EMBO J , IF:9.889 , 2019 Jan , V38 (1) doi: 10.15252/embj.201899819
EGR2 phosphatase regulates OST1 kinase activity and freezing tolerance in Arabidopsis.
State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, China.; College of Horticulture, China Agricultural University, Beijing, China.; Department of Plant Biology, Cornell University, Ithaca, NY, USA.; Institute of Plant Stress Biology, Collaborative Innovation Center of Crop Stress Biology, Henan University, Kaifeng, China.; State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, China yangshuhua@cau.edu.cn.
OST1 (open stomata 1) protein kinase plays a central role in regulating freezing tolerance in Arabidopsis; however, the mechanism underlying cold activation of OST1 remains unknown. Here, we report that a plasma membrane-localized clade-E growth-regulating 2 (EGR2) phosphatase interacts with OST1 and inhibits OST1 activity under normal conditions. EGR2 is N-myristoylated by N-myristoyltransferase NMT1 at 22 degrees C, which is important for its interaction with OST1. Moreover, myristoylation of EGR2 is required for its function in plant freezing tolerance. Under cold stress, the interaction of EGR2 and NMT1 is attenuated, leading to the suppression of EGR2 myristoylation in plants. Plant newly synthesized unmyristoylated EGR2 has decreased binding ability to OST1 and also interferes with the EGR2-OST1 interaction under cold stress. Consequently, the EGR2-mediated inhibition of OST1 activity is released. Consistently, mutations of EGRs cause plant tolerance to freezing, whereas overexpression of EGR2 exhibits decreased freezing tolerance. This study thus unravels a molecular mechanism underlying cold activation of OST1 by membrane-localized EGR2 and suggests that a myristoyl switch on EGR2 helps plants to adapt to cold stress.
PMID: 30429206
Plant J , IF:6.141 , 2019 Jan , V97 (2) : P240-256 doi: 10.1111/tpj.14110
A gene-stacking approach to overcome the trade-off between drought stress tolerance and growth in Arabidopsis.
Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo, 113-8657, Japan.; Max-Planck-Institut fur Molekulare Pflanzenphysiologie, Am Muhlenberg 1, 14476, Golm, Germany.; RIKEN Center for Sustainable Resource Science, Tsurumi-ku, Yokohama, 230-0045, Japan.; Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, 464-8601, Japan.
The molecular breeding of drought stress-tolerant crops is imperative for stable food and biomass production. However, a trade-off exists between plant growth and drought stress tolerance. Many drought stress-tolerant plants overexpressing stress-inducible genes, such as DEHYDRATION-RESPONSIVE ELEMENT-BINDING PROTEIN 1A (DREB1A), show severe growth retardation. Here, we demonstrate that the growth of DREB1A-overexpressing Arabidopsis plants could be improved by co-expressing growth-enhancing genes whose expression is repressed under drought stress conditions. We used Arabidopsis GA REQUIRING 5 (GA5), which encodes a rate-limiting gibberellin biosynthetic enzyme, and PHYTOCHROME-INTERACTING FACTOR 4 (PIF4), which encodes a transcription factor regulating cell growth in response to light and temperature, for growth improvement. We observed an enhanced biomass and floral induction in the GA5 DREB1A and PIF4 DREB1A double overexpressors compared with those in the DREB1A overexpressors. Although the GA5 DREB1A double overexpressors continued to show high levels of drought stress tolerance, the PIF4 DREB1A double overexpressors showed lower levels of stress tolerance than the DREB1A overexpressors due to repressed expression of DREB1A. A multiomics analysis of the GA5 DREB1A double overexpressors showed that the co-expression of GA5 and DREB1A additively affected primary metabolism, gene expression and plant hormone profiles in the plants. These multidirectional analyses indicate that the inherent trade-off between growth and drought stress tolerance in plants can be overcome by appropriate gene-stacking approaches. Our study provides a basis for using genetic modification to improve the growth of drought stress-tolerant plants for the stable production of food and biomass.
PMID: 30285298
Int J Mol Sci , IF:4.556 , 2019 Jan , V20 (2) doi: 10.3390/ijms20020355
Quantitative Proteomic Analysis of Castor (Ricinus communis L.) Seeds During Early Imbibition Provided Novel Insights into Cold Stress Response.
College of Life Science, Inner Mongolia University for Nationalities, Tongliao 028000, China. xiaoyuwang1987@hotmail.com.; Inner Mongolia Key Laboratory for Castor, Tongliao 028000, China. xiaoyuwang1987@hotmail.com.; Inner Mongolia Industrial Engineering Research Center of Universities for Castor, Tongliao 028000, China. xiaoyuwang1987@hotmail.com.; Inner Mongolia Collaborate Innovation Cultivate Center for Castor, Tongliao 028000, China. xiaoyuwang1987@hotmail.com.; Horqin Plant Stress Biology Research Institute of Inner Mongolia University for Nationalities, Tongliao 028000, China. xiaoyuwang1987@hotmail.com.; College of Agriculture, Inner Mongolia University for Nationalities, Tongliao 028000, China. lm@imun.edu.cn.; College of Life Science, Inner Mongolia University for Nationalities, Tongliao 028000, China. liuxuminggenes@hotmail.com.; Inner Mongolia Key Laboratory for Castor, Tongliao 028000, China. liuxuminggenes@hotmail.com.; Inner Mongolia Industrial Engineering Research Center of Universities for Castor, Tongliao 028000, China. liuxuminggenes@hotmail.com.; Inner Mongolia Collaborate Innovation Cultivate Center for Castor, Tongliao 028000, China. liuxuminggenes@hotmail.com.; Horqin Plant Stress Biology Research Institute of Inner Mongolia University for Nationalities, Tongliao 028000, China. liuxuminggenes@hotmail.com.; College of Life Science, Inner Mongolia University for Nationalities, Tongliao 028000, China. 15334940513@163.com.; Inner Mongolia Key Laboratory for Castor, Tongliao 028000, China. 15334940513@163.com.; Inner Mongolia Industrial Engineering Research Center of Universities for Castor, Tongliao 028000, China. 15334940513@163.com.; Inner Mongolia Collaborate Innovation Cultivate Center for Castor, Tongliao 028000, China. 15334940513@163.com.; Horqin Plant Stress Biology Research Institute of Inner Mongolia University for Nationalities, Tongliao 028000, China. 15334940513@163.com.; College of Life Science, Inner Mongolia University for Nationalities, Tongliao 028000, China. duanqiong0209@163.com.; Inner Mongolia Key Laboratory for Castor, Tongliao 028000, China. duanqiong0209@163.com.; Inner Mongolia Industrial Engineering Research Center of Universities for Castor, Tongliao 028000, China. duanqiong0209@163.com.; Inner Mongolia Collaborate Innovation Cultivate Center for Castor, Tongliao 028000, China. duanqiong0209@163.com.; Horqin Plant Stress Biology Research Institute of Inner Mongolia University for Nationalities, Tongliao 028000, China. duanqiong0209@163.com.; College of Life Science, Inner Mongolia University for Nationalities, Tongliao 028000, China. zhangjixing@imun.edu.cn.; Inner Mongolia Key Laboratory for Castor, Tongliao 028000, China. zhangjixing@imun.edu.cn.; Inner Mongolia Industrial Engineering Research Center of Universities for Castor, Tongliao 028000, China. zhangjixing@imun.edu.cn.; Inner Mongolia Collaborate Innovation Cultivate Center for Castor, Tongliao 028000, China. zhangjixing@imun.edu.cn.; Horqin Plant Stress Biology Research Institute of Inner Mongolia University for Nationalities, Tongliao 028000, China. zhangjixing@imun.edu.cn.
