Biotechnological Strategies for Improvement of Withania somnifera (L.) Dunal

  • Kulwinder Kaur
  • Ashok Kumar Panigrahi
  • Pratap Kumar Pati
Chapter

Abstract

Withania somnifera (L.) Dunal is one of the most reputed medicinal plants belonging to the family Solanaceae. The plant contains various steroidal lactones, alkaloids, polyphenols, flavanol glycosides, and sterols, which are responsible for its medicinal properties such as antitumor, neuroprotective, cardioprotective, antiaging, and anti-inflammatory. The main focus of research in W. somnifera is to increase the production of withanolides, the pharmaceutically important constituents. Various in vitro strategies like organ and cell culture and genetic manipulation and molecular biology tools such as metabolic engineering have been adapted to enhance secondary metabolite production. The present chapter provides a brief overview of challenges in W. somnifera and discusses some of the latest developments to address these challenges.

Keywords

Secondary metabolites Genetic transformation In vitro propagation Withanolide Withania somnifera 

Abbreviations

2,4-D

2,4-Dichlorophenoxyacetic acid

BAP

6-Benzylaminopurine

IAA

Indole-3-acetic acid

IBA

Indole-3-butyric acid

KN

Kinetin

MeJ

Methyl jasmonate

NAA

1-Naphthalene acetic acid

PGRs

Plant growth regulators

SA

Salicylic acid

References

  1. Abraham A, Kirson I, Glotter E, Lavie D (1968) A chemotaxonomic study of Withania somnifera (L.) dun. Phytochemistry 7(6):957–962CrossRefGoogle Scholar
  2. Abraham A, Kirson I, Lavie D, Glotte E (1975) The withanolides of Withania somnifera chemotypes I and II. Phytochemistry 14(1):189–194CrossRefGoogle Scholar
  3. Ahmad Baba I, Alia A, Saxena RC, Itoo A, Kumar S, Ahmad M (2013) In vitro propagation of Withania Somnifera (L.) Dunal (Ashwagandha) an endangered medicinal plant. Int J Pharm Sci Invent 2:6–11Google Scholar
  4. Alam N, Hossain M, Khalil MI, Moniruzzaman M, Sulaiman SA, Gan SH (2011) High catechin concentrations detected in Withania somnifera (ashwagandha) by high performance liquid chromatography analysis. BMC Complement Altern Med 11:65–73CrossRefGoogle Scholar
  5. Anjaneyulyu ASR, Satyanarayana Rao D (1997) A new withanolide from the leaves of Withania somnifera. Indian J Chem B Org Chem, Including Med Chem 36(2):161–165Google Scholar
  6. Arumugam A, Gopinath K (2011) Micro propagation and tissue culture of the endangered medicinal plant Withania somnifera by the direct shoot and root initiation method. Int J Appl Biol Pharm Technol 2:315–321Google Scholar
  7. Arumugam A, Gopinath K (2013) In vitro regeneration of an endangered medicinal plant Withania somnifera using four different explants. Plant Tissue Cult Biotechnol 23:79–85CrossRefGoogle Scholar
  8. Bhatia A, Bharti SK, Tewari SK, Sidhu OP, Roy R (2013) Metabolic profiling for studying chemotype variations in Withania somnifera (L.) Dunal fruits using GC-MS and NMR spectroscopy. Phytochemistry 93:105–115CrossRefGoogle Scholar
  9. Cai Z, Kastell A, Knorr D, Smetanska I (2012) Exudation: an expanding technique for continuous production and release of secondary metabolites from plant cell suspension and hairy root cultures. Plant Cell Rep 31(3):461–477CrossRefGoogle Scholar
  10. Chakraborty N, Banerjee D, Ghosh M, Pradhan P, Gupta NS, Acharya K, Banerjee M (2013) Influence of plant growth regulators on callus mediated regeneration and secondary metabolite synthesis in Withania somnifera (L.) Dunal. Physiol Mol Biol Plants 19:117–125CrossRefGoogle Scholar
