Control of glycerol biosynthesis under high salt stress in Arabidopsis
Ahmed Bahieldin A B G , Jamal S. M. Sabir A , Ahmed Ramadan A C , Ahmed M. Alzohairy D , Rania A. Younis B , Ahmed M. Shokry A C , Nour O. Gadalla A E , Sherif Edris A B F , Sabah M. Hassan A B , Magdy A. Al-Kordy A E , Khalid B. H. Kamal A , Samar Rabah A , Osama A. Abuzinadah A and Fotouh M. El-Domyati A BA Department of Biological Sciences, Faculty of Science, King Abdulaziz University (KAU), PO Box 80141, Jeddah 21589, Saudi Arabia.
B Department of Genetics, Faculty of Agriculture, Ain Shams University, Cairo, Egypt.
C Agricultural Genetic Engineering Research Institute (AGERI), Agriculture Research Center (ARC), Giza, Egypt.
D Genetics Department, Faculty of Agriculture, Zagazig University, Zagazig 44511, Egypt.
E Genetics and Cytology Department, Genetic Engineering and Biotechnology Division, National Research Center, Dokki, Egypt.
F Princess Al-Jawhara Al-Brahim Centre of Excellence in Research of Hereditary Disorders (PACER-HD), Faculty of Medicine, King Abdulaziz University (KAU), Jeddah, Saudi Arabia.
G Corresponding author. Email: bahieldin55@gmail.com
Functional Plant Biology 41(1) 87-95 https://doi.org/10.1071/FP13005
Submitted: 4 February 2013 Accepted: 27 June 2013 Published: 24 July 2013
Abstract
Loss-of-function and gain-of-function approaches were utilised to detect the physiological importance of glycerol biosynthesis during salt stress and the role of glycerol in conferring salt tolerance in Arabidopsis. The salt stress experiment involved wild type (WT) and transgenic Arabidopsis overexpressing the yeast GPD1 gene (analogue of Arabidopsis GLY1 gene). The experiment also involved the Arabidopsis T-DNA insertion mutants gly1 (for suppression of glycerol 3-phosphate dehydrogenase or G3PDH), gli1 (for suppression of glycerol kinase or GK), and act1 (for suppression of G3P acyltransferase or GPAT). We evaluated salt tolerance levels, in conjunction with glycerol and glycerol 3-phosphate (G3P) levels and activities of six enzymes (G3PDH, ADH (alcohol dehydrogenase), ALDH (aldehyde dehydrogenase), GK, G3PP (G3P phosphatase) and GLYDH (glycerol dehydrogenase)) involved in the glycerol pathway. The GPD1 gene was used to overexpress G3PDH, a cytosolic NAD+-dependent key enzyme of cellular glycerol biosynthesis essential for growth of cells under abiotic stresses. T2 GPD1-transgenic plants and those of the two mutants gli1 and act1 showed enhanced salt tolerance during different growth stages as compared with the WT and gly1 mutant plants. These results indicate that the participation of glycerol, rather than G3P, in salt tolerance in Arabidopsis. The results also indicate that the gradual increase in glycerol levels in T2 GPD1-transgenic, and gli1 and act1 mutant plants as NaCl level increases whereas they dropped at 200 mM NaCl. However, the activities of the G3PDH, GK, G3PP and GLYDH at 150 and 200 mM NaCl were not significantly different. We hypothesise that mechanism(s) of glycerol retention/efflux in the cell are affected at 200 mM NaCl in Arabidopsis.
Additional keywords: abiotic stress, osmoprotection, T-DNA insertion.
References
Albertyn J, Hohmann S, Prior BA (1994) Characterization of the osmotic stress response in Saccharomyces cerevisiae, osmotic stress and glucose repression regulate glycerol-3-phosphate dehydrogenase independently. Current Genetics 25, 12–18.| Characterization of the osmotic stress response in Saccharomyces cerevisiae, osmotic stress and glucose repression regulate glycerol-3-phosphate dehydrogenase independently.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2cXksFWnt7s%3D&md5=f1160e84686a42345aaae049ff61711bCAS | 8082159PubMed |
Bechtold N, Ellis J, Pelletier G (1993) In planta Agrobacterium mediated gene transfer by infiltration of adult Arabidopsis thaliana plants. Comptes Rendus de l’Académie des Sciences 316, 1194–1199.
