BAG8 positively regulates cold stress tolerance by modulating photosystem, antioxidant system and protein protection in Solanum lycopersicum.

The B-cell lymphoma 2 (Bcl-2)-associated athanogene (BAG) family is a relatively conserved and multifunctional co-chaperones in animals and plants, which can flexibly interact with a variety of proteins and regulate various processes from growth and development to stress response. However, compared with animals, the function of BAG family in plant remains largely unknown, especially in response to cold stress. In this study, we have found that the expression of BAG8 was significantly induced in tomato under cold stress. Results showed that bag8 mutants exhibit significantly reduced tolerance towards cold stress, while BAG8 overexpressing lines were relatively resistant as reflected by the phenotype and membrane peroxidation. Measuring of gas exchange parameters, photosystem I (PSI) and photosystem II (PSII) of tomato leaves under cold stress further revealed that BAG8 mitigated cold-induced damage in photosynthetic system. Additionally, bag8 mutants exhibited more cold-induced reactive oxygen species, which were substantially normalized in BAG8 overexpressing plants. Nevertheless, the activities of antioxidant enzymes which were compromised in bag8 mutants were improved in BAG8 overexpressing plants facing cold stress. Additionally, BAG8 interacted with heat shock protein Hsp70 and protein phosphatase PP2A both in vitro and in vivo. Our results demonstrate that BAG8 plays a positive role in cold tolerance in tomato probably by the improvement of photosystems and antioxidant systems, and by interacting with Hsp70 involved in photosynthesis and PP2A involved in stomatal development.

Autophagy promotes jasmonate-mediated defense against nematodes.

Autophagy, as an intracellular degradation system, plays a critical role in plant immunity. However, the involvement of autophagy in the plant immune system and its function in plant nematode resistance are largely unknown. Here, we show that root-knot nematode (RKN; Meloidogyne incognita) infection induces autophagy in tomato (Solanum lycopersicum) and different atg mutants exhibit high sensitivity to RKNs. The jasmonate (JA) signaling negative regulators JASMONATE-ASSOCIATED MYC2-LIKE 1 (JAM1), JAM2 and JAM3 interact with ATG8s via an ATG8-interacting motif (AIM), and JAM1 is degraded by autophagy during RKN infection. JAM1 impairs the formation of a transcriptional activation complex between ETHYLENE RESPONSE FACTOR 1 (ERF1) and MEDIATOR 25 (MED25) and interferes with transcriptional regulation of JA-mediated defense-related genes by ERF1. Furthermore, ERF1 acts in a positive feedback loop and regulates autophagy activity by transcriptionally activating ATG expression in response to RKN infection. Therefore, autophagy promotes JA-mediated defense against RKNs via forming a positive feedback circuit in the degradation of JAMs and transcriptional activation by ERF1.

HsfA1a confers pollen thermotolerance through upregulating antioxidant capacity, protein repair, and degradation in Solanum lycopersicum L.

The heat shock transcription factors (Hsfs) play critical roles in plant responses to abiotic stresses. However, the mechanism of Hsfs in the regulation of pollen thermotolerance and their specific biological functions and signaling remain unclear. Herein, we demonstrate that HsfA1a played a key role in tomato pollen thermotolerance. Pollen thermotolerance was reduced in hsfA1a mutants but was increased by hsfA1a overexpression, based on pollen viability and germination. Analyzing the whole transcriptome by RNA-seq data, we found that HsfA1a mainly regulated the genes involved in oxidative stress protection, protein homeostasis regulation and protein modification, as well as the response to biological stress in anthers under heat stress. The accumulation of reactive oxygen species in anthers was enhanced in hsfA1a mutants but decreased in HsfA1a-overexpressing lines. Furthermore, HsfA1a bound to the promoter region of genes involved in redox regulation (Cu/Zn-SOD, GST8, and MDAR1), protein repair (HSP17.6A, HSP70-2, HSP90-2, and HSP101) and degradation (UBP5, UBP18, RPN10a, and ATG10) and regulated the expression of these genes in tomato anthers under heat stress. Our findings suggest that HsfA1a maintains pollen thermotolerance and cellular homeostasis by enhancing antioxidant capacity and protein repair and degradation, ultimately improving pollen viability and fertility.

