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Journal Article•DOI•

Hairy Root Culture for Mass-Production of High-Value Secondary Metabolites

Smita Srivastava1, Ashok K. Srivastava1•
Indian Institutes of Technology1
01 Jan 2007-Critical Reviews in Biotechnology (Crit Rev Biotechnol)-Vol. 27, Iss: 1, pp 29-43
TL;DR: The present review highlights the nature, applications, perspectives and scale up of hairy root cultures for the production of valuable secondary metabolites.
Abstract: Plant cell cultivations are being considered as an alternative to agricultural processes for producing valuable phytochemicals. Since many of these products (secondary metabolites) are obtained by direct extraction from plants grown in natural habitat, several factors can alter their yield. The use of plant cell cultures has overcome several inconveniences for the production of these secondary metabolites. Organized cultures, and especially root cultures, can make a significant contribution in the production of secondary metabolites. Most of the research efforts that use differentiated cultures instead of cell suspension cultures have focused on transformed (hairy) roots. Agrobacterium rhizogenes causes hairy root disease in plants. The neoplastic (cancerous) roots produced by A. rhizogenes infection are characterized by high growth rate, genetic stability and growth in hormone free media. These genetically transformed root cultures can produce levels of secondary metabolites comparable to that of intact plants. Hairy root cultures offer promise for high production and productivity of valuable secondary metabolites (used as pharmaceuticals, pigments and flavors) in many plants. The main constraint for commercial exploitation of hairy root cultivations is the development and scaling up of appropriate reactor vessels (bioreactors) that permit the growth of interconnected tissues normally unevenly distributed throughout the vessel. Emphasis has focused on designing appropriate bioreactors suitable to culture the delicate and sensitive plant hairy roots. Recent reactors used for mass production of hairy roots can roughly be divided as liquid-phase, gas-phase, or hybrid reactors. The present review highlights the nature, applications, perspectives and scale up of hairy root cultures for the production of valuable secondary metabolites.
Citations
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Journal Article•DOI•
Carbon flow in the rhizosphere: carbon trading at the soil–root interface

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Davey L. Jones1, Christophe Nguyen2, Roger D. Finlay•
Bangor University1, Institut national de la recherche agronomique2
25 Feb 2009-Plant and Soil
TL;DR: Due to the importance of rhizodeposition in regulating ecosystem functioning, it is critical that future research focuses on resolving the quantitative importance of the different C and N fluxes operating in the rhizosphere and the ways in which these vary spatially and temporally.
Abstract: The loss of organic and inorganic carbon from roots into soil underpins nearly all the major changes that occur in the rhizosphere. In this review we explore the mechanistic basis of organic carbon and nitrogen flow in the rhizosphere. It is clear that C and N flow in the rhizosphere is extremely complex, being highly plant and environment dependent and varying both spatially and temporally along the root. Consequently, the amount and type of rhizodeposits (e.g. exudates, border cells, mucilage) remains highly context specific. This has severely limited our capacity to quantify and model the amount of rhizodeposition in ecosystem processes such as C sequestration and nutrient acquisition. It is now evident that C and N flow at the soil–root interface is bidirectional with C and N being lost from roots and taken up from the soil simultaneously. Here we present four alternative hypotheses to explain why high and low molecular weight organic compounds are actively cycled in the rhizosphere. These include: (1) indirect, fortuitous root exudate recapture as part of the root’s C and N distribution network, (2) direct re-uptake to enhance the plant’s C efficiency and to reduce rhizosphere microbial growth and pathogen attack, (3) direct uptake to recapture organic nutrients released from soil organic matter, and (4) for inter-root and root–microbial signal exchange. Due to severe flaws in the interpretation of commonly used isotopic labelling techniques, there is still great uncertainty surrounding the importance of these individual fluxes in the rhizosphere. Due to the importance of rhizodeposition in regulating ecosystem functioning, it is critical that future research focuses on resolving the quantitative importance of the different C and N fluxes operating in the rhizosphere and the ways in which these vary spatially and temporally.

1,247 citations

Cites background from "Hairy Root Culture for Mass-Product..."

