Cold-induced Genes Research Articles

Molecular mechanisms of low-temperature sensitivity in tropical/subtropical plants: a case study of Casuarina equisetifolia.

Low temperature is a limiting factor affecting plant growth and development. Casuarina equisetifolia, a typical tropical and subtropical tree important for the ecological restoration of coastal beaches, is sensitive to cold stress. By comparing cold tolerance between C. equisetifolia and Arabidopsis, we investigated the molecular basis underlying the cold sensitivity of C. equisetifolia. Transcriptomic analysis showed that the number of cold-induced genes in C. equisetifolia was significantly less than that in Arabidopsis, and notably, the response of cold-induced genes was also delayed in C. equisetifolia. Among the cold-induced genes, C-repeat binding factors (CBFs), the major transcription factors in cold acclimation in Arabidopsis, showed a delayed cold-induced expression in C. equisetifolia, despite that C. equisetifolia CBFs could restore the low temperature-sensitive phenotype of Arabidopsis cbfs triple mutants. Interestingly, some key cold-responsive genes (e.g., COR15A and COR15B) targeted by Arabidopsis CBF were absent in the C. equisetifolia genome and many cold-responsive genes in C. equisetifolia lacked the DRE element (i.e., CBF binding cis-element). Moreover, like in C. equisetifolia, many COR genes in other tropical/subtropical plants lacked the DRE element or were directly missing. These two factors could be the underlying reasons for the low-temperature sensitivity of C. equisetifolia and other tropical/subtropical plants.

A WRKY Transcription Factor PmWRKY57 from Prunus mume Improves Cold Tolerance in Arabidopsis thaliana.

Prunus mume, a woody perennial tree, is valued for its ornamental traits and has been cultivated for a long history. Low temperature is the main environmental factor restricting the distribution and affecting the growth of P. mume. In plants, some WRKY transcription factors have been reported to participate in regulating cold tolerance. However, there were few researches about functional characterization of WRKYs involving in P. mume cold response. Here, a cold-induced WRKY gene named as PmWRKY57 was cloned from a P. mume cultivar 'Guhong Zhusha.' PmWRKY57 protein harboring a WRKY domain and a C2H2 zinc finger motif belongs to Group IIc of WRKY family. The PmWRKY57 protein was located to the nucleus and has transcriptional activation activity. PmWRKY57-overexpresing Arabidopsis thaliana lines showed improved cold tolerance, compared to wild-type plants. Under cold treatment, the leaves of transgenic lines contained significantly lower malondialdehyde content, and higher levels of superoxide dismutase activity, peroxidase activity, and proline content than wild-type plants. Furthermore, the expression levels of cold-response genes such as AtCOR6.6, AtCOR47, AtKIN1, and AtRCI2A were up-regulated in leaves of transgenic A. thaliana compared to those in wild-type plants. This study characterized the function of PmWRKY57 in improving cold tolerance of plants.

Whole genome analysis of Flavobacterium aziz-sancarii sp. nov., isolated from Ardley Island (Antarctica), revealed a rich resistome and bioremediation potential

Despite being one of the most isolated regions in the world, Antarctica is at risk of increased contamination with potentially toxic elements and other toxic chemicals through anthropogenic interventions. In this study, a psychrotolerant bacterium was isolated using the lake water collected from Ardley Island (Antarctica), which can grow at temperatures between 4 and 30 °C and pH values between 6.0 and 9.0. The isolate, named AC, had protease, amylase, and lipase activities with no NaCl tolerance and could degrade 1–5% diesel fuel. Multilocus sequence analysis (MLSA) using 16S rRNA, gyrB, tuf, and rpoD genes resulted in 92.91–98.6% sequence similarities between the isolate AC and other Flavobacterium spp. Whole genome analysis indicated that the genome length of Flavobacterium sp. AC is 5.8 Mbp with a GC content of 34.04% and 1274 genes predicted. The strain AC branched independently from other Flavobacterium spp. in the phylogenetic and phylogenomic trees and ranked a new species named Flavobacterium aziz-sancarii. Genome mining identified several cold-inducible genes, including stress-associated genes such as cold-shock proteins, chaperones, carotenoid biosynthetic genes, or oxidative-stress response genes. In addition, virulence, gliding motility, and biofilm-related genes were determined. Its genome contains 35 and 88 open-reading frames related to potentially toxic element and antibiotic resistance, respectively. F. aziz-sancarii showed a remarkable tolerance of Cr and Ni, with minimal inhibitory concentration values of 2.88 and 2.81 mM, respectively. Pb, Cu, and Zn exposure resulted in moderate toxicity (2.14–2.41 mM), while Cd showed the highest inhibitory effect in bacterial growth (0.74 mM). Antibiotic susceptibility testing indicated multidrug-resistant phenotype in correlation to in silico prediction of antibiotic resistance genes. Overall, our results contribute to biodiversity of Antarctica and provide new insights into resistome profile of Antarctic microorganisms. Additionally, the diesel degradation feature of F. aziz-sancarii offers potential use for the bioremediation of hydrocarbon-contaminated polar ecosystems.

