Chilling Stress Research Articles

Integrative multi-omics analysis of chilling stress in pumpkin (Cucurbita moschata).

Pumpkin (Cucurbita moschata) is an important vegetable crop that often suffers from low-temperature stress during growth. However, the molecular mechanism involved in its response to chilling stress remains unknown. In this study, we comprehensively investigated the effect of chilling stress in pumpkin seedlings by conducting physiological, transcriptomic, and metabolomic analyses. Under chilling stress, there was an overall increase in relative electrical conductivity, along with malondialdehyde, soluble sugar, and soluble protein contents, but decreased superoxide dismutase and peroxidase activities and chlorophyll contents in seedling leaves compared with controls. Overall, 5,780 differentially expressed genes (DEGs) and 178 differentially expressed metabolites (DEMs) were identified under chilling stress. Most DEGs were involved in plant hormone signal transduction and the phenylpropanoid biosynthesis pathway, and ERF, bHLH, WRKY, MYB, and HSF transcription factors were induced. Metabolomic analysis revealed that the contents of salicylic acid (SA), phenylalanine, and tyrosine increased in response to chilling stress. The findings indicated that the SA signaling and phenylpropanoid biosynthesis pathways are key to regulating the responses to chilling stress in pumpkins. Overall, our study provides valuable insights into the comprehensive response of C. moschata to chilling stress, enriching the theoretical basis of this mechanism and facilitating the development of molecular breeding strategies for pumpkin tolerance to chilling stress.

Influence of Foliar Application of Nanoparticles on Low Temperature Resistance of Rice Seedlings

Chilling stress, a common abiotic factor, adversely affects the growth and biomass of rice seedlings during the early stages, ultimately reducing the yield. Effective strategies to mitigate these negative impacts are essential for improving rice productivity. The application of nanotechnology in agriculture, particularly nanoparticles (NPs), has shown a promising effect in alleviating chilling stress in plants. This study evaluates the effects of various nanoparticles, ZnO (0, 50, 100, and 200 mg/L), Fe2O3 (0, 50, 75, and 100 mg/L), TiO2 (0, 50, 75, and 100 mg/L), and CeO2 (0, 50, 75, and 100 mg/L) on the chilling resistance with one control (a water spray) under a normal temperature. Four rice cultivars: LLY-7108 and XZX-6 (Low-temperature-tolerant), and LLY-32 and ZJZ-17 (Low-temperature-susceptible) were tested in this experiment. Rice seedlings were subjected to low temperature conditions (12 h light 14 °C/12 h dark, at 10 °C) for five days, followed by seven days of recovery. The results of this study demonstrate that NPs significantly enhanced seedling height fresh/dry weight and root length compared to untreated controls under chilling stress. NP treatment also reduced the reactive oxygen species (ROS), malondialdehyde (MDA), and proline content, while enhancing superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities, thereby mitigating oxidative damage. The four rice varieties exhibited clear signs of rapid growth recovery and positive physiological changes due to NPs’ application. Among the tested cultivars, LLY-7108 showed the most substantial recovery and physiological responses, while ZJZ-17 exhibited the least. The findings of this study indicate that the foliar application of ZnO (100 mg/L), Fe2O3 (50 mg/L), TiO2 (50 mg/L), and CeO2 (75 mg/L) NPs effectively mitigates chilling stress in rice seedlings, likely by enhancing the antioxidant enzymatic activity while reducing the oxidative damage. This study highlights the potential of NPs as effective agents in reducing the adverse effects of chilling stress on rice.

Light-chilling stress causes hyper-accumulation of iron in shoot, exacerbating leaf oxidative damage in cucumber.

Iron availability within the root system of plants fluctuates depending on various soil factors, which directly impacts plant growth. Simultaneously, various environmental stressors, such as high/low temperatures and high light intensity, affect plant photosynthesis in the leaves. However, the combined effects of iron nutrient conditions and abiotic stresses have not yet been clarified. In this study, we analyzed how iron nutrition conditions impact the chilling-induced damage on cucumber leaves (Cucumis sativus L.). When cucumbers were grown under different iron conditions and then exposed to chilling stress, plants grown under a high-iron condition exhibited more severe chilling-induced damage than the control plants. Conversely, plants grown under a low iron condition showed an alleviation of the chilling-induced damages. These differences were observed in a light-dependent manner, indicating that iron intensified the toxicity of reactive oxygen species generated by photosynthetic electron transport. In fact, plants grown under the low iron condition showed less accumulation of malondialdehyde derived from lipid peroxidation after chilling stress. Notably, the plants grown under the high-iron condition displayed a significant accumulation of iron and an increase in lipid peroxidation in the shoot, specifically after light-chilling stress, but not after dark-chilling stress. This indicated that increased root-to-shoot iron translocation, driven by light and low temperature, exacerbated leaf oxidative damage during chilling stress. These findings also highlight the importance of managing iron nutrition in the face of chilling stress and will facilitate crop breeding and cultivation strategies.

