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Tomato is one of the most popular horticultural crops, and many commercial tomato cultivars are particularly susceptible to Botrytis cinerea. Non-expressor of pathogenesis-related gene 1 (NPR1) is a critical component of the plant defense mechanisms. However, our understanding of how SlNPR1 influences disease resistance in tomato is still limited. In this study, two independent slnpr1 mutants were used to study the role of SlNPR1 in tomato resistance against B. cinerea. Compared to (WT), slnpr1 leaves exhibited enhanced resistance against B. cinerea with smaller lesion sizes, higher activities of chitinase (CHI), β-1, 3-glucanases (GLU) and phenylalanine ammonia-lyase (PAL), and significantly increased expressions of pathogenesis-related genes (PRs). The increased activities of peroxidase (POD), ascorbate peroxidase (APX) and decreased catalase (CAT) activities collectively regulated reactive oxygen species (ROS) homeostasis in slnpr1 mutants. The integrity of the cell wall in slnpr1 mutants was maintained. Moreover, the enhanced resistance was further reflected by induction of defense genes involved in jasmonic acid (JA) and ethylene (ET) signaling pathways. Taken together, these findings revealed that knocking out SlNPR1 resulted in increased activities of defense enzymes, changes in ROS homeostasis and integrity of cell walls, and activation of JA and ET pathways, which confers resistance against B. cinerea in tomato plants., (© 2020 Scandinavian Plant Physiology Society.)

Background: High temperature is a major environmental stress that limits plant growth and agriculture productivity. Mitogen-activated protein kinases (MAPKs) are highly conserved serine and threonine protein kinases that participate in response to diverse environmental stresses in plants. A total of 16 putative SlMAPK genes are identified in tomato, and SlMAPK3 is one of the most extensively studied SlMAPKs. However, the role of SlMAPK3 in response to heat stress is not clearly understood in tomato plants. In this study, we performed functional analysis of SlMAPK3 for its possible role in response to heat stress., Results: qRT-PCR analyses revealed that SlMAPK3 relative expression was depressed by heat stress. Here, wild-type (WT) tomato plants and CRISPR/Cas9-mediated slmapk3 mutant lines (L8 and L13) were used to investigate the function of SlMAPK3 in response to heat stress. Compared with WT plants, slmapk3 mutants exhibited less severe wilting and less membrane damage, showed lower reactive oxygen species (ROS) contents, and presented higher both activities and transcript levels of antioxidant enzymes, as well as elevated expressions of genes encoding heat stress transcription factors (HSFs) and heat shock proteins (HSPs)., Conclusions: CRISPR/Cas9-mediated slmapk3 mutants exhibited more tolerance to heat stress than WT plants, suggesting that SlMAPK3 was a negative regulator of thermotolerance. Moreover, antioxidant enzymes and HSPs/HSFs genes expression were involved in SlMAPK3-mediated heat stress response in tomato plants.

Main Conclusion: Trehalose increased drought tolerance of tomato plants, accompanied by reduced water loss and closed stomata, which was associated with the upregulated ABA signaling-related genes expression, but not in ABA accumulation. Drought is one of the principal abiotic stresses that negatively influence the growth of plant and yield. Trehalose has great agronomic potential to improve the stress tolerance of plants. However, little information is available on the role of ABA and its signaling components in trehalose-induced drought tolerance. The aim of this study is to elucidate the potential mechanism by which trehalose regulates ABA in response to drought stress. In this study, 6-week-old tomato (Solanum lycopersicum cv. Ailsa Craig) plants were treated with 0 or 15.0 mM trehalose solution. Results showed that trehalose treatment significantly enhanced drought tolerance of tomato plants, accompanied by encouraged stomatal closure and protected chloroplast ultrastructure. Compared with controls, trehalose-treated plants showed lower hydrogen peroxide content and higher antioxidant enzymes activities, which contributed to alleviate oxidative damage caused by drought. Moreover, trehalose treatment decreased ABA content, which was followed by the downregulation of ABA biosynthesis genes expression and the upregulation of ABA catabolism genes expression. In contrast, exogenous trehalose upregulated transcript levels of ABA signaling-related genes, including SlPYL1/3/4/5/6/7/9, SlSnRK2.3/4, SlAREB1/2, and SlDREB1. These results suggested that trehalose treatment enhanced drought tolerance of tomato plants, and it's ABA signaling rather than ABA metabolism that was involved in trehalose-induced drought tolerance in tomato plants. These findings provide evidence for the physiological role of trehalose and bring about a new understanding of the possible relationship between trehalose and ABA.

