An Efficient Hairy Root System for Withanolide Production in Withania somnifera (L.) Dunal

Shajahan, Appakan; Thilip, Chandrasekaran; Faizal, Kunnampalli; Mehaboob, Valiyaparambath Musfir; Raja, Palusamy; Aslam, Abubakker; Kathiravan, K.

doi:10.1007/978-3-319-69769-7_7

Cited by 13 publications

(3 citation statements)

References 28 publications

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“…In this case, the plants selected are those species which naturally produce the compound of interest. Some examples of specialized metabolites already produced in wild-type HRCs are azadirachtin (biopesticide) (Thakore and Srivastava, 2017), betalain (red pigments for food industry) (Pavlov et al, 2005), camptothecin (antitumor agent in the treatment of ovarian and colorectal cancers treatment) (Wetterauer et al, 2018), and whitanolide A (brain regenerative compound) (Shajahan et al, 2017). A transgene can also be introduced in wild type HRs in order to increase the amount of a particular specialized metabolite (Jouhikainen et al, 1999;Moyano et al, 2003;Zhang et al, 2004;Ritala et al, 2014).…”

Section: Production Of Specialized Metabolitesmentioning

confidence: 99%

Hairy Root Cultures—A Versatile Tool With Multiple Applications

Gutierrez-Valdes

Häkkinen

Lemasson³

et al. 2020

Front. Plant Sci.

189

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Hairy roots derived from the infection of a plant by Rhizobium rhizogenes (previously referred to as Agrobacterium rhizogenes) bacteria, can be obtained from a wide variety of plants and allow the production of highly diverse molecules. Hairy roots are able to produce and secrete complex active glycoproteins from a large spectrum of organisms. They are also adequate to express plant natural biosynthesis pathways required to produce specialized metabolites and can benefit from the new genetic tools available to facilitate an optimized production of tailor-made molecules. This adaptability has positioned hairy root platforms as major biotechnological tools. Researchers and industries have contributed to their advancement, which represents new alternatives from classical systems to produce complex molecules. Now these expression systems are ready to be used by different industries like pharmaceutical, cosmetics, and food sectors due to the development of fully controlled large-scale bioreactors. This review aims to describe the evolution of hairy root generation and culture methods and to highlight the possibilities offered by hairy roots in terms of feasibility and perspectives.

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Section: Production Of Specialized Metabolitesmentioning

confidence: 99%

Hairy Root Cultures—A Versatile Tool With Multiple Applications

Gutierrez-Valdes

Häkkinen

Lemasson³

et al. 2020

Front. Plant Sci.

189

View full text Add to dashboard Cite

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“…Based on this mechanism, a valuable biotechnological application was exploited, known as hairy root culture (HRC). In the past decade, HRC was extensively applied to a wide variety of plant species (Shajahan et al ., ; Veena and Taylor, ) for a diverse range of biotechnological studies, such as phytochemical and recombinant protein production, analyses of rhizosphere physiology and biochemistry and molecular breeding. For example, overexpression of the geraniol 10‐hydroxylase (G10H) gene or a jasmonate‐responsive APETALA2 (AP2)‐domain transcript factor in HRC, improves catharanthine production in Catharanthus roseus (Paul et al ., ).…”

Section: Introductionmentioning

confidence: 99%

Development of an efficient root transgenic system for pigeon pea and its application to other important economically plants

Meng

Yang

Dong

et al. 2019

Plant Biotechnology Journal

101

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Summary For non‐model plants, functional characterization of genes is still hampered by lack of efficient stable transformation procedures. Here, we report a simple, fast and efficient transformation technique with Agrobacterium rhizogenes for generating stable transgenic roots in living plants to facilitate functional studies in vivo . We showed that injection of A . rhizogenes into stems of various plant species lead to stable transgenic root generation, which can sustain plant growth after the original, non‐transgenic roots were cut off. A transformation system was established for pigeon pea, a major woody food crop, after optimizing the selection of A. rhizogenes strains, bacterium concentration, injection position and seedling age. RT ‐ PCR and fluorescence observation indicated a transgenic root induction efficiency of about 39% in pigeon pea. Furthermore, induction of hairy roots was achieved in nine out of twelve tested economically important plants at an efficiency of 15–39%. As proof of concept, bimolecular fluorescence complementation (Bi FC ) assay was applied to test the interaction between Cc CIPK 14 and Cc CBL 1/2 in pigeon pea. Additionally, ectopic expression of the bZIP transcription factor Md HY 5 from apple confirmed the utility of the transformation technique for engineering anthocyanin synthesis in roots. Taken together, we show that this method allows fast in vivo studies of gene function in a wide range of plant species.

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“…Upon infection, A. rhizogenes transfers its Ri plasmid into host nuclei, by which the hairy roots can be genetically modified. The root genetic transformation system mediated by A. rhizogenes involves a simple and rapid procedure, which avoids tissue culture and has been widely employed for transgenic studies in various plant species over the past decade [ 11 – 15 ]. Although the hairy root system has proven valuable for studying gene function, its utility in tracking organelle activities within plant cells has been largely unexplored.…”

Section: Introductionmentioning

confidence: 99%

Tracking organelle activities through efficient and stable root genetic transformation system in woody plants

Gong,

Chen,

et al. 2023

Horticulture Research

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Due to the protracted transgenic timeline and low efficiency in stable genetic transformation of woody plants, there has been limited exploration of real-time organelle imaging within stable transgenic woody plant cells. Here, we established an efficient in vivo genetic transformation system for woody plants using Agrobacterium rhizogenes-mediated approach. This system was successfully validated in multiple perennial woody species. Using citrus as a model, we introduced organelle-targeted fluorescent reporters via genetic transformation and investigated their subcellular localization and dynamics using advanced imaging techniques, such as confocal microscopy and live-cell imaging. Moreover, we subjected transgenic MT-GFP-labeled mitochondria in root cells to stress conditions simulating agricultural adversities faced by fruit crops. The stress-induced experiments revealed notable alterations in mitochondrial morphology. Our study contributes novel insights into membrane trafficking processes, protein localization dynamics, and cellular physiology in woody plants, while also providing stable and efficient genetic transformation methods for perennial woody species.

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An Efficient Hairy Root System for Withanolide Production in Withania somnifera (L.) Dunal

Cited by 13 publications

References 28 publications

Hairy Root Cultures—A Versatile Tool With Multiple Applications

Hairy Root Cultures—A Versatile Tool With Multiple Applications

Development of an efficient root transgenic system for pigeon pea and its application to other important economically plants

Tracking organelle activities through efficient and stable root genetic transformation system in woody plants

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