Molecular Biology Reports

, Volume 46, Issue 2, pp 1895–1908 | Cite as

Innate endophytic fungus, Aspergillus terreus as biotic elicitor of withanolide A in root cell suspension cultures of Withania somnifera

  • Ramesh Kumar Kushwaha
  • Sucheta Singh
  • Shiv Shanker Pandey
  • Alok Kalra
  • Chikkarasanahalli Shivegowda Vivek BabuEmail author
Original Article


In the present study, root cell suspension cultures of W. somnifera were elicited with mycelial extract (1% w/v) and culture filtrate (5% v/v) of their native endophytic fungus Aspergillus terreus 2aWF in shake flask. Culture filtrate of A. terreus 2aWF significantly elicits withanolide A at 6H (12.20 ± 0.52 µg/g FCB). However, with A. terreus 2aWF mycelial extract, withanolide A content was higher at 24H (10.29 µg/g FCB). Withanolide A content was maximum with salicylic acid (0.1 mM) treatment at 24H (8.3 ± 0.20 µg/g FCB). Further, expression analysis of withanolide pathway genes, hydrogen peroxide production, and lipid peroxidation was carried out after 48H of elicitation with 2aWF mycelial extract and culture filtrate. The expression levels of withanolides biosynthetic pathway genes, viz. HMGR, DXR, FPPS, SQS, SQE, CAS, SMT1, STE1 and CYP710A1 were quantified by real time PCR at 48H of elicitation. In all the treatments, the expression levels of key genes were significantly upregulated as compared to untreated suspension cells. Hydrogen peroxide was noticeably enhanced in SA, mycelia extract and culture filtrate, at 20% (115 ± 4.40 nM/g FCB), 42% (137.5 ± 3.62 nM/g FCB), and 27% (122.8 ± 1.25 nM/g FCB) respectively; however, lipid peroxidation was 0.288 ± 0.014, 0.305 ± 0.041 and 0.253 ± 0.007 (µM/gm FCB) respectively, higher than the control (0.201 ± 0.007 µM/gm FCB).


Withania somnifera Withanolide A Endophyte Elicitor Suspension culture 



This work was supported by grant NWP BSC0117 (XII 5 Year Plan Network Project) from the Council of Scientific and Industrial Research (CSIR), India. Sincere thanks to Director, CSIR-Central Institute of Medicinal and Aromatic Plants, Lucknow, India for his encouragement. R. K. Kushwaha greatly acknowledges, ICMR for financial assistance in the form of fellowship and contingency grant for research activity. SSP acknowledges CSIR, India for financial assistance in the form of Senior Research Associateship (SRA). C. S. Vivek Babu and R. K. Kushwaha greatly acknowledges Dr. Dinesh A. Nagegowda for providing withanolides standards and primers and Dr. D. K. Venkata Rao for sharing his lab facilities.

Author contributions

RKK performed all the experiments and CSV analyzed and prepared the manuscript. All authors read and approved the manuscript.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11033_2019_4641_MOESM1_ESM.docx (586 kb)
Supplementary material 1 (DOCX 585 KB)


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© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Microbial Technology LaboratoryCSIR-Central Institute of Medicinal and Aromatic Plants, Research CentreBangaloreIndia
  2. 2.Microbial Technology DivisionCSIR-Central Institute of Medicinal and Aromatic PlantsLucknowIndia
  3. 3.Academy of Scientific and Innovative Research, (AcSIR)GhaziabadIndia

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