  11. 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 80:496–505CrossRefGoogle Scholar
  12. Chaurasiya ND, Sangwan NS, Sabir F, Misra L, Sangwan RS (2012) Withanolide biosynthesis recruits both mevalonate and DOXP pathways of isoprenogenesis in Ashwagandha Withania somnifera L.(Dunal). Plant Cell Rep 31:1889–1897CrossRefGoogle Scholar
  13. Dewir YH, Chakrabarty D, Lee SH, Hahn EJ, Paek KY (2010) Indirect regeneration of Withania somnifera and comparative analysis of withanolides in in vitro and greenhouse grown plants. Biol Plantarum 54:357–360CrossRefGoogle Scholar
  14. Dharmar K, De-Britto AJ (2011) RAPD analysis of genetic variability in wild populations of Withania somnifera (L.) Dunal. Int J Biol Technol 2(1):21–25Google Scholar
  15. Fatima N, Anis M (2012) Role of growth regulators on in vitro regeneration and histological analysis in Indian ginseng (Withania somnifera L.) Dunal. Physiol Mol Biol Plants 18(1):59–67CrossRefGoogle Scholar
  16. Fatima N, Ahmad N, Anis M (2011) Enhanced in vitro regeneration and change in photosynthetic pigments, biomass and proline content in Withania somnifera L. (Dunal) induced by copper and zinc ions. Plant Physiol Biochem 49:1465–1471CrossRefGoogle Scholar
  17. Fatima N, Ahmad N, Anis M (2016) In vitro propagation and conservation of Withania somnifera (Dunal) L. In: Protocols for in vitro cultures and secondary metabolite analysis of aromatic and medicinal plants. Methods in molecular biology, vol 1391, 2nd edn. Springer, New YorkGoogle Scholar
  18. Furmanowa M, Gajdzis-Kuls D, Ruszkowska J, Czarnocki Z, Obidoska G, Sadowska A, Rani R, Upadhyay SN (2001) In vitro propagation of Withania somnifera and isolation of withanolides with immunosuppressive activity. Planta Med 67:146–149CrossRefGoogle Scholar
  19. Ghimire BK, Seong ES, Kim EH, Lamsal K, Yu CY, Chung IM (2010) Direct shoot organogenesis from petiole and leaf discs of Withania somnifera (L.) Dunal. Afr J Biotechnol 9:7453–7461CrossRefGoogle Scholar
  20. Grover A, Samuel G, Bisaria VS, Sundar D (2013) Enhanced withanolide production by overexpression of squalene synthase in Withania somnifera. J Biosci Bioeng 115:680–685CrossRefGoogle Scholar
  21. Gupta S, Sahu PK (2015) In vitro micro propagation of Withania somnifera (L.) Dunal. Saudi J Biol Sci.  https://doi.org/10.1016/j.sjbs.2015.02.002
  22. Herrera-Estrella L, Simpson J, MartÃnez-Trujillo M (2004) Transgenic plants: an historical perspective. Transgenic Plants Meth Protoc 286:3–31Google Scholar
  23. Hunter WN (2007) The non-mevalonate pathway of isoprenoid precursor biosynthesis. J Biol Chem 282:21573–21577CrossRefGoogle Scholar
  24. Jain R, Kachhwaha S, Kothari SL (2016) In vitro shoot cultures and analysis of steroidal lactones in Withania coagulans (Stocks) Dunal. In: Protocols for in vitro cultures and secondary metabolite analysis of aromatic and medicinal plants. Methods in molecular biology, vol 1391, 2nd edn. Springer, New YorkGoogle Scholar
  25. Jayaprakasam B, Zhang Y, Seeram NP, Nair MG (2003) Growth inhibition of human tumor cell lines by withanolides from Withania somnifera leaves. Life Sci 74:125–132CrossRefGoogle Scholar
  26. Joshi AG, Padhya MA (2010) Shoot regeneration from leaf explants of Withania somnifera (L.) Dunal. Notulae Scientia Biologicae 2(1):63Google Scholar
  27. Kanungo S, Sahoo SL (2011) Direct organogenesis of Withania somnifera L. from apical bud. Int Res J Biotechnol 2:58–61Google Scholar
  28. Kaur K, Singh P, Guleri R, Singh B, Kaur K, Singh V, Pati PK (2017) Biotechnological approaches in propagation and improvement of Withania somnifera (L.) Dunal. In: Science of Ashwagandha: preventive and therapeutic potentials. Springer, Cham.  https://doi.org/10.1007/978-3-319-59192-6_22 CrossRefGoogle Scholar
