Abstract
Sterol glycosyltransferases (SGTs) catalyze the transfer of sugar molecules to diverse sterol molecules, leading to a change in their participation in cellular metabolism. Withania somnifera is a medicinal plant rich in sterols, sterol glycosides and steroidal lactones. Sterols and their modified counterparts are medicinally important and play a role in adaptation of the plant to stress conditions. We have identified 3 members of SGT gene family through RACE (Rapid Amplification of cDNA Ends) in addition to sgtl1 reported earlier. The amino acid sequence deduced from the ORF’s showed homology (45–67 %) to the reported plant SGTs. The expression of the genes was differentially modulated in different organs in W. somnifera and in response to external stimuli. Salicylic acid and methyl jasmonate treatments showed up to 10 fold increase in the expression of sgt genes suggesting their role in defense. The level of expression increased in heat and cold stress indicating the role of sterol modifications in abiotic stress. One of the members, was expressed in E. coli and the enzyme assay showed that the crude enzyme glycosylated stigmasterol. W. somnifera expresses a family of sgt genes and there is a functional recruitment of these genes under stress conditions. The genes which are involved in sterol modification are important in view of medicinal value and understanding stress.
Similar content being viewed by others
References
Matsuda H, Murakami T, Kishi A, Yoshikawa M (2001) Structures of withanosides I, II, III, IV, V, VI, and VII, new withanolide glycosides, from the roots of Indian Withania somnifera DUNAL and inhibitory activity for tachyphylaxis to clonidine in isolated guinea-pig ileum. Bioorg Med Chem 9:1499–1507
Zhao J, Nakamura N, Hattori M, Kuboyama T, Tohda C, Komatsu K (2002) Withanolide derivatives from the roots of Withania somnifera and their neurite outgrowth activities. Chem Pharm Bull (Tokyo) 50:760–765
Lim EK, Bowles DJ (2004) A class of plant glycosyltransferases involved in cellular homeostasis. EMBO J 23:2915–2922
Bowles D, Isayenkova J, Lim EK, Poppenberger B (2005) Glycosyltransferases: managers of small molecules. Curr Opin Plant Biol 8:254–263
Bowles EL, Poppenberger B, Vaistij F (2006) Glycosyltransferases of lipophilic small molecules. Annu Rev Plant Biol 57:567–597
Ikan R (1999) Naturally occurring glycosides. Wiley, Chichester, p 444
Pflugmacher S, Sandermann H (1998) Taxonomic distribution of plant glucosyltransferases acting on xenobiotics. Phytochemistry 49:507–511
Vogt T, Jones P (2000) Glycosyltransferases in plant natural product synthesis: characterization of a supergene family. Trends Plant Sci 5:380–386
Bartholomew D, Lau S, O’Keefe D, Rea P, Viitanen P (2002) Alternate energy-dependent pathways for the vacuolar uptake of glucose and glutathione conjugates. Plant Physiol 130:1562–1572
Lim D, Hou B, Jackson R, Bowles D (2004) Arabidopsis glycosyltransferases as biocatalysts in fermentation for regioselective synthesis of diverse quercetin glucosides. Biotechnol Bioeng 87:623–631
Sharma LK, Madina BR, Chaturvedi P, Sangwan RS, Tuli R (2007) Molecular cloning and characterization of one member of 3beta-hydroxy sterol glucosyltransferase gene family in Withania somnifera. Arch Biochim Biophys 460:48–55
Madina BR, Sharma LK, Chaturvedi P, Sangwan RS, Tuli R (2007) Purification and physico-kinetic characterization of 3beta-hydroxy specific sterol glucosyltransferase from Withania somnifera (L) and its stress response. Biochim Biophys Acta 1774:392–402
Glombitza S, Dubuis PH, Thulke O, Welzl G, Bovet L, Gotz M, Affenzeller M, Geist B, Hehn A, Asnaghi C, Ernst D, Seidlitz HK, Gundlach H, Mayer KF, Martinoia E, Werck-Reichhart D, Mauch F, Schaffner AR (2004) Crosstalk and differential response to abiotic and biotic stressors reflected at the transcriptional level of effector genes from secondary metabolism. Plant Mol Biol 54:817–835