BAG9 Confers Thermotolerance by Regulating Cellular Redox Homeostasis and the Stability of Heat Shock Proteins in Solanum lycopersicum.

The Bcl-2-associated athanogene (BAG) family, a group of co-chaperones that share conservative domains in flora and fauna, is involved in plant growth, development, and stress tolerance. However, the function of tomato BAG genes on thermotolerance remains largely unknown. Herein, we found that the expression of BAG9 was induced during heat stress in tomato plants. Knockout of the BAG9 gene by CRISPR/Cas9 reduced, while its overexpression increased thermotolerance in tomato plants as reflected by the phenotype, photosynthesis rate, and membrane peroxidation. Heat-induced reactive oxygen species and oxidative/oxidized proteins were further increased in bag9 mutants and were normalized in BAG9 overexpressing plants. Furthermore, the activities of antioxidant enzymes, ascorbic acid (AsA)/dehydroascorbic acid (DHA), and reduced glutathione (GSH)/oxidized glutathione (GSSG) were reduced in bag9 mutants and were increased in BAG9 overexpressing plants under heat stress. Additionally, BAG9 interacted with Hsp20 proteins in vitro and in vivo. Accumulation of Hsp proteins induced by heat showed a reduction in bag9 mutants; meanwhile, it was increased in BAG9 overexpressing plants. Thus, BAG9 played a crucial role in response to heat stress by regulating cellular redox homeostasis and the stability of heat shock proteins.

Functions of Redox Signaling in Pollen Development and Stress Response.

Cellular redox homeostasis is crucial for normal plant growth and development. Each developmental stage of plants has a specific redox mode and is maintained by various environmental cues, oxidants, and antioxidants. Reactive oxygen species (ROS) and reactive nitrogen species are the chief oxidants in plant cells and participate in cell signal transduction and redox balance. The production and removal of oxidants are in a dynamic balance, which is necessary for plant growth. Especially during reproductive development, pollen development depends on ROS-mediated tapetal programmed cell death to provide nutrients and other essential substances. The deviation of the redox state in any period will lead to microspore abortion and pollen sterility. Meanwhile, pollens are highly sensitive to environmental stress, in particular to cell oxidative burst due to its peculiar structure and function. In this regard, plants have evolved a series of complex mechanisms to deal with redox imbalance and oxidative stress damage. This review summarizes the functions of the main redox components in different stages of pollen development, and highlights various redox protection mechanisms of pollen in response to environmental stimuli. In continuation, we also discuss the potential applications of plant growth regulators and antioxidants for improving pollen vigor and fertility in sustaining better agriculture practices.

Phytonanotechnology applications in modern agriculture.

With the rapidly changing global climate, the agricultural systems are confronted with more unpredictable and harsh environmental conditions than before which lead to compromised food production. Thus, to ensure safer and sustainable crop production, the use of advanced nanotechnological approaches in plants (phytonanotechnology) is of great significance. In this review, we summarize recent advances in phytonanotechnology in agricultural systems that can assist to meet ever-growing demands of food sustainability. The application of phytonanotechnology can change traditional agricultural systems, allowing the target-specific delivery of biomolecules (such as nucleotides and proteins) and cater the organized release of agrochemicals (such as pesticides and fertilizers). An amended comprehension of the communications between crops and nanoparticles (NPs) can improve the production of crops by enhancing tolerance towards environmental stresses and optimizing the utilization of nutrients. Besides, approaches like nanoliposomes, nanoemulsions, edible coatings, and other kinds of NPs offer numerous selections in the postharvest preservation of crops for minimizing food spoilage and thus establishing phtonanotechnology as a sustainable tool to architect modern agricultural practices.

Comparative Physiological and Proteomic Analyses Reveal the Mechanisms of Brassinolide-Mediated Tolerance to Calcium Nitrate Stress in Tomato.

