COMT1 Silencing Aggravates Heat Stress-Induced Reduction in Photosynthesis by Decreasing Chlorophyll Content, Photosystem II Activity, and Electron Transport Efficiency in Tomato

Despite a range of initiatives to reduce global carbon emission, the mean global temperature is increasing due to climate change. Since rising temperatures pose a serious threat of food insecurity, it is important to further explore important biological molecules that can confer thermotolerance to plants. Recently, melatonin has emerged as a universal abiotic stress regulator that can enhance plant tolerance to high temperature. Nonetheless, such regulatory roles of melatonin were unraveled mainly by assessing the effect of exogenous melatonin on plant tolerance to abiotic stress. Here, we generated melatonin deficient tomato plants by silencing of a melatonin biosynthetic gene, CAFFEIC ACID O-METHYLTRANSFERASE 1 (COMT1), to unveil the role of endogenous melatonin in photosynthesis under heat stress. We examined photosynthetic pigment content, leaf gas exchange, and a range of chlorophyll fluorescence parameters. The results showed that silencing of COMT1 aggravated heat stress by inhibiting both the light reactions and the carbon fixation reactions of photosynthesis. The photosynthetic pigment content, light absorption flux, trapped energy flux, energy dissipation, density of active reaction center per photosystem II (PSII) cross-section, the photosynthetic electron transport rate, the maximum photochemical efficiency of PSII photochemistry, and the rate of CO2 assimilation all decreased in COMT1-silenced plants compared with that of non-silenced plants particularly under heat stress. However, exogenous melatonin alleviated heat-induced photosynthetic inhibition in both genotypes, indicating that melatonin is essential for maintaining photosynthetic capacity under stressful conditions. These findings provide genetic evidence on the vital role of melatonin in photosynthesis and thus may have useful implication in horticultural crop management in the face of climate change

Brassinosteroids Attenuate Moderate High Temperature-Caused Decline in Tea Quality by Enhancing Theanine Biosynthesis in Camellia sinensis L.

Temperature is a major environmental signal that governs plant growth and development. A moderately high ambient temperature alters plant metabolism without significant induction of heat–stress responses. Despite ancillary reports on the negative effect of warmer climate on tea quality, information on specific effect of sub high temperature (SHT) on theanine accumulation is scanty. L-Theanine is the most abundant free amino acid in tea (Camellia sinensis L.) leaves that contributes to the unique umami flavor of green tea infusion. Tea harvested in warmer months lacks distinctive umami taste due to low theanine content. In this study, we showed that SHT (35°C) gradually decreased theanine concentration over time, which was closely associated with the SHT-induced suppression in theanine biosynthetic genes. 24-epibrassinolide (BR), a bioactive brassinosteroids, attenuated the SHT-induced reduction in theanine concentration by upregulating the transcript levels of theanine biosynthetic genes, such as ARGININE DECARBOXYLASE (CsADC), GLUTAMINE SYNTHETASE (CsGS), GLUTAMATE SYNTHASE (CsGOGAT) and THEANINE SYNTHASE (CsTS). Furthermore, time-course analysis of the activity of theanine biosynthetic enzyme reveals that BR-induced regulation of GS and GOGAT activity plays essential role in maintaining theanine content in tea leaves under SHT, which is consistent with the central position of GOGAT in theanine biosynthetic pathway. Therefore, it is convincing to propose that exogenous BR treatment can be advocated to improve summer tea quality by enhancing in vivo accumulation of theanine. However, a future challenge is to use this information on the role of BR in theanine biosynthesis and thermotolerance to further understand how BR may be tuned to benefit plant fitness for enhancing tea quality

Combined Inoculation with Multiple Arbuscular Mycorrhizal Fungi Improves Growth, Nutrient Uptake and Photosynthesis in Cucumber Seedlings