M5C and m6A modifications regulate the mobility of pumpkin CHOLINE KINASE 1 mRNA under chilling stress.

Mobile mRNAs serve as crucial long-distance signaling molecules, responding to environmental stimuli in plants. Although many mobile transcripts have been identified, only a limited subset has been characterized as functional long-distance signals within specific plant species, raising an intriguing question about whether the prevalence of species specificity in mobile transcripts implies a divergence in the mechanisms governing mRNA mobility across distinct plant species. Our study delved into the notable case of CHOLINE KINASE 1 (CK1), an extensively studied instance of mobile mRNAs regulated by a tRNA-like sequence (TLS) in Arabidopsis (Arabidopsis thaliana). We established an association between mRNA mobility and length, independent of TLS numbers. Notably, neither the mobile mRNAs nor the mechanisms underpinning their mobility proved to be conserved across different plant species. The exclusive mobility of pumpkin CK1 mRNA under chilling stress was pivotal in enhancing the chilling tolerance of cucumber/pumpkin heterografts. Distinct from the TLS-mediated mobility of AtCK1 mRNA, the mobility of CmoCK1 mRNA is orchestrated by both m5C and m6A modifications, adding dimensions to our understanding of mRNA transport mechanisms.

Time-specific lipid and gene expression responses to chilling stress in panicoid grass.

This article comments on: Kenchanmane Raju SK, Zhang Y, Mahboub S, Ngu DW, Qiu Y, Harmon FG, Schnable JC, Roston RL. 2024. Rhythmic lipid and gene expression responses to chilling in panicoid grasses. Journal of Experimental Botany 75, https://doi.org/10.1093/jxb/erae247

2-Keto-L-Gulonic Acid Enhances Cold Stress Tolerance in Arabidopsis thaliana via Augmentation of Ascorbic Acid Biosynthesis and Antioxidant Defense Systems

Cold stress severely impairs plant growth, development, and yields. L-ascorbic acid (ASA), a crucial antioxidant, is pivotal in mitigating stress-induced damage. Previous research found that 2-keto-L-gulonic acid (2KGA), a precursor of ASA in its industrial production, effectively enhances the endogenous ASA content in plants. We hypothesized that 2KGA might alleviate chilling stress and tried to verify it through a cultivation experiment of Arabidopsis thaliana. The results demonstrate that the application of 2KGA significantly increased ASA content (24.58%) and up-regulated ASA biosynthetic genes in Arabidopsis at 4 °C for 24 h. Furthermore, 2KGA alleviated the decrease in fresh weight (17.05%) and total chlorophyll content (15.85%) caused by low temperatures. The contents of proline, soluble sugar (SS), soluble protein (SP), and the activities of antioxidant enzymes including superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX) were significantly increased under the 2KGA treatment at low temperatures, while the malondialdehyde (MDA) content was reduced. Moreover, 2KGA up-regulated the ICE-CBF-COR signaling pathway in response to cold stress. These collective findings strongly support the involvement of 2KGA in enhancing cold tolerance in Arabidopsis, presenting an innovative approach for agricultural practices aimed at enhancing crop resilience to environmental stresses.

Comparison of transcriptome and metabolome analysis revealed cold-resistant metabolic pathways in cucumber roots under low-temperature stress in root zone.