  29. Khanna PK, Kumar A, Chandra R, Verma V (2013) Germination behaviour of seeds of Withania somnifera (L.) Dunal: a high value medicinal plant. Physiol Mol Biol Plants 19:449–454CrossRefGoogle Scholar
  30. Kirson I, Glotter E, Abraham A, Lavie D (1970) Constituents of Withania somnifera dun – XI: the structure of three new withanolides. Tetrahedron 26(9):2209–2219CrossRefGoogle Scholar
  31. Kothari SK, Singh CP, Vijay Kumar Y, Singh K (2003) Morphology, yield and quality of ashwagandha (Withania somnifera L. Dunal) roots and its cultivation economics as influenced by tillage depth and plant population density. J Hortic Sci Biotechnol 78(3):422–425CrossRefGoogle Scholar
  32. Kuboyama T, Tohda C, Komatsu K (2005) Neuritic regeneration and synaptic reconstruction induced by withanolide A. Br J Pharmacol 144:961–971CrossRefGoogle Scholar
  33. Kulkarni AA, Thengane SR, Krishnamurthy KV (1996) Direct in vitro regeneration of leaf explants of Withania somnifera (L.) Dunal. Plant Sci 119(1–2):163–168CrossRefGoogle Scholar
  34. Kulkarni AA, Thengane SR, Krishnamurthy KV (2000) Direct shoot regeneration from node, internode, hypocotyl and embryo explants of Withania somnifera. Plant Cell Tiss Org 62:203–209CrossRefGoogle Scholar
  35. Kumar A, Kaul MK, Bhan MK, Khanna PK, Suri KA (2007) Morphological and chemical variation in 25 collections of the Indian medicinal plant, Withania somnifera (L.) Dunal (Solanaceae). Genet Resour Crop Evol 54:655–660CrossRefGoogle Scholar
  36. Kumar A, Mir BA, Sehgal D, Dar TH, Koul S, Kaul MK, Raina SN, Qazi GN (2011) Utility of a multidisciplinary approach for genome diagnostics of cultivated and wild germplasm resources of medicinal Withania somnifera, and the status of new species, W. ashwagandha, in the cultivated taxon. Plant Syst Evol 291:141–151CrossRefGoogle Scholar
  37. Kurapati KRV, Samikkannu T, Atluri VSR, Kaftanovskaya E, Yndart A, Nair MP (2014) β-amyloid 1-42, HIV-1, Ba-L (Clade B) infection and drugs of abuse induced degeneration in human neuronal cells and protective effects of Ashwagandha (Withania somnifera) and its constituent Withanolide A. PLoS One 9(11):e112818CrossRefGoogle Scholar
  38. Manickam VS, Mathavan RE, Antonisamy R (2000) Regeneration of Indian ginseng plantlets from stem callus. Plant Cell Tiss Org 62:181–185CrossRefGoogle Scholar
  39. Manivel P, Reddy RRN, Deore HB (2017) Genetic diversity for root yield and its component traits in Ashwagandha (Withania somnifera (L) Dunal) pure lines derived from JA134 population. Int J Curr Microbiol Appl Sci 6(4):1694–1710CrossRefGoogle Scholar
  40. Mir BA, Koul S, Kumar A, Kaul MK, Soodan AS, Raina SN (2011) Assessment and characterization of genetic diversity in Withania somnifera (L.) Dunal using RAPD and AFLP markers. Afr J Biotechnol 10(66):14746–14756CrossRefGoogle Scholar
  41. Mir BA, Khazir J, Hakeem KR, Koul S, Cowan DA (2014) Enhanced production of withaferin-A in shoot cultures of Withania somnifera (L) Dunal. J Plant Biochem Biotechnol 23:430–434CrossRefGoogle Scholar
  42. Mirjalili MH, Moyano E, Bonfill M, Cusido RM, Palazón J (2009) Steroidal lactones from Withania somnifera, an ancient plant for novel medicine. Molecules 14:2373–2393CrossRefGoogle Scholar
  43. Mishra S, Bansal S, Mishra B, Sangwan RS, Jadaun JS, Sangwan NS (2016) RNAi and homologous over-expression based functional approaches reveal triterpenoid synthase gene-cycloartenol synthase is involved in downstream withanolide biosynthesis in Withania somnifera. PLoS One 11(2):e0149691CrossRefGoogle Scholar
