Chilling Stress Research Articles

Thioredoxin h2 inhibits the MPKK5-MPK3 cascade to regulate the CBF-COR signaling pathway in Citrullus lanatus suffering chilling stress.

Thioredoxins (TRXs) are ubiquitous oxidoreductases and present as a multigenic family. TRXs determine the thiol redox balance, which is crucial for plants in the response to cold stress. However, limited knowledge is available about the role of TRXs in watermelon (Citrullus lanatus), which is highly sensitive to chilling stress in agricultural practice. Here, we identified 18 genes encoding 14 typical and 4 atypical TRXs from the watermelon genome, and found that ClTRXh2 localized at the plasma membrane was largely induced by chilling. Virus-induced gene silencing of ClTRXh2 resulted in watermelon plants that were more sensitive to chilling stress. We further found that ClTRXh2 physically interacted with mitogen-activated protein kinase kinase 5 (ClMPKK5), which was confirmed to phosphorylate and activate ClMPK3 in vitro, and the activation of ClMPK3 by ClMPKK5 was blocked by a point mutation of the Cys-229 residue to Ser in ClMPKK5. Additionally, ClTRXh2 inhibited the chilling-induced activation of ClMPK3, suggesting that the ClMPKK5-ClMPK3 cascade is regulated in a redox-dependent manner. We showed that ClMPK3-silenced plants had increased tolerance to chilling, as well as enhanced transcript abundances of the C-repeat/DREB binding factor (ClCBF) and cold-responsive (ClCOR) genes. Taken together, our results indicate that redox status mediated by ClTRXh2 inhibits ClMPK3 phosphorylation through the interaction between ClTRXh2 and ClMPKK5, which subsequently regulates the CBF-COR signaling pathway when submitted to chilling stress. Hence, our results provide a link between thiol redox balance and MAPK cascade signaling, revealing a conceptual framework to understand how TRX regulates chilling stress tolerance in watermelon.

A tomato chloroplast-targeted DnaJ protein, SlDnaJ20 maintains the stability of photosystem I/II under chilling stress

ABSTRACT DnaJ proteins are key molecular chaperones that act as a part of the stress response to stabilize plant proteins, thereby maintaining protein homeostasis under stressful conditions. Herein we used transgenic plants to explore the role of the tomato (Solanum lycopersicum) SlDnaJ20 chloroplast DnaJ protein in to the resistance of these proteins to cold. When chilled, transgenic plants exhibited superior cold resistance, with reduced growth inhibition and cellular damage and increased fresh mass and chlorophyll content relative to control. These transgenic plants further exhibited increased Fv/Fm, P700 oxidation, φRo, and δRo relative to control plants under chilling conditions. Under these same cold conditions, these transgenic plants also exhibited higher levels of core proteins in the photosystem I (PSI) and II (PSII) complexes (PsaA and PsaB; D1 and D2) relative to control wild-type plants. Together these results suggested that the overexpression of SlDnaJ20 is sufficient to maintain PSI and PSII complex stability and to alleviate associated photoinhibition of these complexes, thereby increasing transgenic plant resistance to cold stress.

Comprehensive analysis of the NAC transcription factor gene family in Kandelia obovata reveals potential members related to chilling tolerance.

Kandelia obovata is an important mangrove species extensively distributed in Eastern Asia that is susceptible to low-temperature stress. NAC (NAM, ATAF1/2 and CUC2) domain proteins are transcription factors (TFs) that play various roles in plant growth and development and in the plant response to environmental stresses. Nevertheless, genome-wide analyses of K. obovata NAC genes (KoNACs) and their responses to chilling stress have rarely been studied. The KoNAC gene family was identified and characterized using bioinformatic analysis, the subcellular location of some NAC proteins was confirmed using confocal microscopy analysis, and the KoNACs that responded to chilling stress were screened using RNA-seq and qRT-PCR analysis. A total of 79 KoNACs were identified, and they were unequally distributed across all 18 chromosomes of K. obovata. The KoNAC proteins could be divided into 16 subgroups according to the phylogenetic tree based on NAC family members of Arabidopsis thaliana. The KoNACs exhibited greater synteny with A. thaliana sequences than with Oryza sativa sequences, indicating that KoNACs underwent extensive evolution after the divergence of dicotyledons and monocotyledons. Segmental duplication was the main driving force of the expansions of KoNAC genes. Confocal microscopy analysis verified that the four randomly selected KoNACs localized to the nucleus, indicating the accuracy of the bioinformatic predictions. Tissue expression pattern analysis demonstrated that some KoNAC genes showed tissue-specific expression, suggesting that these KoNACs might be important for plant development and growth. Additionally, the expression levels of 19 KoNACs were significantly (15 positively and 4 negatively) induced by cold treatment, demonstrating that these KoNACs might play important roles during cold stress responses and might be candidate genes for the genetic engineering of K. obovata with enhanced chilling stress tolerance. Coexpression network analysis revealed that 381 coexpressed pairs (between 13 KoNACs and 284 other genes) were significantly correlated. Seventy-nine KoNACs were identified in K. obovata, nineteen of which displayed chilling-induced expression patterns. These genes may serve as candidates for functional analyses of KoNACs engaged in chilling stress. Our results lay the foundation for evolutionary analyses of KoNACs and their molecular mechanisms in response to environmental stress.