Low ground temperature is a major factor limiting overwintering in cucumber cultivation facilities in northern alpine regions. Lower temperatures in the root zone directly affect the physiological function of the root system, which in turn affects the normal physiological activity of plants. However, the importance of the ground temperature in facilities has not attracted sufficient attention. Therefore, this study tested the cucumber variety Jinyou 35 under three root zone temperatures (room temperature, 20-22°C; suboptimal temperature, 13- 15°C; and low temperature, 8-10°C) to investigated possible cold resistance mechanisms in the root of cucumber seedlings through hormone, metabolomics, and transcriptomics analyses. The results showed that cucumber roots were subjected to chilling stress at different temperatures. Hormone analysis indicated that auxin content was highest in the roots. Jasmonic acid and strigolactone participated in the low-temperature stress response. Auxin and jasmonate are key hormones that regulate the response of cucumber roots to low temperatures. Phenolic acid was the most abundant metabolite in cucumber roots under chilling stress. Additionally, triterpenes may play an important role in chilling resistance. Differentially expressed genes and metabolites were significantly enriched in benzoxazinoid biosynthesis in the room temperature vs. suboptimal temperature groups and the room temperature vs. low temperature groups. Most differentially expressed transcription factor genes in AP2/ERF were strongly induced in cucumber roots by both suboptimal and low-temperature stress conditions. These results provide guidance for the cultivation of cucumber in facilities.

Identification and Characterization of Shaker Potassium Channel Gene Family and Response to Salt and Chilling Stress in Rice.

Shaker potassium channel proteins are a class of voltage-gated ion channels responsible for K+ uptake and translocation, playing a crucial role in plant growth and salt tolerance. In this study, bioinformatic analysis was performed to identify the members within the Shaker gene family. Moreover, the expression patterns of rice Shaker(OsShaker) K+ channel genes were analyzed in different tissues and salt treatment by RT-qPCR. The results revealed that there were eight OsShaker K+ channel genes distributed on chromosomes 1, 2, 5, 6 and 7 in rice, and their promoters contained a variety of cis-regulatory elements, including hormone-responsive, light-responsive, and stress-responsive elements, etc. Most of the OsShaker K+ channel genes were expressed in all tissues of rice, but at different levels in different tissues. In addition, the expression of OsShaker K+ channel genes differed in the timing, organization and intensity of response to salt and chilling stress. In conclusion, our findings provide a reference for the understanding of OsShaker K+ channel genes, as well as their potential functions in response to salt and chilling stress in rice.

Pararhizobium sp. strains enhancing chilling stress tolerance and yield in tomato plants

In temperate climates, low temperatures represent a significant stressor that adversely affects crop yield and production. Tomato (Solanum lycopersicum L.) is a subtropical crop cultivated in temperate regions. However, most tomato cultivars are sensitive to chilling temperatures, which limit their cultivation in colder regions. Some microorganism-based plant biostimulants have been reported to enhance abiotic stress tolerance in crops. In this study, we isolated two Pararhizobium sp. strains (44 and 128) and tested their potential to trigger chilling stress tolerance in tomato. Through transcriptional, metabolic and biochemical analyses we demonstrate that inoculation with strains 44 and 128 enhance chilling stress tolerance by stimulating the ICE1-CBF-COR cold stress signaling pathway at transcriptional level, improving reactive oxygen species (ROS) detoxifying capacity and boosting the biosynthesis of stress-protective metabolites, such as polyamines and reduced glutathione (GSH). Treatment of tomato plants with these strains under non-stress conditions also increased tomato fruit weight and quality attributes. These findings suggest that Pararhizobium strains 44 and 128 could be valuable biostimulants for improving chilling stress tolerance and crop yield.

Auxin as a downstream signal positively participates in melatonin-mediated chilling tolerance of cucumber.

Here, we elucidate the interaction between IAA and melatonin (MT) in response to chilling in cucumber. The results showed that chilling stress induced the increase of endogenous MT and IAA, and the application of MT promoted the synthesis of IAA, while IAA could not affect endogenous MT content under chilling stress. Moreover, MT and IAA application both remarkably increased the chilling tolerance of cucumber seedlings in terms of lower contents of MDA and ROS, higher mRNA abundance of cold response genes, net photosynthetic rate (Pn), maximum regeneration rate of ribulose-1,5-diphosphate (Jmax), Rubisco maximum carboxylation efficiency (Vcmax), the activities and gene expression of RCA and Rubisco, as well as the content of active P700 (I/I0) and photosynthetic electron transport, compared with the plants in H2O treatment. Further analysis revealed that the inhibition of IAA transportation significantly reduced the chilling tolerance induced by MT, whereas the inhibition of endogenous MT did not affect the chilling tolerance induced by IAA. Meanwhile, we found that overexpression of the MT biosynthesis gene CsASMT increased the chilling tolerance, which was blocked by inhibition of endogenous IAA, and the silence of IAA biosynthesis gene CsYUCCA10 decreased the chilling tolerance of cucumber, which could not be alleviated by MT. These data implied IAA acted as a downstream signal to participate in the MT-induced chilling tolerance of cucumber seedlings. The study has implications for the production of greenhouse cucumber in winter seasons.

