Production of camptothecin by hairy roots and regenerated transformed shoots ofOphiorrhiza rugosavar.decumbens
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Camptothecin (CPT), the derivatives of which are used clinically for the treatment of metastatic colon cancer, is isolated from intact plants that can be subjected to environmental fluctuations. In vitro cultures may be an alternate and continuous source for year-round production of CPT. Since CPT production by undifferentiated cell cultures is low, differentiated tissues such as root cultures may be a viable alternate source for CPT production. Hairy roots were induced in Ophiorriza rugosa, a source of CPT, using Agrobacterium rhizogenes strain LBA9402. The hairy roots, when cultured in light, showed spontaneous regeneration of shoots. Analysis of CPT levels in the hairy roots and in vitro-grown transformed shoots revealed 0.009% d.w. and 0.012% d.w., respectively.Keywords:
Camptothecin
Hairy Root Culture
Rugosa
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Objective To induce hairy roots of Echinacea angustifolia by two strains of Agrobacterium rhizogenes A4 and R1000 and establish an in vitro culture system of the hairy roots.Methods Hairy roots were induced by coculture Effects of explants,Agrobacterium rhizogenes,preculture time along with infecting time on the induction rate and effects of different basic media on growth of hairy root were studied.Results The highest induction rate was obtained from leaves with 48h preculture which were induced by R1000 for 10min.The growth of hairy root could be raised by 1 /2MS medium with 0.5 mg / L IBA.Conclusion Establishment of Echchina angustifolia hairy root culture can provide a foundation for the industrial production of active drug component.
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Echinacea
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As a natural genetically engineered strain,Agrobacterium rhizogenes can infect more than 160 plant species,which can lead to special morphological changes of plant and induce a lot of hairy roots production.During the past several decades,people pay more and more attention to research work on hairy roots system,because hairy roots can produce a lot of important secondary metabolites.Mechanization of hairy roots production induced by Agrobacterium rhizogenes was briefly described in this paper.And the recent progress of hairy roots cultivation and application of secondary metabolites were also emphatically introduced.
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An efficient and rapid protocol for the establishment of Artemisia tilesii "hairy" root culture is reported. Leaf explants of aseptically growing plants were cocultured with Agrobacterium rhizogenes A4 wild strain or A. rhizogenes carrying the plasmids with nptII and ifn-α2b genes. Root formation on the explants started in 5-6 days after their cocultivation with bacterial suspension. Prolongation of explant cultivation time on the medium without cefotaxime led to stimulation of root growth. The effects of sucrose concentration as well as of the levels of synthetic indole-3-butyric acid (IBA) and native growth regulator Emistim on the stimulation of A. tilesii "hairy" root growth were studied. Maximum stimulating effect both for the control and for transgenic roots was observed in case of root cultivation on the media supplemented with IBA-up to 7.95- and 9.1-fold biomass increase, respectively. Cultivation on the medium with 10 μl/L Emistime has also led to the control roots growth stimulation (up to 2.75-fold). Emistime at 5 μl/L concentration led to 5.46-fold mass increase in only one "hairy" root line. Higher sucrose content (40 g/L) stimulated growth of two hairy root lines but had no effect on growth of the control roots.