Benz BR, Rhode JM, Cruzan MB (2007) Aerenchyma development and elevated alcohol dehydrogenase activity as alternative responses to hypoxic soils in the Piriqueta caroliniana complex. American Journal of Botany 94, 542–550.
| Aerenchyma development and elevated alcohol dehydrogenase activity as alternative responses to hypoxic soils in the Piriqueta caroliniana complex.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXlt1Ggs70%3D&md5=d2c898638a929fbbfc2b0cef9a43178dCAS | 21636424PubMed |
Blomberg A, Adler L (1989) Roles of glycerol and glycerol-3-phosphate dehydrogenase (NAD+) in acquired osmotolerance of Saccharomyces cerevisiae. Journal of Bacteriology 171, 1087–1092.
Bohnert HJ, Su H, Shen B (1999) Molecular mechanisms of salinity tolerance. In ‘Cold, drought, heat, and salt stress: molecular responses in higher plants’. (Ed. K Shinozaki) pp. 29–60. (RG Landes: Austin, TX)
Bradford MM (1976) A rapid and sensitive for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248–254.
| A rapid and sensitive for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaE28XksVehtrY%3D&md5=bead43c1adb6f57157bea3eab849e63cCAS | 942051PubMed |
Brewster JL, de Valoir T, Dwyer ND, Winter E, Gustin MC (1993) An osmosensing signal transduction pathway in yeast. Science 259, 1760–1763.
| An osmosensing signal transduction pathway in yeast.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK3sXksVCqs7Y%3D&md5=dbad38c95cfc1cf288a3fcfecdf82400CAS | 7681220PubMed |
Cabello-Hurtado F, Ramos J (2004) Isolation and functional analysis of the glycerol permease activity of two new nodulin-like intrinsic proteins from salt stressed roots of the halophyte Atriplex nummularia. Plant Science 166, 633–640.
| Isolation and functional analysis of the glycerol permease activity of two new nodulin-like intrinsic proteins from salt stressed roots of the halophyte Atriplex nummularia.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXhtFWgsLc%3D&md5=c17b3ffa3b349b3a25379745232e711fCAS |
Chanda B, Venugopal SC, Kulshrestha S, Navarre DA, Downie B, Vaillancourt L, Kachroo A, Kachroo P (2008) Glycerol-3-phosphate levels are associated with basal resistance to the emibiotrophic fungus Colletotrichum higginsianum in Arabidopsis. Plant Physiology 147, 2017–2029.
| Glycerol-3-phosphate levels are associated with basal resistance to the emibiotrophic fungus Colletotrichum higginsianum in Arabidopsis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXhtVSrtrrI&md5=d419fb0cdd1c9e34948cad7b8eb67826CAS | 18567828PubMed |
Chen H, Lu Y, Jiang J-G (2012) Comparative analysis on the key enzymes of the glycerol cycle metabolic pathway in Dunaliella salina under osmotic stresses. PLoS ONE 7, e37578
| Comparative analysis on the key enzymes of the glycerol cycle metabolic pathway in Dunaliella salina under osmotic stresses.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XosFWrt7g%3D&md5=61b3b62809e622bcbf0225b5a9d92f08CAS | 22675484PubMed |
Christensen AH, Quail P (1996) Ubiquitin promoter-based vectors for high level expression of selectable and/or screenable marker genes in monocotyledonous plant. Transgenic Research 5, 213–218.
| Ubiquitin promoter-based vectors for high level expression of selectable and/or screenable marker genes in monocotyledonous plant.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK28XjsFamsbo%3D&md5=b42afa42c807b08971648de7a2bc4008CAS | 8673150PubMed |
Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation, Version II. Plant Molecular Biology Reporter 1, 19–21.
| A plant DNA minipreparation, Version II.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL2cXksFWhtrk%3D&md5=cb2ccf6382903bcf5f3af1622173bd02CAS |
Eastmond PJ (2004) Glycerol-insensitive Arabidopsis mutants, gli seedlings lack glycerol kinase, accumulate glycerol and are more resistant to abiotic stress. The Plant Journal 37, 617–625.
| Glycerol-insensitive Arabidopsis mutants, gli seedlings lack glycerol kinase, accumulate glycerol and are more resistant to abiotic stress.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXisVWhtr8%3D&md5=b15e74b493d1c531dee00e63943d6d13CAS | 14756771PubMed |
Edris S, Mutwakil MHZ, Sabir JSM, Ramadan AM, Gadalla NO, Shokry AM, Al-Kordy MA, Abo-Aba SEM, El-Domyati FM, Bahieldin A (2012) Identification of GPD1 gene from yeast via fluorescence differential display-polymerase chain reaction (FDD-PCR). African Journal of Biotechnology 11, 8653–8664.