Price KR, Johnson IT, Fenwick GR (1987) The chemistry and biological significance of saponins in foods and feedingstuffs. Crit Rev Food Sci Nutr 26:27–135
Hostettmann K, Marston A (1995) Saponins: chemistry and pharmacology of natural products. Cambridge University Press, Cambridge
Osbourn A (1996) Saponins and plant defense—a soap story. Trends Plant Sci 1:4–9
Bouarab RM, Peart J, Baulcombe D, Osbourn A (2002) A saponin-detoxifying enzyme mediates suppression of plant defenses. Nature 418:889–892
Ghosal S, Kaur R, Bhattacharya SK (1988) Chemistry and bioactivity of sitoindosides IX and X. Planta Med 54:561
Yamada H, Nishizawa M, Katayama C (1992) Osladin, a sweet principle of Polypodium vulgare, structure revision. Tetrahedron Lett 33:4009–4010
Osbourn A (1996) Preformed antimicrobial compounds and plant defense against fungal attack. Plant Cell 8:1821–1831
Kohara A, Nakajima C, Hashimoto K, Ikenaga T, Tanaka H, Shoyama Y, Yoshida S, Muranaka T (2005) A novel glucosyltransferase involved in steroid saponin biosynthesis in Solanum aculeatissimum. Plant Mol Biol 57:225–239
Moehs CP, Allen PV, Friedman M, Belknap WR (1997) Cloning and expression of solanidine UDP-glucose glucosyltransferase from potato. Plant J 11:227–236
Chong J, Baltz R, Schmitt C, Beffa R, Fritig B, Saindrenan P (2002) Downregulation of a pathogen-responsive tobacco UDP-Glc: phenylpropanoid glucosyltransferase reduces scopoletin glucoside accumulation, enhances oxidative stress, and weakens virus resistance. Plant Cell 14:1093–1107
Jayaprakasam B, Nair M (2003) Cyclooxygenase-2 enzyme inhibitory withanolides from Withania somnifera leaves. Tetrahedron 59:841–849
Ghosal S, Lal J, Radheyshyam S (1989) Immunomodulatory and CNS effect of sitonidosides IX and X, two new glycowithanolides from Withania somnifera. Phytother Res 3:201–206
Bhattacharya SK, Satyan KS, Ghosal S (1997) Antioxidant activity of glycowithanolides from Withania somnifera. Indian J Exp Biol 35:236–239
Bhattacharya A, Ramanathan M, Ghosal S, Bhattacharya SK (2000) Effect of Withania somnifera glycowithanolides on iron-induced hepatotoxicity in rats. Phytother Res 14:568–570
Madina BR, Sharma LK, Chaturvedi P, Sangwan RS, Tuli R (2007) Purification and characterization of a novel glucosyltransferase specific to 27beta-hydroxy steroidal lactones from Withania somnifera and its role in stress responses. Biochim Biophys Acta 1774:1199–1207
Achnine L, Huhman DV, Farag MA, Sumner LW, Blount JW, Dixon RA (2005) Genomics-based selection and functional characterization of triterpene glycosyltransferases from the model legume Medicago truncatula. Plant J 41:875–887
Walters D, Hampson M, Mcpherson A (1993) The induction of systemic resistance in barley to powdery mildew infection using salicylates and various phenolic acids. Ann Appl Biol 122:451–456
Gonzalez-Aguilar GA, Tiznado-Hernandez ME, Zavaleta-Gatica R, Martinez-Tellez MA (2004) Methyl jasmonate treatments reduce chilling injury and activate the defense response of guava fruits. Biochem Biophys Res Commun 313:694–701
Warnecke D, Erdmann R, Fahl A, Hube B, Muller F, Zank T, Zahringer U, Heinz E (1999) Cloning and functional expression of UGT genes encoding sterol glucosyltransferases from Saccharomyces cerevisiae, Candida albicans, Pichia pastoris, and Dictyostelium discoideum. J Biol Chem 274:13048–13059
Rieu I, Powers SJ (2009) Real-time quantitative RT-PCR: design, calculations, and statistics. Plant Cell 21:1031–1033
Warnecke DC, Baltrusch M, Buck F, Wolter FP, Heinz E (1997) UDP-glucose:sterol glucosyltransferase:cloning and functional expression in Escherichia coli. Plant Mol Biol 35:597–603