Physiological and Proteomic Analysis of Various Priming on Rice Seed under Chilling Stress.

Rice (Oryza sativa L.) cultivation using direct seeding is susceptible to chilling stress, particularly during seed germination and early seedling growth in the early season of a double cropping system. Alternatively, seed priming with various plant growth-promoting hormones is an effective technique to promote rapid and uniform emergence under chilling stress. Therefore, we evaluated the impact of gibberellin A3 (GA3) and brassinolide (BR) priming on rice seed emergence, examining their proteomic responses under low-temperature conditions. Results indicated that GA3 and BR increased the seed germination rate by 22.67% and 7.33% at 72 h and 35% and 15% at 96 h compared to the control (CK), respectively. Furthermore, proteomic analysis identified 2551, 2614, and 2592 differentially expressed proteins (DEPs) in GA, BR, and CK, respectively. Among them, GA exhibited 84 upregulated and 260 downregulated DEPs, while BR showed 112 upregulated and 102 downregulated DEPs, and CK had 123 upregulated and 81 downregulated DEPs. Notably, under chilling stress, both GA3 and BR are involved in peroxide metabolism, phenylpropanoid biosynthesis, and inositol phosphate metabolism, enhancing antioxidant capacity and providing energy substances for germination. In addition, GA3 triggers the specific regulation of stress responsive protein activation, GTP activation, and ascorbic acid biosynthesis and promotes the stability and integrity of cell membranes, as well as the synthesis of cell walls, providing physical defense for seeds to resist low temperatures. At the same time, BR triggers specific involvement in ribosome synthesis and amino acid synthesis, promoting biosynthetic ability and metabolic regulation to maintain plant life activities under low-temperature stress. Furthermore, the various genes' expression (OsJ_16716, OsPAL1, RINO1) confirmed GA3 and BR involved in peroxide metabolism, phenylpropanoid biosynthesis, and inositol phosphate metabolism, enhancing antioxidant capacity and providing energy substances for germination. This study provides valuable insights into how rice seed embryo responds to and tolerates chilling stress with GA3 seed priming.

The CpCOR1 gene enhances cold tolerance and antioxidant activity of papaya fruit in response to postharvest chilling stress

Inappropriate low temperatures can induce harmful effects on postharvest papaya fruit. Based on the effects of cold shock treatment (CST, 3 ℃ ice-water for 0.5 h) on reducing the chilling injury index (CI index) of papaya fruit, this study focuses on elucidating the role of CpCOR1, a gene encoding a cold-regulated protein, in the manifestation of low temperature-induced chilling injury in papaya fruit. Our results demonstrate that CST significantly reduced the CI index, concurrently preserving the cell microstructure and mitigating the increase in cell membrane permeability during both 5 ℃ and 25 ℃ storage. Transcriptome analysis and qRT-PCR revealed a substantial up-regulation of the CpCOR1 gene in CST papaya fruit compared to the control fruit. Moreover, bioinformatics and sub-cellular localization assays revealed that CpCOR1 has a coding sequence of 606 bp and an amino acid sequence of 201 aa, and is localized to the cell membrane. Furthermore, overexpression of CpCOR1 in tobacco leaves and papaya fruit significantly alleviated CI symptoms, suppressed yellowing, reduced malondialdehyde (MDA) accumulation, and maintained higher activities of antioxidant enzymes, including catalase (CAT) and ascorbate peroxidase (APX). In summary, our findings suggest that cold shock treatment is an effective strategy to retard the onset of low-temperature damage, with CpCOR1 playing a pivotal role in enhancing cold resistance and antioxidant activity in papaya fruit.