A Pan-Transcriptome Analysis Indicates Efficient Downregulation of the FIB Genes Plays a Critical Role in the Response of Alfalfa to Cold Stress.

Alfalfa (Medicago sativa L.) is a perennial forage legume that is widely distributed throughout the world, and cold stress is an important environmental factor limiting the growth and production of alfalfa in cold regions. However, little is known of the molecular mechanisms regarding cold tolerance in alfalfa. Here, we conducted physiological metabolism assays and pan-transcriptome sequencing on eight cultivars of alfalfa under cold stress conditions. The results of the RNA-seq analysis showed that the genes are "oxidoreductase activity" and "transcription regulator activity", suggesting that genes with such functions are more likely to play important roles in the response to cold stress by alfalfa. In addition, to identify specific gene modules and hub genes in response to alfalfa cold stress, we applied weighted gene co-expression network (WGCNA) analyses to the RNA-seq data. Our results indicate that the modules of genes that focus on the ATPase complex, ribosome biogenesis, are more likely to be involved in the alfalfa response to cold stress. It is important to note that we identified two fibronectin (FIB) genes as hub genes in alfalfa in response to cold stress and that they negatively regulate alfalfa response to chilling stress, and it is possible that dormant alfalfa is more effective at down-regulating FIB expression and therefore more resistant to cold stress.

MaMYB13 is involved in response to chilling stress via activating expression of VLCFAs and phenylpropanoids biosynthesis-related genes in postharvest banana fruit

Fruit chilling injury is the result of physiological dysfunction due to membrane lipid phase change, oxidative damage of biomacromolecules and respiratory metabolism abnormality. However, the involvement of transcription factors in response to fruits chilling tolerance remains largely unclear. Here, MaMYB13 was identified to participate in banana fruit response to chilling stress. MaMYB13 has transcriptional activation activity. When exposed to low temperature, expression of MaMYB13 was enormously induced. Moreover, MaMYB13 promoter was activated by chilling stress. MaMYB13 bound to the promoters of several important very-long-chain fatty acids (VLCFAs) and phenylpropanoids biosynthesis-related genes, including MaKCS11, Ma4CL6 and MaAAE1, and activated their transcription. Furthermore, MaKIN10 X1/3 interacted with MaMYB13 and enhanced MaMYB13-mediated transcriptional activation possibly via phosphorylation. Altogether, our results unravel the mechanism of MaMYB13-MaKIN10 X1/3 interaction regulating banana fruit chilling tolerance through activating the expression of MaKCS11, Ma4CL6 and MaAAE1, providing new insights into the regulatory network of MYB transcription factor.

Genome-Wide Identification and Expression Analysis of the NAC Gene Family in Kandelia obovata, a Typical Mangrove Plant.

