Increased Cold Tolerance Research Articles

Brassinosteroid signaling regulator BIM1 integrates brassinolide and jasmonic acid signaling during cold tolerance in apple.

Although brassinolide (BR) and jasmonic acid (JA) play essential roles in the regulation of cold stress responses, the molecular basis of their crosstalk remains elusive. Here, we show a key component of BR signaling in apple (Malus × domestica), BR INSENSITIVE1 (BRI1)-EMS-SUPPRESSOR1 (BES1)-INTERACTING MYC-LIKE PROTEIN1 (MdBIM1), increases cold tolerance by directly activating expression of C-REPEAT BINDING FACTOR1 (MdCBF1) and forming a complex with C-REPEAT BINDING FACTOR2 (MdCBF2) to enhance MdCBF2-activated transcription of cold-responsive genes. Two repressors of JA signaling, JAZMONATE ZIM-DOMAIN1 (MdJAZ1) and JAZMONATE ZIM-DOMAIN2 (MdJAZ2), interact with MdBIM1 to integrate BR and JA signaling under cold stress. MdJAZ1 and MdJAZ2 reduce MdBIM1-promoted cold stress tolerance by attenuating transcriptional activation of MdCBF1 expression by MdBIM1 and interfering with the formation of the MdBIM1-MdCBF2 complex. Furthermore, the E3 ubiquitin ligase ARABIDOPSIS TÓXICOS en LEVADURA73 (MdATL73) decreases MdBIM1-promoted cold tolerance by targeting MdBIM1 for ubiquitination and degradation. Our results not only reveal crosstalk between BR and JA signaling mediated by a JAZ-BIM1-CBF module, but also provide insights into the post-translational regulatory mechanism of BR signaling.

Transcriptomic analysis revealed the candidate metabolic pathways and genes associated with cold tolerance in a mutant without anthocyanin accumulation in common vetch (Vicia sativa L.)

Common vetch (Vicia sativa L.) is a leguminous crop used to feed livestock with vegetative organs or fertilize soils by returning to the field. Survival of fall-seeded plants is often affected by freezing damage during overwintering. This study aims to investigate the transcriptomic profiling in response to cold in a mutant with reduced accumulation of anthocyanins under normal growth and low-temperature conditions for understanding the underlying mechanisms. The mutant had increased cold a tolerance with higher survival rate and biomass during overwintering compared to the wild type, which led to increased forage production. Transcriptomic analysis in combination with qRT-PCR and physiological measurements revealed that reduced anthocyanins accumulation in the mutant resulted from reduced expression of serial genes involving in anthocyanin biosynthesis, which led to the altered metabolism, with an increased accumulation of free amino acids and polyamines. The higher levels of free amino acids and proline in the mutant under low temperature were associated with improved cold tolerance. The altered expression of some genes involved in ABA and GA signaling was also associated with increased cold tolerance in the mutant.

Karrikin increases tomato cold tolerance via strigolactone and the abscisic acid signaling network

As a class of biostimulants, karrikins (KARs) were first identified from plant-derived smoke to regulate plant growth, development, and stress tolerance. However, the roles of KARs in plant cold tolerance and their crosstalk with strigolactones (SLs) and abscisic acid (ABA) remain elusive. We studied the interaction among KAR, SLs, and ABA in cold acclimatization with KAI2-, MAX1-, SnRK2.5-silenced, or cosilenced plant materials. KAI2 is involved in smoke-water- (SW-) and KAR-mediated cold tolerance. MAX1 acts downstream of KAR in cold acclimation. ABA biosynthesis and sensitivity are regulated by KAR and SLs, which improve cold acclimation through the SnRK2.5 component. The physiological mechanisms of SW and KAR in improving growth, yield, and tolerance under a long-term sublow temperature environment were also studied. SW and KAR were shown to improve tomato growth and yield under sublow temperature conditions by regulating nutritional uptake, leaf temperature control, photosynthetic defense, ROS scavenging, and CBF transcriptional activation. Together, SW, which functions via the KAR-mediated SL and ABA signaling network, has potential application value for increasing cold tolerance in tomato production.

