Abstract
Withania somnifera (L.) Dunal (Family, Solanaceae), commonly known as Ashwagandha is one of the most valuable medicinal plants synthesizing large number of pharmacologically active secondary metabolites known as withanolides. Though the plant has been well characterized in terms of phytochemical profiles as well as pharmaceutical activities, not much is known about the genes responsible for biosynthesis of these compounds. In this study, we have characterized a gene encoding farnesyl diphosphate synthase (FPPS; EC 2.5.1.10), a key enzyme in the pathway of biosynthesis of isoprenoids, from W. somnifera. The full-length cDNA of Withania somnifera FPPS (WsFPPS) of 1,253 bps encodes a polypeptide of 343 amino acids. The amino acid sequence homology and phylogenetic analysis suggest that WsFPPS has close similarity to its counterparts from tomato (SlFPPS) and capsicum (CaaFPPS). Using semi quantitative RT–PCR, the expression pattern of the WsFPPS gene was analyzed in different tissues of Withania chemotypes (NMITLI-101, NMITLI-108, NMITLI-118 and NMITLI-135) as well as in response to elicitors (salicylic acid and methyl jasmonate) and mechanical wounding. The expression analysis suggests that WsFPPS expression varies in different tissues (with maximal expression in flower and young leaf) and chemotypes (with highest level in NMITLI-101) and was significantly elevated in response to salicylic acid, methyl jasmonate and mechanical injury. This is the first report on characterization of an isoprenoid pathway gene involved in withanolide biosynthesis.
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References
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410
Cervantes MC, Gallagher CE, Zhu C, Wurtzel ET (2006) Maize cDNAs expressed in endosperm encode functional farnesyl diphosphate synthase with geranylgeranyl diphosphate synthase. Plant Physiol 141:220–231
Chappell J (1995) The biochemistry and molecular biology of isoprenoid metabolism. Plant Physiol 107:1–6
Chatterjee S, Srivastava S, Khalid A, Singh N, Sangwan RS, Sidhu OP, Roy R, Khetrapal CL, Tuli R (2010) Comprehensive metabolic fingerprinting of Withania somnifera leaf and root extracts. Phytochemistry 71:1085–1094
Chaurasiya ND, Gupta VK, Sangwan RS (2007) Leaf ontogenic phase related dynamics of withaferin A and withanone biogenesis in Ashwagandha (Withania somnifera)- An important medicinal herb. J Plant Biol 50:508–513
Chaurasiya ND, Uniyal GC, Lal P, Misra L, Sangwan NS, Tuli R, Sangwan RS (2008) Analysis of withanolides in root and leaf of Withania somnifera by HPLC with photodiode array and evaporative light scattering detection. Phytochem Anal 19:148–154
Chaurasiya ND, Sangwan RS, Misra LN, Tuli R, Sangwan NS (2009) Metabolic clustering of a core collection of Indian ginseng Withania somnifera Dunal through DNA, isoenzyme, polypeptide and withanolide profile diversity. Fitoterapia 80:496–505
Closa M, Vranová E, Bortolotti C, Bigler L, Arró M, Ferrer A, Gruissem W (2010) The Arabidopsis thaliana FPP synthase isozymes have overlapping and specific functions in isoprenoid biosynthesis, and complete loss of FPP synthase activity causes early developmental arrest. Plant J 63:512–525
Felsenstein J (1989) PHYLIP—phylogeny inference package (Version 3.2). Cladistics 5:164–166
Kaileh M, Berghe WV, Boone E, Essawi T, Haegeman G (2007) Screening of indigenous Palestinian medicinal plants for potential anti-inflammatory and cytotoxic activity. J Ethn Pharmacol 113:510–516
Kim OT, Kim SH, Ohyama K, Muranaka T, Choi YE, Lee HY, Kim MY, Hwang B (2010) Upregulation of phytosterol and triterpene biosynthesis in Centella asiatica hairy roots overexpressed ginseng farnesyl diphosphate synthase. Plant Cell Rep 29:403–411
Koduru S, Kumar R, Srinivasan S, Evers MB, Damodaran C (2010) Notch-1 inhibition by Withaferin-A: a therapeutic target against colon carcinogenesis. Mol Cancer Ther 9:202–210
Lal P, Misra L, Sangwan RS, Tuli R (2006) New withanolides from fresh berries of Withania somnifera. Z Naturforsch 61b:1143–1147
