A Critical Review on Biotechnological Interventions for Production and Yield Enhancement of Secondary Metabolites in Hairy Root Cultures

Hairy Roots ◽  
2018 ◽  
pp. 21-44 ◽  
Author(s):  
Mihir Halder ◽  
Dipasree Roychowdhury ◽  
Sumita Jha
Keyword(s):  
Secondary Metabolites ◽  
Hairy Root ◽  
Critical Review ◽  
Yield Enhancement ◽  
Hairy Root Cultures ◽  
Root Cultures

Hairy Roots

2021 ◽  
pp. 219-252
Keyword(s):  
Secondary Metabolites ◽  
Hairy Root ◽  
Hairy Roots ◽  
High Growth ◽  
Root Culture ◽  
Hairy Root Culture ◽  
High Growth Rate ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Transformed Root Cultures

Agrobacterium rhizogenes induces hairy root disease in plants. The neoplastic (cancerous) roots produced by A. rhizogenes infection, when cultured in hormone free medium, show high growth rate and genetic stability. These genetically transformed root cultures can produce levels of secondary metabolites comparable to that of intact plants. Several elicitation methods can be used to further enhance the production and accumulation of secondary metabolites. Thus, hairy root culture offer promise for high production and productivity of valuable secondary metabolites in many plants. Hairy roots can also produce recombinant proteins from transgenic roots, and thereby hold immense potential for pharmaceutical industry. Hairy root cultures can be used to elucidate the intermediates and key enzymes involved in the biosynthesis of secondary metabolites, and for phytoremediation due to their abundant neoplastic root proliferation property. Various applications of hairy root cultures and potential problems associated with them are discussed in this chapter.

Enhanced accumulation of secondary metabolites in hairy root cultures of Scutellaria lateriflora following elicitation

2012 ◽  
Vol 34 (9) ◽  
pp. 1757-1763 ◽  
Author(s):  
Agata Wilczańska-Barska ◽  
Aleksandra Królicka ◽  
Daniel Głód ◽  
Magdalena Majdan ◽  
Anna Kawiak ◽  
...  
Keyword(s):  
Secondary Metabolites ◽  
Hairy Root ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Scutellaria Lateriflora

Secondary Metabolites in Green Hairy Root Cultures

Planta Medica ◽  
1991 ◽  
Vol 57 (S 2) ◽  
pp. A10-A11
Author(s):  
M. Sauerwein ◽  
K. Shimomura
Keyword(s):  
Secondary Metabolites ◽  
Hairy Root ◽  
Hairy Root Cultures ◽  
Root Cultures

scholarly journals Genetic transformation of Ceratotheca triloba for the production of anthraquinones from hairy root cultures

2012 ◽  
Author(s):  
◽  
Leeann Naicker
Keyword(s):  
Secondary Metabolites ◽  
Cell Suspension ◽  
Hairy Root ◽  
Hairy Roots ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Control Culture ◽  
Hairy Root Cultures ◽  
Shoot Cultures ◽  
Root Cultures