Early planting is one of the strategies used to increase grain yield in temperate regions. However, poor cold tolerance in castor inhibits seed germination, resulting in lower seedling emergence and biomass. Here, the elite castor variety Tongbi 5 was used to identify the differential abundance protein species (DAPS) between cold stress (4 degrees C) and control conditions (30 degrees C) imbibed seeds. As a result, 127 DAPS were identified according to isobaric tag for relative and absolute quantification (iTRAQ) strategy. These DAPS were mainly involved in carbohydrate and energy metabolism, translation and posttranslational modification, stress response, lipid transport and metabolism, and signal transduction. Enzyme-linked immunosorbent assays (ELISA) demonstrated that the quantitative proteomics data collected here were reliable. This study provided some invaluable insights into the cold stress responses of early imbibed castor seeds: (1) up-accumulation of all DAPS involved in translation might confer cold tolerance by promoting protein synthesis; (2) stress-related proteins probably protect the cell against damage caused by cold stress; (3) up-accumulation of key DAPS associated with fatty acid biosynthesis might facilitate resistance or adaptation of imbibed castor seeds to cold stress by the increased content of unsaturated fatty acid (UFA). The data has been deposited to the ProteomeXchange with identifier PXD010043.
PMID: 30654474
Int J Mol Sci , IF:4.556 , 2019 Jan , V20 (2) doi: 10.3390/ijms20020243
Quantitative Proteomic Analysis of the Response to Cold Stress in Jojoba, a Tropical Woody Crop.
College of Life and Environmental Sciences, Minzu University of China, Beijing 100081, China. gaofei@muc.edu.cn.; Beijing Key Laboratory of Gene Resource and Molecular Development, Beijing 100875, China. gaofei@muc.edu.cn.; College of Life and Environmental Sciences, Minzu University of China, Beijing 100081, China. s161088@muc.edu.cn.; College of Life and Environmental Sciences, Minzu University of China, Beijing 100081, China. 16051039@muc.edu.cn.; College of Life and Environmental Sciences, Minzu University of China, Beijing 100081, China. zhouyijun@muc.edu.cn.; Beijing Key Laboratory of Gene Resource and Molecular Development, Beijing 100875, China. gfzh@bnu.edu.cn.; College of Life Sciences, Beijing Normal University, Beijing 100875, China. gfzh@bnu.edu.cn.
Jojoba (Simmondsia chinensis) is a semi-arid, oil-producing industrial crop that have been widely cultivated in tropical arid region. Low temperature is one of the major environmental stress that impair jojoba's growth, development and yield and limit introduction of jojoba in the vast temperate arid areas. To get insight into the molecular mechanisms of the cold stress response of jojoba, a combined physiological and quantitative proteomic analysis was conducted. Under cold stress, the photosynthesis was repressed, the level of malondialdehyde (MDA), relative electrolyte leakage (REL), soluble sugars, superoxide dismutase (SOD) and phenylalanine ammonia-lyase (PAL) were increased in jojoba leaves. Of the 2821 proteins whose abundance were determined, a total of 109 differentially accumulated proteins (DAPs) were found and quantitative real time PCR (qRT-PCR) analysis of the coding genes for 7 randomly selected DAPs were performed for validation. The identified DAPs were involved in various physiological processes. Functional classification analysis revealed that photosynthesis, adjustment of cytoskeleton and cell wall, lipid metabolism and transport, reactive oxygen species (ROS) scavenging and carbohydrate metabolism were closely associated with the cold stress response. Some cold-induced proteins, such as cold-regulated 47 (COR47), staurosporin and temperature sensitive 3-like a (STT3a), phytyl ester synthase 1 (PES1) and copper/zinc superoxide dismutase 1, might play important roles in cold acclimation in jojoba seedlings. Our work provided important data to understand the plant response to the cold stress in tropical woody crops.
PMID: 30634475
Int J Mol Sci , IF:4.556 , 2019 Jan , V20 (1) doi: 10.3390/ijms20010144
Hydrogen Peroxide and Nitric Oxide Crosstalk Mediates Brassinosteroids Induced Cold Stress Tolerance in Medicago truncatula.