The NAC (NAM, ATAF1/2, and CUC2) gene family, one of the largest transcription factor families in plants, acts as positive or negative regulators in plant response and adaption to various environmental stresses, including cold stress. Multiple reports on the functional characterization of NAC genes in Arabidopsis thaliana and other plants are available. However, the function of the NAC genes in the typical woody mangrove (Kandelia obovata) remains poorly understood. Here, a comprehensive analysis of NAC genes in K. obovata was performed with a pluri-disciplinary approach including bioinformatic and molecular analyses. We retrieved a contracted NAC family with 68 genes from the K. obovata genome, which were unevenly distributed in the chromosomes and classified into ten classes. These KoNAC genes were differentially and preferentially expressed in different organs, among which, twelve up-regulated and one down-regulated KoNAC genes were identified. Several stress-related cis-regulatory elements, such as LTR (low-temperature response), STRE (stress response element), ABRE (abscisic acid response element), and WUN (wound-responsive element), were identified in the promoter regions of these 13 KoNAC genes. The expression patterns of five selected KoNAC genes (KoNAC6, KoNAC15, KoNAC20, KoNAC38, and KoNAC51) were confirmed by qRT-PCR under cold treatment. These results strongly implied the putative important roles of KoNAC genes in response to chilling and other stresses. Collectively, our findings provide valuable information for further investigations on the function of KoNAC genes.

Mitigate the reverse effects of frost by cultivate tomato plants under different types of mesh cover for low tunnels

Tomato plants are highly sensitive to chilling stress so this experiment was conducted during successive winter seasons of 2020/2021 and 2021/2022 to investigate the possibility of sustaining production and quality of tomato yield under cold conditions by using different types of mesh cover for low tunnels. Two commercial cultivars, i.e., Super gold F1 and Super streen F1 were used. Seedlings were transplanted under different types of mesh cover for low tunnels. Three types of mesh cover (63%, 65%, 73%) had done to cover the plants of all tested seasons compared to open field condition (control "Without cover"). Results show that significant differences were observed among the two hybrid varieties of tomato studied, in terms of their vegetative growth characteristics (plant height, number of branches, number of leaves, fresh weight, dry weight and leaves area) as well as yield and fruit quality characteristics (Fruit set, total yield, firmness, T.S.S and Vit. C. ‘Super gold’ exhibited significantly higher values for these characteristics, compared to ‘Super strain B’. Chilling reduced the growth parameters (plant height, number of leaves, number of branches, plant fresh weight, plant dry weight) for control plants (without cover) compared to the coverage treatments (63%, 65%, 73%). Significant highest results in the growth, chemical composition of tomato leaves as well as yield and fruit quality parameters were obtained with covering tomato plants by white mesh cover 63% flowed by 65% of treatments which were best than the chilled plants. Concerning the interaction between hybrid varieties and covering treatments, it can be noticed that Cv. Super gold F1 which grown under white mesh cover 63% recorded a highly significant increases in plant height, number of branches, number of leaves, fresh weight, dry weight, leaves area, N, P, K, total chlorophyll, total proline, total sugers contents, fruit set, total yield, firmness, T.S.S and Vit. C. when compared to Cv. Super streen B F1 which grown in open field (without coverage).

Effects of Chilling Treatment on Baicalin, Baicalein, and Wogonin Biosynthesis in Scutellaria baicalensis Plantlets.

When plants are exposed to stressful conditions, they modulate their nutrient balance by regulating their primary and secondary metabolisms to adapt. In this study, changes in primary and secondary metabolites elicited by chilling stress treatment and the effects of treatment duration were examined in roots of Scutellaria baicalensis (S. baicalensis) plantlets. The concentrations of most sugars (maltose, glucose, sucrose, and fructose) and of several amino acids (proline and GABA), which are crucial regarding plant defense mechanisms, increased with increasing duration of chilling stress. Furthermore, salicylic acid levels increased after two-day chilling treatments, which may enhance plant tolerance to cold temperatures. The concentrations of flavones (baicalin, baicalein, and wogonin) increased during chilling stress, and those of phenolic acids (ferulic acid and sinapic acid) increased after two-day chilling treatments. The concentrations of these flavones were positively correlated with sucrose levels which acted as energy sources.