  44. Murthy HN, Dijkstra C, Anthony P, White DA, Davey MR, Power JB, Hahn EJ, Paek KY (2008) Establishment of Withania somnifera hairy root cultures for the production of withanolide A. J Integr Plant Biol 50:975–981CrossRefGoogle Scholar
  45. Nagella P, Murthy HN (2010) Establishment of cell suspension cultures of Withania somnifera for the production of withanolide A. Bioresour Technol 101:6735–6739CrossRefGoogle Scholar
  46. Nagella P, Murthy HN (2011) Effects of macroelements and nitrogen source on biomass accumulation and withanolide-A production from cell suspension cultures of Withania somnifera (L.) Dunal. Plant Cell Tissue Organ Cult 104:119–124CrossRefGoogle Scholar
  47. Nayak SA, Kumar S, Satapathy K, Moharana A, Behera B, Barik DP, Acharya L, Mohapatra PK, Jena PK, Naik SK (2013) In vitro plant regeneration from cotyledonary nodes of Withania somnifera (L.) Dunal and assessment of clonal fidelity using RAPD and ISSR markers. Acta Physiol Plant 35:195–203CrossRefGoogle Scholar
  48. Oksman-Caldentey KM, Inzé D (2004) Plant cell factories in the post-genomic era: new ways to produce designer secondary metabolites. Trends Plant Sci 9(9):433–440CrossRefGoogle Scholar
  49. Pal S, Yadav AK, Singh AK, Rastogi S, Gupta MM, Verma RK, Nagegowda DA, Pal A, Shasany AK (2016) Nitrogen treatment enhances sterols and withaferin A through transcriptional activation of jasmonate pathway, WRKY transcription factors, and biosynthesis genes in Withania somnifera (L.) Dunal. Protoplasma.  https://doi.org/10.1007/s00709-016-0959-x
  50. 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 29:133–141CrossRefGoogle Scholar
  51. Patel N, Patel P, Kendurkar SV, Thulasiram HV, Khan BM (2015) Overexpression of squalene synthase in Withania somnifera leads to enhanced withanolide biosynthesis. Plant Cell Tissue Org Cult 122:409–420CrossRefGoogle Scholar
  52. Praveen N, Murthy HN (2012) Synthesis of withanolide A depends on carbon source and medium pH in hairy root cultures of Withania somnifera. Ind Crop Prod 35:241–243CrossRefGoogle Scholar
  53. Praveen N, Murthy HN (2013) Withanolide A production from Withania somnifera hairy root cultures with improved growth by altering the concentrations of macro elements and nitrogen source in the medium. Acta Physiol Plant 35:811–816CrossRefGoogle Scholar
  54. Rana S, Dhar N, Bhat WW, Razdan S, Khan S, Dhar RS, Dutt P, Lattoo SK (2012) A 12-deoxywithastramonolide-rich somaclonal variant in Withania somnifera (L.) Dunal – molecular cytogenetic analysis and significance as a chemotypic resource. In Vitro Cell Dev Biol 48:546–554CrossRefGoogle Scholar
  55. Rani G, Grover IS (1999) In vitro callus induction and regeneration studies in Withania somnifera. Plant Cell, Tiss Org Cult 57(1):23–27CrossRefGoogle Scholar
  56. Rani G, Virk GS, Nagpal A (2003) Callus induction and plantlet regeneration in Withania somnifera (L.) Dunal. In Vitro Cell Dev Biol Plant 39:468–474Google Scholar
  57. Ray S, Jha S (1999) Withanolide synthesis in cultures of Withania somnifera transformed with Agrobacterium tumefaciens. Plant Sci 146:1–7CrossRefGoogle Scholar
  58. Ray S, Jha S (2001) Production of withaferin A in shoot cultures of Withania somnifera. Planta Med 67:432–436CrossRefGoogle Scholar
  59. Rodriguez-Concepcion M, Boronat A (2002) Elucidation of the methylerythritol phosphate pathway for isoprenoid biosynthesis in bacteria and plastids. A metabolic milestone achieved through genomics. Plant Physiol 130:1079–1089CrossRefGoogle Scholar