L-Glutamate treatment alleviates chilling injury of prune (Prunus domestica L.) fruit by regulating ROS homeostasis, GABA shunt, and energy metabolism

The impacts of L-glutamate (L-Glu) treatment on chilling injury (CI), Ca2+ signaling, mitochondrial ultrastructure, and metabolisms of reactive oxygen species (ROS), γ-aminobutyric acid (GABA), energy of prune fruit under chilling stress were studied. The results found that the optimal concentration of L-Glu to suppress CI occurrence and maintain quality in prune fruit was 0.1 g L−1, which also enhanced the PdGLRs expression, cytoplasmic Ca2+ concentration, the contents of CaM, and CML under cold stress. Moreover, L-Glu treatment could reduce ROS accumulation and increase GABA content, and energy level, contributing to maintaining the integrity of the mitochondrial structure in cold-stored prune fruit. More importantly, PdGLRs expression and CaM/CML content positively correlated with antioxidant enzyme activities, GABA shunt, and energy status in prune fruit. These results indicated that the enhanced cold resistance of L-Glu-treated prunes might be attributed to the activated Ca2+ signaling, thus improving the antioxidant capacity, GABA, and energy levels.

Regulation of spikelet developmental responses to chilling and freezing stress mediated by differential sugar metabolism in winter wheat

Spring cold stress poses a great threat to wheat reproductive growth, leading to compromised spike development and grain yield. There are two types of cold stress i.e. chilling stress (CS, above zero) and freezing stress (FS, below zero). However, it is unclear whether there is a difference in the mechanism of CS and FS in regulating spikelet development. This study aimed to unravel the underlying regulation in determining the difference for wheat spikelet exposed to CS at 2 °C and FS at −2°C by integrative analyses of transcriptome, metabolome, and physiology. Delayed floret development and shrunken cellular morphology in both CS and FS were observed, even malformation and degradation of anther cells occurred in FS. Kyoto Encyclopedia of gene and genomes (KEGG) analyses revealed that the most abundantly enriched pathways are phytohormone biosynthesis, starch and sucrose metabolism, and phenylpropanoid biosynthesis. Further physiological assays related to the identified pathways were performed. Compared to CS, the signal of abscisic acid (ABA), salicylic acid (SA) and jasmonic acid (JA) was more pronounced, and the signal of auxin (IAA) and gibberellin (GA) was inhibited further in FS. In addition, the contents of glucose, fructose and trehalose were elevated in CS, owing to greater activities of cell wall invertase and sucrose synthase, while the hexose content was decreased owing to lower activities of such enzymes in FS, concomitantly, flavonoid barely changed in CS, but it dramatically amounted in FS. Taken together, the glucose and trehalose pathway, along with induced ABA and SA signal were intensified in CS to maintain growth, while greater flavonoid and promoted JA synthesis were induced in FS for cold survival. Understanding the molecular of growth-defense under cold stress would provide a foundation for the development of breeding strategies.

A comprehensive meta-analysis exploring potential of GABA for postharvest chilling injury mitigation in horticultural produce

γ-Aminobutyric acid (GABA) exhibits anti-chilling properties and significantly mitigates chilling stress in horticultural products. While numerous studies have explored the effects of GABA on fruits, vegetables, and flowers, uncertainties persist regarding the optimal GABA application method, concentration, and dipping duration for suppressing chilling injury (CI) symptoms. To address this gap, we conducted a comprehensive global meta-analysis comprising 157 evaluations from 19 research papers to assess the impact of GABA on horticultural goods. Our study revealed an overall substantial reduction in CI (-51.1 %) with GABA application compared to the control group. Furthermore, all relevant markers associated with the GABA shunt pathway, oxidative stress, antioxidant system (enzymatic and non-enzymatic), and lipid oxidation with the CI mechanism exhibited significance, except for GABA-En. Interestingly, dipping emerged as a more effective GABA application. Regarding horticultural commodities, GABA was found more effective in fruits (-54.65 %) followed by vegetables (-45.46 %) and flowers (-31.30 %). The regression analysis revealed that GABA concentrations exceeding 1 mM, 5 mM, and 15 mM were ineffective in mitigating CI in flowers, vegetables, and fruits, respectively. These results highlight the efficacy of the GABA dipping and dipping time, as well as, the precise GABA concentration in alleviating CI in flowers, vegetables, and fruits.

SlWRKY51 regulates proline content to enhance chilling tolerance in tomato.