Gerbeau P, Güçlü J, Ripoche P, Maurel C (1999) Aquaporin Nt-TIPa can account for the high permeability of tobacco cell vacuolar membrane to small neutral solutes. The Plant Journal 18, 577–587.
| Aquaporin Nt-TIPa can account for the high permeability of tobacco cell vacuolar membrane to small neutral solutes.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXlsFGhsbw%3D&md5=26ae554b9676fbef78954842dee2e344CAS | 10417709PubMed |
Gomez KA, Gomez AA (1984) ‘Statistical procedures for agricultural research.’ 2nd edn. (John Wiley & Sons: New York)
Gunde-Cimerman N, Ramos J, Plemenitas A (2009) Halotolerant and halophilic fungi. Mycological Research 113, 1231–1241.
| Halotolerant and halophilic fungi.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXkvF2nug%3D%3D&md5=da60ec1347e2ea6edb042958a2d255ecCAS | 19747974PubMed |
Hohmann S (2009) Control of high osmolarity signaling in the yeast Saccharomyces cerevisiae. FEBS Letters 583, 4025–4029.
| Control of high osmolarity signaling in the yeast Saccharomyces cerevisiae.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXhsFams7fI&md5=e1e2790318067184296d51efbbe3376cCAS | 19878680PubMed |
Horie T, Kaneko T, Sugimoto G, Sasano S, Panda SK, Shibasaka M, Katsuhara M (2011) Mechanisms of water transport mediated by PIP aquaporins and their regulation via phosphorylation events under salinity stress in barley roots. Plant & Cell Physiology 52, 663–675.
| Mechanisms of water transport mediated by PIP aquaporins and their regulation via phosphorylation events under salinity stress in barley roots.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXksFKju7s%3D&md5=a3127f9bb00c4c30834f87753d5e3ccfCAS |
Hubmann G, Guillouet S, Nevoigt E (2011) Gpd1 and Gpd2 fine-tuning for sustainable reduction of glycerol formation in Saccharomyces cerevisiae. Applied and Environmental Microbiology 77, 5857–5867.
| Gpd1 and Gpd2 fine-tuning for sustainable reduction of glycerol formation in Saccharomyces cerevisiae.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXht1OjurvN&md5=78b0973e161d38a9c3a18eacbd586316CAS | 21724879PubMed |
Klepek YS, Geiger D, Stadler R, Klebl F, Landouar-Arsivaud L, Lemoine R, Hedrich R, Sauer N (2005) Arabidopsis POLYOL TRANSPORTER5, a new member of the monosaccharide transporter-like superfamily, mediates H+-symport of numerous substrates, including myo-inositol, glycerol, and ribose. The Plant Cell 17, 204–218.
| Arabidopsis POLYOL TRANSPORTER5, a new member of the monosaccharide transporter-like superfamily, mediates H+-symport of numerous substrates, including myo-inositol, glycerol, and ribose.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXotlantg%3D%3D&md5=ba67114fb70fcc193280645ed8a92fcfCAS | 15598803PubMed |
Kunst L, Browse J, Somerville C (1988) Altered regulation of lipid biosynthesis in a mutant of Arabidopsis deficient in chloroplast glycerol-3-phosphate acyltransferase activity. Proceedings of the National Academy of Sciences of the United States of America 85, 4143–4147.
| Altered regulation of lipid biosynthesis in a mutant of Arabidopsis deficient in chloroplast glycerol-3-phosphate acyltransferase activity.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL1cXkvV2ru7Y%3D&md5=5fcd1886003fb5bd07c69698af2cc3fbCAS | 16593939PubMed |
Lawrence CL, Botting CH, Antrobus R, Coote PJ (2004) Evidence of a new role for the high-osmolarity glycerol mitogen-activated protein kinase pathway in yeast: regulating adaptation to citric acid stress. Molecular and Cellular Biology 24, 3307–3323.