Jasmonic acid (JA)- and ethylene-mediated signaling pathways are reported to have synergistic effects on inhibiting gray mold. The present study aimed to explain the role of ethylene perception in methyl jasmonate (MeJA)-mediated immune responses. Results showed that exogenous MeJA enhanced disease resistance, accompanied by the induction of endogenous JA biosynthesis and ethylene production, which led to the activation of the phenolic metabolism pathway. Blocking ethylene perception using 1-methylcyclopropene (1-MCP) either before or after MeJA treatment could differently weaken the disease responses induced by MeJA, including suppressing the induction of ethylene production and JA contents and reducing activities of lipoxygenase and allene oxide synthase compared to MeJA treatment alone. Consequently, MeJA-induced elevations in the total phenolic content and the activities of phenylalanine ammonia-lyase, cinnamate 4-hydroxylase, 4-coumarate:coenzyme A ligase, and peroxidase were impaired by 1-MCP. These results suggested that ethylene perception participated in MeJA-mediated immune responses in tomato fruit.

Melatonin acts as a crucial signaling molecule with multiple physiological functions in plant response to abiotic and biotic stresses. However, the impact and regulatory mechanism of melatonin on attenuating tomato fruit fungal decay are unclear. In this study, we investigated the potential roles of melatonin in modulating fruit resistance to Botrytis cinerea and explored related physiological and molecular mechanisms. The results revealed that disease resistance was strongly enhanced by melatonin treatment, and 50 μM was confirmed as the best concentration. Melatonin treatment increased the activities of defense-related enzymes and decreased hydrogen peroxide (H 2 O 2 ) content with enhanced antioxidant enzyme activities. Moreover, we found that melatonin treatment increased methyl jasmonate (MeJA) content; up-regulated the expressions of SlLoxD, SlAOC, and SlPI II; and reduced the expressions of SlMYC2 and SlJAZ1. We postulated that melatonin played a positive role in tomato fruit resistance to Botrytis cinerea through regulating H 2 O 2 level and JA signaling pathway.

Background: NPR1, nonexpressor of pathogenesis-related gene 1, is a master regulator involved in plant defense response to pathogens, and its regulatory mechanism in the defense pathway has been relatively clear. However, information about the function of NPR1 in plant response to abiotic stress is still limited. Tomato is the fourth most economically crop worldwide and also one of the best-characterized model plants employed in genetic studies. Because of the lack of a stable tomato NPR1 (SlNPR1) mutant, little is known about the function of SlNPR1 in tomato response to biotic and abiotic stresses., Results: Here we isolated SlNPR1 from tomato 'Ailsa Craig' and generated slnpr1 mutants using the CRISPR/Cas9 system. Analysis of the cis-acting elements indicated that SlNPR1 might be involved in tomato plant response to drought stress. Expression pattern analysis showed that SlNPR1 was expressed in all plant tissues, and it was strongly induced by drought stress. Thus, we investigated the function of SlNPR1 in tomato-plant drought tolerance. Results showed that slnpr1 mutants exhibited reduced drought tolerance with increased stomatal aperture, higher electrolytic leakage, malondialdehyde (MDA) and hydrogen peroxide (H 2 O 2 ) levels, and lower activity levels of antioxidant enzymes, compared to wild type (WT) plants. The reduced drought tolerance of slnpr1 mutants was further reflected by the down-regulated expression of drought related key genes, including SlGST, SlDHN, and SlDREB., Conclusions: Collectively, the data suggest that SlNPR1 is involved in regulating tomato plant drought response. These results aid in further understanding the molecular basis underlying SlNPR1 mediation of tomato drought sensitivity.