Liao ZH, Chen M, Gong YF, Zuo KJ, Wang P, Tan F, Sun XF, Tang KX (2006) A new Farnesyl diphosphate synthase gene from Taxus media Rehder: cloning, characterization and functional complementation. J Integ Plant Biol 48:692–699
Madina BR, Sharma LK, Chaturvedi P, Sangwan RS, Tuli R (2007) Purification and characterization of a novel glucosyltransferase specific to 27β-hydroxy steroidal lactones from Withania somnifera and its role in stress responses. Biochim Biophys Act 1774:1199–1207
Matsushita Y, Kang W, Charlwood BV (1996) Cloning and analysis of a cDNA encoding farnesyl diphosphate synthase from Artemisia annua. Gene 172:207–209
Mirjalili MH, Moyano E, Bonfil M, Cusido RM, Palazon J (2009) Steroidal lactones from Withania somnifera, an ancient plant for novel medicine. Molecules 14:2373–2393
Misra L, Mishra P, Pandey A, Sangwan RS, Sangwan NS, Tuli R (2008) Withanolides from Withania somnifera roots. Phytochem 69:1000–1004
Misra P, Pandey A, Tiwari SK, Nath P, Trivedi PK (2010a) Characterization of isoflavone synthase gene from Psoralea corylifolia:a medicinal plant. Plant Cell Rep 29:747–755
Misra P, Pandey A, Tiwari M, Chandrashekar K, Sidhu OP, Asif MH, Chakrabarty D, Singh PK, Trivedi PK, Nath P, Tuli R (2010b) Modulation of transcriptome and metabolome of tobacco by Arabidopsis transcription factor, AtMYB12, leads to insect resistance. Plant Physiol 152:2258–2268
Pandey V, Misra P, Chaturvedi P, Mishra MK, Trivedi PK, Tuli R (2010) Agrobacterium tumefaciens-mediated transformation of Withania somnifera (L.) Dunal: an important medicinal plant. Plant Cell Rep 29:133–141
Sangwan RS, Chaurasiya ND, Misra LN, Lal P, Uniyal GC, Sharma R, Sangwan NS, Suri KA, Qazi GN, Tuli R (2004) Phytochemical variability in commercial herbal products and preparations of Withania somnifera (Ashwagandha). Curr Sci 86:461–465
Sangwan RS, Chaurasiya ND, Lal P, Misra L, Tuli R, Sangwan NS (2008) Root-contained withanolide A is inherently de novo synthesized within roots in Ashwagandha (Withania somnifera). Physiol Plant 133:278–287
Sanmiya K, Iwasaki T, Matsuoka M, Miyao M, Yamamoto N (1997) Cloning of a cDNA that encodes farnesyl diphosphate synthase and the blue-light-induced expression of the corresponding gene in the leaves of rice plants. Biochim Biophys Act 1350:240–246
Senthil K, Wasnik NG, Kim YJ, Yang DC (2010) Generation and analysis of expressed sequence tags from leaf and root of Withania somnifera (Ashwgandha). Mol Biol Rep 37:893–902
Shah J (2009) Plant under attack: systemic signals in defence. Curr Opin Plant Biol 12:459–464
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 Biochem Biophys 460:48–55
Szkopinska A, Pochocka D (2005) Farnesyl diphosphate synthase; regulation of product specificity. Acta Biochim Pol 52:45–55
Takaya A, Zhanga YW, Asawatreratanakul K, Wititsuwannakul D, Wititsuwannakul R, Takahashi S, Koyama T (2003) Cloning, expression and characterization of a functional cDNA clone encoding GGPPS of Hevea brassiliensis. Biochem Biophy Act 1625:214–220
Ven Murthy MR, Ranjekar PK, Ramassamy C, Deshpande M (2010) Scientific basis for the use of Indian Ayurvedic medicinal plants in the treatment of neurodegenerative disorders: Ashwagandha. Cent Nerv Syst Agents Med Chem (in press)
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The authors are thankful to the Council of Scientific and Industrial Research, New Delhi, Govt. of India, for providing the financial support to carry out this work under NMITLI scheme and Network Project (NWP-08).
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Gupta, P., Akhtar, N., Tewari, S.K. et al. Differential expression of farnesyl diphosphate synthase gene from Withania somnifera in different chemotypes and in response to elicitors. Plant Growth Regul 65, 93–100 (2011). https://doi.org/10.1007/s10725-011-9578-x
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DOI: https://doi.org/10.1007/s10725-011-9578-x