  • ...The neoplastic (cancerous) transformed roots are genetically stable and can grow rapidly in the absence of shoots in a hormone free medium making them suitable for the controlled excretion and collection of secondary metabolites (Srivastava and Srivastava 2007)....

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Journal Article•DOI•
In vitro plant tissue culture: means for production of biological active compounds

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Claudia A. Espinosa-Leal1, César A. Puente-Garza1, Silverio García-Lara1•
Monterrey Institute of Technology and Higher Education1
07 May 2018-Planta
TL;DR: This review summarizes the important bioactive compounds currently produced by plant tissue culture and the fundamental methods and plants employed for their production.
Abstract: Plant tissue culture as an important tool for the continuous production of active compounds including secondary metabolites and engineered molecules. Novel methods (gene editing, abiotic stress) can improve the technique. Humans have a long history of reliance on plants for a supply of food, shelter and, most importantly, medicine. Current-day pharmaceuticals are typically based on plant-derived metabolites, with new products being discovered constantly. Nevertheless, the consistent and uniform supply of plant pharmaceuticals has often been compromised. One alternative for the production of important plant active compounds is in vitro plant tissue culture, as it assures independence from geographical conditions by eliminating the need to rely on wild plants. Plant transformation also allows the further use of plants for the production of engineered compounds, such as vaccines and multiple pharmaceuticals. This review summarizes the important bioactive compounds currently produced by plant tissue culture and the fundamental methods and plants employed for their production.

281 citations

Cites background from "Hairy Root Culture for Mass-Product..."

  • ...Hybrid systems, which include a liquid- and a gas phase, are reactors that attend the disadvantage of the null distribution uniformity of cells in gas-phase reactors, and the limitations of the mass transfer in liquid-phase reactors; these systems offer the best compromise between the two systems, used in alternate way (Srivastava and Srivastava 2007)....

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  • ...…are reactors that attend the disadvantage of the null distribution uniformity of cells in gas-phase reactors, and the limitations of the mass transfer in liquid-phase reactors; these systems offer the best compromise between the two systems, used in alternate way (Srivastava and Srivastava 2007)....

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  • ...Srivastava and Srivastava (2007) described the different configurations and parameters of bioreactors for hairy roots culture, which include liquid phase, gas phase and hybrid systems....

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Journal Article•DOI•
Recent advances towards development and commercialization of plant cell culture processes for the synthesis of biomolecules.

[...]

Sarah A. Wilson1, Susan C. Roberts•
University of Massachusetts Amherst1
01 Apr 2012-Plant Biotechnology Journal
TL;DR: Recent advancements in plant cell culture processing technology are discussed, focusing on progress towards overcoming the problems associated with commercialization of these production systems and highlighting recent commercial successes.
Abstract: Plant cell culture systems were initially explored for use in commercial synthesis of several high-value secondary metabolites, allowing for sustainable production that was not limited by the low yields associated with natural harvest or the high cost associated with complex chemical synthesis. Although there have been some commercial successes, most notably paclitaxel production from Taxus sp., process limitations exist with regards to low product yields and inherent production variability. A variety of strategies are being developed to overcome these limitations including elicitation, in situ product removal and metabolic engineering with single genes and transcription factors. Recently, the plant cell culture production platform has been extended to pharmaceutically active heterologous proteins. Plant systems are beneficial because they are able to produce complex proteins that are properly glycosylated, folded and assembled without the risk of contamination by toxins that are associated with mammalian or microbial production systems. Additionally, plant cell culture isolates transgenic material from the environment, allows for more controllable conditions over field-grown crops and promotes secretion of proteins to the medium, reducing downstream purification costs. Despite these benefits, the increase in cost of heterologous protein synthesis in plant cell culture as opposed to field-grown crops is significant and therefore processes must be optimized with regard to maximizing secretion and enhancing protein stability in the cell culture media. This review discusses recent advancements in plant cell culture processing technology, focusing on progress towards overcoming the problems associated with commercialization of these production systems and highlighting recent commercial successes.

