Tailoring enhanced production of aervine in Aerva lanata (L.) Juss. Ex Schult by Agrobacterium rhizogenes- mediated hairy root cultures

Boobalan, Selvakumar; Kamalanathan, Desingu

doi:10.1016/j.indcrop.2020.112814

Cited by 27 publications

(7 citation statements)

References 63 publications

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“…As a "natural genetic engineer", A. rhizogenes has the ability to transfer the T-DNA region of its Ri plasmid (root-inducing plasmid) into the plant's genome [28]. The injection of this plasmid into the host cells and stimulation of the production and development of highly branched hairy roots are the consequence of the expression of a group of rol/oncogenes (rolA, rolB, rolC, and rolD) in the bacterial Ri plasmid [29,30]. These rol genes are involved in hairy root syndrome induction through different biochemical functions, such as phytohormone homeostasis, reactive oxygen species (ROS) homeostasis, and ornithine cyclodeaminase.…”

Section: Hairy Root Transformationmentioning

confidence: 99%

CRISPR/Cas9 in Planta Hairy Root Transformation: A Powerful Platform for Functional Analysis of Root Traits in Soybean

Niazian

Belzile

Torkamaneh

2022

Plants

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Sequence and expression data obtained by next-generation sequencing (NGS)-based forward genetics methods often allow the identification of candidate causal genes. To provide true experimental evidence of a gene’s function, reverse genetics techniques are highly valuable. Site-directed mutagenesis through transfer DNA (T-DNA) delivery is an efficient reverse screen method in plant functional analysis. Precise modification of targeted crop genome sequences is possible through the stable and/or transient delivery of clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein (CRISPR/Cas) reagents. Currently, CRISPR/Cas9 is the most powerful reverse genetics approach for fast and precise functional analysis of candidate genes/mutations of interest. Rapid and large-scale analyses of CRISPR/Cas-induced mutagenesis is achievable through Agrobacterium rhizogenes-mediated hairy root transformation. The combination of A. rhizogenes hairy root-CRISPR/Cas provides an extraordinary platform for rapid, precise, easy, and cost-effective “in root” functional analysis of genes of interest in legume plants, including soybean. Both hairy root transformation and CRISPR/Cas9 techniques have their own complexities and considerations. Here, we discuss recent advancements in soybean hairy root transformation and CRISPR/Cas9 techniques. We highlight the critical factors required to enhance mutation induction and hairy root transformation, including the new generation of reporter genes, methods of Agrobacterium infection, accurate gRNA design strategies, Cas9 variants, gene regulatory elements of gRNAs and Cas9 nuclease cassettes and their configuration in the final binary vector to study genes involved in root-related traits in soybean.

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Section: Hairy Root Transformationmentioning

confidence: 99%

CRISPR/Cas9 in Planta Hairy Root Transformation: A Powerful Platform for Functional Analysis of Root Traits in Soybean

Niazian

Belzile

Torkamaneh

2022

Plants

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“…Detection was performed at a wavelength of 254 nm using a UV detector. External instrumental facilities were utilized for the HPLC analysis of the plant samples [30]. The samples were stored at −80 • C for this purpose.…”

Section: Determination Of Aervine Contentmentioning

confidence: 99%

Enhancing Bioactive Metabolite Production in Aerva sanguinolenta Callus Cultures through Silver Nanoparticle and Salicylic Acid Elicitation

