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Identification and characterization of protein composition in Withania somnifera—an Indian ginseng

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Abstract

To have better understanding of the processes that occur in Withania somnifera L. Dunal, proteome analyses were initiated on two tissues (seeds & leaves) of this plant. Protein extracts were separated by two-dimensional gel electrophoresis (2-DE) across a broad 3.0–10.0 immobilized pH gradient (IPG) strip that yielded 434 protein spots. A total of 167 individual spots (82 from seeds and 85 from leaves) were excised from the gel and were characterized by peptide mass fingerprinting. From these analyses, 70 individual proteins from seeds and 74 from leaves were identified by protein sequence database interrogation and were catalogued accordingly to different protein functions. A comparative analysis of the two tissues indicated that some enzymes/proteins involved in housekeeping pathways were common to both, whereas some were exclusively tissue specific with specialized metabolic complement. The knowledge gained by this study towards the tissue specific protein expression in W. somnifera would form the basis for our future endeavor of characterization of proteins to understand the physiology and the associated complex metabolic network during its ontogenetic development.

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Abbreviations

2-DE:

Two-dimensional gel electrophoresis

IPG:

Immobilized pH gradient

SGTL1:

Sterol glucosyltransferases

TCA:

Trichloroacetic acid

CHAPS:

3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate

DTT:

Dithiothreitol

SDS-PAGE:

Sodium dodecyl sulfate–polyacrylamide gel electrophoresis

IEF:

Isoelectric focusing

IAA:

Iodoacetamide

MALDI-TOF-MS:

Matrix-assisted laser desorption/ionization-time of flight mass spectrometry

PMF:

Peptide mass fingerprinting.

References

  • Anonymous (2004) Monograph: Withania somnifera. Altern Med Rev 9:211–214

    Google Scholar 

  • Bak-Jensen KS, Laugesen S, Roepstorff P, Svensson B (2004) Two dimensional electrophoresis pattern (pI 6–11) identification of water soluble barley seed and malt proteins by mass spectrometry. Proteomics 4:728–742

    Article  PubMed  CAS  Google Scholar 

  • Bewley JD (1997) Seed germination and dormancy. Plant Cell 9:1055–1066

    Article  PubMed  CAS  Google Scholar 

  • Cahill DJ (2001) Protein and antibody arrays and their medical applications. J Immunol Methods 250:81–91

    Article  PubMed  CAS  Google Scholar 

  • Chenping Xu, Garrett MW, Sullivan J, Thomas JC, Natarajan S (2006) Separation and identification of soybean leaf proteins by two dimensional gel electrophoresis and mass spectrometry. Phytochemistry 67:2431–2440

    Article  Google Scholar 

  • Damerval C, de Vienne D, Zivy M, Thiellement H (1986) Technical improvements in two-dimensional electrophoresis increase the level of genetic variation detected in wheat-seedling proteins. Electrophoresis 7:52–54

    Article  CAS  Google Scholar 

  • Fu Q, Wang BC, Jin X, Li HB, Han P, Wei KH, Zhang XM, Zhu YX (2005) Proteomic analysis and extensive protein identification from dry, germinating Arabidopsis seeds and young seedlings. J Biochem Mol Biol 38:650–660

    Article  PubMed  CAS  Google Scholar 

  • Fukuda M, Islam N, Woo SH, Yamagishi A, Takaoka M, Hirano H (2003) Assessing matrix assisted laser desorption/ionization-time of flight-mass spectrometry as a means of rapid embryo protein identification in rice. Electrophoresis 24:1319–1329

    Article  PubMed  CAS  Google Scholar 

  • Gallardo K, Job C, Groot S, Puype M, Demol H, Vandekerckhove J, Job D (2002) Proteomics of Arabidopsis seed germination. A comparative study of wild-type and gibberellins-deficient seeds. Plant Physiol 129:823–837

    Article  PubMed  CAS  Google Scholar 

  • Islam N, Lonsdale M, Upadhyaya NM, Higgins TJ, Hirano H, Akhurst R (2004) Protein extraction from mature rice leaves for two-dimensional gel electrophoresis and its application in proteome analysis. Proteomics 4:1903–1908

    Article  PubMed  CAS  Google Scholar 

  • Jacobs DI, Gaspari M, Greef JV, Heijden RV, Verpoorte R (2005) Proteome analysis of the medicinal plant Catharanthus roseus. Planta 221:690–704

    Article  PubMed  CAS  Google Scholar 

  • Jacobs DI, Van der Heijden R, Verpoorte R (2000) Proteomics in plant biotechnology and metabolism research. Phytochem Anal 11:277–287