Understanding the Function and Mechanism of Zebrafish Tmem39b in Regulating Cold Resistance.

Autophagy and endoplasmic reticulum (ER) stress response are among the key pathways regulating cold resistance of fish through eliminating damaged cellular components and facilitating the restoration of cell homeostasis upon exposure to acute cold stress. The transmembrane protein 39A (TMEM39A) was reported to regulate both autophagy and ER stress response, but its vertebrate-specific paralog, the transmembrane protein 39B (TMEM39B), has not been characterized. In the current study, we generate tmem39b-knockout zebrafish lines and characterize their survival ability under acute cold stress. We observed that the dysfunction of Tmem39b remarkably decreased the cold resilience of both the larval and adult zebrafish. Gene transcription in the larvae exposed to cold stress and rewarming were characterized by RNA sequencing (RNA-seq) to explore the mechanisms underlying functions of Tmem39b in regulating cold resistance. The results indicate that the deficiency of Tmem39b attenuates the up-regulation of both cold- and rewarming-induced genes. The cold-induced transcription factor genes bif1.2, fosab, and egr1, and the rewarming-activated immune genes c3a.3, il11a, and sting1 are the representatives influenced by Tmem39b dysfunction. However, the loss of tmem39b has little effect on the transcription of the ER stress response- and autophagy-related genes. The measurements of the phosphorylated H2A histone family member X (at Ser 139, abbreviated as γH2AX) demonstrate that zebrafish Tmem39b protects the cells against DNA damage caused by exposure to the cold-warming stress and facilitates tissue damage repair during the recovery phase. The gene modules underlying the functions of Tmem39b in zebrafish are highly enriched in biological processes associated with immune response. The dysfunction of Tmem39b also attenuates the up-regulation of tissue C-reactive protein (CRP) content upon rewarming. Together, our data shed new light on the function and mechanism of Tmem39b in regulating the cold resistance of fish.

The cold-stress responsive gene DREB1A involved in low-temperature tolerance in Xinjiang wild walnut.

BackgroundLow-temperatures have the potential to be a serious problem for plants and can negatively affect the normal growth and development of walnuts. DREB1/CBF (Dehydration Responsive Element Binding Protein 1/C-repeat Binding Factor), one of the most direct transcription factors in response to low-temperature stress, may improve the resistance of plants to low-temperatures by regulating their functional genes. However, few studies have been conducted in walnut. The Xinjiang wild walnut is a rare wild plant found in China, with a large number of excellent trait genes, and is hardier than cultivated walnuts in Xinjiang.MethodsIn this work, we identified all of the DREB1 members from the walnut genome and analyzed their expression levels in different tissues and during low-temperature stress on the Xinjiang wild walnut. The JfDREB1A gene of the Xinjiang wild walnut was cloned and transformed into Arabidopsis thaliana for functional verification.ResultsThere were five DREB1 transcription factors in the walnut genome. Among them, the relative expression level of the DREB1A gene was significantly higher than other members in the different tissues (root, stem, leaf) and was immediately un-regulated under low-temperature stress. The overexpression of the JfDREB1A gene increased the survival rates of transgenic Arabidopsis lines, mainly through maintaining the stability of cell membrane, decreasing the electrical conductivity and increasing the activities of antioxidant enzymes including superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT). Additionally, the expression levels of cold-inducible genes like AtKIN1, AtERD10, AtRD29A, AtCOR15A and AtCOR47, were significantly increased. These results showed that the JfDREB1A gene may play an important role in the response to cold stress of the Xinjiang wild walnut. This study contributes to our understanding of the molecular mechanism of the Xinjiang wild walnut’s response to low-temperature stress and will be beneficial for developing walnut cultivars with improved cold resistance.