Ministry of Education Key Laboratory for Bio-Resource and Eco-Environment, College of Life Science, State Key Laboratory of Hydraulics and Mountain River Engineering, Sichuan University, Chengdu 610064, China. 2014521230002@stu.scu.edu.cn.; Ministry of Education Key Laboratory for Bio-Resource and Eco-Environment, College of Life Science, State Key Laboratory of Hydraulics and Mountain River Engineering, Sichuan University, Chengdu 610064, China. yuanmiao1892@163.com.; Ministry of Education Key Laboratory for Bio-Resource and Eco-Environment, College of Life Science, State Key Laboratory of Hydraulics and Mountain River Engineering, Sichuan University, Chengdu 610064, China. ljzou66@163.com.; Ministry of Education Key Laboratory for Bio-Resource and Eco-Environment, College of Life Science, State Key Laboratory of Hydraulics and Mountain River Engineering, Sichuan University, Chengdu 610064, China. loss0928@sina.com.; Ministry of Education Key Laboratory for Bio-Resource and Eco-Environment, College of Life Science, State Key Laboratory of Hydraulics and Mountain River Engineering, Sichuan University, Chengdu 610064, China. wenrongtan1112@163.com.; Ministry of Education Key Laboratory for Bio-Resource and Eco-Environment, College of Life Science, State Key Laboratory of Hydraulics and Mountain River Engineering, Sichuan University, Chengdu 610064, China. tzhu@ytu.edu.cn.; Ministry of Education Key Laboratory for Bio-Resource and Eco-Environment, College of Life Science, State Key Laboratory of Hydraulics and Mountain River Engineering, Sichuan University, Chengdu 610064, China. hhlin@scu.edu.cn.
Brassinosteroids (BRs) play pivotal roles in modulating plant growth, development, and stress responses. In this study, a Medicago truncatula plant pretreated with brassinolide (BL, the most active BR), enhanced cold stress tolerance by regulating the expression of several cold-related genes and antioxidant enzymes activities. Previous studies reported that hydrogen peroxide (H(2)O(2)) and nitric oxide (NO) are involved during environmental stress conditions. However, how these two signaling molecules interact with each other in BRs-induced abiotic stress tolerance remain largely unclear. BL-pretreatment induced, while brassinazole (BRZ, a specific inhibitor of BRs biosynthesis) reduced H(2)O(2) and NO production. Further, application of dimethylthiourea (DMTU, a H(2)O(2) and OH(-) scavenger) blocked BRs-induced NO production, but BRs-induced H(2)O(2) generation was not sensitive to 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (PTIO, a scavenger of NO). Moreover, pretreatment with DMTU and PTIO decreased BL-induced mitochondrial alternative oxidase (AOX) and the photosystem capacity. However, pretreatment with PTIO was found to be more effective than DMTU in reducing BRs-induced increases in Valt, Vt, and MtAOX1 gene expression. Similarly, BRs-induced photosystem II efficiency was found in NO dependent manner than H(2)O(2). Finally, we conclude that H(2)O(2) was involved in NO generation, whereas NO was found to be crucial in BRs-induced AOX capacity, which further contributed to the protection of the photosystem under cold stress conditions in Medicago truncatula.
PMID: 30609774
Int J Mol Sci , IF:4.556 , 2019 Jan , V20 (1) doi: 10.3390/ijms20010208
Global Phosphoproteomic Analysis Reveals the Defense and Response Mechanisms of Jatropha Curcas Seedling under Chilling Stress.
Key Laboratory of Plant Resources, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China. huiliu@ibcas.ac.cn.; Key Laboratory of Plant Resources, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China. wangfenfen@ibcas.ac.cn.; Key Laboratory of Plant Resources, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China. pengxianjun@ibcas.ac.cn.; Key Laboratory of Plant Resources, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China. jhhuang@ibcas.ac.cn.; Key Laboratory of Plant Resources, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China. shshen@ibcas.ac.cn.
As a promising energy plant for biodiesel, Jatropha curcas is a tropical and subtropical shrub and its growth is affected by one of major abiotic stress, chilling. Therefore, we adopt the phosphoproteomic analysis, physiological measurement and ultrastructure observation to illustrate the responsive mechanism of J. curcas seedling under chilling (4 degrees C) stress. After chilling for 6 h, 308 significantly changed phosphoproteins were detected. Prolonged the chilling treatment for 24 h, obvious physiological injury can be observed and a total of 332 phosphoproteins were examined to be significantly changed. After recovery (28 degrees C) for 24 h, 291 phosphoproteins were varied at the phosphorylation level. GO analysis showed that significantly changed phosphoproteins were mainly responsible for cellular protein modification process, transport, cellular component organization and signal transduction at the chilling and recovery periods. On the basis of protein-protein interaction network analysis, phosphorylation of several protein kinases, such as SnRK2, MEKK1, EDR1, CDPK, EIN2, EIN4, PI4K and 14-3-3 were possibly responsible for cross-talk between ABA, Ca(2+), ethylene and phosphoinositide mediated signaling pathways. We also highlighted the phosphorylation of HOS1, APX and PIP2 might be associated with response to chilling stress in J. curcas seedling. These results will be valuable for further study from the molecular breeding perspective.
PMID: 30626061
Mar Drugs , IF:4.073 , 2019 Jan , V17 (1) doi: 10.3390/md17010038
Asperlin Stimulates Energy Expenditure and Modulates Gut Microbiota in HFD-Fed Mice.
Pharmacology and Toxicology Research Center, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China. cmwu@implad.ac.cn.; Pharmacology and Toxicology Research Center, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China. xiaoyuzhou5213@sina.com.; Pharmacology and Toxicology Research Center, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China. qi940201@163.com.; State Key Laboratory of Natural and Biomimetic Drugs, Peking University, Beijing 100191, China. liudong_1982@126.com.; Pharmacology and Toxicology Research Center, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China. snow20150@163.com.; Pharmacology and Toxicology Research Center, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China. yujiaqi_2018@163.com.; Pharmacology and Toxicology Research Center, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China. 18302458685@163.com.; State Key Laboratory of Natural and Biomimetic Drugs, Peking University, Beijing 100191, China. whlin@bjmu.edu.cn.; Pharmacology and Toxicology Research Center, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China. pguo@implad.ac.cn.
Asperlin is a marine-derived, natural product with antifungal, anti-inflammatory and anti-atherosclerotic activities. In the present study, we showed that asperlin effectively prevented the development of obesity in high-fat diet (HFD)-fed mice. Oral administration of asperlin for 12 weeks significantly suppressed HFD-induced body weight gain and fat deposition without inhibiting food intake. Hyperlipidemia and liver steatosis were also substantially ameliorated. A respiratory metabolism monitor showed that asperlin efficiently increased energy expenditure and enhanced thermogenic gene expression in adipose tissue. Accordingly, asperlin-treated mice showed higher body temperature and were more tolerant of cold stress. Meanwhile, asperlin also increased the diversity and shifted the structure of gut microbiota. Oral administration of asperlin markedly increased the relative abundance of Bacteroidetes, leading to a higher Bacteroidetes-to-Fimicutes ratio. The HFD-induced abnormalities at both phylum and genus levels were all remarkably recovered by asperlin. These results demonstrated that asperlin is effective in preventing HFD-induced obesity and modulating gut microbiota. Its anti-obesity properties may be attributed to its effect on promoting energy expenditure.