Secondary salinization caused by the overaccumulation of calcium nitrate [Ca(NO3)2] in soils due to excessive fertilization has become one of the major handicaps of protected vegetable production. Brassinolide, a bioactive plant steroid hormone, plays an important role in improving abiotic stress tolerance in plants. However, whether and how brassinolide (BR) can alleviate Ca(NO3)2 stress remains elusive. Here, we investigated the effects of exogenous BR on hydroponically grown tomato (Solanum lycopersicum L.) plants under Ca(NO3)2 stress through proteomics combined with physiological studies. Proteomics analysis revealed that Ca(NO3)2 stress affected the accumulation of proteins involved in photosynthesis, stress responses, and antioxidant defense, however, exogenous BR increased the accumulation of proteins involved in chlorophyll metabolism and altered the osmotic stress responses in tomatoes under Ca(NO3)2 stress. Further physiological studies supported the results of proteomics and showed that the exogenous BR-induced alleviation of Ca(NO3)2 stress was associated with the improvement of photosynthetic efficiency, levels of soluble sugars and proteins, chlorophyll contents, and antioxidant enzyme activities, leading to the reduction in the levels of reactive oxygen species and membrane lipid peroxidation, and promotion of the recovery of photosynthetic performance, energy metabolism, and plant growth under Ca(NO3)2 stress. These results show the importance of applying BR in protected agriculture as a means for the effective management of secondary salinization.

Melatonin promotes metabolism of bisphenol A by enhancing glutathione-dependent detoxification in Solanum lycopersicum L.

Bisphenol A (BPA), a widely distributed organic compound, is toxic to animals and plants. Here we show the mechanism of BPA detoxification by melatonin (MEL) in tomato, which is otherwise poorly understood in plants. BPA treatment decreased the quantum yield of photosystem II (Fv/Fm) and increased the membrane lipid peroxidation and reactive oxygen species (ROS) accumulation dose-dependently, whereas exogenous MEL alleviated the BPA effects on Fv/Fm, lipid peroxidation, ROS accumulation and BPA uptake. Furthermore, BPA elevated the glutathione (GSH) content, activities of glutathione S-transferase (GST), and glutathione reductase (GR), and the transcript levels of GSH1, GR1, GST1 and MEL biosynthesis genes (COMT, T5H, and SNAT), whereas BPA + MEL showed even a more profound induction. Silencing GSH1, GR1 and GST1 genes compromised the BPA detoxification potential of tomato plants as revealed by an increased level of ROS, lipid peroxidation and BPA uptake, and a decreased Fv/Fm and GST activity; these changes were alleviated by MEL application. Under in vitro conditions, BPA was glutathionylated by GSH, which was further catalyzed by GST to cysteine and N-acetylcysteine conjugates. These findings suggest a crucial role for MEL in BPA detoxification via GSH and GST, and can be useful to reduce BPA residue for food safety.

Jasmonic Acid Seed Treatment Stimulates Insecticide Detoxification in Brassica juncea L.

The present study focused on assessing the effects of jasmonic acid (JA) seed treatment on the physiology of Brassica juncea seedlings grown under imidacloprid (IMI) toxicity. It has been observed that IMI application declined the chlorophyll content and growth of seedlings. However, JA seed treatment resulted in the significant recovery of chlorophyll content and seedling growth. Contents of oxidative stress markers like superoxide anion, hydrogen peroxide, and malondialdehyde were enhanced with IMI application, but JA seed treatment significantly reduced their contents. Antioxidative defense system was activated with IMI application which was further triggered after JA seed treatment. Activities of antioxidative enzymes and contents of non-enzymatic antioxidants were enhanced with the application of IMI as well as JA seed treatment. JA seed treatment also regulated the gene expression of various enzymes under IMI stress. These enzymes included respiratory burst oxidase (RBO), Ribulose-1,5-bisphosphate carboxylase/oxygenase (RUBISCO), NADH-ubiquinone oxidoreductase (NADH), carboxylesterase (CXE), chlorophyllase (CHLASE), cytochrome P450 monooxygenase (P450). JA seed treatment up-regulated the expressions of RUBISCO, NADH, CXE, and P450 under IMI toxicity. However, expressions of RBO and CHLASE were down-regulated in seedlings germinated from JA seed treatment and grown in presence of IMI. Seed soaking with JA also resulted in a significant reduction of IMI residues in B. juncea seedlings. The present study concluded that seed soaking with JA could efficiently reduce the IMI toxicity by triggering the IMI detoxification system in intact plants.

