title: Comparative study of withanolide production and the related transcriptional responses of biosynthetic genes in fungi elicited cell suspension culture of Withania somnifera in shake flask and bioreactor.
abstract: Ashwagandha (Withania somnifera) is one of the most reputed medicinal plants in the traditional medicinal system. In this study, cell suspension culture of W. somnifera was elicited with cell homogenates of fungi (A. alternata, F. solani, V. dahliae and P. indica) in shake flask and the major withanolides like withanolide A, withaferin A and withanone were analysed. Simultaneously expression levels of key pathway genes from withanolides biosynthetic pathways were also checked via quantitative PCR in shake flask as well as in bioreactor. The results show that highest gene expression of 10.8, 5.8, 4.9, and 3.3 folds were observed with HMGR among all the expressed genes in cell suspension cultures with cell homogenates of 3% P. indica, 5% V. dahliae, 3% A. alternata and 3% F. solani, respectively, in comparison to the control in shake flask. Optimized concentration of cell homogenate of P. indica (3% v/v) was added to the growing culture in 5.0-l bioreactor under optimized up-scaling conditions and harvested after 22 days. The genes of MVA, MEP and withanolides biosynthetic pathways like HMGR, SS, SE, CAS, FPPS, DXR and DXS were up-regulated by 12.5, 4.9, 2.18, 4.65, 2.34, 1.89 and 1.4 folds, respectively in bioreactor. The enhancement of biomass (1.13 fold) and withanolides [withanolide A (1.7), withaferin A (1.5), and withanone (1.5) folds] in bioreactor in comparison to shake flask was also found to be in line with the up-regulation of genes of withanolide biosynthetic pathways.
authors.affiliation: Department of Biotechnology, Faculty of Science, Centre for Transgenic Plant Development, Jamia Hamdard, New Delhi 110062, India.
authors.forename: Seema
authors.initials: S
authors.lastname: Ahlawat
authors.affiliation: Department of Biotechnology, Faculty of Science, Centre for Transgenic Plant Development, Jamia Hamdard, New Delhi 110062, India.
authors.forename: Parul
authors.initials: P
authors.lastname: Saxena
authors.affiliation: Department of Biotechnology, Faculty of Science, Centre for Transgenic Plant Development, Jamia Hamdard, New Delhi 110062, India.
authors.forename: Athar
authors.initials: A
authors.lastname: Ali
authors.affiliation: Department of Biotechnology, Faculty of Science, Centre for Transgenic Plant Development, Jamia Hamdard, New Delhi 110062, India.
authors.forename: Shazia
authors.initials: S
authors.lastname: Khan
authors.affiliation: Department of Biotechnology, Faculty of Science, Centre for Transgenic Plant Development, Jamia Hamdard, New Delhi 110062, India. Electronic address: mzabdin@rediffmail.com.
authors.forename: Malik Z
authors.initials: MZ
authors.lastname: Abdin
mesh_terms: D018533:Biomass; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D005658:Fungi / Q000502:physiology; D018506:Gene Expression Regulation, Plant; D014315:Triterpenes / Q000032:analysis / Q000378:metabolism; D032341:Withania / Q000166:cytology / Q000235:genetics / Q000378:metabolism* / Q000382:microbiology*; D054358:Withanolides / Q000032:analysis / Q000378:metabolism*
chemical_list: D014315:Triterpenes; D054358:Withanolides; C560597:withanone; C009684:withaferin A
keywords: Bioreactor; Cell suspension cultures; P. indica; W. somnifera; Withanolides