270 citations

Journal Article•DOI•
Pharmaceutically Active Natural Product Synthesis and Supply via Plant Cell Culture Technology

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Martin E. Kolewe1, Vishal Gaurav1, Susan C. Roberts1•
University of Massachusetts Amherst1
07 Apr 2008-Molecular Pharmaceutics
TL;DR: This review focuses on some of the key challenges in utilizing and commercializing plant cell culture suspension technology, with a focus on pharmaceutically active natural products.
Abstract: The chemical diversity of plant-derived natural products allows them to function in a multitude of ways including flavor enhancers, agricultural chemicals, and importantly, human medicinals Supply of pharmaceutically active natural products is often a challenge due to the slow growing nature of some species, low yields found in nature, and unpredictable variability in accumulation Several production options are available including natural harvestation, total chemical synthesis, semisynthesis from isolated precursors, and expression of plant pathways in microbial systems However, for some medicinal natural products, such as the anticancer agent paclitaxel, where low yields in nature, chemical complexity and lack of knowledge of the complete biosynthetic pathway, preclude many of these options, plant cell culture technology is an attractive alternative for supply Plant cell suspension cultures are amenable to scale-up, environmental optimization, and metabolic engineering This review focuses on some of the key challenges in utilizing and commercializing plant cell culture suspension technology, with a focus on pharmaceutically active natural products Recent research has been directed toward application of traditional strategies such as reactor design, cell immobilization, and enzyme elicitation as well as emerging strategies such as characterizing cellular heterogeneity and variability through flow cytometric techniques, metabolic engineering, and system-wide analysis

236 citations

Journal Article•DOI•
Plant cell culture strategies for the production of natural products

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Marisol Ochoa-Villarreal1, Susan Howat1, Sunmi Hong, Mi Ok Jang, Young-Woo Jin, Eun-Kyong Lee, Gary J. Loake1 •
University of Edinburgh1
01 Mar 2016-Journal of Biochemistry and Molecular Biology
TL;DR: There remain significant obstacles to the commercial synthesis of high-value chemicals from these sources but the relatively recent isolation, culturing and characterisation of cambial meristematic cells (CMCs), provides an emerging platform to circumvent many of these potential difficulties.
Abstract: Plants have evolved a vast chemical cornucopia to support their sessile lifestyles. Man has exploited this natural resource since Neolithic times and currently plant-derived chemicals are exploited for a myriad of applications. However, plant sources of most high-value natural products (NPs) are not domesticated and therefore their production cannot be undertaken on an agricultural scale. Further, these plant species are often slow growing, their populations limiting, the concentration of the target molecule highly variable and routinely present at extremely low concentrations. Plant cell and organ culture constitutes a sustainable, controllable and environmentally friendly tool for the industrial production of plant NPs. Further, advances in cell line selection, biotransformation, product secretion, cell permeabilisation, extraction and scale-up, among others, are driving increases in plant NP yields. However, there remain significant obstacles to the commercial synthesis of high-value chemicals from these sources. The relatively recent isolation, culturing and characterisation of cambial meristematic cells (CMCs), provides an emerging platform to circumvent many of these potential difficulties. [BMB Reports 2016; 49(3): 149-158].

216 citations

Cites background from "Hairy Root Culture for Mass-Product..."

  • ...scopolamine, atropine, anthraquinone, indole alkaloids cardiac glycosides (23)....

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References
More filters
Journal Article•DOI•
A revised medium for rapid growth and bio assays with tobacco tissue cultures

[...]

