Plants are a tremendous source for the discovery of new products of medicinal value for drug deve... more Plants are a tremendous source for the discovery of new products of medicinal value for drug development. Today several distinct chemicals derived from plants are important drugs currently used in one or more countries in the world. Many of the drugs sold today are simple synthetic modifications or copies of the naturally obtained substances. The evolving commercial importance of secondary metabolites has in recent years resulted in a great interest in secondary metabolism, particularly in the possibility of altering the production of bioactive plant metabolites by means of tissue culture technology. Plant cell culture technologies were introduced at the end of the 1960's as a possible tool for both studying and producing plant secondary metabolites. Different strategies, using an in vitro system, have been extensively studied to improve the production of plant chemicals. The focus of the present review is the application of tissue culture technology for the production of some i...
A procedure developed for identifying the developmental stage of tobacco microspores is described... more A procedure developed for identifying the developmental stage of tobacco microspores is described. Fixation in Carnoy's solution (ethanol, chloroform, acetic acid, 6: 3: 1) for at least 30 rein is followed by soaking in 70% ethanol. Storage, if necessary, at 5-8℃ is recommended. Maceration in a 2: 1: 1 mixture of 15% CrO3, 10% HNO3, and 5% HCL for 7-10 min, is required to remove spore walls. Hardening is then done in 1: 1 95% ethanol-propionic acid and staining in 1% aceto-carmine for 20-30 min.
Background Fritillaria cirrhosa, an important Chinese medicinal herb, is a Class-III protected an... more Background Fritillaria cirrhosa, an important Chinese medicinal herb, is a Class-III protected and highly exploited species by pharmaceutical industry. Dwindling wild populations of species are unable to meet market demand. Therefore, this study was carried out to develop an in vitro propagation method for bulblet production. Also, the study aimed to carry out LC–MS/MS analysis of tissue culture-derived bulblets and callus for the presence of isosteroidal alkaloids (peimissine, verticine, and verticinone), and compare its quantities with commercially available crude drug samples. Results In vitro seed germination (91%) of F. cirrhosa was achieved on Murashige and Skoog’s basal medium (MSBM) supplemented with 6-benzylaminopurine (1 mg L−1) and α-naphthalene-acetic-acid (0.4 mg L−1). On transfer of germinated seeds from Petri-dishes to glass bottles containing hormone-free MSBM, 37.5% of seedlings developed bulblets after 3 months of incubation. Regeneration and multiplication of bulb...
Current Plant Science and Biotechnology in Agriculture, 1996
Bamboo is a monocotyledonous evergreen plant belonging to the family Gramineae and subfamily Bamb... more Bamboo is a monocotyledonous evergreen plant belonging to the family Gramineae and subfamily Bambusoideae. Its distribution covers tropical, subtropical and temperate zones of the world. Sinocalamus latiflora Munro (Syn. Bambusa oldhamii Munro) is the major bamboo species growing in Taiwan. It can be utilized as vegetable, ornamental plant and paper pulp for various purposes. Bamboo flowers only once during the end of its lifetime, which occurs at the end of its first fruiting season (Nadgauda et al., 1990). The improvement of this species is not possible through conventional breeding methods due to irregular flowering habit, extremely low seed viability, and dependence of seed production on unpredictably long intervals (6–50 years) (Mehta et al., 1982). Propagation through seeds is also undependable on account of the long interval needed to seed (50–60 years in S. latiflora). Vegetative propagation through conventionally used offset is beset with problems such as bulkiness and non-availability of the propagules and difficulties in transport over long distances (Mehta et al., 1982; Rao et al., 1985).
Plants and fungi provide a vast range of natural products including pharmaceuticals with diverse ... more Plants and fungi provide a vast range of natural products including pharmaceuticals with diverse chemical structures and a broad array of biological activities. In the past few decades, there has been a worldwide resurgence of interest in the study and use of medicinal plants and fungi in the health-care system. Several medicinal plants and fungi are involved in the industrial processing of profitable products used in human medicines. However, due to severe constraints on the availability of desired plant and fungus materials, different strategies, including tissue culture, have been extensively studied. The present chapter focuses on the application of various in vitro culture systems in the production of bioactive secondary metabolites in medicinal herbs and fungi in Taiwan. All the species described in this article are sources of traditional Chinese medicines. The review article includes the research work carried out in our laboratory on the production of plant secondary metabolites in callus cultures of Saussurea involucrata, Solanum melongena, and Salvia miltiorrhiza and in cell suspension cultures of Taxus mairei, Gnetiana davidii var. formosana, and Angelica dahurica var. formosana. Also, plant metabolites were obtained from in vitro shoots of Glossogyne tenuifolia, Saussurea involucrata, and Polygonum multiflorum, aerial parts of Scrophularia yoshimurae, tubers of Corydalis yanhusuo, tissue culture plants, and hairy roots of Gentiana scabra. Also, we could achieve the production of terpenoids in Antrodia cinnamomea, a medicinal fungus, and tanshinones in Salvia miltiorrhiza using T-DNA activation-tagging technique (Agrobacterium tumefaciens-mediated transformation).