Phytomelatonin: Recent advances and future prospects.

Melatonin (MEL) has been revealed as a phylogenetically conserved molecule with a ubiquitous distribution from primitive photosynthetic bacteria to higher plants, including algae and fungi. Since MEL is implicated in numerous plant developmental processes and stress responses, the exploration of its functions in plant has become a rapidly progressing field with the new paradigm of involvement in plants growth and development. The pleiotropic involvement of MEL in regulating the transcripts of numerous genes confirms its vital involvement as a multi-regulatory molecule that architects many aspects of plant development. However, the cumulative research in plants is still preliminary and fragmentary in terms of its established functions compared to what is known about MEL physiology in animals. This supports the need for a comprehensive review that summarizes the new aspects pertaining to its functional role in photosynthesis, phytohormonal interactions under stress, cellular redox signaling, along with other regulatory roles in plant immunity, phytoremediation, and plant microbial interactions. The present review covers the latest advances on the mechanistic roles of phytomelatonin. While phytomelatonin is a sovereign plant growth regulator that can interact with the functions of other plant growth regulators or hormones, its qualifications as a complete phytohormone are still to be established. This review also showcases the yet to be identified potentials of phytomelatonin that will surely encourage the plant scientists to uncover new functional aspects of phytomelatonin in plant growth and development, subsequently improving its status as a potential new phytohormone.

Melatonin Alleviates High Temperature-Induced Pollen Abortion in Solanum lycopersicum.

Melatonin is a pleiotropic signal molecule that plays critical roles in regulating plant growth and development, as well as providing physiological protections against various environmental stresses. Nonetheless, the mechanisms for melatonin-mediated pollen thermotolerance remain largely unknown. In this study, we report that irrigation treatment with melatonin (20 µM) effectively ameliorated high temperature-induced inactivation of pollen and inhibition of pollen germination in tomato (Solanum lycopersicum) plants. Melatonin alleviated reactive oxygen species production in tomato anthers under high temperature by the up-regulation of the transcription and activities of several antioxidant enzymes. Transmission electron micrograph results showed that high temperature-induced pollen abortion is associated with a premature degeneration of the tapetum cells and the formation of defective pollen grains with degenerated nuclei at the early uninuclear microspore stage, whilst melatonin protected degradation of organelles by enhancing the expression of heat shock protein genes to refold unfolded proteins and the expression of autophagy-related genes and formation of autophagosomes to degrade denatured proteins. These findings suggest a novel function of melatonin to protect pollen activity under high temperature and support the potential effects of melatonin on reproductive development of plants.

Castasterone confers copper stress tolerance by regulating antioxidant enzyme responses, antioxidants, and amino acid balance in B. juncea seedlings.

The aim of the present study was to explore the effect of exogenous application of castasterone (CS) on physiologic and biochemical responses in Brassica juncea seedlings under copper (Cu) stress. Seeds were pre-soaked in different concentrations of CS and grown for 7 days under various levels of Cu. The exposure of B. juncea to higher levels of Cu led to decrease of morphologic parameters, with partial recovery of length and fresh weight in the CS pre-treated seedlings. Metal content was high in both roots and shoots under Cu exposure while the CS pre-treatment reduced the metal uptake. Accumulation of hydrogen peroxide (H2O2) and superoxide anion radical (O2-) were chosen as stress biomarker and higher levels of H2O2 (88.89%) and O2- (62.11%) showed the oxidative stress in metal treated B. juncea seedlings, however, CS pre-treatment reduced ROS accumulation in Cu-exposed seedlings. The Cu exposures lead to enhance the plant's enzymatic and non-enzymatic antioxidant system. It was observed that enzymatic activities of ascorbate peroxidase (APOX), dehydroascorbate reductase (DHAR), and glutathione reductase (GR), glutathione perxoidase (GPOX) and gultrathione-s-transferase increased while activity of monodehydroascorbate reductase (MDHAR) decreased under Cu stress. The pre-treatment with CS positively affected the activities of enzymes. RT-PCR analysis showed that mRNA transcript levels were correlated with total enzymatic activity of DHAR, GR, GST and GSH. Increase in the gene expression of DHAR (1.85 folds), GR (3.24 folds), GST-1 (2.00 folds) and GSH-S (3.18 folds) was noticed with CS pre-treatment. Overall, the present study shows that Cu exposure induced severe oxidative stress in B. juncea plants and exogenous application of CS improved antioxidative defense system by modulating the ascorbate-glutathione cycle and amino acid metabolism.