| Evidence of a new role for the high-osmolarity glycerol mitogen-activated protein kinase pathway in yeast: regulating adaptation to citric acid stress.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXjtFKjtrc%3D&md5=07550b279555da0e251dfb19b576654cCAS | 15060153PubMed |
Lee HK, Cho SK, Son O, Xu Z, Hwang I, Kim WT (2009) Drought stress-induced Rma1H1, a RING membrane-anchor E3 ubiquitin ligase homolog, regulates aquaporin levels via ubiquitination in transgenic Arabidopsis plants. The Plant Cell 21, 622–641.
| Drought stress-induced Rma1H1, a RING membrane-anchor E3 ubiquitin ligase homolog, regulates aquaporin levels via ubiquitination in transgenic Arabidopsis plants.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXksVKrt7o%3D&md5=a23aa29d71f90027b590a4160762d8deCAS | 19234086PubMed |
Li SY, Gomelsky M, Duan J, Zhang Z, Gomelsky L, Zhang X, Epstein PN, Ren J (2004) Overexpression of Aldehyde Dehydrogenase-2 (ALDH2) Transgene prevents acetaldehyde-induced cell injury in human umbilical vein endothelial cells. The Journal of Biological Chemistry 279, 11 244–11 252.
| Overexpression of Aldehyde Dehydrogenase-2 (ALDH2) Transgene prevents acetaldehyde-induced cell injury in human umbilical vein endothelial cells.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXitFyhsLY%3D&md5=180732e03bc70adeca04bafbaec15266CAS |
Liu WZ, Faber R, Feese M, Remington SJ, Pettigrew DW (1994) Escherichia coli glycerol kinase, role of a tetramer interface in regulation by fructose 1,6-biphosphate and phosphotransferase system regulatory protein IIIglc. Biochemistry 33, 10 120–10 126.
| Escherichia coli glycerol kinase, role of a tetramer interface in regulation by fructose 1,6-biphosphate and phosphotransferase system regulatory protein IIIglc.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2cXltVarsLg%3D&md5=30ffb6bf324c9d4730a7c2f825343dd3CAS |
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2–ΔΔ C t method. Methods 25, 402–408.
| Analysis of relative gene expression data using real-time quantitative PCR and the 2–ΔΔ C t method.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XhtFelt7s%3D&md5=cda22da73d6034ad2e4c74827f633baaCAS | 11846609PubMed |
Luyten K, Albertyn J, Skibbe WF, Prior BA, Ramos J, Thevelein JM, Hohmann S (1995) Fps1, a yeast member of the MIP family of channel proteins, is a facilitator for glycerol uptake and efflux and is inactive under osmotic stress. EMBO Journal 14, 1360–1371.
Mandel MK, Chanda B, Xia Y, Yu K, Sekine K-T, Gao Q-M, Selote D, Kachroo A, Kachroo P (2011) Glycerol-3-phosphate and systemic immunity. Plant Signaling & Behavior 6, 1–4.
Melamed D, Pnueli L, Arava Y (2008) Yeast translational response to high salinity: global analysis reveals regulation at multiple levels. RNA 14, 1337–1351.
| Yeast translational response to high salinity: global analysis reveals regulation at multiple levels.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXot1amtL8%3D&md5=c8314ef0aa8f40af7c39b3a7678f1c2dCAS | 18495938PubMed |
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum 15, 473–497.
| A revised medium for rapid growth and bio assays with tobacco tissue cultures.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaF3sXksFKm&md5=00e945f2329f915ef516a05110061063CAS |
Padamsee M, Kumar TKA, Riley R, Binder M, Boyd A, Calvo AM, Furukawa K, Hesse C, Hohmann S, James TY, LaButti K, Lapidus A, Lindquist E, Lucas S, Miller K, Shantappa S, Grigoriev IV, Hibbett DS, McLaughlin DJ, Spatafora JW, Aime MC (2012) The genome of the xerotolerant mold Wallemia sebi reveals adaptations to osmotic stress and suggests cryptic sexual reproduction. Fungal Genetics and Biology 49, 217–226.