Screening Cold Tolerance in St. Augustinegrass (Stenotaphrum secundatum (Walt.) Kuntze) for USDA Hardiness Zone 7

Among warm-season turfgrasses, St. Augustinegrass [Stenotaphrum secundatum (Walt.) Kuntze] generally has poor cold tolerance yet excellent shade tolerance. As mostly hot summers follow cold winters in USDA Hardiness Zone 7, severely damaging tall fescue [Festuca arundineacea Schreb.] and centipedegrass [Eremochloa ophiuroides (Munro) Hack.], a St. Augustinegrass cultivar cold tolerant enough to be grown for shady lawns would greatly benefit both home owners and sod growers in this and potentially other regions. Eight St. Augustinegrass samples were selected from an initial collection of 30, including industry standards ‘Raleigh’ and ‘Palmetto’, for further testing from an established germplasm collection of material collected from lawns grown in USDA Hardiness Zone 7. Percent regrowth was calculated 4 weeks after samples underwent a 2 week cold acclimation process, followed by freezing at 0°C, -4°C, and -6°C separately for 3 h. The germplasm samples designated at ‘A’, ‘G’, and ‘H’ had greater clipping regrowth percentages compared to the industry cold tolerant standard, ‘Raleigh’ at either -4°C and -6°C indicating possible increased cold tolerance for several selections compared to ‘Raleigh’. An increased cold tolerant St. Augustinegrass would provide a wider selection of desirable turfgrasses in the transition zone growing region of the world, especially shady sites.

OsLUX Confers Rice Cold Tolerance as a Positive Regulatory Factor.

During the early seedling stage, rice (Oryza sativa L.) must overcome low-temperature stress. While a few cold-tolerance genes have been characterized, further excavation of cold-resistance genes is still needed. In this study, we identified a cold-induced transcription factor-LUX ARRHYTHMO (LUX)-in rice. OsLUX was found to be specifically expressed in leaf blades and upregulated by both cold stress and circadian rhythm. The full-length OsLUX showed autoactivation activity, and the OsLUX protein localized throughout the entire onion cell. Overexpressing OsLUX resulted in increased cold tolerance and reduced ion leakage under cold-stress conditions during the seedling stage. In contrast, the knockout of OsLUX decreased seedling cold tolerance and showed higher ion leakage compared to the wild type. Furthermore, overexpressing OsLUX upregulated the expression levels of oxidative stress-responsive genes, which improved reactive oxygen species (ROS) scavenging ability and enhanced tolerance to chilling stress. Promoter analysis showed that the OsLUX promoter contains two dehydration-responsive element binding (DREB) motifs at positions -510/-505 (GTCGGa) and -162/-170 (cCACCGccc), which indicated that OsDREB1s and OsDREB2s probably regulate OsLUX expression by binding to the motif to respond to cold stress. Thus, OsLUX may act as a downstream gene of the DREB pathway. These results demonstrate that OsLUX serves as a positive regulatory factor of cold stress and that overexpressing OsLUX could be used in rice breeding programs to enhance abiotic stress tolerance.

Membrane lipid metabolism and scavenging of reactive oxygen species are essential for cold resistance of Solanum habrochaites (LA1777): Lipidomics and transcriptomes analysis