Toshio Murashige1, Folke Skoog1•
University of Wisconsin-Madison1
01 Jul 1962-Physiologia Plantarum
TL;DR: In vivo redox biosensing resolves the spatiotemporal dynamics of compartmental responses to local ROS generation and provide a basis for understanding how compartment-specific redox dynamics may operate in retrograde signaling and stress 67 acclimation in plants.
Abstract: In experiments with tobacco tissue cultured on White's modified medium (basal meditmi hi Tnhles 1 and 2) supplemenk'd with kiticthi and hidoleacctic acid, a slrikin^' fourlo (ive-told intTease iu yield was ohtaitu-d within a three to Tour week j^rowth period on addition of an aqtteotis exlrarl of tobacco leaves (Fi^'ures 1 and 2). Subse(iueutly it was found Ihiit this jnoniotiou oi' f^rowih was due mainly though nol entirely to inorj^auic rather than organic con.stitttenls in the extract. In the isolation of Rrowth factors from plant tissues and other sources inorj '̂anic salts are fre(|uently carried along with fhe organic fraclioits. When tissue cultures are used for bioassays, therefore, il is necessary lo lake into account increases in growth which may result from nutrient elements or other known constituents of the medium which may he present in the te.st materials. To minimize interference trom rontaminaitis of this type, an altempt has heen made to de\\eh)p a nieditmi with such adequate supplies of all re(iuired tnineral nutrients and cotntnott orgattic cottslitueitls that no apprecial»le change in growth rate or yield will result from the inlroduclion of additional amounts in the range ordinarily expected to be present in tnaterials to be assayed. As a point of referetice for this work some of the culture media in mc)st common current use will he cotisidered briefly. For ease of comparis4)n Iheir mineral compositions are listed in Tables 1 and 2. White's nutrient .solution, designed originally for excised root cultures, was based on Uspeuski and Uspetiskaia's medium for algae and Trelease and Trelease's micronutrieni solution. This medium also was employed successfully in the original cttltivation of callus from the tobacco Iiybrid Nicotiana gtauca x A', tanijadorffii, atitl as further modified by White in 194̂ ^ and by others it has been used for the

63,098 citations

"Hairy Root Culture for Mass-Product..." refers background in this paper

  • ...For example, lawsone normally accumulates only in the aerial part of the plant, but hairy roots of Lawsonia inermis grown in half- or fullstrength MS medium (Murashige and Skoog, 1962) can produce lawsone under dark conditions (Bakkali et al., 1997)....

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  • ...For example, lawsone normally accumulates only in the aerial part of the plant, but hairy roots of Lawsonia inermis grown in half- or fullstrength MS medium (Murashige and Skoog, 1962) can produce lawsone under dark conditions (Bakkali et al....

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Journal Article•DOI•
Plant cell cultures: Chemical factories of secondary metabolites

[...]

S. Ramachandra Rao1, Gokare Aswathanarayana Ravishankar2•
Japan Advanced Institute of Science and Technology1, Central Food Technological Research Institute2
01 May 2002-Biotechnology Advances
TL;DR: The production of the pungent food additive capsaicin, the natural colour anthocyanin and the natural flavour vanillin is described in detail.

1,330 citations

"Hairy Root Culture for Mass-Product..." refers background in this paper

  • ...Other factors like elicitors (Pitta-Alvarez et al., 2000), biotransformations of precursors and genetic manipulations through the Ri plasmid of A. rhizogenes also influence the yield of secondary metabolites from hairy roots (Rao and Ravishankar, 2002)....

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  • ...They often grow as fast as or faster than normal roots (untransformed) due to their extensive branching, resulting in the presence of many meristems (Charlwood and Charlwood, 1991; Flores et al., 1999) and they do not require phytohormones in the medium (Rao and Ravishankar, 2002)....

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Journal Article•DOI•
Production of plant secondary metabolites: a historical perspective

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Frédéric Bourgaud1, Antoine Gravot1, S. Milesi1, Eric Gontier1•
Institut national de la recherche agronomique1
01 Oct 2001-Plant Science
TL;DR: Plant cell culture technologies were introduced at the end of the 1960s as a possible tool for both studying and producing plant secondary metabolites, and it is now possible to manipulate the pathways that lead to secondary plant compounds.

978 citations

"Hairy Root Culture for Mass-Product..." refers background in this paper

  • ...Fast growth, low doubling time, ease of maintenance of hairy roots and their ability to synthesize a large range of chemical compounds offer an additional advantage as a continuous source for the production of valuable secondary metabolites (Bourgaud et al., 2001)....

