Hairy Root Cultures of Gynostemma pentaphyllum (Thunb.) Makino: A Promising Approach for the Production of Gypenosides as an Alternative of Ginseng Saponins

Chang, Chia-Seng; Chang, Ku-Shang; Lin, Yichen; Liu, Sin-Yie; Chen, Chien-Yuan

doi:10.1007/s10529-005-8653-7

Cited by 36 publications

(22 citation statements)

References 10 publications

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“…Some of the medicinal plant species have been reported to induce fast-growing productive hairy root. They include red sage (Salvia miltiorrhiza) (Shi et al 2007), ginseng (Panax ginseng) (Sathiyamoorthy et al 2010), American ginseng (Panax quinquefolius) (Mathur et al 2010;Kochan et al 2013), Jiaogulan (Gynostemma pentaphyllum) (Chang et al 2005), snow lotus (Saussurea involucrate) (Fu et al 2006) and Anisodus luridus (Qin et al 2014). Hairy roots are being explored as a promising alternative for the rapid production of the bioactive components of these plants.…”

Section: Introductionmentioning

confidence: 99%

Transgenic hairy roots of Tetrastigma hemsleyanum: induction, propagation, genetic characteristics and medicinal components

Song

et al. 2015

Plant Cell Tiss Organ Cult

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Tetrastigma hemsleyanum Diels & Gilg ex Diels (T. hemsleyanum) is an important medicinal plant in China. Its bulbous root is the most valuable part for medicine. There is an increasing demand for T. hemsleyanum in the medicinal market. However, the natural slow growth rate of T. hemsleyanum hardly meets the demand of the medicinal market. There is the need of alternative way for rapid and large-scale production of T. hemsleyanum. In this study, we investigated the induction of hairy roots by using Agrobacterium rhizogenes (A. rhizogenes) as an alternative method of increasing production of the secondary metabolites in hairy roots of T. hemsleyanum. Herein, we reported our successful induction of hairy roots of T. hemsleyanum by cucumopine type A. rhizogenes K599 and their rapid propagation. Firstly, we selected the optimal mediums for induction of hairy roots and for subculture of induced hairy roots. We found that when A. rhizogenes K599 carrying both an endogenous plasmid pRi2659 and an exogenous plasmid pRI101-AN-GFP was used to infect the leaves of T. hemsleyanum seedlings, the optimal medium for inducting hairy roots was the medium of MS ? 0.4 mg/L 1-naphthaleneacetic acid. The optimal medium for subculture of hairy roots was the combination of MS ? 1.0 mg/L indole-3-butyric acid ? 0.5 or 1.0 mg/L kinetin. During suspension culture of hairy roots, the rapid growth phase occurred in the period of 15-28 days. Secondly, we employed polymerase chain reaction (PCR) assays to examine the integration of T-DNA of A. rhizogenes. The PCR results revealed the existence of a phenomenon of lacking of integration of one or more of 11 genes within T-DNA of plasmid pRi2659 into the genomic DNA of hairy roots. Thirdly, we conducted histocytological analysis to examine the structural features of the induced hairy roots and observed that xylem structures of the hairy roots were simplified as compared to those of the bulbous roots and ordinary fine roots of non-transgenic plant. Finally, we applied high performance liquid chromatography to analyze the medicinal components in several organs of the T. hemsleyanum plants. The results showed that the contents of kaempferol in all hairy root lines were significantly higher than those in bulbous roots, fine roots, stems and leaves. These results suggest that insertion and integration of T-DNA from A. rhizogenes can significantly increase the kaempferol content in hairy roots of T. hemsleyanum.

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Section: Introductionmentioning

confidence: 99%

Transgenic hairy roots of Tetrastigma hemsleyanum: induction, propagation, genetic characteristics and medicinal components

Song

et al. 2015

Plant Cell Tiss Organ Cult

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“…In vitro culture can offer new opportunities by providing plant material independently of the climatic condition. Due to the fact that usually secondary metabolites are produced only in small quantities in in vitro cultures of non-differentiated plant cells (DiCosmo and Misawa 1985) the culture of fast growing transgenic, hairy roots becomes a promising source of plant material for pharmaceutical use (Altamura 2004, Chang et al 2005, Momčilović et al 1997, Tiwari et al 2008). In the case of R. graveolens this perspective is especially interesting as the chemical synthesis of pharmaceutically valuable furanocoumarins accumulated in its tissues is very expensive and does not always lead to the production of biologically active compounds (Bohlmann andEilert 1994, Bourgaud et al 1994).…”

Section: Introductionmentioning

confidence: 99%

Genetic transformation of Ruta graveolens L. by Agrobacterium rhizogenes: hairy root cultures a promising approach for production of coumarins and furanocoumarins