Low temperature significantly limits the production of tomato (Solanum lycopersicum) in greenhouse. Solanum habrochaites is a wild species of tomato closely related to Solanum lycopersicum and is more tolerant to cold than S. lycopersicum. However, the mechanism by which S. habrochaites resists low temperature is unknown. This study aimed to analyze the physiology, transcriptomics, and lipidomics of LA1777 (S. habrochaites variety) after exposure to cold stress for 3 days to assess its specific mechanism to cold tolerance. A cold-sensitive cultivated tomato variety AC was used as controls. Physiological analysis showed that the two tomato varieties were significantly affected by cold stress. However, LA1777 had more stable physiological characteristics and maintained relatively good photosynthetic capacity than the AC variety. Transcriptome analysis suggested that the stability of cell membrane structure and function might promote cold tolerance of LA1777. Furthermore, 243 differentially expressed genes (DEGs) involved in lipid metabolism were identified. Particularly, the upregulation of temperature-induced lipocalin genes (TILs) and fatty acid desaturase genes (FADs) promoted cold tolerance by inhibiting the excessive accumulation of reactive oxygen species (ROS) and increasing membrane lipid unsaturation, respectively. Meanwhile, lipidomics data indicated that monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG) levels were significantly reduced in both plants under cold stress. MGDG levels decreased by 28% and 54% in LA1777 and AC, respectively, while DGDG decreased by 7% and 29%, respectively. Further results showed that high phosphoinositide (PI) content may increase cold tolerance in LA1777 by maintaining phosphatidylinositol signaling system. Also, results showed that adenosine triphosphate (ATP) produced via β-oxidation of fatty acids in mitochondria can maintain a higher metabolic level of wild tomato under cold stress. A metabolic model of the mechanism of cold tolerance in tomato was also constructed. This study provides a basis for further analysis of the molecular mechanism of cold resistance in tomato, which is crucial for genetic improvement.

Seasonal variation in thermoregulatory capacity of three closely related Afrotropical Estrildid finches introduced to Europe

A species' potential geographical range is largely determined by how the species responds physiologically to its changing environment. It is therefore crucial to study the physiological mechanisms that species use to maintain their homeothermy in order to address biodiversity conservation challenges, such as the success of invasions of introduced species. The common waxbill Estrilda astrild, the orange-cheeked waxbill E. melpoda, and the black-rumped waxbill E. troglodytes are small Afrotropical passerines that have established invasive populations in regions where the climate is colder than in their native ranges. As a result, they are highly suitable species for studying potential mechanisms for coping with a colder and more variable climate. Here, we investigated the magnitude and direction of seasonal variation in their thermoregulatory traits, such as basal (BMR), summit (Msum) metabolic rates and thermal conductance. We found that, from summer to autumn, their ability to resist colder temperatures increased. This was not related to larger body masses or higher BMR and Msum, but instead, species downregulated BMR and Msum toward the colder season, suggesting energy conservation mechanisms to increase winter survival. BMR and Msum were most strongly correlated with temperature variation in the week preceding the measurements. Common waxbill and black-rumped waxbill, whose native ranges encompass the highest degree of seasonality, showed the most flexibility in metabolic rates (i.e., stronger downregulation toward colder seasons). This ability to adjust thermoregulatory traits, combined with increased cold tolerance, may facilitate their establishment in areas characterized by colder winters and less predictable climates.

Suppression of the HOS1 Gene Affects the Level of ROS Depending on Light and Cold.

The E3 ubiquitin-protein ligase HOS1 is an important integrator of temperature information and developmental processes. HOS1 is a negative regulator of plant cold tolerance, and silencing HOS1 leads to increased cold tolerance. In the present work, we studied ROS levels in hos1Cas9Arabidopsis thaliana plants, in which the HOS1 gene was silenced by disruption of the open reading frame via CRISPR/Cas9 technology. Confocal imaging of intracellular reactive oxygen species (ROS) showed that the hos1 mutation moderately increased levels of ROS under both low and high light (HL) conditions, but wild-type (WT) and hos1Cas9 plants exhibited similar ROS levels in the dark. Visualization of single cells did not reveal differences in the intracellular distribution of ROS between WT and hos1Cas9 plants. The hos1Cas9 plants contained a high basal level of ascorbic acid, maintained a normal balance between reduced and oxidized glutathione (GSH and GSSG), and generated a strong antioxidant defense response against paraquat under HL conditions. Under cold exposure, the hos1 mutation decreased the ROS level and substantially increased the expression of the ascorbate peroxidase genes Apx1 and Apx2. When plants were pre-exposed to cold and further exposed to HL, the expression of the NADPH oxidase genes RbohD and RbohF was increased in the hos1Cas9 plants but not in WT plants. hos1-mediated changes in the level of ROS are cold-dependent and cold-independent, which implies different levels of regulation. Our data indicate that HOS1 is required to maintain ROS homeostasis not only under cold conditions, but also under conditions of both low and high light intensity. It is likely that HOS1 prevents the overinduction of defense mechanisms to balance growth.