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  • ...It is the causative agent of hairy root disease in plants (Giri and Narasu, 2000; Bourgaud et al., 2001)....

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Journal Article•DOI•
Natural plant chemicals: sources of industrial and medicinal materials

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Manuel F. Balandrin, James A. Klocke, Eve Syrkin Wurtele, Wm. Hugh Bollinger
07 Jun 1985-Science
TL;DR: In the future, biologically active plant-derived chemicals can be expected to play an increasingly significant role in the commercial development of new products for regulating plant growth and for insect and weed control.
Abstract: Many higher plants produce economically important organic compounds such as oils, resins, tannins, natural rubber, gums, waxes, dyes, flavors and fragrances, pharmaceuticals, and pesticides. However, most species of higher plants have never been described, much less surveyed for chemical or biologically active constituents, and new sources of commercially valuable materials remain to be discovered. Advances in biotechnology, particularly methods for culturing plant cells and tissues, should provide new means for the commercial processing of even rare plants and the chemicals they produce. These new technologies will extend and enhance the usefulness of plants as renewable resources of valuable chemicals. In the future, biologically active plant-derived chemicals can be expected to play an increasingly significant role in the commercial development of new products for regulating plant growth and for insect and weed control.

968 citations

"Hairy Root Culture for Mass-Product..." refers background or methods in this paper

  • ...This activity causes hairy root disease (a number of small roots protrude as fine hairs at the infection site and proliferate rapidly) (Balandrin et al., 1985)....

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  • ...Consequently, in many cases organic synthesis is not cost effective, and extraction from field-grown plants has been the major method used to economically obtain these important secondary metabolites (Balandrin et al., 1985; Dicosmo and Misawa, 1995)....

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  • ...The biggest challenge of producing secondary metabolites from plant cell suspension cultures is that secondary metabolites are usually produced by specialized cells and/or at distinct developmental stages (Balandrin et al., 1985; Mukundan et al., 1997)....

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  • ...as fine hairs at the infection site and proliferate rapidly) (Balandrin et al., 1985)....

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Journal Article•DOI•
Transformation of several species of higher plants by Agrobacterium rhizogenes: sexual transmission of the transformed genotype and phenotype

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David Tepfer1•
Institut national de la recherche agronomique1
01 Jul 1984-Cell
TL;DR: The T-DNA of the Ri plasmid from Agrobacterium rhizogenes is compatible with the regeneration of whole plants from genetically transformed roots and is transmitted through meiosis to the progeny of genetically transformed plants in carrot, tobacco, and morning glory.

705 citations

"Hairy Root Culture for Mass-Product..." refers background in this paper

  • ...Hairy roots are characterized by a high degree of lateral branching, profusion of root hairs and absence of geotropism (Tepfer, 1984)....

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  • ...Since the opine synthesis in A. rhizogenes infected plant cells is encoded by T-DNA of Ri plasmid (White et al., 1982, 1985), its detection serves as an effective biochemical marker in elucidating the transformed nature of the cultured root tissue (Petit et al., 1983; Tepfer, 1984)....

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  • ..., 1982, 1985), its detection serves as an effective biochemical marker in elucidating the transformed nature of the cultured root tissue (Petit et al., 1983; Tepfer, 1984)....

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  • ...As described earlier, transformed roots have an altered phenotype, profusion of laterals, and show lack of geotropism (Tepfer, 1984)....

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A revised medium for rapid growth and bio assays with tobacco tissue cultures

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01 Mar 2000-Biotechnology Advances
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10 Feb 2007-Applied Microbiology and Biotechnology
Milen I. Georgiev, Atanas Pavlov, Thomas Bley
Hairy root research: recent scenario and exciting prospects.

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01 Jun 2006-Current Opinion in Plant Biology
Stéphanie Guillon, Jocelyne Trémouillaux-Guiller, Pratap Kumar Pati, Marc Rideau, Pascal Gantet 
Plant cell cultures: Chemical factories of secondary metabolites

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01 May 2002-Biotechnology Advances
S. Ramachandra Rao, Gokare Aswathanarayana Ravishankar