Investigations were carried out to study the effects of light-emitting diode (LED) lights on grow... more Investigations were carried out to study the effects of light-emitting diode (LED) lights on growth and development of isosteroidal alkaloids in embryogenic calli of Fritillaria cirrhosa D. Don, an important traditional Chinese medicine herb. Calli were cultured in glass bottles, each containing 100 mL of Murashige and Skoog’s basal medium supplemented with 2% sucrose and 0.4% gellan gum powder, a gelling agent. These bottles were incubated in a specially designed plant growth chamber equipped with eight different LED lights consisting of single or combinations of four different light spectra emitting blue (450 nm), green (525 nm), red (660 nm), and far-red (730 nm) light. After three months of incubation, morphological changes in embryogenic calli were recorded, and LC-MS/MS analysis of cultures was carried out for peimisine, sipeimine, peiminine, and peimine. The highest number of somatic embryos and the maximum fresh weight was recorded in calli incubated under red (9R), infrared...
Journal of the American Society for Horticultural Science, 2012
The genus Dendrobium is important in traditional Chinese herbal medicine, and the precise identif... more The genus Dendrobium is important in traditional Chinese herbal medicine, and the precise identification of Dendrobium species is critical for the treatment and for pharmacological research. In the present study, a ribosomal DNA (rDNA) internal transcribed spacer (ITS) region-based analysis was used to ascertain the phylogenetic relationship among 20 Dendrobium species. The lengths of the ITS regions among the 20 species ranged from 636 to 653 bp, and the identities of the rDNA regions among the different species ranged from 75.7% to 99.1%. The results also showed that the ITS1 and ITS2 regions exhibit more variation than the 5.8S rDNA. A phylogenetic tree derived from the ITS sequence indicated that six medicinal Dendrobium species, of which five are common medicinal plants in the Taiwan market, were closely related and shared a common clade. Multiplex polymerase chain reaction (PCR) amplification was successfully performed to identify the six medicinal Dendrobium species, and ampl...
Background Gentiana scabra Bunge commonly known as ‘Long dan cao’ in China has been used in tradi... more Background Gentiana scabra Bunge commonly known as ‘Long dan cao’ in China has been used in traditional Chinese medicines for more than 2000 years. Dry roots and rhizome of the herb have been used for the treatment of inflammation, anorexia, indigestion and gastric infections. Iridoids and secoiridoids are the main bioactive compounds which attribute to the pharmacological properties of this plant. The species is difficult to mass propagate by seed due to the low percentage of germination and limited dormancy period. Wild populations in some locations are considered to be in the endangered category due to over exploitation. Results In the present study, we report an efficient micropropagation system. Shoot apices of six weeks old in vitro grown G. scabra plants were used as explants for the in vitro propagation. Induction of multiple shoots (9.1/explant) was achieved on the culture of shoot apices on half strength Murashige and Skoog’s basal medium (MSBM) containing 2.0 mg/L−1 6-ben...
Plants are a tremendous source for the discovery of new products of medicinal value for drug deve... more Plants are a tremendous source for the discovery of new products of medicinal value for drug development. Today several distinct chemicals derived from plants are important drugs currently used in one or more countries in the world. Many of the drugs sold today are simple synthetic modifications or copies of the naturally obtained substances. The evolving commercial importance of secondary metabolites has in recent years resulted in a great interest in secondary metabolism, particularly in the possibility of altering the production of bioactive plant metabolites by means of tissue culture technology. Plant cell culture technologies were introduced at the end of the 1960's as a possible tool for both studying and producing plant secondary metabolites. Different strategies, using an in vitro system, have been extensively studied to improve the production of plant chemicals. The focus of the present review is the application of tissue culture technology for the production of some i...
A procedure developed for identifying the developmental stage of tobacco microspores is described... more A procedure developed for identifying the developmental stage of tobacco microspores is described. Fixation in Carnoy's solution (ethanol, chloroform, acetic acid, 6: 3: 1) for at least 30 rein is followed by soaking in 70% ethanol. Storage, if necessary, at 5-8℃ is recommended. Maceration in a 2: 1: 1 mixture of 15% CrO3, 10% HNO3, and 5% HCL for 7-10 min, is required to remove spore walls. Hardening is then done in 1: 1 95% ethanol-propionic acid and staining in 1% aceto-carmine for 20-30 min.
Background Fritillaria cirrhosa, an important Chinese medicinal herb, is a Class-III protected an... more Background Fritillaria cirrhosa, an important Chinese medicinal herb, is a Class-III protected and highly exploited species by pharmaceutical industry. Dwindling wild populations of species are unable to meet market demand. Therefore, this study was carried out to develop an in vitro propagation method for bulblet production. Also, the study aimed to carry out LC–MS/MS analysis of tissue culture-derived bulblets and callus for the presence of isosteroidal alkaloids (peimissine, verticine, and verticinone), and compare its quantities with commercially available crude drug samples. Results In vitro seed germination (91%) of F. cirrhosa was achieved on Murashige and Skoog’s basal medium (MSBM) supplemented with 6-benzylaminopurine (1 mg L−1) and α-naphthalene-acetic-acid (0.4 mg L−1). On transfer of germinated seeds from Petri-dishes to glass bottles containing hormone-free MSBM, 37.5% of seedlings developed bulblets after 3 months of incubation. Regeneration and multiplication of bulb...