Many secondary metabolites that have been extracted from medicinal plants have been used as source of clinical drugs. However, the concentration of the active metabolites in plants is generally low. An attractive alternative for producing these important secondary metabolites is via plant tissue culture technology. More particularly, the genetic transformation of a plant tissue by Agrobaterium rhizogenes has been employed for producing high yields of secondary metabolites. In a previous study, three structurally similar anthraquinones: 9,10-Anthracenedione, 1-Hydroxy-4-methylanthraquinone and 5,8-Dimethoxy-2,3,10,10a-tetrahydro-1H,4aH-phenanthrene-4,9-dione, and one steroid; Androst-5-ene-3, 17, 19-triol were isolated from the root extracts of C. triloba. The anthraquinones have shown to exhibit the anticancer mechanism which involves the inhibition of the activity of the human topoisomerase II enzyme that transforms supercoiled DNA to linear DNA. However, these anthraquinones were found in very low concentrations. Therefore, in this study we used plant cell and tissue culture systems (cell suspension, shoot and hairy root cultures) of C. triloba to increase the production of anthraquinones. Since the establishment of C. triloba in vitro plant systems required a source sterile explants, a protocol that involved the use of NaCIO was optimized for the sterilization and subsequent germination of C. triloba seeds which were micro-propagated into shoot cultures. These cultures provided a source explants for the induction of callus and hairy root cultures. The biomass of these plant cell and tissue cultures were subsequently bulked up for the extraction for anthraquinones and the yields were compared followed by fractionation and identification of the major compounds. The bioactivity of the fractions was evaluated by testing their cytotoxicity on cancer cells and anti-topoisomerase activity. The sterilization protocol that provided sterile seeds was found to be a solution of 30% NaCIO at an exposure time of 10 minutes. From the sterilized seeds shoot cultures were established on MS medium. The leaf explants of the shoot cultures were then used to induce callus cultures which subsequently were transferred to liquid medium whereby the total biomass of suspension cultures increased from 4 g to 134.18 g (wet weight). Also hairy roots cultures were established from stem explants with a low cell density inoculum of A. rhizogenes at a transformation efficiency of 73%. The growth of these hairy roots was slow in hormone free medium. This was overcomed with the use NAA and IAA which increased the xvii biomass from 1.03 g in the control culture (without hormone) to 23.91 g and 46.13 g respectively. An evaluation of the anthraquinones in the field root and hairy root, cell suspension and shoot culture extracts was carried out by using their Thin Layer Chromatography profiles and the High Performance Liquid Chromatography profiles as well as the standards, 9,10-Anthracenedione and 1-Hydroxy-4-methylanthaquinone. TLC analysis showed that the RF values of the fractions CT01 and CT02 matched the RF values of anthraquinones standards while HPLC analysis revealed that hairy root cultures supplemented with IAA (125.03 μg.mg-1) or NAA (98.25 μg. mg-1) produced a higher concentration of anthraquinones than the control culture (without hormone) (13.33 μg.mg-1), the field roots (33.51 μg. mg-1) and the shoot (3.23 μg.mg-1) and cell suspension cultures (13.17 μg.mg-1). Due to co-elution of the compounds in HPLC analysis, six fractions were isolated by Preparative Thin Layer Chromatography from the hairy root extract (obtained from the culture supplemented with NAA) and were coded as CT01, CT02, CT03, CT04, CT05 and CT06. The compounds in these fractions were identified by Electron Ionization-Liquid chromatography-Mass Spectroscopy and it was found that the hairy roots produced one acridone derivative; 5-Methoxy-2-nitro-10H-acridin-9-one, one naphthoquinone derivative; 2H-Naphto[2,3-b]pyran-5,10-dione,3,4-dihydro-2,2-dimethyl- and seven anthracenedione derivatives. These were: i) 5,8-Dimethoxy-2,3,10,10a-tetrahydro-1H,4aH-phenanthrene-4,9-dione, ii) 9,10-Anthracenedione, 2-methyl-, iii) 1-Hydroxy-4-methylanthraquinone, iv) 9,10-Anthracenedione, 2-ethyl-, v) 1,5-Diaminoanthraquinone, vi) Phenanthrene, 3,6-dimethoxy-9-methyl-, vii) 9,10-Anthracenedione, 1,4-dimethyl-. Fractions CT01 (5,8-Dimethoxy-2,3,10,10a-tetrahydro-1H,4aH-phenanthrene-4,9-dione, 9,10-Anthracenedione, 2-methyl- and 1-Hydroxy-4-methylanthraquinone) and CT02 (9,10- Anthracenedione, 2-ethyl-) were cytotoxic to the DU-145 cancer cell line at concentrations of 125 μg.mg-1 to 1000 μg.mg-1. These fractions also showed anti-topoisomerase activity as they inhibited the conversion of supercoiled DNA into linear DNA. In conclusion this is the first study that describes the transformation of C. triloba by A. rhizogenes mediated transformation and compares the production of anthraquinones in C. triloba hairy roots to the field roots, shoot and cell suspension cultures. This study has xviii indicated that hairy root cultures is a high-yielding production system for anthraquinones (5,8-Dimethoxy-2,3,10,10a-tetrahydro-1H,4aH-phenanthrene-4,9-dione, 1-Hydroxy-4-methylanthraquinone, 9,10-Anthracenedione, 2-methyl- and 9,10- Anthracenedione, 2-ethyl-) which could have the potential to be used in cancer therapy. In addition the discovery of C. triloba hairy roots having the biosynthetic capacity to synthesize five valuable anthraquinone derivatives that are not found the field roots has also been revealed.

Hairy Roots

2022 ◽  
pp. 735-759
Author(s):  
Pradip Chandra Deka
Keyword(s):  
Secondary Metabolites ◽  
Hairy Root ◽  
Hairy Roots ◽  
High Growth ◽  
Root Culture ◽  
Hairy Root Culture ◽  
High Growth Rate ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Transformed Root Cultures

Agrobacterium rhizogenes induces hairy root disease in plants. The neoplastic (cancerous) roots produced by A. rhizogenes infection, when cultured in hormone free medium, show high growth rate and genetic stability. These genetically transformed root cultures can produce levels of secondary metabolites comparable to that of intact plants. Several elicitation methods can be used to further enhance the production and accumulation of secondary metabolites. Thus, hairy root culture offer promise for high production and productivity of valuable secondary metabolites in many plants. Hairy roots can also produce recombinant proteins from transgenic roots, and thereby hold immense potential for pharmaceutical industry. Hairy root cultures can be used to elucidate the intermediates and key enzymes involved in the biosynthesis of secondary metabolites, and for phytoremediation due to their abundant neoplastic root proliferation property. Various applications of hairy root cultures and potential problems associated with them are discussed in this chapter.

Isoflavone Glycoside Formation in Transformed and Non-Transformed Suspension and Hairy Root Cultures of Lupinus polyphyllus and Lupinus hartwegii

1991 ◽  
Vol 46 (9-10) ◽  
pp. 725-734 ◽  
Author(s):  
J. Berlin ◽  
L. Fecker ◽  
C. Rügenhagen ◽  
C. Sator ◽  
D. Strack ◽  
...  
Keyword(s):  
Secondary Metabolites ◽  
Cell Suspension ◽  
Hairy Root ◽  
Southern Analysis ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Wild Type ◽  
Lupinus Polyphyllus ◽  
Isoflavone Glycoside ◽  
Type Strains

Abstract Transformed cell suspension and hairy root cultures were established by infecting seedlings of Lupinus polyphyllus and L. hartwegii with various wild type strains of Agrobacterium tume-faciens and A. rhizogenes. Transformation of the cultures was confirmed either by their phyto­hormone autotrophy, detection of opines or southern analysis. Glueosides of genistein and 2′-hydroxygenistein, were found to be the main secondary metabolites in normal and trans­formed suspension cultures as well as in hairy root cultures. Although some of the isoflavone glycosides of the cultures were apparently new constituents of Lupinus, they were afterwards also found in young seedlings.

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