Mycorrhizal inoculation stimulates growth, photosynthesis and nutrient uptake in a wide range of host plants. However, the ultimate effects of arbuscular mycorrhyzal (AM) symbiosis vary with the plants and fungal species involved in the association. Therefore, identification of the appropriate combinations of AM fungi (AMF) that interact synergistically to improve their benefits is of high significance. Here, three AM fungal compositions namely VT (Claroideoglomus sp., Funneliformis sp., Diversispora sp., Glomus sp., and Rhizophagus sp.) and BF (Glomus intraradices, G. microageregatum BEG and G. Claroideum BEG 210), and Funneliformis mosseae (Fm) were investigated with respect to the growth, gas exchange parameters, enzymes activities in Calvin cycles and related gene expression in cucumber seedlings. The results showed that VT, BF and Fm could successfully colonize cucumber root to a different degree with the colonization rates 82.38, 74.65, and 70.32% at 46 days post inoculation, respectively. The plant height, stem diameter, dry weight, root to shoot ratio of cucumber seedlings inoculated with AMF increased significantly compared with the non-inoculated control. Moreover, AMF colonization greatly increased the root activity, chlorophyll content, net photosynthetic rate, light saturated rate of the CO2 assimilation (Asat), maximum carboxylation rate (Vcmax) and maximum ribulose-1,5-bis-phosphate (RuBP) regeneration rate (Jmax), which were increased by 52.81, 30.75, 58.76, 47.00, 69.15, and 65.53% when inoculated with VT, respectively. The activities of some key enzymes such RuBP carboxylase/oxygenase (RuBisCO), D-fructose-1,6-bisphosphatase (FBPase), D-fructose-6-phosphatase (F6P) and ribulose-5-phosphate kinase (Ru5PK), and related gene expression involved in the Calvin cycle including RCA, FBPase, FBPA, SBPase, rbcS and rbcL were upregulated by AMF colonization. AMF inoculation also improved macro- and micro nutrient contents such as N, P, K, S, Ca, Cu, Fe, Mn, Mg, and Zn in roots. Further analysis revealed that inoculation with VT had relatively better effect on growth of cucumber seedling followed by BF and Fm, indicating that AMF composition consisting of distant AMF species may have a better effect than a single or closely related AMF spp. This study advances the understanding of plant responses to different AM fungi toward development of strategies on AMF-promoted vegetable production

Table_1_COMT1 Silencing Aggravates Heat Stress-Induced Reduction in Photosynthesis by Decreasing Chlorophyll Content, Photosystem II Activity, and Electron Transport Efficiency in Tomato.doc

<p>Despite a range of initiatives to reduce global carbon emission, the mean global temperature is increasing due to climate change. Since rising temperatures pose a serious threat of food insecurity, it is important to further explore important biological molecules that can confer thermotolerance to plants. Recently, melatonin has emerged as a universal abiotic stress regulator that can enhance plant tolerance to high temperature. Nonetheless, such regulatory roles of melatonin were unraveled mainly by assessing the effect of exogenous melatonin on plant tolerance to abiotic stress. Here, we generated melatonin deficient tomato plants by silencing of a melatonin biosynthetic gene, CAFFEIC ACID O-METHYLTRANSFERASE 1 (COMT1), to unveil the role of endogenous melatonin in photosynthesis under heat stress. We examined photosynthetic pigment content, leaf gas exchange, and a range of chlorophyll fluorescence parameters. The results showed that silencing of COMT1 aggravated heat stress by inhibiting both the light reactions and the carbon fixation reactions of photosynthesis. The photosynthetic pigment content, light absorption flux, trapped energy flux, energy dissipation, density of active reaction center per photosystem II (PSII) cross-section, the photosynthetic electron transport rate, the maximum photochemical efficiency of PSII photochemistry, and the rate of CO₂ assimilation all decreased in COMT1-silenced plants compared with that of non-silenced plants particularly under heat stress. However, exogenous melatonin alleviated heat-induced photosynthetic inhibition in both genotypes, indicating that melatonin is essential for maintaining photosynthetic capacity under stressful conditions. These findings provide genetic evidence on the vital role of melatonin in photosynthesis and thus may have useful implication in horticultural crop management in the face of climate change.</p

Red Light-Induced Systemic Resistance Against Root-Knot Nematode Is Mediated by a Coordinated Regulation of Salicylic Acid, Jasmonic Acid and Redox Signaling in Watermelon

Red light (RL) can stimulate plant defense against foliar diseases; however, its role in activation of systemic defense against root diseases remains unclear. Here, the effect of RL on root knot nematode Meloidogyne incognita (RKN) infestation was investigated in watermelon plants (Citrullus lanatus L.). Plants were exposed to 200 μmol m-2 s-1 photosynthetic photon flux density RL at the canopy level for 21 days using light-emitting photodiodes. The results showed that RL significantly suppressed gall formation and nematode development, which was closely associated with the RL-induced attenuation of oxidative stress in roots. Gene expression analysis showed that RL caused a transient upregulation of PR1 and WRKY70 transcripts at 7 days post inoculation in RKN-infected plants. Further investigation revealed that RL-induced systemic defense against RKN was attributed to increased jasmonic acid (JA) and salicylic acid (SA) content, and transcript levels of their biosynthetic genes in roots. Interestingly, while malondialdehyde content decreased, H2O2 accumulation increased in RL-treated RKN-plants, indicating a potential signaling role of H2O2 in mediating RL-induced systemic defense. Furthermore, analysis of enzymatic and non-enzymatic antidoxidants revealed that RL-induced enhanced defense agaist RKN was also attributed to increased activities of antioxidant enzymes as well as redox homeostasis. Taken together, these findings suggest that RL could enhance systemic resistance against RKN, which is mediated by a coordinated regulation of JA- and SA-dependent signaling, antioxidants, and redox homeostasis in watermelon plants