PMID: 30634484
Plant Cell Physiol , IF:4.062 , 2019 Jan , V60 (1) : P7-18 doi: 10.1093/pcp/pcy209
Cold Influences Male Reproductive Development in Plants: A Hazard to Fertility, but a Window for Evolution.
College of Life Sciences, South-Central University for Nationalities, Wuhan, China.; School of Integrative Plant Science, Cornell University, Ithaca, NY, USA.; Experiment Center of Forestry in North China, Chinese Academy of Forestry, Beijing, China.; School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China.; Department of Plants and Crops, unit HortiCell, Faculty of Bioscience Engineering, University of Ghent, Ghent, Belgium.
Being sessile organisms, plants suffer from various abiotic stresses including low temperature. In particular, male reproductive development of plants is extremely sensitive to cold which may dramatically reduce viable pollen shed and plant fertility. Cold stress disrupts stamen development and prominently interferes with the tapetum, with the stress-responsive hormones ABA and gibberellic acid being greatly involved. In particular, low temperature stress delays and/or inhibits programmed cell death of the tapetal cells which consequently damages pollen development and causes male sterility. On the other hand, studies in Arabidopsis and crops have revealed that ectopically decreased temperature has an impact on recombination and cytokinesis during meiotic cell division, implying a putative role for temperature in manipulating plant genomic diversity and architecture during the evolution of plants. Here, we review the current understanding of the physiological impact of cold stress on the main male reproductive development processes including tapetum development, male meiosis and gametogenesis. Moreover, we provide insights into the genetic factors and signaling pathways that are involved, with putative mechanisms being discussed.
PMID: 30602022
Plant Cell Physiol , IF:4.062 , 2019 Jan , V60 (1) : P152-165 doi: 10.1093/pcp/pcy197
Variation in ICE1 Methylation Primarily Determines Phenotypic Variation in Freezing Tolerance in Arabidopsis thaliana.
Weed Research Laboratory, Nanjing Agricultural University, Nanjing, China.
Cold stress is a major abiotic factor plants face during their life cycle. Although plants often exhibit phenotypic variation in cold tolerance, the underlying mechanism remains poorly understood. In the present study, the 50% lethal temperature (LT50) values of 37 Arabidopsis thaliana accessions at latitudes from 15 degrees to 58 degrees ranged from -13.2 degrees C to -4.9 degrees C and were closely correlated with the cold climates of the collection sites. According to a methylation analysis of all C-repeat (CRT)-binding factor (CBF) pathway genes, the coding and promoter regions of AtICE1, a regulator of CBF genes, exhibited the greatest variability in methylation levels among the accessions and included 5-122 methylated cytosine residues. In contrast, unmethylated or only slightly methylated genes in the CBF pathway showed little variation among the accessions. According to a gene expression analysis of four selected A. thaliana populations with distinct methylation patterns, except for the down-regulated gene AtCBF2, the expression levels of all members of the CBF pathway were negatively correlated with AtICE1 gene methylation levels. Treatment of the four A. thaliana populations with the DNA methylation inhibitory reagent 5-azacytidine resulted in a 30.0-78.3% enhancement of freezing tolerance and decreases in LT50 values of approximately 1.9-3.6 degrees C. Similar effects were observed in drm2 mutants, including 30.0-48.3% increases in freezing tolerance and decreases in LT50 values of approximately 0.7-3.4 degrees C. Thus, the AtICE1 methylation-regulated transcription of CBF pathway genes is responsible for the phenotypic variation in the freezing tolerance observed in A. thaliana.
PMID: 30295898
Sci Rep , IF:3.998 , 2019 Jan , V9 (1) : P863 doi: 10.1038/s41598-018-37165-7
The increase of photosynthetic carbon assimilation as a mechanism of adaptation to low temperature in Lotus japonicus.
Instituto de Investigaciones Biotecnologicas-Instituto Tecnologico de Chascomus (IIB-INTECH), UNSAM-CONICET, Buenos Aires, Argentina.; Institut de Biologie Integrative de la Cellule (I2BC), CNRS, CEA, Universite Paris-Sud, Universite Paris-Saclay, Gif sur Yvette, France.; Departamento de Bioquimica y Biologia Molecular-Universitat de Valencia, Valencia, Spain.; Departamento de Bioquimica y Biologia Molecular-Universitat de Valencia, Valencia, Spain. carrascp@uv.es.; Instituto de Fisiologia y Recursos Geneticos Vegetales (IFRGV) Ing "Victorio S Trippi", Instituto Nacional de Tecnologia Agropecuaria (INTA), Cordoba, Argentina.
Low temperature is one of the most important factors affecting plant growth, it causes an stress that directly alters the photosynthetic process and leads to photoinhibition when severe enough. In order to address the photosynthetic acclimation response of Lotus japonicus to cold stress, two ecotypes with contrasting tolerance (MG-1 and MG-20) were studied. Their chloroplast responses were addressed after 7 days under low temperature through different strategies. Proteomic analysis showed changes in photosynthetic and carbon metabolism proteins due to stress, but differentially between ecotypes. In the sensitive MG-1 ecotype acclimation seems to be related to energy dissipation in photosystems, while an increase in photosynthetic carbon assimilation as an electron sink, seems to be preponderant in the tolerant MG-20 ecotype. Chloroplast ROS generation was higher under low temperature conditions only in the MG-1 ecotype. These data are consistent with alterations in the thylakoid membranes in the sensitive ecotype. However, the accumulation of starch granules observed in the tolerant MG-20 ecotype indicates the maintenance of sugar metabolism under cold conditions. Altogether, our data suggest that different acclimation strategies and contrasting chloroplast redox imbalance could account for the differential cold stress response of both L. japonicus ecotypes.
PMID: 30696867
BMC Genomics , IF:3.594 , 2019 Jan , V20 (1) : P33 doi: 10.1186/s12864-018-5395-1
Banana sRNAome and degradome identify microRNAs functioning in differential responses to temperature stress.