Chilling stress is a major environmental factor that significantly reduces crop production. To adapt to chilling stress, plants activate a series of cellular responses and accumulate an array of metabolites, particularly proline. Here, we report that the transcription factor SlWRKY51 increases proline contents in tomato (Solanum lycopersicum) under chilling stress. SlWRKY51 expression is induced under chilling stress. Knockdown or knockout of SlWRKY51 led to chilling-sensitive phenotypes, with lower photosynthetic capacity and more reactive oxygen species (ROS) accumulation than the wild type (WT). The proline contents were significantly reduced in SlWRKY51 knockdown and knockout lines under chilling stress, perhaps explaining the phenotypes of these lines. D-1-pyrroline-5-carboxylate synthetase (P5CS), which catalyses the rate-limiting step of proline biosynthesis, is encoded by two closely related P5CS genes (P5CS1 and P5CS2). We demonstrate that SlWRKY51 directly activates the expression of P5CS1 under chilling stress. In addition, the VQ (a class of plant-specific proteins containing the conserved motif FxxhVQxhTG) family member SlVQ10 physically interacts with SlWRKY51 to enhance its activation of P5CS1. Our study reveals that the chilling-induced transcription factor SlWRKY51 enhances chilling tolerance in tomato by promoting proline accumulation.

Photosynthesis and Latex Burst Characteristics in Different Varieties of Rubber Trees (Hevea brasiliensis) under Chilling Stress, Combing Bark Tensile Property and Chemical Component Analysis

Rubber trees (Hevea brasiliensis) serve as the primary source of natural rubber. Their native habitat is characterized by warm and humid conditions, so they are particularly sensitive to low temperatures. Under such stress, latex burst can cause severe damage to rubber trees, which is due to the uniqueness of their economically productive parts. In order to establish a correlation between young and mature rubber trees and provide a novel prospective for investigating the mechanisms of latex burst and chilling resistance in rubber trees, the chlorophyll contents, photosynthesis, and chlorophyll fluorescence parameters in four varieties of one-year-old rubber tree seedlings were analyzed under artificially simulated chilling stress. The latex burst characteristics were subsequently recorded. A comprehensive statistical analysis of the chilling-resistance rank was conducted using the membership function method and the combination weighting method. Meanwhile, chemical compositions and tensile properties of barks from two-year-old twigs of mature rubber trees were ascertained. A correlation analysis between chilling resistance, chemical compositions, and tensile properties was performed to identify any interrelationships among them. The results showed that the number and the total area of latex-burst positions in variety Reken628 seedlings were greater than those in other varieties, and the lowest number and total area of latex-burst positions were observed in variety RRIM600 and variety PR107, respectively. With the exception of variety GT1, nectar secretion was noted in all other varieties of rubber tree seedlings under chilling stress. The chilling resistance of the four varieties decreased in the following order: variety GT1 > variety RRIM600 > variety PR107 > variety Reken628. The chilling resistance was strongly (p < 0.001) negatively correlated with cellulose content and acid-insoluble lignin content, respectively. The total area of latex burst was significantly (p < 0.001) and positively correlated with holocellulose content and maximum load, respectively. Furthermore, this study also provides new insights into the mechanism of nectar secretion induced by low temperatures and its association with the chilling resistance of rubber trees.

Salicylic acid improves chilling tolerance via CsNPR1-CsICE1 interaction in grafted cucumbers.

Salicylic acid (SA) plays a role in the regulation of grafting-induced cold tolerance. However, the molecular mechanism behind it is still unknown. Here, we established that the phenylalanine ammonia-lyase (PAL) pathway-dependent elevate in SA content in grafted cucumber leaves was not only synthesized in the leaves but also transported from the roots under chilling stress. RNAi-CsPAL with low SA content as rootstock reduced SA accumulation in grafted seedling leaves while decreasing rootstock-induced cold tolerance, as evidenced by higher electrolyte leakage (EL), hydrogen peroxide (H2O2), and superoxide anion (O2 ·-) contents and lower expression of cold-responsive genes (CsICE1, CsDREB1A, CsDREB1B, and CsCOR47), whereas OE-CsPAL with high SA content as rootstock improved the cold tolerance of grafted plants in comparison with the wild type (WT). In addition, CsNPR1 was significantly upregulated in grafted cucumber under chilling stress, with exogenous and endogenous overexpressed SA inducing its transcriptional expression and protein stability, which exhibited higher expression in grafted plants than in self-root plants. While CsNPR1-overexpression (OE-CsNPR1) seedlings as scions were more tolerant to chilling stress than WT seedlings, CsNPR1-suppression (Anti-CsNPR1) seedlings as scions were more vulnerable to chilling stress. Notably, CsNPR1-CsICE1 interactions alleviated ROS accumulation and activated the expression of CsDREB1A, CsDREB1B, CsCOR47, CsCOR15, CsCOR413, and CsKIN1 to enhance SA-mediated chilling tolerance in grafted cucumber. Overall, our findings reveal that SA enhances chilling tolerance in grafted cucumbers via the model of the CsNPR1-CsICE1 transcriptional regulatory cascade.