Overexpression of E3 Ubiquitin Ligase Gene AdBiL Contributes to Resistance against Chilling Stress and Leaf Mold Disease in Tomato

Ubiquitination is a common regulatory mechanism, playing a critical role in diverse cellular and developmental processes in eukaryotes. However, a few reports on the functional correlation between E3 ubiquitin ligases and reactive oxygen species (ROS) or reactive nitrogen species (RNS) metabolism in response to stress are currently available in plants. In the present study, the E3 ubiquitin ligase gene AdBiL (Adi3 Binding E3 Ligase) was introduced into tomato line Ailsa Craig via Agrobacterium-mediated method. Transgenic lines were confirmed for integration into the tomato genome using PCR. Transcription of AdBiL in various transgenic lines was determined using real-time PCR. Evaluation of stress tolerance showed that T1 generation of transgenic tomato lines showed only mild symptoms of chilling injury as evident by higher biomass accumulation and chlorophyll content than those of non-transformed plants. Compared with wild-type plants, the contents of AsA, AsA/DHA, GSH and the activity of GaILDH, γ-GCS and GSNOR were increased, while H2O2, O2.−, MDA, NO, SNOs, and GSNO accumulations were significantly decreased in AdBiL overexpressing plants in response to chilling stress. Furthermore, transgenic tomato plants overexpressing AdBiL showed higher activities of enzymes such as G6PDH, 6PGDH, NADP-ICDH, and NADP-ME involved in pentose phosphate pathway (PPP). The transgenic tomato plants also exhibited an enhanced tolerance against the necrotrophic fungus Cladosporium fulvum. Tyrosine nitration protein was activated in the plants infected with leaf mold disease, while the inhibition could be recovered in AdBiL gene overexpressing lines. Taken together, our results revealed a possible physiological role of AdBiL in the activation of the key enzymes of AsA–GSH cycle, PPP and down-regulation of GSNO reductase, thereby reducing oxidative and nitrosative stress in plants. This study demonstrates an optimized transgenic strategy using AdBiL gene for crop improvement against biotic and abiotic stress factors

Responses of Plant Proteins to Heavy Metal Stress—A Review

Plants respond to environmental pollutants such as heavy metal(s) by triggering the expression of genes that encode proteins involved in stress response. Toxic metal ions profoundly affect the cellular protein homeostasis by interfering with the folding process and aggregation of nascent or non-native proteins leading to decreased cell viability. However, plants possess a range of ubiquitous cellular surveillance systems that enable them to efficiently detoxify heavy metals toward enhanced tolerance to metal stress. As proteins constitute the major workhorses of living cells, the chelation of metal ions in cytosol with phytochelatins and metallothioneins followed by compartmentalization of metals in the vacuoles as well as the repair of stress-damaged proteins or removal and degradation of proteins that fail to achieve their native conformations are critical for plant tolerance to heavy metal stress. In this review, we provide a broad overview of recent advances in cellular protein research with regards to heavy metal tolerance in plants. We also discuss how plants maintain functional and healthy proteomes for survival under such capricious surroundings

Growth temperature-induced changes in biomass accumulation, photosynthesis and glutathione redox homeostasis as influenced by hydrogen peroxide in cucumber

Hydrogen peroxide (HO) and glutathione (GSH) are involved in the stress response in plants. To elucidate the role of HO in the acclimation of CO assimilation under sub- or supra-optimal growth temperatures, we examined the effect and interaction of HO manipulation on the photosynthetic metabolism of cucumber plants (Cucumis sativus L.) grown under five temperature regimes spanning above and below the optimal growth temperature; 11/9, 18/15, 25/20, 32/27, and 39/33°C (day/night), with or without dimethylthiourea (DMTU) or HO treatment. As expected, exposure to sub- or supra-optimal growth temperatures resulted in decreased plant growth, associated with a decline in CO assimilation (A), Rubisco content, and activities of enzymes involved in the CO assimilation, as well as a decrease in the ratio of reduced (GSH) to oxidized (GSSG) glutathione (GSH/GSSG). Foliar application of HO promoted, whilst DMTU retarded the capacity of plants to acclimate to non-optimal growth temperatures; this was consistently shown in altered activity of redox-sensitive enzymes involved in CO assimilation. These results strongly suggest that the influence of growth temperature on CO assimilation was primarily targeted at the activities of the redox-sensitive enzymes of CO assimilation. Meanwhile, the data suggest that the cellular HO level is an important signal for the glutathione-dependent regulation of redox-sensitive enzymes of CO assimilation in cucumber plants