Vernooij B, Friedrich L, Morse A, Reist R, Kolditz-Jawhar R, Ward E, Uknes S, Kessmann H, Ryals J (1994) Salicylic acid is not the translocated signal responsible for inducing systemic acquired resistance but is required in signal transduction. Plant Cell 6:959–965
Frey S, Carver T (1998) Induction of systemic resistance in pea to pea powdery mildew by exogenous application of salicylic acid. J Phytopathol 146:239–245
Murakami-Murofushi K, Nishikawa K, Hirakawa E, Murofushi H (1997) Heat stress induces a glycosylation of membrane sterol in myxoamoebae of a true slime mold, Physarum polycephalum. J Biol Chem 272:486–489
Kunimoto S, Murofushi W, Kai H, Ishida Y, Uchiyama A, Kobayashi T, Kobayashi S, Murofushi H, Murakami-Murofushi K (2002) Steryl glucoside is a lipid mediator in stress-responsive signal transduction. Cell Struct Funct 27:157–162
Palta J (1980) Alterations in membrane transport properties by freezing injury in herbaceous plants: evidence against the rupture theory. Physiol Plant 50:169–175
Steponkus P (1984) Role of the plasma membrane in freezing injury and cold acclimation. Annu Rev Plant Physiol Plant Mol Biol 35:543–584
Thomashow MF (1998) Role of cold-responsive genes in plant freezing tolerance. Plant Physiol 118:1–8
Martin D, Tholl D, Gershenzon J, Bohlmann J (2002) Methyl jasmonate induces traumatic resin ducts, terpenoid resin biosynthesis, and terpenoid accumulation in develo** xylem of Norway spruce stems. Plant Physiol 129:1003–1018
Kim HJ, Chen F, Wang X, Rajapakse NC (2006) Effect of methyl jasmonate on secondary metabolites of sweet basil (Ocimum basilicum L.). J Agric Food Chem 54:2327–2332
Hayashi H, Huang P, Inoue K (2003) Up-regulation of soyasaponin biosynthesis by methyl jasmonate in cultured cells of Glycyrrhiza glabra. Plant Cell Physiol 44:404–411
Aoyagi H, Kobayashi Y, Yamada K, Yokoyama M, Kusakari K, Tanaka H (2001) Efficient production of saikosaponins in Bupleurum falcatum root fragments combined with signal transducers. Appl Microbiol Biotechnol 57:482–488
Chaturvedi P, Misra P, Tuli R (2011) Sterol glycosyltransferases—the enzymes that modify sterols. Appl Biochem Biotechnol 165:47–68
Horvath DM, Chua NH (1996) Identification of an immediate-early salicylic acid-inducible tobacco gene and characterization of induction by other compounds. Plant Mol Biol 31:1061–1072
Horvath DM, Huang DJ, Chua NH (1998) Four classes of salicylate-induced tobacco genes. Mol Plant Microbe Interact 11:895–905
Fraissinet-Tachet L, Baltz R, Chong J, Kauffmann S, Fritig B, Saindrenan P (1998) Two tobacco genes induced by infection, elicitor and salicylic acid encode glucosyltransferases acting on phenylpropanoids and benzoic acid derivatives, including salicylic acid. FEBS Lett 437:319–323
Taguchi G, Ubukata T, Hayashida N, Yamamoto H, Okazaki M (2003) Cloning and characterization of a glucosyltransferase that reacts on 7-hydroxyl group of flavonol and 3-hydroxyl group of coumarin from tobacco cells. Arch Biochem Biophys 420:95–102
O’Donnell P, Truesdale M, Calvert C, Dorans A, Roberts M, Bowles D (1998) A novel tomato gene that rapidly responds to wound and pathogen-related signals. Plant J 14:137–142
Hlywka JJ, Stephenson GR, Sears MK, Yada RY (1994) Effect of insect damage on glycoalkaloid content in potato (Solanum tuberosum). J Agric Food Chem 42:2545–2550
Keukens EA, de Vrije T, van den Boom C, de Waard P, Plasman HH, Thiel F, Chupin V, Jongen WM, de Kruijff B (1995) Molecular basis of glycoalkaloid induced membrane disruption. Biochim Biophys Acta 1240:216–228
Chen J, Burke JJ, **n Z, Xu C, Velten J (2006) Characterization of the Arabidopsis thermosensitive mutant atts02 reveals an important role for galactolipids in thermotolerance. Plant Cell Environ 29:1437–1448