Comparing time-series transcriptomes between chilling-resistant and -susceptible rice reveals potential transcription factors responding to chilling stress.

Low temperature is one of the most important environmental factors that inhibits rice growth and grain yield. Transcription factors (TFs) play crucial roles in chilling acclimation by regulating gene expression. However, transcriptional dynamics and key regulators responding to low temperature remain largely unclear in rice. In this study, a transcriptome-based comparative analysis was performed to explore genome-wide gene expression profiles between a chilling-resistant cultivar DC90 and a chilling-susceptible cultivar 9311 at a series of time points under low temperature treatment and recovery condition. A total of 3,590 differentially expressed genes (DEGs) between two cultivars were determined and divided into 12 co-expression modules. Meanwhile, several biological processes participating in the chilling response such as abscisic acid (ABA) responses, water deprivation, protein metabolic processes, and transcription regulator activities were revealed. Through weighted gene co-expression network analysis (WGCNA), 15 hub TFs involved in chilling conditions were identified. Further, we used the gene regulatory network (GRN) to evaluate the top 50 TFs, which might have potential roles responding to chilling stress. Finally, five TFs, including a C-repeat binding factor (OsCBF3), a zinc finger-homeodomain protein (OsZHD8), a tandem zinc finger protein (OsTZF1), carbon starved anther (CSA), and indeterminate gametophyte1 (OsIG1) were identified as crucial candidates responsible for chilling resistance in rice. This study deepens our understanding in the gene regulation networks of chilling stress in rice and offers potential gene resources for breeding climate-resilient crops.

Ectopic overexpression of ShCBF1 and SlCBF1 in tomato suggests an alternative view of fruit responses to chilling stress postharvest.

Postharvest chilling injury (PCI) is a physiological disorder that often impairs tomato fruit ripening; this reduces fruit quality and shelf-life, and even accelerates spoilage at low temperatures. The CBF gene family confers cold tolerance in Arabidopsis thaliana, and constitutive overexpression of CBF in tomato increases vegetative chilling tolerance, in part by retarding growth, but, whether CBF increases PCI tolerance in fruit is unknown. We hypothesized that CBF1 overexpression (OE) would be induced in the cold and increase resistance to PCI. We induced high levels of CBF1 in fruit undergoing postharvest chilling by cloning it from S. lycopersicum and S. habrochaites, using the stress-inducible RD29A promoter. Harvested fruit were cold-stored (2.5°C) for up to three weeks, then rewarmed at 20°C for three days. Transgene upregulation was triggered during cold storage from 8.6- to 28.6-fold in SlCBF1-OE, and between 3.1- to 8.3-fold in ShCBF1-OE fruit, but developmental abnormalities in the absence of cold induction were visible. Remarkably, transgenic fruit displayed worsening of PCI symptoms, i.e., failure to ripen after rewarming, comparatively higher susceptibility to decay relative to wild-type (WT) fruit, lower total soluble solids, and the accumulation of volatile compounds responsible for off-odors. These symptoms correlated with CBF1 overexpression levels. Transcriptomic analysis revealed that the ripening and biotic and abiotic stress responses were altered in the cold-stored transgenic fruit. Seedlings grown from 'chilled' and 'non-chilled' WT fruit, in addition to 'non-chilled' transgenic fruit were also exposed to 0°C to test their photosynthetic response to chilling injury. Chilled WT seedlings adjusted their photosynthetic rates to reduce oxidative damage; 'non-chilled' WT seedlings did not. Photosynthetic parameters between transgenic seedlings were similar at 0°C, but SlCBF1-OE showed more severe photoinhibition than ShCBF1-OE, mirroring phenotypic observations. These results suggest that 1) CBF1 overexpression accelerated fruit deterioration in response to cold storage, and 2) Chilling acclimation in fructus can increase chilling tolerance in seedling progeny of WT tomato.