title: Nitrogen treatment enhances sterols and withaferin A through transcriptional activation of jasmonate pathway, WRKY transcription factors, and biosynthesis genes in Withania somnifera (L.) Dunal.
abstract: The medicinal plant Withania somnifera is researched extensively to increase the quantity of withanolides and specifically withaferin A, which finds implications in many pharmacological activities. Due to insufficient knowledge on biosynthesis and unacceptability of transgenic approach, it is preferred to follow alternative physiological methods to increase the yield of withanolides. Prior use of elicitors like salicylic acid, methyl jasmonate, fungal extracts, and even mechanical wounding have shown to increase the withanolide biosynthesis with limited success; however, the commercial viability and logistics of application are debatable. In this investigation, we tested the simple nitrogeneous fertilizers pertaining to the enhancement of withaferin A biosynthesis. Application of ammonium sulfate improved the sterol contents required for the withanolide biosynthesis and correlated to higher expression of pathway genes like FPPS, SMT1, SMT2, SMO1, SMO2, and ODM. Increased expression of a gene homologous to allene oxide cyclase, crucial in jasmonic acid biosynthetic pathway, suggested the involvement of jasmonate signaling. High levels of WRKY gene transcripts indicated transcriptional regulation of the pathway genes. Increase in transcript level could be correlated with a corresponding increase in the protein levels for WsSMT1 and WsWRKY1. The withaferin A increase was also demonstrated in the potted plants growing in the glasshouse and in the open field. These results implicated simple physiological management of nitrogen fertilizer signal to improve the yield of secondary metabolite through probable involvement of jasmonate signal and WRKY transcription factor for the first time, in W. somnifera besides improving the foliage.
authors.affiliation: Biotechnology Division, CSIR-Central Institute of Medicinal and Aromatic Plants, P.O. CIMAP, Lucknow, 226015, Uttar Pradesh, India.|Academy of Scientific and Innovative Research, New Delhi, India.
authors.forename: Shaifali
authors.initials: S
authors.lastname: Pal
authors.affiliation: Analytical Chemistry Division, CSIR-Central Institute of Medicinal and Aromatic Plants, P.O. CIMAP, Lucknow, 226015, Uttar Pradesh, India.
authors.forename: Akhilesh Kumar
authors.initials: AK
authors.lastname: Yadav
authors.affiliation: Biotechnology Division, CSIR-Central Institute of Medicinal and Aromatic Plants, P.O. CIMAP, Lucknow, 226015, Uttar Pradesh, India.
authors.forename: Anup Kumar
authors.initials: AK
authors.lastname: Singh
authors.affiliation: Biotechnology Division, CSIR-Central Institute of Medicinal and Aromatic Plants, P.O. CIMAP, Lucknow, 226015, Uttar Pradesh, India.
authors.forename: Shubhra
authors.initials: S
authors.lastname: Rastogi
authors.affiliation: Analytical Chemistry Division, CSIR-Central Institute of Medicinal and Aromatic Plants, P.O. CIMAP, Lucknow, 226015, Uttar Pradesh, India.
authors.forename: Madan Mohan
authors.initials: MM
authors.lastname: Gupta
authors.affiliation: Soil Science Division, CSIR-Central Institute of Medicinal and Aromatic Plants, P.O. CIMAP, Lucknow, 226015, Uttar Pradesh, India.
authors.forename: Rajesh Kumar
authors.initials: RK
authors.lastname: Verma
authors.affiliation: Biotechnology Division, CSIR-Central Institute of Medicinal and Aromatic Plants, P.O. CIMAP, Lucknow, 226015, Uttar Pradesh, India.
authors.forename: Dinesh A
authors.initials: DA
authors.lastname: Nagegowda
authors.affiliation: Biotechnology Division, CSIR-Central Institute of Medicinal and Aromatic Plants, P.O. CIMAP, Lucknow, 226015, Uttar Pradesh, India.
authors.forename: Anirban
authors.initials: A
authors.lastname: Pal
authors.affiliation: Biotechnology Division, CSIR-Central Institute of Medicinal and Aromatic Plants, P.O. CIMAP, Lucknow, 226015, Uttar Pradesh, India. ak.shasany@cimap.res.in.|Academy of Scientific and Innovative Research, New Delhi, India. ak.shasany@cimap.res.in.
authors.forename: Ajit Kumar
authors.initials: AK
authors.lastname: Shasany
mesh_terms: D000645:Ammonium Sulfate / Q000494:pharmacology; D053898:Biosynthetic Pathways / Q000187:drug effects / Q000235:genetics*; D003517:Cyclopentanes / Q000378:metabolism*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D017343:Genes, Plant; D009584:Nitrogen / Q000494:pharmacology*; D054883:Oxylipins / Q000378:metabolism*; D010758:Phosphorus / Q000494:pharmacology; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D011188:Potassium / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism; D013261:Sterols / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*; D015533:Transcriptional Activation / Q000187:drug effects*; D014508:Urea / Q000494:pharmacology; D032341:Withania / Q000187:drug effects / Q000235:genetics*; D054358:Withanolides / Q000378:metabolism*
chemical_list: D003517:Cyclopentanes; D054883:Oxylipins; D010940:Plant Proteins; D017382:Reactive Oxygen Species; D013261:Sterols; D014157:Transcription Factors; D054358:Withanolides; D010758:Phosphorus; C011006:jasmonic acid; D014508:Urea; C009684:withaferin A; D009584:Nitrogen; D011188:Potassium; D000645:Ammonium Sulfate; D004121:Dimethyl Sulfoxide
keywords: Allene oxide cyclase; Ammonium sulfate; Pathway genes; Secondary metabolites; Stress