307-OR: Impaired Differentiation of Trpv1-Positive Adipose Progenitors Activates Compensatory Thermogenic Mechanisms

Brown and beige thermogenic adipocytes are specialized in energy consumption and linked to metabolic health. Canonically, thermogenic adipocytes are thought to be derived from mesenchymal PdgfRα+ adipocyte progenitor cells (APC) . We recently identified a novel population of cold-recruited APCs from the vascular smooth muscle lineage marked by the expression of the ion channel Trpv1 in mouse adipose tissue. To determine the function of the Trpv1+ APC-derived adipocytes, we impaired their adipogenic capacity by deletion of Pparγ in Trpv1-expressing cells in mice (Trpv1-Cre::Pparγ-flox) . Inhibition of de novo adipogenesis from Trpv1+ APCs resulted in modest changes in brown adipose tissue, but it caused a marked reduction in the expression of cold-induced browning genes, including UCP1, in white adipose tissue (WAT) , indicating impaired beige adipogenesis. Interestingly, Trpv1-Cre::Pparγ-flox mice did not show altered weight gain, glucose tolerance, or insulin sensitivity on either regular chow or high-fat diets. This discrepancy led us to search for potential compensatory mechanisms. We observed increased PdgfRα expression in WAT of Trpv1-Cre::Pparγ-flox animals, suggesting that impaired adipogenic capacity of the Trpv1+ APCs may trigger recruitment of the PdgfRα+ APCs. Additionally, while UCP1 expression was noticeably reduced in the beige adipocytes within WAT, the expression of transcripts involved in the UCP1-independent thermogenic pathways, including Ckmt1, Gatm, and Slc6a8, key components of the creatine futile cycle, was increased in the WAT of the Trpv1-Cre::Pparγ-flox mice upon cold exposure. Together, these data highlight a potential distinction between the thermogenic pathways mediated by adipocytes derived from the Trpv1+ versus those derived from the PdgfRα+ APCs. This functional difference of thermogenic adipocytes could be critical in designing strategies to target these cells as a therapeutic approach for metabolic diseases. Disclosure H.Camara: None. F.Shamsi: None. M.Lynes: None. Y.Tseng: Consultant; Cellarity. Funding National Institutes of Health (R01DK102898)

Biochemical and molecular responses of hot pepper (Capsicum annuum) to cold stress

Four cold tolerant hot pepper (Capsicum annuum L.) genotypes were identified at Division of Vegetable Science,ICAR-IARI, New Delhi which showed significantly better agronomic performance during the cold winter conditions ofDelhi during 2019–20 when most of other genotypes succumbed. Present investigations were carried out to understand the biochemical and molecular basis of cold tolerance in these genotypes for which they were compared with cold sensitive genotypes for various biochemical parameters as well as for expression of some cold inducible genes. The results indicated that the cold tolerant genotypes had inherent biochemical and molecular mechanism which helped them in cold acclimation. These genotypes had strong antioxidant defense and ROS (Reactive Oxygen Species) scavenging system as displayed by significantly high activities of guaiacol-peroxidase and superoxide dismutase and lower levels of lipid peroxidation in response to cold stress. They also accumulated higher concentration of proline to maintain their membrane fluidity and H2O2. Twelve of the fourteen cold inducible genes studied in the present investigations had significantly high expression in cold tolerant genotypes. However the expression profile of these genes differed in different genotypes indicating that mechanism of cold tolerance was not identical in all the genotypes. The confirmation of existence of innate cold tolerance mechanism in our test genotypes can pave a way for future utilization of these genotypes in cold stress breeding.

Genome-Wide Identification of the PMEI Gene Family in Tea Plant and Functional Analysis of CsPMEI2 and CsPMEI4 Through Ectopic Overexpression.