Kunimoto S, Kobayashi T, Kobayashi S, Murakami-Murofushi K (2000) Expression of cholesteryl glucoside by heat shock in human fibroblasts. Cell Stress Chaperones 5:3–7
Houde M, Belcaid M, Ouellet F, Danyluk J, Monroy AF, Dryanova A, Gulick P, Bergeron A, Laroche A, Links MG, MacCarthy L, Crosby WL, Sarhan F (2006) Wheat EST resources for functional genomics of abiotic stress. BMC Genom 7:149
Uemura M, Steponkus PL (1994) A contrast of the plasma membrane lipid composition of oat and rye leaves in relation to freezing tolerance. Plant Physiol 104:479–496
Anderson JV, Li QB, Haskell DW, Guy CL (1994) Structural organization of the spinach endoplasmic reticulum-luminal 70-kilodalton heat-shock cognate gene and expression of 70-kilodalton heat-shock genes during cold acclimation. Plant Physiol 104:1359–1370
Wang CY, Fung RW, Ding CK (2005) Reducing chilling injury and enhancing transcript levels of heat shock proteins, PR-proteins and alternative oxidase by methyl jasmonate and methyl salicylate in tomatoes and peppers. ISHS Acta Horticulturae, 682
Zhang C, Guy CL (2006) In vitro evidence of Hsc70 functioning as a molecular chaperone during cold stress. Plant Physiol Biochem 44:844–850
Suzuki H, Achnine L, Xu R, Matsuda SP, Dixon RA (2002) A genomics approach to the early stages of triterpene saponin biosynthesis in Medicago truncatula. Plant J 32:1033–1048
Suzuki H, Reddy MS, Naoumkina M, Aziz N, May GD, Huhman DV, Sumner LW, Blount JW, Mendes P, Dixon RA (2005) Methyl jasmonate and yeast elicitor induce differential transcriptional and metabolic re-programming in cell suspension cultures of the model legume Medicago truncatula. Planta 220:696–707
Choi DW, Jung J, Ha YI, Park HW, SuIn DS, Chung HJ, Liu JR (2005) Analysis of transcripts in methyl jasmonate-treated ginseng hairy roots to identify genes involved in the biosynthesis of ginsenosides and other secondary metabolites. Plant Cell Rep 23:557–566
Hu FX, Zhong JJ (2007) Role of jasmonic acid in alteration of ginsenoside heterogeneity in elicited cell cultures of Panax notoginseng. J Biosci Bioeng 104:513–516
Chen RJ, Chung TY, Li FY, Lin NH, Tzen JT (2009) Effect of sugar positions in ginsenosides and their inhibitory potency on Na(+)/K(+)-ATPase activity. Acta Pharmacol Sin 30:61–69
Morrissey J, Osbourn A (1999) Fungal resistance to plant antibiotics as a mechanism of pathogenesis. Microbiol Mol Biol Rev 63:708–724
Becker P, Weltring K (1998) Purification and characterization of α-chaconinase of Gibberella pulicaris. FEMS Microbiol Lett 167:197–202
Poppenberger B, Berthiller F, Lucyshyn D, Sieberer T, Schuhmacher R (2003) Detoxification of the Fusarium mycotoxin deoxynivalenol by a UDP-glucosyltransferase from Arabidopsis thaliana. J Biol Chem 278:47905–47914
Debolt S, Scheible WR, Schrick K, Auer M, Beisson F, Bischoff V, Bouvier-Nave P, Carroll A, Hematy K, Li Y, Milne J, Nair M, Schaller H, Zemla M, Somerville C (2009) Mutations in UDP-Glucose:sterol glucosyltransferase in Arabidopsis Cause transparent testa phenotype and suberization defect in seeds. Plant Physiol 151:78–87
Acknowledgments
We are grateful to the Council of Scientific and Industrial Research, Government of India for financial support to the project under New Millennium Indian Technology Leadership Initiative.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Chaturvedi, P., Mishra, M., Akhtar, N. et al. Sterol glycosyltransferases-identification of members of gene family and their role in stress in Withania somnifera . Mol Biol Rep 39, 9755–9764 (2012). https://doi.org/10.1007/s11033-012-1841-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11033-012-1841-3