Key Laboratory of Plant Resources Conservation and Sustainable Utilization, Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou, 510650, China. zhuhong@scbg.ac.cn.; Key Laboratory of Plant Resources Conservation and Sustainable Utilization, Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou, 510650, China.; University of Chinese Academy of Sciences, Beijing, 100049, China.
BACKGROUND: Temperature stress is a major environmental factor affecting not only plant growth and development, but also fruit postharvest life and quality. MicroRNAs (miRNAs) are a class of non-coding small RNAs that play important roles in various biological processes. Harvested banana fruit can exhibit distinct symptoms in response to different temperature stresses, but the underlying miRNA-mediated regulatory mechanisms remained unknown. RESULTS: Here, we profiled temperature-responsive miRNAs in banana, using deep sequencing and computational and molecular analyses. In total 113 known miRNAs and 26 novel banana-specific miRNAs were identified. Of these miRNAs, 42 miRNAs were expressed differentially under cold and heat stresses. Degradome sequencing identified 60 target genes regulated by known miRNAs and half of these targets were regulated by 15 temperature-responsive miRNAs. The correlative expression patterns between several miRNAs and their target genes were further validated via qRT-PCR. Our data showed that miR535 and miR156 families may derive from a common ancestor during evolution and jointly play a role in fine-tuning SPL gene expression in banana. We also identified the miRNA-triggered phased secondary siRNAs in banana and found miR393-TIR1/AFB phasiRNA production displaying cold stress-specific enrichment. CONCLUSIONS: Our results provide a foundation for understanding the miRNA-dependent temperature stress response in banana. The characterized correlations between miRNAs and their response to temperature stress could serve as markers in the breeding programs or tools for improving temperature tolerance of banana.
PMID: 30630418
BMC Genomics , IF:3.594 , 2019 Jan , V20 (1) : P93 doi: 10.1186/s12864-019-5454-2
Genome-wide characterization and expression profiling of SWEET genes in cabbage (Brassica oleracea var. capitata L.) reveal their roles in chilling and clubroot disease responses.
Jiangsu Key Laboratory for Horticultural Crop Genetic Improvement, Institute of Vegetable Crops, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, People's Republic of China.; Zhenjiang Agricultural Research Institute, Jurong, Jiangsu, 212400, People's Republic of China.; Boyce Thompson Institute, Cornell University, Ithaca, NY, 14853, USA.; Boyce Thompson Institute, Cornell University, Ithaca, NY, 14853, USA. zf25@cornell.edu.; Jiangsu Key Laboratory for Horticultural Crop Genetic Improvement, Institute of Vegetable Crops, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, People's Republic of China. jbli@jaas.ac.cn.
BACKGROUND: The SWEET proteins are a group of sugar transporters that play a role in sugar efflux during a range of biological processes, including stress responses. However, there has been no comprehensive analysis of the SWEET family genes in Brassica oleracea (BoSWEET), and the evolutionary pattern, phylogenetic relationship, gene characteristics of BoSWEET genes and their expression patterns under biotic and abiotic stresses remain largely unexplored. RESULTS: A total of 30 BoSWEET genes were identified and divided into four clades in B. oleracea. Phylogenetic analysis of the BoSWEET proteins indicated that clade II formed first, followed by clade I, clade IV and clade III, successively. Clade III, the newest clade, shows signs of rapid expansion. The Ks values of the orthologous SWEET gene pairs between B. oleracea and Arabidopsis thaliana ranged from 0.30 to 0.45, which estimated that B. oleracea diverged from A. thaliana approximately 10 to 15 million years ago. Prediction of transmembrane regions showed that eight BoSWEET proteins contain one characteristic MtN3_slv domain, twenty-one contain two, and one has four. Quantitative reverse transcription-PCR (qRT-PCR) analysis revealed that five BoSWEET genes from clades III and IV exhibited reduced expression levels under chilling stress. Additionally, the expression levels of six BoSWEET genes were up-regulated in roots of a clubroot-susceptible cabbage cultivar (CS-JF1) at 7 days after inoculation with Plasmodiophora brassicae compared with uninoculated plants, indicating that these genes may play important roles in transporting sugars into sink roots associated with P. brassicae colonization in CS-JF1. Subcellular localization analysis of a subset of BoSWEET proteins indicated that they are localized in the plasma membrane. CONCLUSIONS: This study provides important insights into the evolution of the SWEET gene family in B. oleracea and other species, and represents the first study to characterize phylogenetic relationship, gene structures and expression patterns of the BoSWEET genes. These findings provide new insights into the complex transcriptional regulation of BoSWEET genes, as well as potential candidate BoSWEET genes that promote sugar transport to enhance chilling tolerance and clubroot disease resistance in cabbage.
PMID: 30696401
Plant Sci , IF:3.591 , 2019 Jan , V278 : P20-31 doi: 10.1016/j.plantsci.2018.10.009
A DREB1 gene from zoysiagrass enhances Arabidopsis tolerance to temperature stresses without growth inhibition.
College of Life & Environmental Science, Minzu University of China, Beijing, PR China.; College of Life & Environmental Science, Minzu University of China, Beijing, PR China. Electronic address: Wei.s.j@163.com.
The DREB (dehydration-responsive element binding) protein family comprises transcription factors that can increase the survivability of a plant under abiotic stresses by regulating expression of multiple genes and altering downstream metabolism at the cost of growth retardation and developmental delay. In this study, a gene for the DREB1-type transcription factor, designated ZjDREB1.4, was isolated from zoysiagrass (Zoysia japonica Steud.), a popular warm-season turfgrass. This gene contains a conserved AP2/ERF DNA-binding domain flanking the signature sequence of DREB1 and belongs to a DREB1 branch in the grass family that expands in the warm-season species. The expression of ZjDREB1.4 was significantly induced by chilling stress (4-15 degrees C), moderately induced by salt stress, and only slightly induced by drought stress. The product of ZjDREB1.4 was targeted to the nucleus and showed strong transactivation activity but weak binding to the DRE with ACCGAC as the core sequence. The ZjDREB1.4 protein bound to GCCGAC more preferentially than to ACCGAC. Overexpression of ZjDREB1.4 in Arabidopsis induced the expression of multiple genes including a part of the CBF-regulon, and moderately increased the levels of proline and soluble sugars under normal growth conditions. The transgenic Arabidopsis plants showed an increase in tolerance to high and freezing temperature stresses without obvious growth inhibition and with only a few days delay in bolting. ZjDREB1.4 is potentially useful for producing transgenic plants that are tolerant to high temperature and/or cold stresses with few negative effects.
