Hairy Root Cultures: A Novel Way to Mass Produce Plant Secondary Metabolites

Sakthivel, Natarajan; Largia, Muthaiah Joe Virgin; Kumar, Ramakrishnan Ramesh; Satish, Lakkakula; Swamy, Mallappa Kumara; Ramesh, Manikandan

doi:10.1007/978-981-19-5779-6_17

Cited by 7 publications

(3 citation statements)

References 144 publications

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“…Hairy root culture presents an alternative approach for enhancing biochemical properties and specific organic compounds, along with other classes of secondary metabolites [ 202 ]. This method involves transforming root cultures using Agrobacterium rhizogenes , resulting in the formation of hairy roots through the insertion of transfer DNA (T-DNA) [ 203 ].…”

Section: Application Of Plant Tissue Culture Techniques Associated Wi...mentioning

confidence: 99%

From Nature to Lab: A Review of Secondary Metabolite Biosynthetic Pathways, Environmental Influences, and In Vitro Approaches

Reshi,

Ahmad,

Lukatkin

et al. 2023

Metabolites

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Secondary metabolites are gaining an increasing importance in various industries, such as pharmaceuticals, dyes, and food, as is the need for reliable and efficient methods of procuring these compounds. To develop sustainable and cost-effective approaches, a comprehensive understanding of the biosynthetic pathways and the factors influencing secondary metabolite production is essential. These compounds are a unique type of natural product which recognizes the oxidative damage caused by stresses, thereby activating the defence mechanism in plants. Various methods have been developed to enhance the production of secondary metabolites in plants. The elicitor-induced in vitro culture technique is considered an efficient tool for studying and improving the production of secondary metabolites in plants. In the present review, we have documented various biosynthetic pathways and the role of secondary metabolites under diverse environmental stresses. Furthermore, a practical strategy for obtaining consistent and abundant secondary metabolite production via various elicitation agents used in culturing techniques is also mentioned. By elucidating the intricate interplay of regulatory factors, this review paves the way for future advancements in sustainable and efficient production methods for high-value secondary metabolites.

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Section: Application Of Plant Tissue Culture Techniques Associated Wi...mentioning

confidence: 99%

From Nature to Lab: A Review of Secondary Metabolite Biosynthetic Pathways, Environmental Influences, and In Vitro Approaches

Reshi,

Ahmad,

Lukatkin

et al. 2023

Metabolites

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show abstract

“…While extraction and puri cation of the metabolites from these owers have been the norm [11], innovative biotechnological approaches are now being explored. For instance, hairy root (HR) culture [12], enhanced through biotechnological tools like the CRISPR/Cas9 system via Agrobacterium rhizogenes-mediated transformation, offer a potent alternative for metabolite production (Gantait & Mukherjee, 2021). This biotechnological approach possesses remarkable properties that set it apart.…”

Section: Introductionmentioning

confidence: 99%

Mother Transformer: A High-Throughput, Cost-Effective in Planta Hairy Root Transformation Method for Cannabis

Ajdanian,

Torkamaneh

2024

Preprint

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Background Hairy root (HR) transformation assays mediated by Agrobacterium rhizogenes, both in vitro and ex vitro, are essential tools in plant biotechnology and functional genomics. These assays can be significantly influenced by various factors, which ultimately can enhance the efficiency. In this study, we optimized a two-step ex vitro HR transformation method using the actual mother plant combined with the RUBY system and tested with existing methods.Results The two-step ex vitro method proved more efficient than both the one-step ex vitro and in vitro methods, with the highest transformation efficiency of 90% observed in the actual plant. This technique also demonstrated a faster and less complicated approach, reducing time to achieve massive transgenic HR formation by 9–29 days compared to other methods.Conclusions A novel, quicker, less complicated, and more efficient two-step transformation method for cannabis has been established, presenting a significantly lower risk of contamination. This protocol is particularly interesting to produce secondary metabolites using the CRISPR/Cas system in cannabis. We anticipate that this method will facilitate substantial time savings by rapidly producing hundreds of transformed samples.

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“…This approach therefore opens the way to achieving the production levels and developmental capacities in hairy roots necessary for their sustainable application as a biotechnological platform ( Padilla et al., 2021 ). Furthermore, the use of omics techniques to study differentially expressed genes in C. asiatica hairy roots lays the groundwork for further investigation into the transcriptional regulation of centelloside content ( Khan et al., 2023 ; Shilpha et al., 2023 ).…”

Section: Introductionmentioning

confidence: 99%

Insights into enhancing Centella asiatica organ cell biofactories via hairy root protein profiling

Alcalde,

Hidalgo-Martinez,

Bru Martínez

et al. 2023

Front. Plant Sci.

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Recent advancements in plant biotechnology have highlighted the potential of hairy roots as a biotechnological platform, primarily due to their rapid growth and ability to produce specialized metabolites. This study aimed to delve deeper into hairy root development in C. asiatica and explore the optimization of genetic transformation for enhanced bioactive compound production. Previously established hairy root lines of C. asiatica were categorized based on their centelloside production capacity into HIGH, MID, or LOW groups. These lines were then subjected to a meticulous label-free proteomic analysis to identify and quantify proteins. Subsequent multivariate and protein network analyses were conducted to discern proteome differences and commonalities. Additionally, the quantification of rol gene copy numbers was undertaken using qPCR, followed by gene expression measurements. From the proteomic analysis, 213 proteins were identified. Distinct proteome differences, especially between the LOW line and other lines, were observed. Key proteins related to essential processes like photosynthesis and specialized metabolism were identified. Notably, potential biomarkers, such as the Tr-type G domain-containing protein and alcohol dehydrogenase, were found in the HIGH group. The presence of ornithine cyclodeaminase in the hairy roots emerged as a significant biomarker linked with centelloside production capacity lines, indicating successful Rhizobium-mediated genetic transformation. However, qPCR results showed an inconsistency with rol gene expression levels, with the HIGH line displaying notably higher expression, particularly of the rolD gene. The study unveiled the importance of ornithine cyclodeaminase as a traceable biomarker for centelloside production capacity. The strong correlation between this biomarker and the rolD gene emphasizes its potential role in optimizing genetic transformation processes in C. asiatica.

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Hairy Root Cultures: A Novel Way to Mass Produce Plant Secondary Metabolites

Cited by 7 publications

References 144 publications

From Nature to Lab: A Review of Secondary Metabolite Biosynthetic Pathways, Environmental Influences, and In Vitro Approaches

From Nature to Lab: A Review of Secondary Metabolite Biosynthetic Pathways, Environmental Influences, and In Vitro Approaches

Mother Transformer: A High-Throughput, Cost-Effective in Planta Hairy Root Transformation Method for Cannabis

Insights into enhancing Centella asiatica organ cell biofactories via hairy root protein profiling

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