stimulated the growth of plant height after fluorescence. The results of this study point to the BrMYC2 overexpression that may provide a beneficial effect on plant growth and development via plant resistance to the fungal pathogen.

Melatonin elevated Sclerotinia sclerotiorum resistance via modulation of ATP and glucosinolate biosynthesis in Brassica rapa ssp. pekinensis.

Sclerotinia stem rot is a common disease found in Brassica rapa that is caused by the necrotic plant pathogen Sclerotinia sclerotiorum. Melatonin (MT) has known biological activity and effectively relieved this type of Sclerotinia stem rot in B. rapa. To better understand the mechanisms behind MT-induced S. sclerotiorum resistance in B. rapa, we performed both proteomic and metabolomic analysis. Our results showed that during S. sclerotiorum infection, thiamine synthesis was activated and defended against it. In infected leaves, ribosomal synthesis-related proteins responded positively to MT treatment. Integrated proteomic and metabolomic analysis showed that amino acid metabolism was activated by MT treatment. After MT treatment, adenosine-triphosphate (ATP) content and the activity of antioxidant enzymes were both increased in B. rapa infected leaves. Cysteine synthase, sulfur transfer-related proteins, and glucosinolate (GS) were all increased after MT treatment in infected B. rapa leaves. Taken together, these results indicated that B. rapa leaves promoted thiamine formation to defend against S. sclerotiorum infection. Moreover, MT helped further induce antioxidant activation in B. rapa in an ATP-dependent manner and stimulating GS biosynthesis to well inhibit the S. sclerotiorum infection. SIGNIFICANCE: Melatonin (MT) has biological activity and effectively relieved the Sclerotinia stem rot of Brassica rapa caused by the necrotic plant pathogen Sclerotinia sclerotiorum. In order to reveal the molecular mechanisms of MT-induced S. sclerotiorum resistance in B. rapa, comprehensive proteomic and metabolomic analyses were conducted. The integration analysis of omic-data illustrated that the modulation of ATP and glucosinolate biosynthesis induced by MT administration helped to defend the infection of S. sclerotiorum in B. rapa. Our results will provide insights into MT-induced anti-fungal mechanism and therapeutic strategies to mitigate Sclerotinia stem rot of B. rapa, thereby increasing plant yield and decreasing economic losses.