Pectin methylesterase inhibitor (PMEI) inhibits pectin methylesterase (PME) activity at post-translation level, which plays core roles in vegetative and reproductive processes and various stress responses of plants. However, the roles of PMEIs in tea plant are still undiscovered. Herein, a total of 51 CsPMEIs genes were identified from tea plant genome. CsPMEI1-4 transcripts were varied in different tea plant tissues and regulated by various treatments, including biotic and abiotic stresses, sugar treatments, cold acclimation and bud dormancy. Overexpression of CsPMEI4 slightly decreased cold tolerance of transgenic Arabidopsis associated with lower electrolyte leakage, soluble sugars contents and transcripts of many cold-induced genes as compared to wild type plants. Under long-day and short-day conditions, CsPMEI2/4 promoted early flowering phenotypes in transgenic Arabidopsis along with higher expression levels of many flowering-related genes. Moreover, overexpression of CsPMEI2/4 decreased PME activity, but increased sugars contents (sucrose, glucose, and fructose) in transgenic Arabidopsis as compared with wild type plants under short-day condition. These results indicate that CsPMEIs are widely involved in tea plant vegetative and reproductive processes, and also in various stress responses. Moreover, CsPMEI4 negatively regulated cold response, meanwhile, CsPMEI2/4 promoted early flowering of transgenic Arabidopsis via the autonomous pathway. Collectively, these results open new perspectives on the roles of PMEIs in tea plant.

Cold Response Transcriptome Analysis of the Alternative Splicing Events Induced by the Cold Stress in D. catenatum.

Dendrobium catenatum Lindl is a valuable medicinal herb and gardening plant due to its ornamental value and special medical value. Low temperature is a major bottleneck restricting D. catenatum expansion towards the north, which influences the quality and yield of D. catenatum. In this study, we analysed the cold response of D. catenatum by RNA-Seq. A total of 4302 differentially expressed genes were detected under cold stress, which were mainly linked to protein kinase activity, membrane transport and the glycan biosynthesis and metabolism pathway. We also identified 4005 differential alternative events in 2319 genes significantly regulated by cold stress. Exon skipping and intron retention were the most common alternative splicing isoforms. Numerous genes were identified that differentially modulated under cold stress, including cold-induced transcription factors and splicing factors mediated by AS (alternative splicing). GO enrichment analysis found that differentially alternatively spliced genes without differential expression levels were related to RNA/mRNA processing and spliceosomes. DAS (differentially alternative splicing) genes with different expression levels were mainly enriched in protein kinase activity, plasma membrane and cellular response to stimulus. We further identified and cloned DcCBP20 in D. catenatum; we found that DcCBP20 promotes the generation of alternative splicing variants in cold-induced genes under cold stress via genetic experiments and RT–PCR. Taken together, our results identify the main cold-response pathways and alternative splicing events in D. catenatum responding to cold treatment and that DcCBP20 of D. catenatum get involved in regulating the AS and gene expression of cold-induced genes during this process. Our study will contribute to understanding the role of AS genes in regulating the cold stress response in D. catenatum.

Multigene Transformation Through Cre-lox Mediated Site-Specific Integration in Rice.

Plant transformation with multiple genes is a major challenge, rendering multi-trait engineering extremely difficult in crop plants. One of the hurdles in multigene transformation is the uncontrolled integration process that leads to low quality transgenic lines that are unsuitable for practical application. Recombinase-mediated site-specific integration has been tested and validated for developing high quality transgenic lines expressing one, two, or multiple genes. Of the numerous recombinase systems tested, Cre-lox and FLP-FRT show high efficiency in plants. Recently, Cre-lox system was successfully used to stack a set of 3 constitutive, 1 heat-induced, and 1 cold-induced gene. A number of transgenic lines were obtained through a relatively small effort, and the resulting transgenic lines all expressed the genes properly as determined by their promoter-specificity. Here, a method of Cre-lox mediated stacking of a multigene construct is described using rice as a model crop.

Elucidation of molecular mechanisms of cold-inducible gene expression in higher plants

Elucidation of molecular mechanisms of cold-inducible gene expression in higher plants

The CBL-interacting protein kinase CaCIPK13 positively regulates defence mechanisms against cold stress in pepper.

