Comparison of transcriptome and metabolome analysis revealed differences in cold resistant metabolic pathways in different apple cultivars under low temperature stress

Freezing injury in winter is an important abiotic stress that seriously affects plant growth and development.Deciduous fruit trees resist freezing injury by inducing dormancy.However,different cultivars of the same species have different cold resistance strategies.Little is known about the molecular mechanism of apple trees in response to freezing injury during winter dormancy.Therefore,in this study,1-year-old branches of the cold-resistant cultivar‘Hanfu’(HF)and the cold-sensitive cultivar‘Changfuji No.2’(CF)were used to explore their cold resistance through physiological,biochemical,transcriptomics,and metabolomics analyses.Combining physiological and biochemical data,we found that HF had a stronger osmotic regulation ability and antioxidant enzyme activity than CF,as well as stronger cold resistance.The functional enrichment analysis showed that both cultivars were significantly enriched in pathways related to signal transduction,hormone regulation,and sugar metabolism under freezing stress.In addition,the differentially expressed genes(DEGs)encoding galactinol synthase,raffinose synthase,and stachyose synthetase in raffinose family oligosaccharides(RFOs)metabolic pathways were upregulated in HF,and raffinose and stachyose were accumulated,while their contents in CF were lower.HF accumulated 4-aminobutyric acid,spermidine,and ascorbic acid to scavenge reactive oxygen species(ROS).While the contents of oxidized glutathione,vitamin C,glutathione,and spermidine in CF decreased under freezing stress,consequently,the ability to scavenge ROS was low.Furthermore,the transcription factors apetala 2/ethylene responsive factor(AP2/ERF)and WRKY were strongly induced under freezing stress.In summary,the difference in key metabolic components of HF and CF under freezing stress is the major factor affecting their difference in cold resistance.The obtained results deepen our understanding of the cold resistance mechanism in apple trees in response to freezing injury during dormancy.

Theoretical and numerical studies of crack initiation and propagation in rock masses under freezing pressure and far-field stress

Water-bearing rocks exposed to freezing temperature can be subjected to freezeethaw cycles leading tocrack initiation and propagation, which are the main causes of frost damage to rocks. Based on theGriffith theory of brittle fracture mechanics, the crack initiation criterion, propagation direction, andcrack length under freezing pressure and far-field stress are analyzed. Furthermore, a calculation methodis proposed for the stress intensity factor （SIF） of the crack tip under non-uniformly distributed freezingpressure. The formulae for the crack/fracture propagation direction and length of the wing crack underfreezing pressure are obtained, and the mechanism for coalescence of adjacent cracks is investigated.In addition, the necessary conditions for different coalescence modes of cracks are studied. Using thetopology theory, a new algorithm for frost crack propagation is proposed, which has the capability todefine the crack growth path and identify and update the cracked elements. A model that incorporatesmultiple cracks is built by ANSYS and then imported into FLAC3D. The SIFs are then calculated using aFISH procedure, and the growth path of the freezing cracks after several calculation steps is demonstratedusing the new algorithm. The proposed method can be applied to rocks containing fillings such asdetritus and slurry. 2014 Institute of Rock and Soil Mechanics, Chinese Academy of Sciences. Production and hosting byElsevier B.V. All rights reserved.

Reciprocal regulation between the negative regulator PP2CG1 phosphatase and the positive regulator OST1 kinase confers cold response in Arabidopsis

Protein phosphorylation and dephosphorylation have been reported to play important roles in plant cold responses.In addition,phospho-regulatory feedback is a conserved mechanism for biological processes and stress responses in animals and plants.However,it is less well known that a regulatory feedback loop is formed by the protein kinase and the protein phosphatase in plant responses to cold stress.Here,we report that OPEN STOMATA 1(OST1)and PROTEIN PHOSPHATASE 2C G GROUP 1(PP2CG1)reciprocally regulate the activity during the cold stress response.The interaction of PP2CG1 and OST1 is inhibited by cold stress,which results in the release of OST1 at the cytoplasm and nucleus from suppression by PP2CG1.Interestingly,cold-activated OST1 phosphorylates PP2CG1 to suppress its phosphatase activity,thereby amplifying cold signaling in plants.Mutations of PP2CG1 and its homolog PP2CG2 enhance freezing tolerance,whereas overexpression of PP2CG1 decreases freezing tolerance.Moreover,PP2CG1 negatively regulates protein levels of C-REPEAT BINDING FACTORs(CBFs)under cold stress.Our results uncover a phosphor/dephosphor-regulatory feedback loop mediated by PP2CG1 phosphatase and OST1 protein kinase in plant cold responses.