Sidwa-Gorycka

Królicka

Orlita

et al. 2009

Plant Cell Tiss Organ Cult

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Ruta graveolens L. is a source of pharmacologically active compounds such as coumarins, furanocoumarins and furoquinolone alkaloids. Hypocotyls, callus and shoots of R. graveolens were inoculated with bacteria from two Agrobacterium rhizogenes strains. Hairy root cultures were established after inoculation of hypocotyls with wild A. rhizogenes strain LBA 9402. The transgenic nature of the regenerated tissue was confirmed by PCR amplification. Coumarins, furanocoumarins and alkaloids present in the hairy root tissue were identified by GC and GC-MS and compared with those present in in vitro shoot cultures. The level of pinnarin and rutacultin, bergapten, isopimpinelin and xanthotoxin was approximately twofold higher in hairy root than in shoot cultures. Two additional coumarins: osthole and osthenol, never been found in R. graveolens, were identified in hairy root tissue. Besides coumarins, alkaloids were identified: dictamnine, skimmianine, kokusaginine, rybalinine and an isomer of rybalinine. The levels of nearly all coumarins and alkaloids in hairy roots cultured in the darkness were higher than those accumulated under a photoperiod mode.

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“…pentaphyllum is an important plant due to having such important bioactive molecule, and that is why its genetic transformation is necessary for improving its quality and quantity. G. pentaphyllum leaf discs were cocultured with A. rhizogenes by Chang et al ( 2005 ). Hairy roots produced high levels of gypenosides in comparison to native roots of G. pentaphyllum.…”

Section: Gynostemma Pentaphyllum (Thunb) Makinomentioning

confidence: 99%

Contribution of Biotechnological Tools in the Enhancement of Secondary Metabolites in Selected Medicinal Climbers

Srivastava

Singh

Misra

2015

Biotechnological Strategies for the Conservation of Medicinal and Ornamental Climbers

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Plants by nature protect themselves by producing secondary metabolites which often have pronounced bioactivities. The presence of these metabolites is responsible for the active interaction of plants with their environment defending themselves against a variety of herbivores and pathogenic microorganisms as well as various kinds of abiotic stresses. Climber plants contain large number of useful secondary metabolites. Besides their pharmacological role, secondary metabolites also contain some other properties. Artabotrys hexapetalus , a climbing herb, secretes oil which is used in perfume industry. Capparis zeylanica is a thorny stout climbing shrub, used in the treatment of snake bite and to cure small pox, cholera, etc. Stem barks and roots of Tinospora cordifolia , a woody climber, are used in dysentery and diarrhoea. Toddalia asiatica , an evergreen climber, produced nitidine secondary metabolite which contains anti-HIV and antimalarial and anticancerous properties, but its production is not enough for commercial supply. Tissue culture technology has emerged as a supplementary branch to fulfi l demands for this valuable secondary metabolite. Various in vitro methods for enhancement of secondary metabolites are available such as hairy root culture, treatment of elicitors and use of precursors and introduction of any foreign gene via bacterial transformation.Current progresses have been made in the fi eld of molecular biology through the alteration in metabolic skeleton of plant secondary metabolism. With the use of various genes (involved in the synthesis of enzymes and their regulatory proteins), diverse pathways have been traced and being transformed. Antisense technology has been emerged as an additional alternative for enhancement of secondary metabolites. In Tylophora indica enhancement in kaempferol, an antioxidant compound was observed by using precursors like salicylic acid, ornithine, cinnamic acid, tyrosine and phenylalanine. The total dry weight of 2-hydroxy-4-methoxy benzaldehyde was enhanced in hairy root cultures of Decalepis hamiltonii . The present chapter provides an insight on the use of biotechnological techniques for the enhancement of secondary metabolites in medicinal climbers.

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Hairy Root Cultures of Gynostemma pentaphyllum (Thunb.) Makino: A Promising Approach for the Production of Gypenosides as an Alternative of Ginseng Saponins

Abstract: Hairy root cultures of Gynostemma pentaphyllum were established by infecting leaf discs with Agrobacterium rhizogenes. The dry biomass of hairy roots grown in MS medium for 49 days was 7.3 g l(-1) with a gypenoside content of 38 mg g(-1) dry wt.

Cited by 36 publications

References 10 publications

Transgenic hairy roots of Tetrastigma hemsleyanum: induction, propagation, genetic characteristics and medicinal components

Transgenic hairy roots of Tetrastigma hemsleyanum: induction, propagation, genetic characteristics and medicinal components

Genetic transformation of Ruta graveolens L. by Agrobacterium rhizogenes: hairy root cultures a promising approach for production of coumarins and furanocoumarins

Contribution of Biotechnological Tools in the Enhancement of Secondary Metabolites in Selected Medicinal Climbers

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