PMID: 30471726
BMC Plant Biol , IF:3.497 , 2019 Jan , V19 (1) : P43 doi: 10.1186/s12870-019-1646-9
Physiological and iTRAQ-based proteomic analyses reveal the function of exogenous gamma-aminobutyric acid (GABA) in improving tea plant (Camellia sinensis L.) tolerance at cold temperature.
College of Horticulture, Nanjing Agricultural University, No.1 Weigang, Nanjing, Jiangsu Province, 210095, People's Republic of China.; Wuxi NextCODE Genomics, 288 Fute Zhong Road, Shanghai, 200131, People's Republic of China.; College of Horticulture, Nanjing Agricultural University, No.1 Weigang, Nanjing, Jiangsu Province, 210095, People's Republic of China. fangwp@njau.edu.cn.
BACKGROUND: Internal gamma-Aminobutyric Acid (GABA) interacting with stress response substances may be involved in the regulation of differentially abundant proteins (DAPs) associated with optimum temperature and cold stress in tea plants (Camellia sinensis (L.) O. Kuntze). RESULTS: Tea plants supplied with or without 5.0 mM GABA were subjected to optimum or cold temperatures in this study. The increased GABA level induced by exogenous GABA altered levels of stress response substances - such as glutamate, polyamines and anthocyanins - in association with improved cold tolerance. Isobaric tags for relative and absolute quantification (iTRAQ) - based DAPs were found for protein metabolism and nucleotide metabolism, energy, amino acid transport and metabolism other biological processes, inorganic ion transport and metabolism, lipid metabolism, carbohydrate transport and metabolism, biosynthesis of secondary metabolites, antioxidant and stress defense. CONCLUSIONS: The iTRAQ analysis could explain the GABA-induced physiological effects associated with cold tolerance in tea plants. Analysis of functional protein-protein networks further showed that alteration of endogenous GABA and stress response substances induced interactions among photosynthesis, amino acid biosynthesis, and carbon and nitrogen metabolism, and the corresponding differences could contribute to improved cold tolerance of tea plants.
PMID: 30700249
Planta , IF:3.39 , 2019 Jan , V249 (1) : P235-249 doi: 10.1007/s00425-018-3054-z
Overexpression of geraniol synthase induces heat stress susceptibility in Nicotiana tabacum.
Department of Biological Science and Technology, Faculty of Industrial Science and Technology, Tokyo University of Science, 6-3-1 Niijuku, Katsushika-ku, Tokyo, 125-8585, Japan.; Iwate Biotechnology Research Center, Kitakami, Iwate, 024-0003, Japan.; Department of Life Sciences and Systems Biology, Plant Physiology Unit, University of Turin, Via Quarello15/A, I-10135, Turin, Italy.; College of Agriculture, Academic Institute, Shizuoka University, Shizuoka, 422-8529, Japan.; Research Institute of Green Science and Technology, Shizuoka University, Shizuoka, 422-8529, Japan.; Center for Ecological Research, Kyoto University, Otsu, 520-2113, Japan.; Department of Biological Science and Technology, Faculty of Industrial Science and Technology, Tokyo University of Science, 6-3-1 Niijuku, Katsushika-ku, Tokyo, 125-8585, Japan. garimura@rs.tus.ac.jp.
MAIN CONCLUSION: Transgenic tobacco plants overexpressing the monoterpene alcohol geraniol synthase exhibit hypersensitivity to thermal stress, possibly due to suppressed sugar metabolism and transcriptional regulation of genes involved in thermal stress tolerance. Monoterpene alcohols function in plant survival strategies, but they may cause self-toxicity to plants due to their hydrophobic and highly reactive properties. To explore the role of these compounds in plant stress responses, we assessed transgenic tobacco plants overexpressing the monoterpene alcohol geraniol synthase (GES plants). Growth, morphology and photosynthetic efficiency of GES plants were not significantly different from those of control plants (wild-type and GUS-transformed plants). While GES plants' direct defenses against herbivores or pathogens were similar to those of control plants, their indirect defense (i.e., attracting herbivore enemy Nesidiocoris tenuis) was stronger compared to that of control plants. However, GES plants were susceptible to cold stress and even more susceptible to extreme heat stress (50 degrees C), as shown by decreased levels of sugar metabolites, invertase activity and its products (Glc and Fru), and leaf starch granules. Moreover, GES plants showed decreased transcription levels of the WRKY33 transcription factor gene and an aquaporin gene (PIP2). The results of this study show that GES plants exhibit enhanced indirect defense ability against herbivores, but conversely, GES plants exhibit hypersensitivity to heat stress due to suppressed sugar metabolism and gene regulation for thermal stress tolerance.
PMID: 30478473
J Plant Physiol , IF:3.013 , 2019 Jan , V232 : P188-199 doi: 10.1016/j.jplph.2018.12.001
Seasonal changes in cold hardiness and carbohydrate metabolism in four garden rose cultivars.
Department of Plants and Crops, Ghent University, Coupure links 653, 9000 Gent, Belgium.; ILVO, Plant Sciences Unit, Caritasstraat 39, 9090 Melle, Belgium.; Department of Plants and Crops, Ghent University, Coupure links 653, 9000 Gent, Belgium. Electronic address: MarieChristine.VanLabeke@UGent.be.