Effects of chilling at different growth stages on rice photosynthesis, plant growth, and yield

Chilling is the main limiting factor for rice production in high-altitude and high-latitude regions. Many studies have reported the impacts of chilling on leaf photosynthesis and biomass accumulation, but few of them explored the response of photosynthesis to chilling at different growth stages of rice and whether the decrease of photosynthesis at each stage is caused by stomatal or non-stomatal limiting factor. In order to achieve high grain yield for rice, it is critical to understand how chilling affects leaf photosynthesis and plant growth. In this study, we conducted a two-year field experiment (in 2018 and 2019) to investigate the effects of chilling at booting, flowering, and grain filling stages on leaf photosynthesis, plant growth, and grain yield for a japonica rice cultivar (Songjing6). At each target growth stage, the treatment included four chilling temperatures (12, 15, 18, and 21 ℃, respectively) and four chilling durations (2, 4, 6, and 8 days, respectively). The results showed that rice photosynthesis gradually decreased as the chilling temperature was decreasing and the chilling duration was increasing. The net photosynthetic rate decreased the most at the booting stage, followed by the flowering stage, and the grain filling stage. Compared with the chilling treatment at 2 days, the chilling treatment at 12, 15, and 18 ℃ for 4, 6, and 8 days reduced net photosynthetic rate by 15.1–36.1 %, 18.7–27.2 %, and 15.2–17.2 % at the booting, flowering, and grain filling stages, respectively. The chilling-induced decrease in net photosynthetic rate was mainly attributed to stomatal limiting factor at the booting and flowering stages but non-stomatal limiting factor at the grain filling stage. During the chilling treatment, the available light for rice plants was reduced and the rice plants were impaired in their ability to use weak light. The decrease in photosynthesis lead to reduced leaf source capacity and crop growth rate, hence the reduced aboveground biomass and increased source and sink increment ratio, and ultimately decreased the grain yield for rice. Our results indicate that maintaining efficient photosynthetic functioning at the booting and flowering stages is the key to combat chilling stress for rice. • Chilling decreased the net photosynthetic rate of rice most at the booting stage, followed by the flowering stage and the grain filling stage. • Chilling-induced decrease in the net photosynthetic rate was mainly attributed to stomatal limiting factor at the booting and flowering stages, but non-stomatal limiting factor at the grain filling stage. • The simulation of light response curve showed chilling at the booting and flowering stages decreased photosynthetic capacity and photosynthetic effective time，while chilling mainly affects the ability of using strong light at the grain filling stage.

Transcriptomic Responses to Chilling Reveal Potential Chilling Tolerance Mechanisms in Cucumber.

Chilling is a devastating stress that has led to a crisis of production for cucumber (Cucumis sativus L.). To determine the molecular mechanisms underlying chilling responses in cucumber, we investigated physiological changes and transcriptomic responses to chilling stress in the chilling-tolerant inbred line CC and chilling-susceptible inbred line R1461. Physiological analysis showed that CC had a higher survival rate, lower H2O2 accumulation, and ion leakage than R1461 after chilling treatment. RNA-seq analysis identified 938 differentially expressed genes (DEGs) in response to chilling and revealed that chilling stress regulated the transcript levels of genes related to hormones, including auxin, salicylic acid (SA), jasmonic acid (JA), and ethylene. RT-qPCR and pharmacological analysis suggested that cucumber chilling tolerance was associated with variation in the gene expression involved in ethylene biosynthesis and signaling. Exogenously applying 1-aminocyclopropane-1-carboxylic acid (ACC), the precursor of ethylene, improved the chilling tolerance of cucumber, while the exogenous application of the ethylene inhibitor AgNO3 impaired the chilling tolerance of cucumber. After ACC treatment, the difference in chilling tolerance between CC and R1461 disappeared, suggesting that the different chilling tolerance level between CC and R1461 is dependent on the ethylene biosynthesis and signaling pathway. In addition, a comparison of cucumber lines with different chilling tolerances revealed that chilling tolerance is highly associated with the up-regulation of C-repeat binding factor (CBF) genes, while natural variation in the promoter of CsCBF1 is associated with chilling response. This study thus provides information on transcriptomic responses in different varieties of chilling-tolerant cucumber and reveals potential chilling tolerance mechanisms that could be used to improve chilling tolerance in cucumber.

Hydrogen-rich water alleviates chilling injury-induced lignification of kiwifruit by inhibiting peroxidase activity and improving antioxidant system.