Involvement of Asada-Halliwell Pathway During Phytoremediation of Chromium (VI) in Brassica juncea L. Plants.

Brassica juncea (Indian mustard) L. plants were exposed to different concentrations (0.0, 0.1, 0.3 and 0.5 mM) of Chromium (Cr) and harvested after 30 and 60 days of sowing for the analysis of growth parameters, metal uptake and oxidative stress markers. Significant accumulation of Cr (VI) by B. juncea L. plants resulted in the reduced growth and modulations in the pool of various biochemical stress markers. The toxic effects of Cr (VI) on growth and other stress markers (protein content, lipid peroxidation and antioxidative enzymes viz.SOD, CAT, POD, APOX, GR, DHAR and MDHAR) in B. juncea L. were observed to be concentration and time dependent. Effect of Cr (VI) on biochemical parameters was differential and their maximum activities of SOD, POD, APX, GR, DHAR and lipid peroxidation were recorded at 0.5 mM concentration in 30 days old plants. Whereas, trend in the activities of most of the stress markers was reversed in 60 days old plants. The results obtained from the study suggested that Cr (VI) stress inhibited growth of B. juncea L. plants is directly interrelated with its accumulation and resulted in the modulation in activities of various stress markers.

Arsenic induced modulation of antioxidative defense system and brassinosteroids in Brassica juncea L.

Brassica juncea (Indian mustard) L. plants were exposed to different concentrations (0.0, 0.1, 0.2 and 0.3mM) of arsenic (V) and harvested after 30 and 60 days of sowing for the analysis of growth parameters, metal uptake, brassinosteroids (BRs) synthesis and oxidative stress markers. As (V) significantly hampered the growth of B. juncea plants and triggered the modulations of various stress markers like proteins, antioxidative enzymes (SOD, CAT, POD, APX, GR, MDHAR and DHAR) and MDA content. Furthermore, As (V) induced the synthesis of 4 BRs, castasterone, teasterone, 24-epibrassinolide, and typhasterol, which were isolated and characterized by gas chromatography-mass spectrometry (GC-MS). The study further highlig5895hted the significant uptake of arsenic ions by mustard plants.

Plant steroid hormones produced under Ni stress are involved in the regulation of metal uptake and oxidative stress in Brassica juncea L.

Brassinosteroids (BRs) are involved in the amelioration of various biotic and abiotic stresses. With an aim to explore the role of BRs under heavy metal stress, plants of Brassica juncea L. were grown in pots. The plants were subjected to various concentrations of Nickel metal (0.0, 0.2, 0.4 and 0.6 mM) and harvested on 60th day in order to observe the expression of these hormones. The isolated BRs from the leaves of Brassica plants characterized by GC-MS include 24-Epibrassinolide (24-EBL), Castasterone, Dolicholide and Typhasterole. The effect of isolated 24-EBL was studied on Ni metal uptake and antioxidative defense system in 60 d old plants of Brassica. It was observed that 24-EBL significantly increased the activities of stress ameliorating enzymes and lowered the metal uptake in plants. This is the first report in B. juncea L. plants showing the expression of BRs under metal treatments and effect of the isolated 24-EBL on metal uptake and in oxidative stress management.