A natural promoter variation of SlBBX31 confers enhanced cold tolerance during tomato domestication.

Cold stress affects crop growth and productivity worldwide. Understanding the genetic basis of cold tolerance in germplasms is critical for crop improvement. Plants can coordinate environmental stimuli of light and temperature to regulate cold tolerance. However, it remains unknown which gene in germplasms could have such function. Here, we utilized genome-wide association study (GWAS) to investigate the cold tolerance of wild and cultivated tomato accessions and discovered that increased cold tolerance is accompanied with tomato domestication. We further identified a 27-bp InDel in the promoter of the CONSTANS-like transcription factor (TF) SlBBX31 is significantly linked with cold tolerance. Coincidentally, a key regulator of light signalling, SlHY5, can directly bind to the SlBBX31 promoter to activate SlBBX31 transcription while the 27-bp InDel can prevent S1HY5 from transactivating SlBBX31. Parallel to these findings, we observed that the loss of function of SlBBX31 results in impaired tomato cold tolerance. SlBBX31 can also modulate the cold-induced expression of several ERF TFs including CBF2 and DREBs. Therefore, our study has uncovered that SlBBX31 is possibly selected during tomato domestication for cold tolerance regulation, providing valuable insights for the development of hardy tomato varieties.

Rapid cold hardening response of the phytoseiid mite Neoseiulus striatus: increased cold tolerance but not reduced predation

The predatory mite Neoseiulus striatus (Wu) (Acari: Phytoseiidae), which has been found on maize plants in Inner Mongolia, is regarded as a promising biological control agent of small sucking pests, especially Tetranychid mites. Temperature is an important abiotic factor, and a sudden drop in temperature may affect its performance when released in areas with big circadian temperature differences. Rapid cold hardening is a type of phenotypic plasticity that allows ectotherms to quickly enhance their cold tolerance in response to a brief chilling exposure. However, it is not clear whether N. striatus possesses such plasticity. To understand how this species of phytoseiid mite copes with short-term low-temperature stress, its rapid cold hardening response was studied in the laboratory by first exploring its critical temperature. Then, the effects of exposure to a series of temperatures (0, 5, 10, 15, and 20°C) for different durations on the survival of N. striatus were investigated to determine the optimal temperature and duration for cold hardening. Moreover, the effect of cold hardening on the consumption of Tetranychus urticae by N. striatus was also investigated, as was the response of immature stages. The critical temperature for N. striatus was - 15°C, at which its survival dropped below 10%. An exposure to 5°C for 2h was optimal for rapid cold hardening, efficiently increasing survival in N. striatus. Rapid cold hardening was found in all developmental stages of N. striatus, and there was no significant effect of exposure on the consumption of spider mites. Our study shows that exposure to low temperatures during a limited period can enhance the cold hardiness but not reduce predation by N. striatus, which will be advantageous to its field applications during a period of large diurnal temperature fluctuations.

Variability in Cold Tolerance of Food and Clinical Listeria monocytogenes Isolates.