  60. Rout JR, Sahoo SL, Das R (2011) An attempt to conserve Withania somnifera (L.) Dunal-A highly essential medicinal plant, through in vitro callus culture. Pak J Bot 43:1837–1842Google Scholar
  61. Sabir F, Sangwan NS, Chaurasiya ND, Misra LN, Tuli R, Sangwan RS (2008) Rapid micropropagation of Withania somnifera L. accessions from axillary meristems. J Herbs, Spices Med Plants 13:123–133CrossRefGoogle Scholar
  62. Sangwan RS, Chaurasiya ND, Lal P, Misra L, Uniyal GC, Tuli R, Sangwan NS (2007) Withanolide A biogeneration in in vitro shoot cultures of Ashwagandha (Withania somnifera Dunal), a main medicinal plant in Ayurveda. Chem Pharm Bull 55:1371–1375CrossRefGoogle Scholar
  63. Sangwan RS, Chaurasiya 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 133:278–287CrossRefGoogle Scholar
  64. Saravanakumar A, Aslam A, Shajahan A (2012) Development and optimization of hairy root culture systems in Withania somnifera (L.) Dunal for withaferin-A production. Afr J Biotechnol 11:16412Google Scholar
  65. Saritha KV, Naidu CV (2007) In vitro flowering of Withania somnifera Dunal. – an important antitumor medicinal plant. Plant Sci 172:847–851CrossRefGoogle Scholar
  66. Sen J, Sharma AK (1991) Micropropagation of Withania somnifera from germinating seeds and shoot tips. Plant Cell Tiss Org Cult 26:71–73CrossRefGoogle Scholar
  67. Shanks JV, Morgan J (1999) Plant ‘hairy root’culture. Curr Opin Biotechnol 10(2):151–155CrossRefGoogle Scholar
  68. Sharma A, Pati PK (2011a) First report of Withania somnifera (L.) Dunal, as a new host of cowbug (Oxyrachis tarandus, Fab.) in plains of Punjab, northern India. World Appl Sci J 14:1344–1346Google Scholar
  69. Sharma A, Pati PK (2011b) First record of 28-spotted ladybird beetle, Henosepilachna vigintioctopunctata (F.) infesting Withania somnifera (L.) Dunal in Punjab province of Northern India. Pest Technol 5:91–92Google Scholar
  70. Sharma A, Pati PK (2012a) First record of Ashwagandha as a new host to the invasive mealybug (Phenacoccus solenopsis Tinsley) in India. Entomol News 123:59–62CrossRefGoogle Scholar
  71. Sharma A, Pati PK (2012b) First record of the carmine spider mite, Tetranychus urticae, infesting Withania somnifera in India. J Insect Sci 12:50CrossRefGoogle Scholar
  72. Shinde A, Gahunge P, Rath SK (2015) Conservation and sustainability of ashwagandha: a medicinal plant. J Biol Sci Opin 3:94–99CrossRefGoogle Scholar
  73. Siddique NA, Bari MA, Shahnewaz S, Rahman MH, Hasan MR, Khan MST, Islam MS (2004) Plant regeneration of Withania somnifera (L.) Dunal (Ashwagandha) from nodal segments derived callus an endangered medicinal plant in Bangladesh. J Biol Sci 4:219–223CrossRefGoogle Scholar
  74. Singh S, Kumar S (1998) Withania somnifera: The Indian ginseng ashwagandha. Central Institute of Medicinal and Aromatic Plants, LucknowGoogle Scholar
  75. Singh AK, Varshney R, Sharma M, Agarwal SS, Bansal KC (2006) Regeneration of plants from alginate-encapsulated shoot tips of Withania somnifera (L.) Dunal, a medicinally important plant species. J Plant Physiol 163:220–223CrossRefGoogle Scholar
  76. Singh S, Pal S, Shanker K, Chanotiya CS, Gupta MM, Dwivedi UN, Shasany AK (2014) Sterol partitioning by HMGR and DXR for routing intermediates toward withanolide biosynthesis. Physiol Plant 152:617–633CrossRefGoogle Scholar
  77. Singh P, Guleri R, Singh V, Kaur G, Kataria H, Singh B, Kaur G, Kaul SC, Wadhwa R, Pati PK (2015a) Biotechnological interventions in Withania somnifera (L.) Dunal. Biotechnol Genet Eng Rev.  https://doi.org/10.1080/02648725.2015.1020467 CrossRefGoogle Scholar