Kiwifruit is prone to chilling stress and displays chilling injury (CI) such as lignification; however, the underlying physicochemical mechanism remains largely unknown. Here, the changes in levels of quality attributes, lignin biosynthesis, antioxidant system and sugars were compared in kiwifruit between control and hydrogen-rich water (HRW) treatments during cold storage for 90 days at 0°C. The results reveal that HRW is an effective measure for CI alleviation, as indicated by the decrease in lignification level with repressed peroxidase activity but enhanced polyphenol oxidase activity. The amelioration of membrane peroxidation was suggested by the repressed levels of H2 O2 and malondialdehyde. They were accompanied by the improvement of antioxidant system, which is supported by the enhancement of sugars including fructose and glucose. In conclusion, HRW can enhance chilling tolerance, as suggested by the alleviation of lignification through inhibiting peroxidase activity and elevating the antioxidant system to attenuate membrane peroxidation. © 2022 Society of Chemical Industry.

Metabolomic analysis of rapeseed priming with H2O2 in response to germination under chilling stress

Metabolomic analysis of rapeseed priming with H2O2 in response to germination under chilling stress

Nitric oxide is involved in melatonin-induced cold tolerance in postharvest litchi fruit

Previous studies indicate that melatonin, as an important bioactive molecule, appreciably improves cold tolerance in postharvest litchi fruit. However, whether other signaling molecules are involved in melatonin-induced cold tolerance in litchi fruit has not been clarified. In this study, the effect of the interplay between melatonin and nitric oxide (NO) on chilling stress tolerance in relation to redox homeostasis in litchi fruit was investigated. In the first experiment, exogenous applications of melatonin and sodium nitroprusside (SNP) showed similar efficacy in decelerating the development of chilling injury (CI), as indicated by lower CI index and higher retention of pericarp redness in litchi fruit during refrigeration, whereas the opposite effect was found with NO scavenger 2-(4-carboxyphenyl)− 4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO). The ameliorating effect of melatonin on CI was evidently impaired under cotreatment with melatonin and cPTIO (melatonin + cPTIO). In the second experiment, the biochemical response of litchi fruit to melatonin and melatonin + cPTIO during cold storage was analyzed. The results exhibited that melatonin triggered NO accumulation by activating nitrate reductase (NR) and nitric oxide synthase (NOS) in refrigerated litchi fruit. Melatonin-induced NO accumulation was reduced under treatment with melatonin + cPTIO, but this combined treatment did not affect the NR and NOS activities when compared to melatonin alone. Reactive oxygen species (ROS) in melatonin-treated fruit were markedly quenched by activating antioxidant enzymes and increasing antioxidant metabolite contents, thus alleviating membrane lipid peroxidation and maintaining membrane integrity. Melatonin also led to increased expression of oxidized protein repair-related genes, including LcMsrA1, LcMsrA2, LcMsrB1, LcMsB2, LcTrx2 and LcNTR1. Moreover, an enhanced level of protein S-nitrosylation was observed in fruit upon melatonin treatment. However, the above physiological effect associated with improved fruit cold tolerance afforded by melatonin was partially counteracted by cPTIO. The results suggest that endogenous NO could mediate melatonin-induced cold tolerance in litchi fruit via regulation of redox status.

Genome-wide identification of RsGRAS gene family reveals positive role of RsSHRc gene in chilling stress response in radish (Raphanus sativus L.)

Radish (Raphanus sativus L.) is an important worldwide root vegetable crop. Little information of the GRAS gene family was available in radish. Herein, a total of 51 GRAS family members were firstly identified from radish genome, and unevenly located onto nine radish chromosomes. Expression analysis of RsGRAS genes in taproot displayed that RsSCL15a and RsSHRc were highly expressed in the radish cambium, and its expression level was increased with the taproot thickening. Comparative transcriptome analysis revealed that the expression patterns of RsGRAS genes varied upon exposure to different abiotic stresses including heavy metals, salt and heat. The expression level of six RsGRAS genes including RsSHRc was increased under chilling stress in two radish genotypes with different cold tolerance. Further analysis indicated that RsGRAS genes could respond to cold stress rapidly and the expression of RsSHRc was up-regulated at different development stages (cortex splitting and thickening stages) under long-term cold treatment. Transient expression of RsSHRc gene in radish showed that RsSHRc possessed the reliable function of eliminating reactive oxygen species (ROS), inhibiting the formation of malondialdehyde (MDA) and promoting to accumulate proline under cold stress. Together, these findings provided insights into the function of RsGRAS genes in taproot development and chilling stress response in radish.