The aim of this study was to investigate the level of strain variability amongst food and clinical Listeria monocytogenes isolates growing at low temperatures (4 and 7 °C) in both laboratory media and real food matrices. Isolates (n = 150) grown in laboratory media demonstrated a large variation in growth profiles measured using optical density. Overall, it was noted that clinical isolates exhibited a significantly higher growth rate (p ≤ 0.05) at 7 °C than the other isolates. Analysis of variance (ANOVA) tests of isolates grouped using Multi Locus Sequence Typing (MLST) revealed that clonal complex 18 (CC18) isolates were significantly (p ≤ 0.05) faster growing at 4 °C than other CC-type isolates while CC101, CC18, CC8, CC37 and CC14 were faster growing than other CC types at 7 °C. Euclidean distance and Ward method-based hierarchical clustering of mean growth rates classified 33.33% of isolates as faster growing. Fast and slow growing representative isolates were selected from the cluster analysis and growth rates were determined using plate count data in laboratory media and model food matrices. In agreement with the optical density experiments, CC18 isolates were faster and CC121 isolates were slower than other CC types in laboratory media, UHT milk and fish pie. The same trend was observed in chocolate milk but the differences were not statistically significant. Moreover, pan-genome analysis (Scoary) of isolate genome sequences only identified six genes of unknown function associated with increased cold tolerance while failing to identify any known cold tolerance genes. Overall, an association that was consistent in laboratory media and real food matrices was demonstrated between isolate CC type and increased cold tolerance.

Cloning of TaeRF1 gene from Caucasian clover and its functional analysis responding to low-temperature stress.

Low temperature (LT) is an important threat to the normal growth of plants. In this study, based on the full-length transcriptome sequencing results, the cold resistance genes were cloned from Caucasian clover with strong cold resistance. We cloned the CDS of TaeRF1, which is 1311 bp in length and encodes 436 amino acids. The molecular weight of the protein is 48.97 kDa, which had no transmembrane structure, and its isoelectric point (pI) was 5.42. We predicted the structure of TaeRF1 and found 29 phosphorylation sites. Subcellular localization showed that TaeRF1 was localized and expressed in cell membrane and chloroplasts. The TaeRF1 gene was induced by stress due to cold, salt, alkali and drought and its expression level was higher in roots and it was more sensitive to LT. Analysis of transgenic A. thaliana plants before and after LT treatment showed that the TaeRF1 gene enhanced the removal of excess H2O2, and increased the activity of antioxidant enzymes, thus improving the plant's ability to resist stress. Additionally, the OE lines showed increased cold tolerance by upregulating the transcription level of cold-responsive genes (CBF1, CBF2, COR15B, COR47, ICE1, and RD29A). This study demonstrates that TaeRF1 is actively involved in the responses of plants to LT stress. We also provide a theoretical basis for breeding and a potential mechanism underlying the responses of Caucasian clover to abiotic stress.

Comparative transcriptome analysis of Liriomyza trifolii (Burgess) and Liriomyza sativae (Blanchard) (Diptera: Agromyzidae) in response to rapid cold hardening.

The ability of insets to react efficiently to fluctuation in temperature is crucial for them to survive in variable surroundings. Rapid cold hardening (RCH) is a process that increase cold tolerance in most insect species. The molecular mechanisms of RCH remain largely unknown, and whether it is associated with transcriptional changes is unclear. In this study, we compared the transcriptomes of Liriomyza trifolii and L. sativae exposed to RCH to investigate the transcript abundance due to RCH in both species. RNA-seq revealed 93,166 assembled unigenes, and 34,303 of these were annotated in the L. trifolii and L. sativae transcriptome libraries. After a 4-h treatment at 1°C (RCH) compared with control, 268 and 606 unigenes were differentially expressed in L. trifolii and L. sativae, respectively. When comparing pupae exposed to 2h cold shock directly with pupae went through 4h acclimation prior to 2h cold shock, 60 and 399 unigenes were differentially expressed in L trifolii and L sativae, respectively. Genes that were commonly expressed in both L. trifolii and L. sativae, included cytochrome P450, cuticular protein, glucose dehydrogenase, solute carrier family 22 and cationic amino acid transporter. Additionally, several pathways including galactose metabolism and peroxisome were significantly enriched during RCH. Our results show that the transcriptional response is correlated with RCH in the pupal stage of the two Liriomyza species, but more transcriptional changes were identified in L sativae than in L. trifolii.

