Overexpression of PtHMGR enhances the drought and salt tolerance of poplar.

BACKGROUND AND AIMS Soil salinization and aridification are swiftly engulfing the limited land resources on which humans depend, restricting agricultural production. HMGR is important in terpenoid biosynthesis, and terpenoids are involved in plant growth, development, and responses to environmental stresses. This study aimed providing some guidance for obtaining salt and drought resistant poplar. METHODS Protein expression system was used to obtain PtHMGR protein, and high performance liquid chromatography was used to detect the activity of PtHMGR protein in vitro. In addition, a simplified version of the leaf infection method was used for transformation of Nanlin895 poplar (Populus× euramericana cv.). qRT-PCR was used to identify expression levels of genes. KEY RESULTS PtHMGR catalyzed a reaction involving HMG-CoA and NADPH to form mevalonate. Overexpression of PtHMGR in Populus × euramericana cv Nanlin895 improved drought and salinity tolerance. In the presence of NaCl and PEG6000, the rates of rooting and survival of PtHMGR-overexpressing poplars were higher than those of wild type poplars. The transgenic lines also exhibited higher proline content, and peroxidase and superoxide dismutase activities, and a lower malondialdehyde level under osmotic stress. In addition, the expression of reactive oxygen species scavenging and formation-related genes was altered by osmotic stress. Moreover, the effect of osmotic stress on the transcript levels of stress-related genes differed between the transgenic and wild type poplars. CONCLUSION PtHMGR catalyzed a reaction involving HMG-CoA and NADPH to form mevalonate in vitro. Overexpression of PtHMGR promoted root development, increased the expression of ROS scavenging-related genes, decreased the expression of ROS formation-related genes, and increased the activity of antioxidant enzymes in the transgenic poplars, enhancing their tolerance to osmotic stress. In addition, overexpression of PtHMGR increased expression of the stress-related genes KIN1, COR15, and AAO3, and decreased that of ABI, MYB, MYC2, and RD22, enhancing the stress resistance of poplar.

[1]  Kezhong Zhang,et al.  Analysis of Key Enzyme Genes in Carotenoid Metabolism Pathway of Lilium and Cloning of LoLcyB , 2019, Molecular Plant Breeding.

[2]  T. Yin,et al.  Functional analyses of NDPK2 in Populus trichocarpa and overexpression of PtNDPK2 enhances growth and tolerance to abiotic stresses in transgenic poplar. , 2017, Plant physiology and biochemistry : PPB.

[3]  R. M. Rivero,et al.  Reactive oxygen species, abiotic stress and stress combination. , 2017, The Plant journal : for cell and molecular biology.

[4]  V. Singh,et al.  Microbial modulation of bacoside A biosynthetic pathway and systemic defense mechanism in Bacopa monnieri under Meloidogyne incognita stress , 2017, Scientific Reports.

[5]  A. Hofmann,et al.  Targeted and Untargeted Approaches Unravel Novel Candidate Genes and Diagnostic SNPs for Quantitative Resistance of the Potato (Solanum tuberosum L.) to Phytophthora infestans Causing the Late Blight Disease , 2016, PloS one.

[6]  L. Zheng,et al.  Cloning and characterization of an elicitor-responsive gene encoding 3-hydroxy-3-methylglutaryl coenzyme A reductase involved in 20-hydroxyecdysone production in cell cultures of Cyanotis arachnoidea. , 2014, Plant physiology and biochemistry : PPB.

[7]  Jörg-Peter Schnitzler,et al.  Plant volatiles and the environment. , 2014, Plant, cell & environment.

[8]  Xue-duan Liu,et al.  Cloning and characterization of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) gene from Paris fargesii Franch. , 2014, Indian Journal of Biochemistry & Biophysics.

[9]  Wei Liu,et al.  Species-Specific Expansion and Molecular Evolution of the 3-hydroxy-3-methylglutaryl Coenzyme A Reductase (HMGR) Gene Family in Plants , 2014, PloS one.

[10]  R. Finkelstein,et al.  Abscisic Acid Synthesis and Response , 2013, The arabidopsis book.

[11]  P. Trivedi,et al.  Cloning and functional characterization of 3-hydroxy-3-methylglutaryl coenzyme A reductase gene from Withania somnifera: an important medicinal plant , 2013, Protoplasma.

[12]  W. Boland,et al.  Plant defense against herbivores: chemical aspects. , 2012, Annual review of plant biology.

[13]  M. J. Chalmers,et al.  Molecular Mimicry Regulates ABA Signaling by SnRK2 Kinases and PP2C Phosphatases , 2012, Science.

[14]  Jie Ren,et al.  Suppression of 9-cis-Epoxycarotenoid Dioxygenase, Which Encodes a Key Enzyme in Abscisic Acid Biosynthesis, Alters Fruit Texture in Transgenic Tomato1[W][OA] , 2012, Plant Physiology.

[15]  M. J. Chalmers,et al.  Structural basis for basal activity and autoactivation of abscisic acid (ABA) signaling SnRK2 kinases , 2011, Proceedings of the National Academy of Sciences.

[16]  R. K. Kar.  Plant responses to water stress: Role of reactive oxygen species , 2011, Plant signaling & behavior.

[17]  A. Ferrer,et al.  Modulation of plant HMG-CoA reductase by protein phosphatase 2A , 2011, Plant signaling & behavior.

[18]  H. Miziorko.  Enzymes of the mevalonate pathway of isoprenoid biosynthesis. , 2011, Archives of biochemistry and biophysics.

[19]  Shufeng Zhou,et al.  Cloning and characterization of a novel 3-hydroxy-3-methylglutaryl coenzyme A reductase gene from Salvia miltiorrhiza involved in diterpenoid tanshinone accumulation. , 2011, Journal of plant physiology.