Cold stress is one of the main factors limiting growth and development in pepper. Calcineurin B-like proteins (CBLs) are specific calcium sensors with non-canonical EF-hands to capture calcium signals, and interact with CBL-interacting protein kinases (CIPKs) in the regulation of various stresses. In this study, we isolated a cold-induced CIPK gene from pepper named CaCIPK13, which encodes a protein of 487 amino acids. In silico analyses indicated that CaCIPK13 is a typical CIPK family member with a conserved NAF motif, which consists of the amino acids asparagine, alanine, and phenylalanine. The CaCIPK13 protein was located in the nucleus and plasma membrane. Knock down of CaCIPK13 resulted in enhanced sensitivity to cold stress in pepper, with increased malondialdehyde content, H2O2 accumulation, and electrolyte leakage, while the catalase, peroxidase, superoxide dismutase activities and anthocyanin content were decreased. The transcript level of cold and anthocyanin-related genes was substantially decreased in CaCIPK13-silenced pepper leaves relative to the empty vector control. On the contrary, overexpression of CaCIPK13 in tomato improved cold tolerance via increasing anthocyanin content and activities of reactive oxygen species scavenging enzymes. Furthermore, the interaction of CaCIPK13 with CaCBL1/6/7/8 was Ca2+-dependent. These results indicate that CaCIPK13 plays a positive role in cold tolerance mechanism via CBL-CIPK signalling.

Genome-wide analysis of Dof transcription factors and their response to cold stress in rice (Oryza sativa L.)

BackgroundRice (Oryza sativa L.) is a food crop for humans worldwide. However, temperature has an effect during the vegetative and reproductive stages. In high-latitude regions where rice is cultivated, cold stress is a major cause of yield loss and plant death. Research has identified a group of plant-specific transcription factors, DNA binding with one zinc fingers (DOFs), with a diverse range of functions, including stress signaling and stress response during plant growth. The aim of this study was to identify Dof genes in two rice subspecies, indica and japonica, and screen for Dof genes that may be involved in cold tolerance during plant growth.ResultsA total of 30 rice Dofs (OsDofs) were identified using bioinformatics and genome-wide analyses and phylogenetically analyzed. The 30 OsDOFs were classified into six subfamilies, and 24 motifs were identified based on protein sequence alignment. The chromosome locations of OsDofs were determined and nine gene duplication events were identified. A joint phylogenetic analysis was performed on DOF protein sequences obtained from four monocotyledon species to examine the evolutionary relationship of DOF proteins. Expression profiling of OsDofs from two japonica cultivars (Longdao5, which is cold-tolerant, and Longjing11, which is cold-sensitive) revealed that OsDof1 and OsDof19 are cold-inducible genes. We examined the seed setting rates in OsDof1- and OsDof19-overexpression and RNAi lines and found that OsDof1 showed a response to cold stress.ConclusionsOur investigation identified OsDof1 as a potential target for genetic breeding of rice with enhanced cold tolerance.

Overexpression of IbFAD8 Enhances the Low-Temperature Storage Ability and Alpha-Linolenic Acid Content of Sweetpotato Tuberous Roots.

Sweetpotato is an emerging food crop that ensures food and nutrition security in the face of climate change. Alpha-linoleic acid (ALA) is one of the key factors affecting plant stress tolerance and is also an essential nutrient in humans. In plants, fatty acid desaturase 8 (FAD8) synthesizes ALA from linoleic acid (LA). Previously, we identified the cold-induced IbFAD8 gene from RNA-seq of sweetpotato tuberous roots stored at low-temperature. In this study, we investigated the effect of IbFAD8 on the low-temperature storage ability and ALA content of the tuberous roots of sweetpotato. Transgenic sweetpotato plants overexpressing IbFAD8 (TF plants) exhibited increased cold and drought stress tolerance and enhanced heat stress susceptibility compared with non-transgenic (NT) plants. The ALA content of the tuberous roots of TF plants (0.19 g/100 g DW) was ca. 3.8-fold higher than that of NT plants (0.05 g/100 g DW), resulting in 8–9-fold increase in the ALA/LA ratio in TF plants. Furthermore, tuberous roots of TF plants showed better low-temperature storage ability compared with NT plants. These results indicate that IbFAD8 is a valuable candidate gene for increasing the ALA content, environmental stress tolerance, and low-temperature storage ability of sweetpotato tuberous roots via molecular breeding.