Internal freezing and heat loss of apple (Malus domestica Borkh.) and sweet cherry (Prunus avium L.) reproductive buds are decreased with cellulose nanocrystal dispersions.

Cold damage has caused more economic losses to fruit crop growers in the U.S. than any other weather hazard, making it a perennial concern for producers. Cellulose nanocrystals (CNCs) represent a new generation of renewable bio-nanomaterials, with many unique physical and chemical properties, including their low thermal conductivity. Our team has developed a process for creating CNC dispersions that can be sprayed onto woody perennial crops, forming a thin insulating film around buds which has been shown to increase cold tolerance. Using digital scanning calorimetry (DSC) on dormant apple (Malus domestica Borkh.) reproductive buds, we investigated the thermodynamic properties of plant materials treated with CNC dispersion at lower temperatures. Scanning electron microscopy (SEM) was used to evaluate the thickness of the CNC films and their deposition on the sweet cherry bud surface. Apple buds treated with 3% CNC exhibited lethal freezing at temperatures 3.2°C and 5.5°C lower than the untreated control when sampled 1 and 3 days after application, respectively. Additionally, the latent heat capacity (J/g) of the 3% CNC-treated buds was 46% higher compared with untreated buds 1 day after application, and this difference increased 3 days after application to 168% higher. The emissivity of cherry buds treated with 3% CNC was reduced by an average of 16% compared with the untreated buds. SEM was able to detect the dried films on the surface of the buds 3 days after application. Film thickness measured with SEM increased with material concentration. The emissivity, HTE, and LTE results show that CNC-treated reproductive buds released thermal energy at a slower rate than the untreated buds and, consequently, exhibited internal ice nucleation events at temperatures as much as 5.5°C lower. The increased enthalpy during the LTE in the CNC-treated apple buds shows more energy released at lethal internal freezing, indicating that CNC coatings are increasing the amount of supercooled water. The effects of CNC shown during the DSC tests were increased by CNC concentration and time post-application. These results suggest that CNC dispersions dry into nanofilms on the bud surface, which affects their thermodynamic processes at low temperatures.

Recent insights into cell responses to cold stress in plants: Signaling, defence, and potential functions of phosphatidic acid

Upon exposure to cold stress (CS), crop plants face an array of detrimental effects on growth, development, and final yield. Seed germination, seedling emergence, leaf number, root morphology, photosynthetic activity and metabolism are all disrupted by CS. Plants have evolved various mechanisms to avoid or induce cold tolerance from the cellular level to the whole plant level. Phosphatidic acid (PA) is considered a prime signaling phospholipid in plant membranes that plays an important role in signal transduction in stress avoidance responses. Recent research studies have advanced our understanding of the functions of PA in plant response to abiotic stress. As a second messenger, PA can bind to target proteins to enhance plant cold response and adaption. However, there is a paucity of information on PA-mediated alterations in lipid metabolism as a plant cold tolerance mechanism. Firstly, this review presents plant response to CS at morphological, physiological, biochemical, and molecular levels. The authors also discuss the roles of protective proteins, osmo-protectants, antioxidant systems and lipid metabolisms in plant cold tolerance mechanisms. Furthermore, the current review documents state of the art literature on the effects of CS on plant PA biosynthesis pathways (PLD and PLC-DGK pathways) in plants and its promising role in improving cold tolerance. Finally, we summarized the PA signaling in plant cold tolerance and suggested research directions in CS studies in the future. • Cold stress (CS) significantly reduces seedling emergence, photosynthetic rate, plant biomass and enhances ROS accumulation. • Plants increase cold tolerance by triggering COR genes and producing protective proteins, osmo-protectants, and antioxidants. • Lipid remodeling is a key strategy to maintain membrane stability and functions in plants under CS. • Phosphatidic acid (PA) is an emerging phospholipid that plays an essential role in lipid metabolism and CS response. • Under CS conditions, PA accumulates rapidly through Phospholipase D and Phospholipase C – Diacylglycerol kinase pathways. • PA can bind to target proteins to enhance plant CS response and adaptation.