  78. Singh AK, Dwivedi V, Rai A, Pal S, Reddy SGE, Rao DKV, Shasany AK, Nagegowda DA (2015b) Virus-induced gene silencing of Withania somnifera squalene synthase negatively regulates sterol and defence-related genes resulting in reduced withanolides and biotic stress tolerance. Plant Biotechnol J 13:1287–1299CrossRefGoogle Scholar
  79. Singh V, Singh B, Sharma A, Kaur K, Gupta AP, Salar RK, Hallan V, Pati PK (2016a) Leaf spot disease adversely affects human health promoting constituents and withanolide biosynthesis in Withania somnifera (L.) Dunal. J Appl Microbiol.  https://doi.org/10.1111/jam.13314 CrossRefGoogle Scholar
  80. Singh G, Tiwari M, Singh SP, Singh S, Trivedi PK, Misra P (2016b) Silencing of sterol glycosyltransferases modulates the withanolide biosynthesis and leads to compromised basal immunity of Withania somnifera. Sci Rep.  https://doi.org/10.1038/srep25562
  81. Singh P, Guleri R, Pati PK (2016c) In Vitro propagation of Withania somnifera (L.) Dunal. In: Protocols for in vitro cultures and secondary metabolite analysis of aromatic and medicinal plants. Methods in molecular biology, vol 1391, 2nd edn. Springer, New YorkGoogle Scholar
  82. Singh P, Guleri R, Angurala A, Kaur K, Kaur K, Kaul SC, Wadhwa R, Pati PK (2017) Addressing challenges to enhance the bioactives of Withania Somnifera through organ, tissue, and cell culture based approaches. Biomed Res Int.  https://doi.org/10.1155/2017/3278494
  83. Sivanandhan G, Mariashibu TS, Arun M, Rajesh M, Kasthurirengan S, Selvaraj N, Ganapathi A (2011) The effect of polyamines on the efficiency of multiplication and rooting of Withania somnifera (L.) Dunal and content of some withanolides in obtained plants. Acta Physiol Plant 33:2279–2288CrossRefGoogle Scholar
  84. Sivanandhan G, Rajesh M, Arun M, Jeyaraj M, Dev GK, Arjunan A, Manickavasagam M, Muthuselvam M, Selvaraj N, Ganapathi A (2012) Effect of culture conditions, cytokinins, methyl jasmonate and salicylic acid on the biomass accumulation and production of withanolides in multiple shoot culture of Withania somnifera (L.) Dunal using liquid culture. Acta Physiol Plant 35:715–728CrossRefGoogle Scholar
  85. Sivanandhan G, Dev GK, Jeyaraj M, Rajesh M, Arjunan A, Muthuselvam M, Manickavasagam M, Selvaraj N, Ganapathi A (2013a) Increased production of withanolide A, withanone, and withaferin A in hairy root cultures of Withania somnifera (L.) Dunal elicited with methyl jasmonate and salicylic acid. Plant Cell Tissue Organ Cult 114:121–129CrossRefGoogle Scholar
  86. Sivanandhan G, Dev GK, Jeyaraj M, Rajesh M, Muthuselvam M, Selvaraj N, Manickavasagam M, Ganapathi A (2013b) A promising approach on biomass accumulation and withanolides production in cell suspension culture of Withania somnifera (L.) Dunal. Protoplasma 250:885–898CrossRefGoogle Scholar
  87. Sivanandhan G, Dev GK, Theboral J, Selvaraj N, Ganapathi A, Manickavasagam M (2015) Sonication, vacuum infiltration and thiol compounds enhance the Agrobacterium-mediated transformation frequency of Withania somnifera (L.) Dunal. PLoS One 10:e0124693CrossRefGoogle Scholar
  88. Sivanesan I (2007) Direct regeneration from apical bud explants of Withania somnifera Dunal. Indian J Biotechnol 6:125Google Scholar
  89. Sivanesan I, Murugesan K (2008) An efficient regeneration from nodal expiants of Withania somnifera Dunal. Asian J Plant Sci 7:551–556CrossRefGoogle Scholar
  90. Soni P, Bahadur AN, Tiwari U, Kanungo VK (2011) Micropropagation of a medicinal plant Withania somnifera (L.) Dunal by shoot bud culture. Int Q J Life Sci 6:135–137Google Scholar
