TY - JOUR
T1 - Differential Gene Expression and Withanolides Biosynthesis During in vitro and ex vitro Growth of Withania somnifera (L.) Dunal
AU - Thorat, Sachin Ashok
AU - Kaniyassery, Arya
AU - Poojari, Poornima
AU - Rangel, Melissa
AU - Tantry, Shashikala
AU - Kiran, Kodsara Ramachandra
AU - Joshi, Manjunath B.
AU - Rai, Padmalatha S.
AU - Botha, Anna Maria
AU - Muthusamy, Annamalai
N1 - Funding Information:
We thank the Manipal Academy of Higher Education (MAHE), Manipal, Karnataka, India, TIFAC-CORE and FIST, DST, New Delhi, and K-FIST, VGST, Govt. of Karnataka, for the facilities. We are grateful to K. Satyamoorthy, Director, Manipal School of Life Sciences, MAHE for their encouragement and support. We are thankful to MAHE for the T.M.A. Pai Ph.D. scholarship to SAT, AK, and KK. We are indebted to the International Cooperation Division, Department of Science and Technology (DST), Govt. of India, New Delhi, for the financial assistance through the Indo-South Africa Joint Collaborative Project (DST/INT/South Africa/P-10/2016 dated 15 September 2016) to AM and as Women Scientist [Ref. No. SR/WOS-A/LS-564/2011 (G) dated 26 July 2012], DST, New Delhi, to PP. We would like to thank M. Radhakrishna Rao for his suggestions.
Funding Information:
We thank the Manipal Academy of Higher Education (MAHE), Manipal, Karnataka, India, TIFAC-CORE and FIST, DST, New Delhi, and K-FIST, VGST, Govt. of Karnataka, for the facilities. We are grateful to K. Satyamoorthy, Director, Manipal School of Life Sciences, MAHE for their encouragement and support. We are thankful to MAHE for the T.M.A. Pai Ph.D. scholarship to SAT, AK, and KK. We are indebted to the International Cooperation Division, Department of Science and Technology (DST), Govt. of India, New Delhi, for the financial assistance through the Indo-South Africa Joint Collaborative Project (DST/INT/South Africa/P-10/2016 dated 15 September 2016) to AM and as Women Scientist [Ref. No. SR/WOS-A/LS-564/2011 (G) dated 26 July 2012], DST, New Delhi, to PP. We would like to thank M. Radhakrishna Rao for his suggestions.
Publisher Copyright:
Copyright © 2022 Thorat, Kaniyassery, Poojari, Rangel, Tantry, Kiran, Joshi, Rai, Botha and Muthusamy.
PY - 2022/6/14
Y1 - 2022/6/14
N2 - Ashwagandha (Withania somnifera L. Dunal) is a medicinally important plant with withanolides as its major bioactive compounds, abundant in the roots and leaves. We examined the influence of plant growth regulators (PGRs) on direct organogenesis, adventitious root development, withanolide biosynthetic pathway gene expression, withanolide contents, and metabolites during vegetative and reproductive growth phases under in vitro and ex vitro conditions. The highest shooting responses were observed with 6-benzylaminopurine (BAP) (2.0 mg L–1) + Kinetin (KIN) (1.5 mg L–1) supplementation. Furthermore, BAP (2.0 mg L–1) + KIN (1.5 mg L–1) + gibberellic acid (GA3) (0.5 mg L–1) exhibited better elongation responses with in vitro flowering. Half-strength MS medium with indole-3-butyric acid (IBA) (1.5 mg L–1) exhibited the highest rooting responses and IBA (1.0 mg L–1) with highest fruits, and overall biomass. Higher contents of withaferin A (WFA) [∼8.2 mg g–1 dry weight (DW)] were detected in the reproductive phase, whereas substantially lower WFA contents (∼1.10 mg g–1 DW) were detected in the vegetative phase. Cycloartenol synthase (CAS) (P = 0.0025), sterol methyltransferase (SMT) (P = 0.0059), and 1-deoxy-D-xylulose-5-phosphate reductase (DXR) (P = 0.0375) genes resulted in a significant fold change in expression during the reproductive phase. The liquid chromatography-mass spectrometry (LC-MS) analysis revealed metabolites that were common (177) and distinct in reproductive (218) and vegetative (167) phases. Adventitious roots cultured using varying concentrations of indole-3-acetic acid (IAA) (0.5 mg L–1) + IBA (1.0 mg L–1) + GA3 (0.2 mg L–1) exhibited the highest biomass, and IAA (0.5 mg L–1) + IBA (1.0 mg L–1) exhibited the highest withanolides content. Overall, our findings demonstrate the peculiarity of withanolide biosynthesis during distinct growth phases, which is relevant for the large-scale production of withanolides.
AB - Ashwagandha (Withania somnifera L. Dunal) is a medicinally important plant with withanolides as its major bioactive compounds, abundant in the roots and leaves. We examined the influence of plant growth regulators (PGRs) on direct organogenesis, adventitious root development, withanolide biosynthetic pathway gene expression, withanolide contents, and metabolites during vegetative and reproductive growth phases under in vitro and ex vitro conditions. The highest shooting responses were observed with 6-benzylaminopurine (BAP) (2.0 mg L–1) + Kinetin (KIN) (1.5 mg L–1) supplementation. Furthermore, BAP (2.0 mg L–1) + KIN (1.5 mg L–1) + gibberellic acid (GA3) (0.5 mg L–1) exhibited better elongation responses with in vitro flowering. Half-strength MS medium with indole-3-butyric acid (IBA) (1.5 mg L–1) exhibited the highest rooting responses and IBA (1.0 mg L–1) with highest fruits, and overall biomass. Higher contents of withaferin A (WFA) [∼8.2 mg g–1 dry weight (DW)] were detected in the reproductive phase, whereas substantially lower WFA contents (∼1.10 mg g–1 DW) were detected in the vegetative phase. Cycloartenol synthase (CAS) (P = 0.0025), sterol methyltransferase (SMT) (P = 0.0059), and 1-deoxy-D-xylulose-5-phosphate reductase (DXR) (P = 0.0375) genes resulted in a significant fold change in expression during the reproductive phase. The liquid chromatography-mass spectrometry (LC-MS) analysis revealed metabolites that were common (177) and distinct in reproductive (218) and vegetative (167) phases. Adventitious roots cultured using varying concentrations of indole-3-acetic acid (IAA) (0.5 mg L–1) + IBA (1.0 mg L–1) + GA3 (0.2 mg L–1) exhibited the highest biomass, and IAA (0.5 mg L–1) + IBA (1.0 mg L–1) exhibited the highest withanolides content. Overall, our findings demonstrate the peculiarity of withanolide biosynthesis during distinct growth phases, which is relevant for the large-scale production of withanolides.
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U2 - 10.3389/fpls.2022.917770
DO - 10.3389/fpls.2022.917770
M3 - Article
C2 - 35774803
AN - SCOPUS:85133507703
SN - 1664-462X
VL - 13
SP - 917770
JO - Frontiers in Plant Science
JF - Frontiers in Plant Science
M1 - 917770
ER -