Cold induced genes (CIGs) regulate flower development and dormancy in Prunus avium L.

The flower buds continue to develop during the whole winter in tree fruit species, which is affected by environmental factors and hormones. However, little is known about the molecular mechanism of flower development during dormancy phase of sweet cherry in response to light, temperature and ABA. Therefore, we identified two cold induced gene (CIG) PavCIG1 and PavCIG2 from sweet cherry, which were closely to PpCBF and PyDREB from Prunus persica and Prunus yedoensis by using phylogenetic analysis, suggesting conserved functions with these evolutionarily closer DREB subfamily genes. Subcellular localization analysis indicated that, PavCIG1 and PavCIG2 were both localized in the nucleus. The seasonal expression levels of PavCIG1 and PavCIG2 were higher at the stage of endodormancy in winter, and induced by low temperature. Ectopic expression of PavCIG1 and PavCIG2 resulted in a delayed flowering in Arabidopsis. Furthermore, PavCIG2 increased light-responsive gene PavHY5 transcriptional activity by binding to its promoter, meanwhile, PavHY5-mediated positive feedback regulated PavCIG2. Moreover, ABA-responsive protein PavABI5-like could also increase transcriptional activity of PavCIG and PavCIG2. In addition, PavCIG and PavCIG2 target gene PavCAL-like was involved in floral initiation, demonstrated by ectopic expression in Arabidopsis. These findings provide evidences to better understand the molecular mechanism of CIG-mediated flower development and dormancy in fruit species, including sweet cherry.

Genomic Variation Landscape of the Model Salt Cress Eutrema salsugineum.

Eutrema salsugineum has long been used as the model for examining salt and other abiotic stress in plants. In addition to the forward genetics approaches widely used in the lab, natural variations undoubtedly will provide a rich genetic resource for studying molecular mechanisms underlying the stress tolerance and local adaptation of this species. We used 90 resequencing whole genomes of natural populations of this species across its Asian and North American distributions to detect the selection signals for genes involved in salt and other stresses at the species-range level and local distribution. We detected selection signals for genes involved in salt and other abiotic tolerance at the species level. In addition, several cold-induced and defense genes showed selection signals due to local adaptation in North America-NE Russia or northern China, respectively. These variations and findings provide valuable resources for further deciphering genetic mechanisms underlying the stress tolerance and local adaptations of this model species.

Transcriptome and metabolomic analysis to reveal the browning spot formation of \u2018Huangguan\u2019 pear

BackgroundBrowning spot (BS) disorders seriously affect the appearance quality of ‘Huangguan’ pear and cause economic losses. Many studies on BS have mainly focused on physiological and biochemical aspects, and the molecular mechanism remains unclear.ResultsIn the present study, the structural characteristics of ‘Huangguan’ pear with BS were observed via scanning electron microscopy (SEM), the water loss and brown spots were evaluated, and transcriptomic and metabolomics analyses were conducted to reveal the molecular mechanism underlying ‘Huangguan’ pear skin browning disorder. The results showed that the occurrence of BS was accompanied by a decrease in the wax layer and an increase in lignified cells. Genes related to wax biosynthesis were downregulated in BS, resulting in a decrease in the wax layer in BS. Genes related to lignin were upregulated at the transcriptional level, resulting in upregulation of metabolites related to phenylpropanoid biosynthesis. Expression of calcium-related genes were upregulated in BS. Cold-induced genes may represent the key genes that induce the formation of BS. In addition, the results demonstrated that exogenous NaH2PO4·2H2O and ABA treatment could inhibit the incidence of BS during harvest and storage time by increasing wax-related genes and calcium-related genes expression and increasing plant resistance, whereas the transcriptomics results indicated that GA3 may accelerate the incidence and index of BS.ConclusionsThe results of this study indicate a molecular mechanism that could explain BS formation and elucidate the effects of different treatments on the incidence and molecular regulation of BS.