Identification of Optimal Reference Genes for Normalization of qPCR Analysis during Pepper Fruit Development

Due to its high sensitivity and reproducibility, quantitative real-time PCR (qPCR) is practiced as a useful research tool for targeted gene expression analysis. For qPCR operations, the normalization with suitable reference genes (RGs) is a crucial step that eventually determines the reliability of the obtained results. Although pepper is considered an ideal model plant for the study of non-climacteric fruit development, at present no specific RG have been developed or validated for the qPCR analyses of pepper fruit. Therefore, this study aimed to identify stably expressed genes for their potential use as RGs in pepper fruit studies. Initially, a total of 35 putative RGs were selected by mining the pepper transcriptome data sets derived from the PGP (Pepper Genome Platform) and PGD (Pepper Genome Database). Their expression stabilities were further measured in a set of pepper (Capsicum annuum L. var. 007e) fruit samples, which represented four different fruit developmental stages (IM: Immature; MG: Mature green; B: Break; MR: Mature red) using the qPCR analysis. Then, based on the qPCR results, three different statistical algorithms, namely geNorm, Normfinder, and boxplot, were chosen to evaluate the expression stabilities of these putative RGs. It should be noted that nine genes were proven to be qualified as RGs during pepper fruit development, namely CaREV05 (CA00g79660); CaREV08 (CA06g02180); CaREV09 (CA06g05650); CaREV16 (Capana12g002666); CaREV21 (Capana10g001439); CaREV23 (Capana05g000680); CaREV26 (Capana01g002973); CaREV27 (Capana11g000123); CaREV31 (Capana04g002411); and CaREV33 (Capana08g001826). Further analysis based on geNorm suggested that the application of the two most stably expressed genes (CaREV05 and CaREV08) would provide optimal transcript normalization in the qPCR experiments. Therefore, a new and comprehensive strategy for the identification of optimal RGs was developed. This strategy allowed for the effective normalization of the qPCR analysis of the pepper fruit development at the whole pepper genome level. This study not only explored the optimal RGs specific for studying pepper fruit development, but also introduced a referable strategy of RG mining which could potentially be implicated in other plant species

Table_2_Red Light-Induced Systemic Resistance Against Root-Knot Nematode Is Mediated by a Coordinated Regulation of Salicylic Acid, Jasmonic Acid and Redox Signaling in Watermelon.DOCX

<p>Red light (RL) can stimulate plant defense against foliar diseases; however, its role in activation of systemic defense against root diseases remains unclear. Here, the effect of RL on root knot nematode Meloidogyne incognita (RKN) infestation was investigated in watermelon plants (Citrullus lanatus L.). Plants were exposed to 200 μmol m^-2 s^-1 photosynthetic photon flux density RL at the canopy level for 21 days using light-emitting photodiodes. The results showed that RL significantly suppressed gall formation and nematode development, which was closely associated with the RL-induced attenuation of oxidative stress in roots. Gene expression analysis showed that RL caused a transient upregulation of PR1 and WRKY70 transcripts at 7 days post inoculation in RKN-infected plants. Further investigation revealed that RL-induced systemic defense against RKN was attributed to increased jasmonic acid (JA) and salicylic acid (SA) content, and transcript levels of their biosynthetic genes in roots. Interestingly, while malondialdehyde content decreased, H₂O₂ accumulation increased in RL-treated RKN-plants, indicating a potential signaling role of H₂O₂ in mediating RL-induced systemic defense. Furthermore, analysis of enzymatic and non-enzymatic antidoxidants revealed that RL-induced enhanced defense agaist RKN was also attributed to increased activities of antioxidant enzymes as well as redox homeostasis. Taken together, these findings suggest that RL could enhance systemic resistance against RKN, which is mediated by a coordinated regulation of JA- and SA-dependent signaling, antioxidants, and redox homeostasis in watermelon plants.</p