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Catharanthus roseus
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Abstract Agrobacterium rhizogenes Conn. causes hairy root disease in plants. Hairy root‐infected A. rhizogenes is characterized by a high growth rate and genetic stability. Hairy root cultures have been proven to be an efficient means of producing secondary metabolites that are normally biosynthesized in roots of differentiated plants. Furthermore, a transgenic root system offers tremendous potential for introducing additional genes along with the Ri plasmid, especially with modified genes, into medicinal plant cells with A. rhizogenes vector systems. The cultures have turned out to be a valuable tool with which to study the biochemical properties and the gene expression profile of metabolic pathways. Moreover, the cultures can be used to elucidate the intermediates and key enzymes involved in the biosynthesis of secondary metabolites. The present article discusses various applications of hairy root cultures in plant genetic engineering and potential problems associated with them. (Managing editor: Wei Wang)
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Gene engineering
Plant cell
Metabolic Engineering
Root (linguistics)
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Hairy roots, transformed with Agrobacterium rhizogenes, have been found to be suitable for the production of secondary metabolites because of their stable and high productivity in hormone-free culture conditions. A number of plant species including many medicinal plants have been successfully transformed with Agrobacterium rhizogenes. Transformed root cultures have also been found to be a potential source of high-value pharmaceuticals. In this article the most important alkaloids produced by hairy roots are summarised. Several different methods have been used to increase the alkaloid accumulation in hairy root cultures. The selection of high productive root lines based on somaclonal variation offers an interesting option to enhance the productivity. Elicitors and modification of culture conditions have been shown to increase the growth and the alkaloid production in some cases. Genetic engineering is a modern tool to regulate the secondary metabolism also in hairy roots. However, our knowledge on biosynthesis of many alkaloids is still poor. Only a limited number of enzymes and their respective genes which regulate the biosynthetic pathways are fully characterised.
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Secondary metabolism
Somaclonal variation
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Agrobacterium rhizogenes-mediated transformation results in the development of hairy roots at the site of infection. The production of hairy roots involves cocultivation of explants with A. rhizogenes and the subsequent selection of hairy roots on hormone-free medium. Hairy roots have many applications for research including secondary product production and for the study of biochemical pathways. In addition, transgenic plants regenerated from hairy roots often show an altered phenotype due to the presence of the rol genes. In this chapter we describe how to produce and grow hairy root cultures, how to regenerate shoots from these hairy roots, and how to conduct molecular analysis of these cultures.
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Pharmacognosy Magazine,2014,10,37,77-82.DOI:10.4103/0973-1296.126671Published:February 2014Type:Original ArticleAuthors:Bing Huang, Huanjie Lin, Chuanyan Yan, Hongyan Qiu, Lipeng Qiu, and Rongmin Yu Author(s) affiliations:Bing Huang1, Huanjie Lin1, Chuanyan Yan1, Hongyan Qiu1, Lipeng Qiu2, Rongmin Yu2 1 Department of Natural Medicinal Chemistry, College of Pharmacy, Guangdong Pharmaceutical University, Guangzhou, China 2 Biotechnology Institute of Chinese Materia Medica, Jinan University, Guangzhou, China Abstract:Background: Polygonum multiflorum is an important medicinal plant. Hairy roots systems obtained by transforming plant tissues with the natural genetic engineer Agrobacterium rhizogenes can produce valuable biological active substances, which have immense potential in the pharmaceutical industry. Objective: To optimize the inductive and cultural conditions of P. multiflorum hairy roots and to identify the major active secondary metabolites in hairy roots. Materials and Methods: P. multiflorum hairy root were mediated with A. rhizogenes R1601 to induce hairy roots. Four combinations, including Murashige-Skoog (MS), 1/2 MS, B 5, and White, were investigated to optimize the culture medium. MS medium was selected for the growth measurement. The qualitative and quantitative determinations of free anthraquinone in hairy roots were compared with the calli and aseptic plantlets using high-performance liquid chromatography. Results: The inductive rates of hairy roots by leaves were higher than for any other explants. The presence of agropine in the P. multiflorum hairy roots confirmed that they were indeed transgenic. MS medium was the most suitable of the four media for hairy root growth. Meanwhile, the growth kinetics and nutrient consumption results showed that the hairy roots displayed a sigmoidal growth curve and that their optimal inoculation time was 18-21 days. The determination of the anthraquinone constituents indicated that the rhein content of the hairy roots reached 2.495 μg g−1 and was 2.55-fold higher than that of natural plants. Conclusion: Transgenic hairy roots of P. multiflorum could be one of the most potent materials for industrial-scale production of bioactive anthraquinone constituents. Keywords:Agrobacterium rhizogenes R1601, anthraquinones, Polygonum multiflorum, transgenic hairy rootsView:PDF (817.18 KB) Full Text
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Anthraquinones
Polygonum
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