[20]  Robert D Hall,et al.  Plant molecular stress responses face climate change. , 2010, Trends in plant science.

[21]  F. Loreto,et al.  Abiotic stresses and induced BVOCs. , 2010, Trends in plant science.

[22]  C. Dai,et al.  Molecular cloning, characterization and function analysis of the gene encoding HMG-CoA reductase from Euphorbia Pekinensis Rupr , 2010, Molecular Biology Reports.

[23]  S. Cutler,et al.  In vitro Reconstitution of an ABA Signaling Pathway , 2009, Nature.

[24]  Kazuki Saito,et al.  Functional genomics for plant natural product biosynthesis. , 2009, Natural product reports.

[25]  P. Leng,et al.  Cloning and functional analysis of 9-cis-epoxycarotenoid dioxygenase (NCED) genes encoding a key enzyme during abscisic acid biosynthesis from peach and grape fruits. , 2009, Journal of plant physiology.

[26]  R. Utsumi,et al.  Diversity, regulation, and genetic manipulation of plant mono- and sesquiterpenoid biosynthesis , 2009, Cellular and Molecular Life Sciences.

[27]  P. McCourt,et al.  Abscisic Acid Inhibits Type 2C Protein Phosphatases via the PYR/PYL Family of START Proteins , 2009, Science.

[28]  Claudia E Vickers,et al.  A unified mechanism of action for volatile isoprenoids in plant abiotic stress. , 2009, Nature chemical biology.

[29]  Hongyan Yao,et al.  Molecular cloning and functional analysis of the gene encoding 3-hydroxy-3-methylglutaryl coenzyme A reductase from hazel (Corylus avellana L. Gasaway). , 2007, Journal of biochemistry and molecular biology.

[30]  Jianhua Zhang,et al.  Abscisic acid is a key inducer of hydrogen peroxide production in leaves of maize plants exposed to water stress. , 2006, Plant & cell physiology.

[31]  M. Gribskov,et al.  The Genome of Black Cottonwood, Populus trichocarpa (Torr. & Gray) , 2006, Science.

[32]  E. Grill,et al.  Generation of Active Pools of Abscisic Acid Revealed by In Vivo Imaging of Water-Stressed Arabidopsis1 , 2005, Plant Physiology.

[33]  J. Pereira,et al.  Understanding plant responses to drought - from genes to the whole plant. , 2003, Functional plant biology : FPB.

[34]  Jian-Kang Zhu,et al.  Cell Signaling during Cold, Drought, and Salt Stress Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.000596. , 2002, The Plant Cell Online.

[35]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[36]  S. Ha,et al.  Molecular characterization of Hmg2 gene encoding a 3-hydroxy-methylglutaryl-CoA reductase in rice. , 2001, Molecules and cells.

[37]  P. León,et al.  1-Deoxy-d-xylulose-5-phosphate Synthase, a Limiting Enzyme for Plastidic Isoprenoid Biosynthesis in Plants* , 2001, The Journal of Biological Chemistry.

[38]  J. Schwender,et al.  Chlorophyta exclusively use the 1-deoxyxylulose 5-phosphate/2-C-methylerythritol 4-phosphate pathway for the biosynthesis of isoprenoids , 2001, Planta.

[39]  B. M. Lange,et al.  Isoprenoid biosynthesis: the evolution of two ancient and distinct pathways across genomes. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[40]  J. Zhu,et al.  Genetic analysis of plant salt tolerance using Arabidopsis. , 2000, Plant physiology.

[41]  Y. Leu,et al.  Tetranortriterpenoid insect antifeedants from Severinia buxifolia , 1997 .

[42]  Y. Yukimune,et al.  Methyl jasmonate-induced overproduction of paclitaxel and baccatin III in Taxus cell suspension cultures , 1996, Nature Biotechnology.

[43]  A. Boronat,et al.  Targeting and topology in the membrane of plant 3-hydroxy-3-methylglutaryl coenzyme A reductase. , 1995, The Plant cell.

[44]  D. Hardie,et al.  Bacterial expression of the catalytic domain of 3-hydroxy-3-methylglutaryl-CoA reductase (isoform HMGR1) from Arabidopsis thaliana, and its inactivation by phosphorylation at Ser577 by Brassica oleracea 3-hydroxy-3-methylglutaryl-CoA reductase kinase. , 1995, European journal of biochemistry.

[45]  C. Lamb,et al.  Isolation of a monocot 3-hydroxy-3-methylglutaryl coenzyme A reductase gene that is elicitor-inducible , 1994, Plant Molecular Biology.

[46]  I. Maldonado-Mendoza,et al.  Nucleotide Sequence of a cDNA Encoding 3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase from Catharanthus roseus. , 1992, Plant physiology.

[47]  N. Chua,et al.  Characterization of cDNA and genomic clones encoding 3-hydroxy-3-methylglutaryl-coenzyme A reductase from Hevea brasiliensis , 1991, Plant Molecular Biology.

[48]  F. Hegardt,et al.  Isolation and structural characterization of a cDNA encoding Arabidopsis thaliana 3-hydroxy-3-methylglutaryl coenzyme A reductase , 1989, Plant Molecular Biology.

[49]  E. Ábrahám,et al.  Duplicated P5CS genes of Arabidopsis play distinct roles in stress regulation and developmental control of proline biosynthesis. , 2008, The Plant journal : for cell and molecular biology.

[50]  T. Koshiba,et al.  Complex regulation of ABA biosynthesis in plants. , 2002, Trends in plant science.

[51]  D. Wititsuwannakul,et al.  3-Hydroxy-3-methylglutaryl coenzyme A reductase from the latex of Hevea brasiliensis. , 1990 .