Effects of the Protein Hydrolysate Pretreatment on Cucumber Plants Exposed to Chilling Stress

This study aimed to evaluate the effects of the protein hydrolysate Naturamin WSP on the antioxidant defense system and oxidation-related damage of young cucumber plants exposed to chilling stress. Low positive temperatures have a negative effect on plant growth and performance, and besides visible alterations, such as inhibited growth, significant changes occur at the cellular level. Plants grown at low temperature typically suffer from oxidative damage, which leads to increased lipid peroxidation. Moreover, chilling-stressed plants accumulate more proline to protect their cell membranes. The application of biostimulants such as the protein hydrolysate Naturamin WSP can alleviate some of the adverse effects caused by low temperature. Our results indicated an increased activity of guaiacol peroxidase (GPOD) in all plants treated with the biostimulant regardless of the temperature of cultivation. The mitigation of damages caused by chilling stress might be explained by an enhanced anti-oxidative defense, as demonstrated by the activity of guaiacol peroxidases and increased proline concentrations in Naturamin WSP-treated plants.

Nitric Oxide Confers Chilling Stress Tolerance by Regulating Carbohydrate Metabolism and the Antioxidant Defense System in Melon (Cucumis melo L.) Seedlings

The objective of the present study was to consider the regulatory role of exogenous nitric oxide (NO) supplementation in response to chilling stress impose alterations on different physiological parameters in melon seedlings. Melon seedlings were treated with sodium nitroprusside (SNP, an NO donor), hemoglobin (a NO scavenger), NG-nitro-L-arginine methyl ester (an NO synthase inhibitor), and tungstate (a nitrite reductase inhibitor) under chilling stress conditions. The results showed that exogenous SNP improves the growth of melon seedlings under chilling stress conditions and ameliorates the harmful effects of chilling stress by increasing the levels of chlorophyll and soluble solutes, elevating the activity of sucrose phosphate synthase by enhancing the expression level of CmSPS. Moreover, exogenous NO significantly enhances the expression of genes and activities of antioxidant enzymes under chilling stress, resulting in lower reactive oxygen species accumulation. However, the protective effects of SNP are reversed by both NO scavenging and inhibition. Collectively, our results reveal that NO has the ability to ameliorate the harmful effects of chilling stress on melon seedlings by regulating carbohydrate metabolism and the antioxidant defense system.

SEED PRIMING EFFECTS ON MORPHOLOGICAL TRAITS OF AMARANTHUS HYPOCHONDRIACUS UNDER OPTIMAL AND LOW TEMPERATURES

The recent research evaluated the effects of growth stimulant seed treatment on the morphological traits of Amaranthus hypochondriacus L. cv. ‘Krepysh’ grown under optimal and low positive temperature conditions. The seeds were continuously soaked for 4 h in five different solutions of growth stimulants, i.e., salicylic acid (SAA - 138 mg/L), hydrogen peroxide (H2O2 - 10 and 50 mmol/L), calcium chloride (CaCl2 - 3000 mg/L), succinic acid (SUA - 500 mg/L), and control (distilled water). The stimulant-primed seeds were germinated at optimal temperature (23°C) and continued to germinate at low temperatures of 10°C (T10) and 23°C (T23). The results showed that seed germination rates viz., germination potential (GP), germination rate (GR), germination index (GI), viability index (VI), and seed vigor index (SVI), were significantly improved with seed quality potential compared with the control. Under low positive temperature, seeds treated with succinic acid, H2O2, and CaCl2 had the most significant effects on improving seed quality and induced cold resistance in the seeds. The morphological indicators of amaranth seedlings, i.e., biomass, hypocotyl, and root length, were also significantly improved with seed treatment by growth stimulants. Priming of amaranth seeds with hydrogen peroxide and succinic acid showed a greater increase in seedlings’ biomass at room (23°C) and low (10°C) temperatures. The seed treatment with SUA and SAA significantly contributed to enhancing the hypocotyl length. The amaranth roots achieved maximum length after seed treatment with SUA and CaCl2. In general, the seed treatment effects on seedling's biomass under chilling stress were associated with the potential of inducing cold tolerance in seedlings.

Exogenous Ca2+ priming can improve peanut photosynthetic carbon fixation and pod yield under early sowing scenarios in the field.