Methyl jasmonate (MeJA) and ethylene play important roles in mediating defense responses against Botrytis cinerea. Ethylene response factors (ERFs) are the final components of ethylene signal transduction; whether SlERF2 participates in disease resistance against Botrytis cinerea is unclear. The objective of this study was to investigate the role of SlERF2 in MeJA-mediated defense response by using both sense and antisense SlERF2 tomato fruit. Our results showed that both MeJA treatment and pathogen infection upregulated SlERF2 expression level. Overexpression of SlERF2 enhanced tomato fruit resistance against Botrytis cinerea. MeJA treatment increased ethylene production, promoted the activities of chitinase, β-1,3-glucanase, phenylalanine ammonia-lyase, and peroxidase, and elevated pathogenesis-related protein content and total phenolic content. Moreover, the effects of MeJA on disease response were reinforced in sense SlERF2 tomato fruit, while they were weakened in antisense SlERF2 tomato fruit. These results indicated that SlERF2 was involved in MeJA-mediated disease resistance against Botrytis cinerea in tomato fruit.

Mitogen-activated protein kinases (MAPKs) play an important role in defense responses to biotic and abiotic stresses. In order to investigate the role of SlMAPK3 in tomato plant resistance to Botrytis cinerea, two lines of slmapk3 mutants and wild-type (WT) tomato plants were used. The results showed that slmapk3 mutants were more susceptible to B. cinerea and that knockout of SlMAPK3 reduced the activities of defense enzymes and enhanced the accumulation of reactive oxygen species (ROS). Furthermore, we detected the expressions of salicylic acid (SA) and jasmonic acid (JA) signaling-related genes and found that knockout of SlMAPK3 enhanced the expressions of SlPR1, SlPAD4 and SlEDS1, whereas reduced the expressions of SlLoxC, SlPI I and SlPI II and enhanced the expressions of SlJAZ1 and SlMYC2. We postulate that SlMAPK3 plays a positive role in tomato plant resistance to B. cinerea through regulating ROS accumulation and SA and JA defense signaling pathways.

Chilling stress is the main constraint in tomato ( Solanum lycopersicum) production, as this is a chilling-sensitive horticultural crop. The highly conserved C-repeat binding factors (CBFs) are cold-response-system components found in many species. In this study, we generated slcbf1 mutants using the CRISPR-Cas9 system and investigated the role of SlCBF1 in tomato-plant chilling tolerances. The slcbf1 mutants exhibited more severe chilling-injury symptoms with higher electrolyte leakage and malondialdehyde levels than wild-type (WT) plants. Additionally, slcbf1 mutants showed lower proline and protein contents and higher hydrogen peroxide contents and activities of antioxidant enzymes than WT plants. Knockout of SlCBF1 significantly increased indole acetic acid contents but decreased methyl jasmonate, abscisic acid, and zeatin riboside contents. The reduced chilling tolerance of the slcbf1 mutants was further reflected by the down-regulation of CBF-related genes. These results contribute to a better understanding of the molecular basis underlying SlCBF1 mediation of tomato chilling sensitivity.

Drought stress is one of the most destructive environmental factors that affect tomato plants adversely. Mitogen-activated protein kinases (MAPKs) are important signaling molecules that respond to drought stress. In this study, SlMAPK3 was induced by drought stress, and the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (CRISPR/Cas9) system was utilized to generate slmapk3 mutants. Two independent T1 transgenic lines and wild-type (WT) tomato plants were used for analysis of drought tolerance. Compared with WT plants, slmapk3 mutants exhibited more severe wilting symptom, higher hydrogen peroxide content, lower antioxidant enzymes activities, and suffered more membrane damage under drought stress. Furthermore, knockout of SlMAPK3 led to up- or down-regulated expressions of drought stress-responsive genes including SlLOX, SlGST, and SlDREB. The results suggest that SlMAPK3 is involved in drought response in tomato plants by protecting cell membranes from oxidative damage and modulating transcription of stress-related genes.