Functional Characterization of Salt‑Stress Induced Rare Cold Inducible Gene from Camelina sativa (CsRCI2D)

Rare-cold-inducible 2 (RCI2) genes are involved in plant response to abiotic stresses. In this study, we report the functional role of a Camelina RCI2, CsRCI2D, in plant salt stress response. The localization of CsRCI2D was observed in the plasma membrane and intracellular membranes by confocal analysis in tobacco leaf and western blot analysis in Camelina. The full length CsRCI2D cDNA clone was not able to complement the salt sensitivity of △spmp3 lacking the PMP3 gene. However, a C-terminal tail deleted CsRCI2D cDNA was able to restore the level of salt tolerance to that of WT. CsRCI2D-overexpressing Camelina showed better germination rate and seedling growth. CsRCI2D overexpression decreased Na+ accumulation in both roots and shoots but increased K+ accumulation in the shoots under salt stress. Furthermore, CsRCI2D-overexpressing Camelina displayed lower H2O2 and malondialdehyde (MDA) upon salt stress. Under normal growth condition, CsRCI2D-overexpressing Camelina showed higher transcript levels of all antioxidant genes (CsCuSODs, CsMnSOD1, CsFeSODs, CsCATs, CsAPX1, and CsGR), whereas salt stress significantly induced all antioxidant genes in WT. These results indicate that salt-upregulated CsRCI2D plays a positive role in Camelina seed germination and seedling growth under salt stress by maintenance of ion homeostasis and modulating the expression of antioxidant-related genes.

Xenogeneic silencing relies on temperature-dependent phosphorylation of the host H-NS protein in Shewanella.

Lateral gene transfer (LGT) plays a key role in shaping the genome evolution and environmental adaptation of bacteria. Xenogeneic silencing is crucial to ensure the safe acquisition of LGT genes into host pre-existing regulatory networks. We previously found that the host nucleoid structuring protein (H-NS) silences prophage CP4So at warm temperatures yet enables this prophage to excise at cold temperatures in Shewanella oneidensis. However, whether H-NS silences other genes and how bacteria modulate H-NS to regulate the expression of genes have not been fully elucidated. In this study, we discovered that the H-NS silences many LGT genes and the xenogeneic silencing of H-NS relies on a temperature-dependent phosphorylation at warm temperatures in S. oneidensis. Specifically, phosphorylation of H-NS at Ser42 is critical for silencing the cold-inducible genes including the excisionase of CP4So prophage, a cold shock protein, and a stress-related chemosensory system. By contrast, nonphosphorylated H-NS derepresses the promoter activity of these genes/operons to enable their expression at cold temperatures. Taken together, our results reveal that the posttranslational modification of H-NS can function as a regulatory switch to control LGT gene expression in host genomes to enable the host bacterium to react and thrive when environmental temperature changes.

Posttranslational regulation of multiple clock-related transcription factors triggers cold-inducible gene expression in Arabidopsis

Cold stress is an adverse environmental condition that affects plant growth, development, and crop productivity. Under cold stress conditions, the expression of numerous genes that function in the stress response and tolerance is induced in various plant species, and the dehydration-responsive element (DRE) binding protein 1/C-repeat binding factor (DREB1/CBF) transcription factors function as master switches for cold-inducible gene expression. Cold stress strongly induces these DREB1 genes. Therefore, it is important to elucidate the mechanisms of DREB1 expression in response to cold stress to clarify the perception and response of cold stress in plants. Previous studies indicated that the central oscillator components of the circadian clock, CIRCADIAN CLOCK-ASSOCIATED 1 (CCA1) and LATE ELONGATED HYPOCOTYL (LHY), are involved in cold-inducible DREB1 expression, but the underlying mechanisms are not clear. We revealed that the clock-related MYB proteins REVEILLE4/LHY-CCA1-Like1 (RVE4/LCL1) and RVE8/LCL5 are quickly and reversibly transferred from the cytoplasm to the nucleus under cold stress conditions and function as direct transcriptional activators of DREB1 expression. We found that CCA1 and LHY suppressed the expression of DREB1s under unstressed conditions and were rapidly degraded specifically in response to cold stress, which suggests that they act as transcriptional repressors and indirectly regulate the cold-inducible expression of DREB1s We concluded that posttranslational regulation of multiple clock-related transcription factors triggers cold-inducible gene expression. Our findings clarify the complex relationship between the plant circadian clock and the regulatory mechanisms of cold-inducible gene expression.