Harnessing cold-resilient and calcium-enriched peanut production technology are crucial for high-yielding peanut cultivation in high-latitude areas. However, there is limited field data about how exogenous calcium (Ca2+) application would improve peanut growth resilience during exposure to chilling stress at early sowing (ES). To help address this problem, a two-year field study was conducted to assess the effects of exogenous foliar Ca2+ application on photosynthetic carbon fixation and pod yield in peanuts under different sowing scenarios. We measured plant growth indexes, leaf photosynthetic gas exchange, photosystems activities, and yield in peanuts. It was indicated that ES chilling stress at the peanut seedling stage led to the reduction of Pn, gs, Tr, Ls, WUE, respectively, and the excessive accumulation of non-structural carbohydrates in leaves, which eventually induced a chilling-dependent feedback inhibition of photosynthesis due mainly to weaken growth/sink demand. While exogenous Ca2+ foliar application improved the export of nonstructural carbohydrates, and photosynthetic capacity, meanwhile activated cyclic electron flow, thereby enhancing growth and biomass accumulation in peanut seedlings undergoing ES chilling stress. Furthermore, ES combined with exogenous Ca2+ application can significantly enhance plant chilling resistance and peanut yield ultimately in the field. In summary, the above results demonstrated that exogenous foliar Ca2+ application restored the ES-linked feedback inhibition of photosynthesis, enhancing the growth/sink demand and the yield of peanuts.

CaCl2 mitigates chilling injury in loquat fruit via the CAMTA5-mediated transcriptional repression of membrane lipid degradation genes

Loquat fruit is vulnerable to chilling injury (CI) under long-term low temperature storage. The application of calciumchloride (CaCl2) can alleviate CI in loquat fruit, however, the molecular mechanisms remain unclear. In this study, the loquat fruit were immersed in 1 % CaCl2 solution for 10 min and then stored at 1 ± 1 ℃ for 35 d. The results showed that CaCl2 treatment suppressed the increases in internal browning index, electrolyte leakage, and malonaldehyde content in loquat fruit under chilling stress. The internal browning index in CaCl2-treated loquat fruit was 33.34 % lower than that in the control at the end of storage. Meanwhile, CaCl2 treatment delayed the phospholipids degradation, and retained high level of unsaturated/saturated fatty acid ratio, which was 15.21 % higher than that in the control fruit at the end of storage. The CaCl2 treatment also decreased the enzymatic activities and gene expressions of phospholipase D (PLD), phospholipase C (PLC), and lipoxygenase (LOX) in loquat fruit during cold storage. Moreover, a calmodulin-binding transcription activator (CAMTA) transcription factor (TF), EjCAMTA5, was cloned from loquat fruit. The EjCAMTA5 expression was up-regulated by cold stress and CaCl2 treatment. Further investigation revealed that EjCAMTA5 directly bound to the promoters of EjPLC6-like and EjLOX5 to repress their transcription. Taken together, these findings implied that CaCl2 application could activate the EjCAMTA5-mediated transcriptional repression of EjPLC6-like and EjLOX5 genes, which contributed to delaying the degradation of membrane lipid and maintaining the integrity of cell membrane, thereby inhibiting the occurrence of browning in loquat fruit under chilling stress.

Genome-wide identification of the CDPK gene family reveals the CDPK-RBOH pathway potential involved in improving chilling tolerance in peach fruit

Calcium-dependent protein kinase (CDPK), as an essential calcium receptor, plays a major role in the perception and decoding of intracellular calcium signaling in plant development and stress responses. Here, the CDPK gene family was analyzed at the genome-wide level in peach. This study identified 17 PpCDPK gene members from the peach genome, and systematically investigated phylogenetic relationships, conserved motifs, exon-intron structures, chromosome distribution, and cis-acting elements of each PpCDPK gene using bioinformatics. Furthermore, the expression levels of most PpCDPK genes were significantly changed under the CaCl2, EBR, GB, cold shock, hot water treatments, and various temperatures in the cold storage of peach fruits. RNA-seq data showed that PpCDPK2, PpCDPK7, PpCDPK10, and PpCDPK13 were related to postharvest chilling stress in peach. The interaction network of PpCDPK7 protein showed that the interaction between PpCDPK7 and PpRBOH may be the intersectional point of Ca2+ and ROS signal transmission during cold storage of peach fruits. These systematic analyses are helpful to further characterize the regulation of PpCDPKs and CDPK-RBOH mediated signal cascades in postharvest chilling injury in peach fruit.