Gene expression analysis of the withanolide biosynthetic pathway in hairy root cultures of Withania somnifera elicited with methyl jasmonate and the fungus Piriformospora indica

Saxena, Pravin; Ahlawat, Seema; Ali, Athar; Khan, Shazia; Abdin, Malik Zainul

doi:10.1007/s13199-016-0416-9

Cited by 24 publications

(11 citation statements)

References 41 publications

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“…In this study, HMGR exhibited the highest fold change difference with respect to the vegetative phase, followed by a significant fold change difference in SMT , CAS , and DXR expression. Similar results were observed with the overexpression of CAS (3.08-fold) in the hairy root culture of W. somnifera , which enhanced withanolide production compared to the control groups ( Saxena et al, 2017 ). CAS regulates the synthesis of cycloartenol from the precursor squalene 2,3-epoxidases, while the increased expression of DXR and HMGR promotes the production of withanolide contents in leaf tissues during drought stress conditions ( Singh et al, 2014 ; Singh et al, 2018 ).…”

Section: Discussionsupporting

confidence: 80%

Differential Gene Expression and Withanolides Biosynthesis During in vitro and ex vitro Growth of Withania somnifera (L.) Dunal

et al. 2022

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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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Section: Discussionsupporting

confidence: 80%

Differential Gene Expression and Withanolides Biosynthesis During in vitro and ex vitro Growth of Withania somnifera (L.) Dunal

et al. 2022

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“…Different concentrations (1, 3, and 5%) of cell homogenates of root endophytic fungus Piriformospora indica (cell homogenate of Piriformospora indica [CHP]), used as biofertilizer, bioregulator, and bioprotector against stress conditions, was used as biotic elicitor in HRC of Withania somnifera for varying time periods . Treatment of W. somnifera HRCs for 48 h with 3% CHP enhanced biomass as well as withanolides viz., withanolide A, withaferin A, withanoside IV, and withanoside V production (1.15‐, 2.7‐, 2.5‐, 2.34‐, and 2.9‐fold, respectively) as compared to untreated hairy roots.…”

Section: Elicitors and Their Effects On Hrcsmentioning

confidence: 99%

“…Treatment of W. somnifera HRCs for 48 h with 3% CHP enhanced biomass as well as withanolides viz., withanolide A, withaferin A, withanoside IV, and withanoside V production (1.15‐, 2.7‐, 2.5‐, 2.34‐, and 2.9‐fold, respectively) as compared to untreated hairy roots. Further, gene expression studies revealed that the application of 3% CHP for 48 h upregulated the expression of HMGR (3.2‐fold), farnesyl pyrophosphate synthase (FPPS) (3.55‐fold), squalene synthase (SS) (2.87‐fold), squalene epoxidase (SE) (3.25‐fold), cycloartenol synthase (CAS) (3.08‐fold), obtusifoliol‐14‐demethylase (4.42‐fold), sterol methyltransferase 1 (4.81‐fold), and sterol‐22‐desaturase (5.02‐fold) genes than in untreated hairy roots .…”

Section: Elicitors and Their Effects On Hrcsmentioning

confidence: 99%

Elicitation: A biotechnological tool for enhanced production of secondary metabolites in hairy root cultures

Halder

Sarkar

Jha

2019

Engineering in Life Sciences

194

116

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Elicitation is a possible aid to overcome various difficulties associated with the large‐scale production of most commercially important bioactive secondary metabolites from wild and cultivated plants, undifferentiated or differentiated cultures. Secondary metabolite accumulation in vitro or their efflux in culture medium has been elicited in the undifferentiated or differentiated tissue cultures of several plant species by the application of a low concentration of biotic and abiotic elicitors in the last three decades. Hairy root cultures are preferred for the application of elicitation due to their genetic and biosynthetic stability, high growth rate in growth regulator‐free media, and production consistence in response to elicitor treatment. Elicitors act as signal, recognized by elicitor‐specific receptors on the plant cell membrane and stimulate defense responses during elicitation resulting in increased synthesis and accumulation of secondary metabolites. Optimization of various parameters, such as elicitor type, concentration, duration of exposure, and treatment schedule is essential for the effectiveness of the elicitation strategies. Combined application of different elicitors, integration of precursor feeding, or replenishment of medium or in situ product recovery from the roots/liquid medium with the elicitor treatment have showed improved accumulation of secondary metabolites due to their synergistic effect. This is a comprehensive review about the progress in the elicitation approach to hairy root cultures from 2010 to 2019 and the information provided is valuable and will be of interest for scientists working in this area of plant biotechnology.

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“…Cai et al 142 reported that the number of flavonoids in the Avena sativa L. was markedly increased by the fermentation by the Aspergillus niger. In this context, different concentrations of a root endophytic fungus Piriformospora indica cell homogenates were utilized as an abiotic elicitor in Withania somnifera hairy root for different periods 143 . When the hairy root cultures of Withania somnifera were treated with 3% cell homogenates of Piriformospora indica for 48 hrs, it enhanced the biomass and the production of withanolides, viz., withaferin A, withanolide A, withanoside V and withanoside IV as compared to control (untreated hairy root).…”

Section: Enhancement Of Plant Nutraceuticals Production Bymentioning

confidence: 99%

In vitro Production of Plant Nutraceuticals

Bekheet¹,

Sota²

2022

Biotechnology

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Biotechnology methods particularly tissue culture techniques offer new approaches for the in vitro production of valuable plant nutraceuticals. The large-scale production of secondary metabolites in a year-round system without seasonal constraints is one of the advantages of plant tissue culture techniques. Furthermore, enhancement of in vitro nutraceuticals production can be realized by a selection of high-producing cell lines and medium optimization. In this respect, several techniques have been adopted to improve the production of plant-derived nutraceuticals such as mutation, elicitation, precursor feeding, genetic transformation and metabolic engineering. In vitro mutagenesis of cultured cells and tissues by irradiation or chemical mutagens represents a feasible method for the improvement of nutraceuticals production. Likewise, transformation is another strategy that is used to enhance nutraceuticals in plant cells or organ tissues. At this point, hairy root culture induced by Agrobacterium rhizogenes is the most promising transformation technique used for in vitro production of valuable plant compounds. The benefits of hairy roots are high metabolite productivity, high growth rates and inherent genetic stability. On the other hand, due to its effectiveness and practical feasibility, elicitation is considered the most applied strategy for enhancing the production of desired compounds in plant biotechnology. Biotic (biological origin) and abiotic (non-biological origin) elicitors are used to increase the production of plant metabolites by different effects on the cellular processes in the plant system. Furthermore, bioreactors developed for in vitro industrial-scale production of desired compounds can be used for continuous and scaling-up plant nutraceuticals production. This article discusses different aspects of biotechnology used for the selection and enhancement production of nutraceuticals from plant cultures grown cells, in vitro.

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Gene expression analysis of the withanolide biosynthetic pathway in hairy root cultures of Withania somnifera elicited with methyl jasmonate and the fungus Piriformospora indica

Cited by 24 publications

References 41 publications

Differential Gene Expression and Withanolides Biosynthesis During in vitro and ex vitro Growth of Withania somnifera (L.) Dunal

Differential Gene Expression and Withanolides Biosynthesis During in vitro and ex vitro Growth of Withania somnifera (L.) Dunal

Elicitation: A biotechnological tool for enhanced production of secondary metabolites in hairy root cultures

In vitro Production of Plant Nutraceuticals

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