Hydrogen peroxide (H 2 O 2 ) acts as a signaling molecule in response to cold stress. Mitogen-activated protein kinases (MAPKs) and C-repeat/dehydration-responsive factor (CBF) play important roles in cold response regulation. To investigate the roles of MAPKs and CBF in H 2 O 2 -induced chilling tolerance, tomato (Solanum lycopersicum cv. Ailsa Craig) plants were treated with 1 mM H 2 O 2 before chilling treatment. The results showed that H 2 O 2 treatment protected subcellular structure, increased concentrations of abscisic acid (ABA), zeatin riboside (ZR), and methyl jasmonate (MeJA), but decreased the concentration of gibberellic acid (GA 3 ). Furthermore, 1 mM H 2 O 2 treatment enhanced the activities of antioxidant enzymes. Meanwhile, relative expressions of SlMAPK1/2/3 and SlCBF1 in H 2 O 2 -treated plants were higher than those in the control. Our findings suggest that H 2 O 2 treatment might enhance the chilling tolerance of tomato plants by activating SlMAPK1/2/3 and SlCBF1 gene expression and by regulating phytohormone concentrations and antioxidant enzyme activities.

Effects of salicylic acid (SA) on gibberellin (GA) homeostasis, C-repeat/dehydration-responsive element binding factor (CBF) pathway, and antioxidant enzyme systems linked to chilling- and oxidative-stress tolerance in tomato fruit were investigated. Mature green tomatoes (Solanum lycopersicum L. cv. Moneymaker) were treated with 0, 0.5, and 1 mM SA solution for 15 min before storage at 4 °C for 28 days. In comparison to 0 or 0.5 mM SA, 1 mM SA significantly decreased the chilling injury (CI) index in tomato fruit. In the SA-treated fruit, the upregulation of GA biosynthetic gene (GA3ox1) expression was followed by gibberellic acid (GA 3 ) surge and DELLA protein degradation. CBF1 participated in the SA-modulated tolerance and stimulated the expression of GA catabolic gene (GA2ox1). Furthermore, 1 mM SA enhanced activities of antioxidant enzymes and, thus, reduced reactive oxygen species accumulation. Our findings suggest that SA might protect tomato fruit from CI and oxidative damage through regulating GA metabolism, CBF1 gene expression, and antioxidant enzyme activities.

Background: Mitogen-activated protein kinase (MAPK, EC 2.7.11.24) cascade from several plant species has been shown to be activated during response to abiotic stress. Ethylene plays an important role in fruit tolerance to environmental stress. However, the mechanisms by which MAPK regulates defence systems in fruit and the relationship between MAPK and ethylene remain to be determined., Results: MAPK inhibitor significantly decreased the chilling tolerance of tomato (Lycopersicon esculentum cv. Lichun) fruit during cold storage. Moreover, decreases in ethylene content, LeACS2 expression and activities of 1-aminocyclopropane-1-carboxylic acid (ACC) synthase (ACS, EC 4.4.1.14) and ACC oxidase (ACO, EC 1.14.17.4) due to MAPK inhibitor occurred within 24 h after cold treatment. Upon treatment with cold and ethephon, the ethylene content, activities of ACS and ACO and expression of LeACS2, LeACO1 and LeMAPK4 increased., Conclusion: The results showed the regulation of MAPK in ethylene biosynthesis to protect tomato fruit from cold stress. In addition, the participation of LeMAPK4 in cold-induced ethylene biosynthesis in tomato fruit was indicated., (© 2013 Society of Chemical Industry.)

L-arginine is the precursor of nitric oxide (NO). In order to examine the influence of L-arginine on tomato fruit resistance, preharvest green mature tomato fruits (Solanum lycopersicum cv. No. 4 Zhongshu) were treated with 0.5, 1, and 5 mM L-arginine. The reduced lesion size (in diameter) on fruit caused by Botrytis cinerea, as well as activities of phenylalanine ammonia-lyase (PAL), Chitinase (CHI), β-1,3-glucanase (GLU), and polyphenoloxidase (PPO), was compared between L-arginine treated fruits and untreated fruits. We found that induced resistance increased and reached the highest level at 3-6 days after treatment. Endogenous NO concentrations were positively correlated with PAL, PPO, CHI, and GLU activities after treatment with Pearson coefficients of 0.71, 0.94, 0.97, and 0.87, respectively. These results indicate that arginine induces disease resistance via its effects on NO biosynthesis and defensive enzyme activity.