We studied metabolic adaptations to cold stress in roses and identified genes in the carbohydrate pathway during acclimation and deacclimation. A field experiment with four rose cultivars belonging to different USDA plant hardiness zones was set up in Melle, Belgium (51 degrees 0' N, 3 degrees 48' E). The more cold-hardy cultivars ('Dagmar Hastrup' and 'John Cabot') reached their lowest LT50 value in December, indicating a rapid acclimation after the first occurrence of frost. Less cold-hardy cultivars ('Abraham Darby' and 'Chandos Beauty') reached their lowest LT50 in January/February when exposed to prolonged freezing temperatures. A cell dehydration pattern was found in the less cold-hardy cultivars 'Abraham Darby' and 'Chandos Beauty'. The expression of dehydrins (RhDHN5 and RhDNH6) was up-regulated during November-January. Carbohydrate metabolism is highly involved in cold acclimation in roses. Starch decreased from November towards January in all four cultivars and the hydrolysis of starch by the beta-amylolytic pathway (BAM, DPE2) was identified in 'Dagmar Hastrup' from November to January. Oligosaccharides correlated with cold hardiness in three cultivars although no significant upregulation in RhMIPS and RhRS6, key genes in their biosynthesis, was found. Higher sucrose levels were found during acclimation in hardy cultivars, although transcript levels of RhINV2 was more prominent in 'Chandos Beauty'.
PMID: 30537606
Biochem Biophys Res Commun , IF:2.985 , 2019 Jan , V509 (1) : P148-153 doi: 10.1016/j.bbrc.2018.12.092
A wheat GTP-binding protein like gene reduces tolerance to low temperature in Arabidopsis.
School of Life Science, Qufu Normal University, Qufu, Shandong, 273165, PR China.; School of Life Science, Qufu Normal University, Qufu, Shandong, 273165, PR China. Electronic address: SYG-0423@163.com.
Low temperature adversely affects plant growth and crop yield. The studies largely focus on cold stress (<4 degrees C), while the response upon low temperature higher than 4 degrees C is rarely documented so far. Here, we isolate a GTP-binding protein beta subunit like gene TaGPBL. TaGPBL is responsive to low temperature of 16 degrees C, and its ectopic overexpression in Arabidopsis results in more remarkable growth restriction under 16 degrees C, but has no effect under 22 degrees C. TaGBPL overexpression reduces the induction of cold-inducible genes and the activities of ROS scavengers and producers in lower temperature dependent manner. The data indicate that TaGBPL participates in the response to low temperature, which provides evidence for deepening our insight into the role of G-protein in temperature perception and signaling transduction.
PMID: 30579594
Free Radic Res , IF:2.839 , 2019 Jan , V53 (1) : P45-56 doi: 10.1080/10715762.2018.1548767
Cold stress on Araucaria angustifolia embryogenic cells results in oxidative stress and induces adaptation: implications for conservation and propagation.
a Department of Biochemistry and Molecular Biology , Federal University of Parana , Curitiba , Brazil.
Araucaria angustifolia (Bert.) O. Kuntze is a species critically endangered of extinction and its development and propagation is strongly affected by abiotic stress. We have previously shown the activation of uncoupling protein in A. angustifolia embryogenic stem cells subjected to cold stress. Now, we have furthered those studies by exposing these cells to cold stress (4 +/- 1 degrees C for either 24 or 48 h) and evaluating parameters associated with oxidative stress and alterations in the cellular and mitochondrial responses. Cold stress affect the H2O2 levels and lipid peroxidation increased after both stress condition, an effect associated with the decrease in the activities of peroxidases, catalase and ascorbate/dehydroascorbate ratio. On the other hand, the activities of ascorbate peroxidase, monodehydroascorbate and dehydroascorbate reductases increased as an indication of adaptation. Another important impact of cold stress conditions was the decrease of external alternative NAD(P)H dehydrogenases activity and the increase of mitochondrial mass. These results show that cold stress induces oxidative stress in A. angustifolia embryogenic cells, which results in activation of the glutathione-ascorbate cycle as a compensation for the decrease in the activities of catalase, peroxidases, and external NAD(P)H dehydrogenases. Our results contribute to the understanding of the pathways that gymnosperms employ to overcome oxidative stress, which must be explored in order to improve the methods of conservation and propagation of A. angustifolia.
PMID: 30764670
Funct Plant Biol , IF:2.617 , 2019 Jan , V47 (1) : P80-90 doi: 10.1071/FP19163
A new morphological method to identify cold tolerance of melon at seedling stage.
Key Laboratory of Protected Horticulture of Education Ministry and Liaoning Province, College of Horticulture, Shenyang Agricultural University, National and Local Joint Engineering Research Centre of Northern Horticultural, Facilities Design and Application Technology (Liaoning), Shenyang, 110866, Liaoning, PR China.; Key Laboratory of Protected Horticulture of Education Ministry and Liaoning Province, College of Horticulture, Shenyang Agricultural University, National and Local Joint Engineering Research Centre of Northern Horticultural, Facilities Design and Application Technology (Liaoning), Shenyang, 110866, Liaoning, PR China; and Corresponding author. Email: qihongyan@syau.edu.cn.
Low temperature restrains the growth and development of melons, as well as severely impairing the yield and quality. To obtain a rapid and accurate method for evaluating cold tolerance of melon, 10 genotypes were selected to investigate their cold tolerance at seedling stage. Chilling stress (15 degrees C/6 degrees C, day/night) increased leaf angles and caused leaves wilted: the phenotypes of the 10 genotypes were obviously different. Thus, a new predicted method for chilling injury index (CII) of melon was constructed based on the change of leaf angle and leaf state. The CII showed significant correlation with survival rate, maximum photochemical quantum yield of PSII (Fv/Fm) and changes of SPAD value. Moreover, the validity of the method was further verified by seedlings growth, photosynthesis, membrane permeability and metabolites accumulation of four screened genotypes. Taken together, this work provides a morphological and accurate method for evaluating cold tolerance in melon.
PMID: 31813411
Plant Biol (Stuttg) , IF:2.167 , 2019 Jan , V21 Suppl 1 : P77-83 doi: 10.1111/plb.12893
The Arabidopsis THADA homologue modulates TOR activity and cold acclimation.
Centre for Organismal Studies (COS), University of Heidelberg, Heidelberg, Germany.; German Cancer Research Center (DKFZ), Heidelberg, Germany.; Forschungszentrum Julich, Julich, Germany.