A calcium-dependent protein kinase gene SpCPK33 from Solanum pennellii associated with increased cold tolerance in tomato

Calcium-dependent protein kinases (CDPKs, CPKs) represent a vital class of calcium sensors, which play a crucial role in plant growth, development and adaption to complex environmental stresses. Wild species tend to exhibit greater tolerance than cultivated species under environmental stress. Here, we isolated a calcium-dependent protein kinase gene SpCPK33 located primarily on the plasma membrane of abiotic-resistant species (Solanum pennellii LA0716). It was highly expressed in stems and leaves and was also induced by cold stress. Compared with WT plants, the overexpression of SpCPK33 in cultivated tomato (cv M82) enhanced its tolerance to cold stress. Transgenic lines demonstrated strong vitality under low temperature treatment. Moreover, the levels of malondialdehyde (MDA) and reactive oxygen species (ROS) were decreased in SpCPK33-overexpressing plants. The activities of antioxidant enzymes and the levels of osmotic regulatory substances were higher. The transcript levels of cold stress-related genes were up-regulated. In summary, the results indicate that SpCPK33-overexpressing transgenic plants experience less severe chilling injury under cold stress, and improved tomato cold tolerance by scavenging ROS accumulation and modulating the expression of stress-related genes.

MYPT1-PP1β phosphatase negatively regulates both chromatin landscape and co-activator recruitment for beige adipogenesis

Protein kinase A promotes beige adipogenesis downstream from β-adrenergic receptor signaling by phosphorylating proteins, including histone H3 lysine 9 (H3K9) demethylase JMJD1A. To ensure homeostasis, this process needs to be reversible however, this step is not well understood. We show that myosin phosphatase target subunit 1- protein phosphatase 1β (MYPT1-PP1β) phosphatase activity is inhibited via PKA-dependent phosphorylation, which increases phosphorylated JMJD1A and beige adipogenesis. Mechanistically, MYPT1-PP1β depletion results in JMJD1A-mediated H3K9 demethylation and activation of the Ucp1 enhancer/promoter regions. Interestingly, MYPT1-PP1β also dephosphorylates myosin light chain which regulates actomyosin tension-mediated activation of YAP/TAZ which directly stimulates Ucp1 gene expression. Pre-adipocyte specific Mypt1 deficiency increases cold tolerance with higher Ucp1 levels in subcutaneous white adipose tissues compared to control mice, confirming this regulatory mechanism in vivo. Thus, we have uncovered regulatory cross-talk involved in beige adipogenesis that coordinates epigenetic regulation with direct activation of the mechano-sensitive YAP/TAZ transcriptional co-activators.

WRKY41/WRKY46-miR396b-5p-TPR module mediates abscisic acid-induced cold tolerance of grafted cucumber seedlings.