| The genome of the xerotolerant mold Wallemia sebi reveals adaptations to osmotic stress and suggests cryptic sexual reproduction.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XjslemsLs%3D&md5=3d828d2d3527a693b629e38cc161ce6cCAS | 22326418PubMed |
Påhlman A-K, Granath K, Ansell R, Hohmann S, Adler L (2001) The yeast glycerol 3-phosphatases Gpp1p and Gpp2p are required for glycerol biosynthesis and differentially involved in the cellular responses to osmotic, anaerobic, and oxidative stress. Journal of Biological Chemistry 276, 3555–3563.
| The yeast glycerol 3-phosphatases Gpp1p and Gpp2p are required for glycerol biosynthesis and differentially involved in the cellular responses to osmotic, anaerobic, and oxidative stress.Crossref | GoogleScholarGoogle Scholar | 11058591PubMed |
Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Research 29, e45
| A new mathematical model for relative quantification in real-time RT-PCR.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD38nis12jtw%3D%3D&md5=aa80c9eea3a1151f965b4bfe903672daCAS | 11328886PubMed |
Rawls KS, Martin JH, Maupin-Furlow JA (2011) Activity and transcriptional regulation of bacterial protein-like glycerol-3-phosphate dehydrogenase of the haloarchaea in Haloferax volcanii. Journal of Bacteriology 193, 4469–4476.
| Activity and transcriptional regulation of bacterial protein-like glycerol-3-phosphate dehydrogenase of the haloarchaea in Haloferax volcanii.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXhtV2gtbrK&md5=9f4b0a83750a7f3baa39f91806602818CAS | 21725010PubMed |
Ruzheinikov SN, Burke J, Sedelnikova S, Baker PJ, Taylor R, Bullough PA, Muir NM, Gore MG, Rice DW (2001) Glycerol dehydrogenase, structure, specificity, and mechanism of a family III polyol dehydrogenase. Structure 9, 789–802.
| Glycerol dehydrogenase, structure, specificity, and mechanism of a family III polyol dehydrogenase.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXntVajsrY%3D&md5=56116602632265f85b7691285b5ae209CAS | 11566129PubMed |
Schubert D, Lechtenberg B, Forsbach A, Gils M, Bahadur S, Schmidt R (2004) Silencing in Arabidopsis T-DNA transformants, the predominant role of a gene-specific RNA sensing mechanism versus position effects. The Plant Cell 16, 2561–2572.
| Silencing in Arabidopsis T-DNA transformants, the predominant role of a gene-specific RNA sensing mechanism versus position effects.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXptVSiu7o%3D&md5=898d022c275dda1543852e6b0c9d74d3CAS | 15367719PubMed |
Shen W, Wei Y, Dauk M, Tan Y, Taylor DC, Selvaraj G, Zou J (2006) Involvement of a glycerol-3-phosphate dehydrogenase in modulating the NADH/NAD+ ratio provides evidence of a mitochondrial glycerol-3-phosphate shuttle in Arabidopsis. The Plant Cell 18, 422–441.
| Involvement of a glycerol-3-phosphate dehydrogenase in modulating the NADH/NAD+ ratio provides evidence of a mitochondrial glycerol-3-phosphate shuttle in Arabidopsis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28Xhs1ensr4%3D&md5=b5314890660680a171580d9f7ff8ec48CAS | 16415206PubMed |
Venugopal SC, Chanda B, Vaillancourt L, Kachroo A, Kachroo P (2009) The common metabolite glycerol-3-phosphate is a novel regulator of plant defense signaling. Plant Signaling & Behavior 4, 746–749.
| The common metabolite glycerol-3-phosphate is a novel regulator of plant defense signaling.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXhs1Knsbc%3D&md5=4d2e63645c86056130c7bba475b3ca92CAS |
Wei Y, Shen W, Dauk M, Wang F, Selvaraj G, Zou J (2004) Targeted gene disruption of glycerol-3-phosphate dehydrogenase in Colletotrichum gloeosporioides reveals evidence that glycerol is a significant transferred nutrient from host plant to fungal pathogen. Journal of Biological Chemistry 279, 429–435.
| Targeted gene disruption of glycerol-3-phosphate dehydrogenase in Colletotrichum gloeosporioides reveals evidence that glycerol is a significant transferred nutrient from host plant to fungal pathogen.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXhtVSqt7%2FL&md5=630027f9446ebaf8cc141fc2e4f7c2a2CAS | 14563847PubMed |