    Article  CAS  Google Scholar 

  • Kim S II, Kim JY, Kim EA, Kwoon KH, Kim KW, Cho K, Hwa Lee J, Nam MH, Yang DC, Shin Yoo JS, Park YM (2003) Proteome analysis of hairy root from Panax ginseng C. A. Meyer using peptide fingerprinting, internal sequencing and expressed sequence tag data. Proteomics 3:2379–2392

    Article  PubMed  CAS  Google Scholar 

  • Koller A, Washburn MP, Markus-Lange B, Andon NL, Deciu C, Haynes PA, Hays L, Schieltz D, Ulaszek R, Wei J, Wolters D, Yates JR III (2002) Proteomic survey of metabolic pathways of rice. Proc Natl Acad Sci USA 99:11969–11974

    Article  PubMed  CAS  Google Scholar 

  • Koomneef M, Bentsink L, Hilhorst H (2002) Seed dormancy and germination. Curr Opin Plant Biol 5:33–36

    Article  Google Scholar 

  • Lal P, Misra LN, Sangwan RS, Tuli R (2006) New withanolides from fresh berries of Withania somnifera. Z Naturforsch B 61b:1143–1147

    Google Scholar 

  • Mak Y, Skylas DJ, Willows R, Connolly A, Cordwell SJ, Wrigley CW, Sharp PJ, Copeland L (2006) A proteomic approach to the identification and characterization of protein composition in wheat germ. Funct Integr Genom 6:322–337

    Article  CAS  Google Scholar 

  • Misra L, Mishra P, Pandey A, Sangwan RS, Sangwan NS, Tuli R (2008) Withanolides from Withania somnifera roots. Phytochemistry 69:1000–1004

    Article  PubMed  CAS  Google Scholar 

  • Misra LN, Siddiqi SA (2005) Biologically active inositol, sterols and lipid derivatives from Sesbania bispinosa. Indian J Chem 44B:1915–1921

    CAS  Google Scholar 

  • Sangwan RS, Chaurasiyaa ND, Lal P, Misra L, Tuli R, Sangwan NS (2008) Withanolide A is inherently de novo biosynthesized in roots of the medicinal plant Ashwagandha. Withania somnifera. Physiol Plant 133:278–287

    Article  PubMed  CAS  Google Scholar 

  • Schuler M (1996) Plant cytochrome P450 monooxygenases. Crit Rev Plant Sci 15:235–284

    CAS  Google Scholar 

  • Sharma LK, Madina BR, Chaturvedi P, Sangwan RS, Tuli R (2007) Molecular cloning and characterization of one member of 3b-hydroxy sterol glucosyltransferase gene family in Withania somnifera. Arch Biochem Biophys 460:48–55

    Article  PubMed  CAS  Google Scholar 

  • Skylas DJ, Cordwell SJ, Hains PG, Larsen MR, Basseal DJ, Walsh BJ, Blumenthal CW, Rathmell LC, Wrigley CW (2002) Heat shock of wheat during grain filling: proteins associated with heat-tolerance. J Cereal Sci 35:175–188

    Article  CAS  Google Scholar 

  • Touzet P, Riccardi F, Morin C, Damerval C, Huet JC, Pernollet JC, Zivy M, de Vienne D (1996) The maize two-dimensional gel protein database: towards an integrated genome analysis programme. Theor Appl Genet 93:997–1005

    Article  CAS  Google Scholar 

  • Wu FS, Wang MY (1984) Extraction of proteins for sodium dodecyl-sulfate Polyacrylamide-gel electrophoresis from protease-rich plant-tissues. Anal Biochem 139:100–103

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

Authors are thankful to the Council of Scientific and Industrial Research (CSIR), New Delhi, Government of India, for funding the present research work under Network Programme NWP 0008.

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Authors and Affiliations

  1. Plant Biotechnology, Indian Institute of Integrative Medicine (CSIR), Canal Road, Jammu, Tawi, 180001, India

    Rekha Sapru Dhar, Suphla Bajpai Gupta, Sumeer Razdan, Wajid Waheed Bhat, Satiander Rana, Surrinder Kumar Lattoo & Shabnam Khan

  2. Medicinal Chemistry, Indian Institute of Integrative Medicine (CSIR), Canal Road, Jammu, Tawi, 180001, India

    Parvinder Pal Singh

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  1. Rekha Sapru Dhar
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  2. Suphla Bajpai Gupta
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  3. Parvinder Pal Singh
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  5. Wajid Waheed Bhat
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  6. Satiander Rana
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  7. Surrinder Kumar Lattoo
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Correspondence to Rekha Sapru Dhar.

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Dhar, R.S., Gupta, S.B., Singh, P.P. et al. Identification and characterization of protein composition in Withania somnifera—an Indian ginseng. J. Plant Biochem. Biotechnol. 21, 77–87 (2012). https://doi.org/10.1007/s13562-011-0083-0

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  • DOI: https://doi.org/10.1007/s13562-011-0083-0

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