Low temperature is one of the most important environmental factors that affect global survival of humans and animals and equally importantly the distribution of plants and crop productivity. Survival of metazoan cells under cold stress requires regulation of the sensor-kinase Target Of Rapamycin (TOR). TOR controls growth of eukaryotic cells by adjusting anabolic and catabolic metabolism. Previous studies identified the Thyroid Adenoma Associated (THADA) gene as the major effect locus by positive selection in the evolution of modern human adapted to cold. Here we investigate the role of THADA in TOR signaling and cold acclimation of plants. We applied BLAST searches and homology modeling to identify the AtTHADA (AT3G55160) in Arabidopsis thaliana as the highly probable orthologue protein. Reverse genetics approaches were combined with immunological detection of TOR activity and metabolite profiling to address the role of the TOR and THADA for growth regulation and cold acclimation. Depletion of the AtTHADA gene caused complete or partial loss of full-length mRNA, respectively, and significant retardation of growth under non-stressed conditions. Furthermore, depletion of AtTHADA caused hypersensitivity towards low-temperatures. Atthada displayed a lowered energy charge. This went along with decreased TOR activity, which offers a molecular explanation for the slow growth phenotype of Atthada. Finally, we used TOR RNAi lines to identify the de-regulation of TOR activity as one determinant for sensitivity towards low-temperatures. Taken together our results provide evidence for a conserved function of THADA in cold acclimation of eukaryotes and suggest that cold acclimation in plants requires regulation of TOR.
PMID: 30098100
Plant Direct , IF:1.725 , 2019 Jan , V3 (1) : Pe00104 doi: 10.1002/pld3.104
Classifying cold-stress responses of inbred maize seedlings using RGB imaging.
Department of Plant and Microbial Biology University of Minnesota St. Paul Minnesota.; Donald Danforth Plant Science Center St. Louis Missouri.; Present address: Bayer U.S. Crop Science St. Louis Missouri.; Department of Botany University of Wisconsin-Madison Madison Wisconsin.; Department of Plant Pathology University of Minnesota St. Paul Minnesota.
Increasing the tolerance of maize seedlings to low-temperature episodes could mitigate the effects of increasing climate variability on yield. To aid progress toward this goal, we established a growth chamber-based system for subjecting seedlings of 40 maize inbred genotypes to a defined, temporary cold stress while collecting digital profile images over a 9-daytime course. Image analysis performed with PlantCV software quantified shoot height, shoot area, 14 other morphological traits, and necrosis identified by color analysis. Hierarchical clustering of changes in growth rates of morphological traits and quantification of leaf necrosis over two time intervals resulted in three clusters of genotypes, which are characterized by unique responses to cold stress. For any given genotype, the set of traits with similar growth rates is unique. However, the patterns among traits are different between genotypes. Cold sensitivity was not correlated with the latitude where the inbred varieties were released suggesting potential further improvement for this trait. This work will serve as the basis for future experiments investigating the genetic basis of recovery to cold stress in maize seedlings.
PMID: 31245751
Gene Expr Patterns , IF:0.897 , 2019 Jan , V31 : P7-17 doi: 10.1016/j.gep.2018.10.002
Analysis of Brassica napus dehydrins and their Co-Expression regulatory networks in relation to cold stress.
Department of Plant Biotechnology, Faculty of Agriculture, University of Guilan, Rasht, Iran.; Department of Plant Biotechnology, Faculty of Agriculture, University of Guilan, Rasht, Iran. Electronic address: hsamizadeh@guilan.ac.ir.; Department of Cell and Molecular Biology, Faculty of Biological Sciences and Biotechnology, Shahid Beheshti University. G.C., Evin, Tehran, Iran. Electronic address: n_farrokhi@sbu.ac.ir.
Dehydrins (DHNs) are plant specific cold and drought stress-responsive proteins that belong to late embryogenesis abundant (LEA) protein families. B. napus DHNs (BnDHNs) were computationally analyzed to establish gene regulatory- and protein-protein interaction networks. Promoter analyses suggested functionality of phytohormones in BnDHNs gene network. The relative expressions of some BnDHNs were analyzed using qRT-PCR in seedling leaves of both cold-tolerant (Zarfam) and -sensitive (Sari Gul) canola treated/untreated by cold. Our expression data were indicative of the importance of BnDHNs in cold tolerance in Zarfam. BnDHNs were classified into three classes according to the expression pattern. Moreover, expression of three BnDHN types, SKn (BnLEA10 and BnLEA18), YnKn (BnLEA90) and YnSKn (BnLEA104) were significantly high in the tolerant cultivar at 12h of cold treatment. Our findings put forward the possibility of considering these genes as screening biomarker to determine cold-tolerant breeding lines; something that needs to be further corroborated. Furthermore, these genes may have some implications in developing such tolerant lines via transgenesis.
PMID: 30408599
Zhongguo Zhong Yao Za Zhi , 2019 Jan , V44 (2) : P293-297 doi: 10.19540/j.cnki.cjcmm.20181106.005
[Cloning and function analysis of promoter of DcCDPK8 from Dendrobium catenatum].
Dendrobium State Forestry Engineering Research Center,State Key Laboratory of Subtropical Silviculture,Zhejiang A & F Universiy Lin'an 311300,China.
DcCDPK8 involved in abiotic stress such as low temperature and signal transduction of hormones ABA and MeJA,but the transcriptional regulation is still unclear. In order to study the core promoter region of DcCDPK8 gene in Dendrobium catenatum and explore its transcriptional regulation mechanism,the DcCDPK8 gene promoter sequence was cloned by PCR from D. catenatum. Promoter sequence function was studied by fusion of 5 'terminal deletion and GUS gene. The results showed that the promoter sequence of DcCDPK8 gene has a low-temperature responsive element( LTR) between~(-1) 749 bp and-614 bp,two MeJA responsive elements between~(-1) 749 bp and-230 bp,and one ABA responsive elements between-614 bp and-230 bp. Three 5'-end different deletion fragments were constructed to fuse the eukaryotic expression vectors p BI121 with GUS,which were transformed into tobacco leaves. The GUS activity under cold stress treatment was DcCDPK8-p1>DcCDPK8-p2>DcCDPK8-p3. GUS activity under exogenous ABA induction was DcCDPK8-p1>DcCDPK8-p2>DcCDPK8-p3,and GUS activity under exogenous MeJA induction was DcCDPK8-p1>DcCDPK8-p2>DcCDPK8-p3. It is speculated that the ABA response element( ARE) in the promoter sequences of DcCDPK8 is positive regulatory role in response to exogenous ABA,the MeJA cis-acting element plays a negative role in response to exogenous MeJA.
PMID: 30989948