Grafting is one of the key agronomic measures to enhance the tolerance to environmental stresses in horticultural plants, but the specific molecular regulation mechanism in this tolerance largely remains unclear. Here, we found that cucumber grafted onto figleaf gourd rootstock increased cold tolerance through abscisic acid (ABA) activating WRKY41/WRKY46-miR396b-5p-TPR (tetratricopeptide repeat-like superfamily protein) module. Cucumber seedlings grafted onto figleaf gourd increased cold tolerance and induced the expression of miR396b-5p. Furthermore, overexpression of cucumber miR396b-5p in Arabidopsis improved cold tolerance. 5’ RNA ligase-mediated rapid amplification of cDNA ends (5’ RLM-RACE) and transient transformation experiments demonstrated that TPR was the target gene of miR396b-5p, while TPR overexpression plants were hypersensitive to cold stress. The yeast one-hybrid and dual-luciferase assays showed that both WRKY41 and WRKY46 bound to MIR396b-5p promoter to induce its expression. Furthermore, cold stress enhanced the content of ABA in the roots and leaves of figleaf gourd grafted cucumber seedlings. Exogenous application of ABA induced the expression of WRKY41 and WRKY46, and cold tolerance of grafted cucumber seedlings. However, figleaf gourd rootstock-induced cold tolerance was compromised when plants were pretreated with ABA biosynthesis inhibitor. Thus, ABA mediated figleaf gourd grafting-induced cold tolerance of cucumber seedlings through activating the WRKY41/WRKY46-miR396b-5p-TPR module.

Vvi-miPEP172b and vvi-miPEP3635b increase cold tolerance of grapevine by regulating the corresponding MIRNA genes

As a kind of small molecular weight proteins, many peptides have been discovered, including peptides encoded by pri-miRNA (miPEPs). Similar as traditional phytohormone or signaling molecular, these peptides participate in numerous plant growth processes. MicroRNAs (miRNAs) play an important regulatory role in plant stress response. While the roles of miPEPs in response to abiotic stress has not been studied now. In this study, to explore whether miPEPs could contribute to low temperature (4ºC) tolerance of plants, the expression pattern of 23 different vvi-MIRs were analyzed by qRT-PCR in 'Thompson Seedless' (Vitis vinifera) plantlets under cold stress (4ºC) firstly, and vvi-MIR172b and vvi-MIR3635b which showed an elevated expression levels were selected to identify miPEPs. Through transient expression, one small open reading frame (sORF) in each of the two pri-miRNAs could increase the expression of corresponding vvi-MIR, and the amino acid sequences of sORFs were named vvi-miPEP172b and vvi-miPEP3635b, respectively. The synthetic vvi-miPEP172b and vvi-miPEP3635b were applied to the grape plantlets, and the tissue culture plantlets exhibited a higher cold tolerance compared with the control groups. These results revealed the effective roles of miPEPs in plant cold stress resistance for the first time, providing a theoretical basis for the future application of miPEPs to agricultural production.

IsoSeq and RNA-Seq analyses uncover the molecular response of Dalbergia odorifera T. Chen to low temperature

In previous studies, we found young plants propagated from acclimated D. odorifera T. Chen trees displayed increased cold tolerance. To understand the molecular and physiological mechanisms underlying this phenomenon, we prepared acclimated and non-acclimated young D. odorifera T. Chen trees for physiological, RNA-Seq, and IsoSeq analyses. First, both the acclimated and non-acclimated young trees were grown in 10 ℃ condition, then treated at −3 ℃ for one day, and finally recovered at 10 ℃. The measurement of physiological parameters, including superoxide dismutase, peroxidase, malondialdehyde, thiobarbituric acid, and soluble sugar, showed that the physiological change of acclimated plants is smoother than non-acclimated plants. The RNA-Seq analyses pointed out that cold acclimation already fixed the different gene expression patterns of D. odorifera T. Chen trees. The hormone-related, secondary metabolic, and signal transduction related biological genes tend to show different expressions between the acclimated and non-acclimated D. odorifera T. Chen trees. Moreover, the change of gene expression for some biological processes, such as alpha-Linolenic acid metabolism and its response to hydrogen peroxide, seems to occur earlier in non-acclimated than acclimated plants. The ISOSeq analyses pointed out that alternative splicing (AS) of some genes was also found, and these AS events were predicted to play important roles in regulating different expression patterns between non-acclimated and acclimated plants. Therefore, according to the performance of this study, we are able to provide some novel understanding of cold tolerance enhanced by cold acclimation in perennial trees.