et al. 2023

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Callus elicitation is advantageous for metabolite production due to its ability to increase yield, provide controllable conditions, and allow for genetic manipulation, offering a sustainable and scalable alternative to traditional plant-based extraction methods for the production of bioactive substances. In this research, in vitro callus cultures (CCs) of the wild medicinal plant Aerva sanguinolenta were used to evaluate the efficacy of various elicitation regimes of silver nanoparticles (AgNPs) and salicylic acid (SA) to evoke an increased production of secondary metabolites, such as aervine and antioxidant metabolites. Three concentrations of SA (i.e., 20, 50, and 100 µM) and three concentrations of AgNPs (i.e., 30, 60, and 90 µg/L) were used on shoot explant cultures using MS (Murashige and Skoog) media. All the SA and AgNP elicitation treatments significantly increased the production of antioxidant metabolites, total phenolic contents (TPCs), and total flavonoid contents (TFCs) compared to the control treatment experiments. The contents of aervine were increased significantly upon elicitation compared to the control trial. Furthermore, the antioxidant potential of the test extract was enhanced compared to the control treatment. Comparatively, the AgNPs were more beneficial as elicitors than the SA treatments. The elicitation treatments with about 90 µg/L AgNPs and 100 µM SA were the best among all elicitation regimes. Callus elicitation with SA and AgNPs can stimulate increased metabolite production and be used as a sustainable practice in the welfare and service industries for drug development and drug discovery.

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“…Many published review articles and a wide range of scientific reports notify that the solution for most of the obstacles related to plant biomass harvesting for secondary metabolites production comes with in vitro bioprocessing of hairy roots [ 6 , 7 , 8 , 9 ]. Due to stable genetic alterations induced by Rhizobium rhizogenes , hairy root features intensified growth in a culture media free of plant growth regulators, with the propensity for lateral branching and enhanced biosynthesis of secondary metabolites specific for root tissue differentiation [ 10 ]. Such a beneficial phenotype of transgenic hairy roots fits well for industrial applications of in vitro plant biomass cultures, making it possible to conduct validated scaled-up bioprocesses in bioreactors (e.g., disposable aero- or hydroponic systems) and providing the tool to overcome ecological or economic limitations [ 11 , 12 ].…”

Section: Introductionmentioning

confidence: 99%

Selective Impact of MTMS-Based Xerogel Morphology on Boosted Proliferation and Enhanced Naphthoquinone Production in Cultures of Rindera graeca Transgenic Roots

Wierzchowski

Nowak

Kawka

et al. 2022

IJMS

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In situ extraction is a method for separating plant secondary metabolites from in vitro systems of plant biomass cultures. The study aimed to investigate the MTMS-based xerogels morphology effect on the growth kinetics and deoxyshikonin productivity in xerogel-supported in vitro culture systems of Rindera graeca hairy root. Cultures were supplemented with three types of xerogel, i.e., mesoporous gel, microporous gel, and agglomerated precipitate, in the disintegrated or monolithic form. Structure, oil sorption capacity, and SEM analyses for xerogel-based additives were performed. Application of monolithic macroporous xerogel resulted in the highest biomass proliferation, i.e., 5.11-fold fresh biomass increase after four weeks of the screening culture. The highest deoxyshikonin production (i.e., 105.03 µg) was noted when hairy roots were maintained with particles of disintegrated mesoporous xerogel. The detailed kinetics investigations (6-week culture) revealed the highest growth of hairy root biomass and secondary metabolite production, equaling 9.46-fold fresh weight biomass and 204.08 µg deoxyshikonin, respectively. MTMS-based xerogels have been recognized as selective biocompatible scaffolds for boosting the proliferation of transgenic roots or for productivity enhancement of naphthoquinones without detrimental effects on biomass growth, and their successful applicability in in situ removal of secondary plant metabolites has been experimentally confirmed.

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Tailoring enhanced production of aervine in Aerva lanata (L.) Juss. Ex Schult by Agrobacterium rhizogenes- mediated hairy root cultures

Cited by 27 publications

References 63 publications

CRISPR/Cas9 in Planta Hairy Root Transformation: A Powerful Platform for Functional Analysis of Root Traits in Soybean

CRISPR/Cas9 in Planta Hairy Root Transformation: A Powerful Platform for Functional Analysis of Root Traits in Soybean

Enhancing Bioactive Metabolite Production in Aerva sanguinolenta Callus Cultures through Silver Nanoparticle and Salicylic Acid Elicitation

Selective Impact of MTMS-Based Xerogel Morphology on Boosted Proliferation and Enhanced Naphthoquinone Production in Cultures of Rindera graeca Transgenic Roots

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