  91. Srivastava S, Sangwan RS, Tripathi S, Mishra B, Narnoliya LK, Misra LN, Sangwan NS (2015) Light and auxin responsive cytochrome P450s from Withania somnifera Dunal: cloning, expression and molecular modelling of two pairs of homologue genes with differential regulation. Protoplasma 252(6):1421–1437CrossRefGoogle Scholar
  92. Supe U, Dhote F, Roymon MG (2006) In vitro plant regeneration of Withania somnifera. Plant Tiss Cult Biotechnol 16:111–115Google Scholar
  93. Suttipanta N, Pattanaik S, Kulshrestha M, Patra B, Singh SK, Yuan L (2011) The transcription factor CrWRKY1 positively regulates the terpenoid indole alkaloid biosynthesis in Catharanthus roseus. Plant Physiol 157(4):2081–2093CrossRefGoogle Scholar
  94. Tang K, Shen Q, Yan T, Fu X (2014) Transgenic approach to increase artemisinin content in Artemisia annua L. Plant Cell Rep 33(4):605–615CrossRefGoogle Scholar
  95. Teli NP, Patil NM, Pathak HM, Bhalsing SR, Maheshwari VL (1999) Withania somnifera (Ashwagandha): regeneration through meristem culture. J Plant Biotechnol Biochem 8:109–111CrossRefGoogle Scholar
  96. Udayakumar R, Choi CW, Kim KT, Kim SC, Kasthurirengan S, Mariashibu TS, SahayaRayan JJ, Ganapathi A (2013) In vitro plant regeneration from epicotyl explant of Withania somnifera (L) Dunal. J Med Plants Res 7:43–52Google Scholar
  97. Udayakumar R, Kasthurirengan S, Mariashibu TS, Rayan JJS, Ganapathi A, Kim SC, Kim JJ, Choi CW (2014) Agrobacterium-mediated genetic transformation of Withania somnifera using nodal explants. Acta Physiol Plant 36:1969–1980CrossRefGoogle Scholar
  98. Vakeswaran V, Krishnasamy V (2003) Improvement in storability of Ashwagandha (Withania somnifera Dunal) seeds through pre-storage treatments by triggering their physiological and biochemical properties [abstract]. Seed Technol 25:203–203Google Scholar
  99. Verma S, Srivastava SK (2014) A wonder plant Withania: pharmacological and chemical review. CSIR-CIMAP 11:72Google Scholar
  100. Verpoorte R, Van der Heijden R, Ten Hoopen HJG, Memelink J (1999) Metabolic engineering of plant secondary metabolite pathways for the production of fine chemicals. Biotechnol Lett 21(6):467–479CrossRefGoogle Scholar
  101. Wadhwa R, Singh R, Gao R, Shah N, Widodo N, Nakamoto T et al (2013) Water extract of Ashwagandha leaves has anticancer activity: identification of an active component and its mechanism of action. PLoS One 8(10):e77189CrossRefGoogle Scholar
  102. Widodo N, Kaur K, Shrestha BG, Takagi Y, Ishii T, Wadhwa R, Kaul SC (2007) Selective killing of cancer cells by leaf extract of Ashwagandha: identification of a tumorinhibitory factor and the first molecular insights to its effect. Clin Cancer Res 13:2298–2306CrossRefGoogle Scholar
  103. Widodo N, Takagi Y, Shrestha BG, Ishii T, Kaul SC, Wadhwa R (2008) Selective killing of cancer cells by leaf extract of Ashwagandha: components, activity and pathway analyses. Cancer Lett 262:37–47CrossRefGoogle Scholar
  104. Wu S, Schalk M, Clark A, Miles RB, Coates R (2006) Redirection of cytosolic or plastidic isoprenoid precursors elevates terpene production in plants. Nat Biotechnol 24(11):1441–1447CrossRefGoogle Scholar

Copyright information

© Springer Nature Singapore Pte Ltd. 2018

Authors and Affiliations

  • Kulwinder Kaur
    • 1
  • Ashok Kumar Panigrahi
    • 2
  • Pratap Kumar Pati
    • 1
  1. 1.Department of BiotechnolgyGuru Nanak Dev UniversityAmritsarIndia
  2. 2.Department of PharmacologyVeer Surendra Sai Institute of Medical Sciences and ResearchBurla, SambalpurIndia

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