Current Plant Science and Biotechnology in Agriculture, 1996
Bamboo is a monocotyledonous evergreen plant belonging to the family Gramineae and subfamily Bamb... more Bamboo is a monocotyledonous evergreen plant belonging to the family Gramineae and subfamily Bambusoideae. Its distribution covers tropical, subtropical and temperate zones of the world. Sinocalamus latiflora Munro (Syn. Bambusa oldhamii Munro) is the major bamboo species growing in Taiwan. It can be utilized as vegetable, ornamental plant and paper pulp for various purposes. Bamboo flowers only once during the end of its lifetime, which occurs at the end of its first fruiting season (Nadgauda et al., 1990). The improvement of this species is not possible through conventional breeding methods due to irregular flowering habit, extremely low seed viability, and dependence of seed production on unpredictably long intervals (6–50 years) (Mehta et al., 1982). Propagation through seeds is also undependable on account of the long interval needed to seed (50–60 years in S. latiflora). Vegetative propagation through conventionally used offset is beset with problems such as bulkiness and non-availability of the propagules and difficulties in transport over long distances (Mehta et al., 1982; Rao et al., 1985).
Plants and fungi provide a vast range of natural products including pharmaceuticals with diverse ... more Plants and fungi provide a vast range of natural products including pharmaceuticals with diverse chemical structures and a broad array of biological activities. In the past few decades, there has been a worldwide resurgence of interest in the study and use of medicinal plants and fungi in the health-care system. Several medicinal plants and fungi are involved in the industrial processing of profitable products used in human medicines. However, due to severe constraints on the availability of desired plant and fungus materials, different strategies, including tissue culture, have been extensively studied. The present chapter focuses on the application of various in vitro culture systems in the production of bioactive secondary metabolites in medicinal herbs and fungi in Taiwan. All the species described in this article are sources of traditional Chinese medicines. The review article includes the research work carried out in our laboratory on the production of plant secondary metabolites in callus cultures of Saussurea involucrata, Solanum melongena, and Salvia miltiorrhiza and in cell suspension cultures of Taxus mairei, Gnetiana davidii var. formosana, and Angelica dahurica var. formosana. Also, plant metabolites were obtained from in vitro shoots of Glossogyne tenuifolia, Saussurea involucrata, and Polygonum multiflorum, aerial parts of Scrophularia yoshimurae, tubers of Corydalis yanhusuo, tissue culture plants, and hairy roots of Gentiana scabra. Also, we could achieve the production of terpenoids in Antrodia cinnamomea, a medicinal fungus, and tanshinones in Salvia miltiorrhiza using T-DNA activation-tagging technique (Agrobacterium tumefaciens-mediated transformation).
Investigations were carried out to study the effects of light-emitting diode (LED) lights on grow... more Investigations were carried out to study the effects of light-emitting diode (LED) lights on growth and development of isosteroidal alkaloids in embryogenic calli of Fritillaria cirrhosa D. Don, an important traditional Chinese medicine herb. Calli were cultured in glass bottles, each containing 100 mL of Murashige and Skoog’s basal medium supplemented with 2% sucrose and 0.4% gellan gum powder, a gelling agent. These bottles were incubated in a specially designed plant growth chamber equipped with eight different LED lights consisting of single or combinations of four different light spectra emitting blue (450 nm), green (525 nm), red (660 nm), and far-red (730 nm) light. After three months of incubation, morphological changes in embryogenic calli were recorded, and LC-MS/MS analysis of cultures was carried out for peimisine, sipeimine, peiminine, and peimine. The highest number of somatic embryos and the maximum fresh weight was recorded in calli incubated under red (9R), infrared...
Journal of the American Society for Horticultural Science, 2012
The genus Dendrobium is important in traditional Chinese herbal medicine, and the precise identif... more The genus Dendrobium is important in traditional Chinese herbal medicine, and the precise identification of Dendrobium species is critical for the treatment and for pharmacological research. In the present study, a ribosomal DNA (rDNA) internal transcribed spacer (ITS) region-based analysis was used to ascertain the phylogenetic relationship among 20 Dendrobium species. The lengths of the ITS regions among the 20 species ranged from 636 to 653 bp, and the identities of the rDNA regions among the different species ranged from 75.7% to 99.1%. The results also showed that the ITS1 and ITS2 regions exhibit more variation than the 5.8S rDNA. A phylogenetic tree derived from the ITS sequence indicated that six medicinal Dendrobium species, of which five are common medicinal plants in the Taiwan market, were closely related and shared a common clade. Multiplex polymerase chain reaction (PCR) amplification was successfully performed to identify the six medicinal Dendrobium species, and ampl...
Background Gentiana scabra Bunge commonly known as ‘Long dan cao’ in China has been used in tradi... more Background Gentiana scabra Bunge commonly known as ‘Long dan cao’ in China has been used in traditional Chinese medicines for more than 2000 years. Dry roots and rhizome of the herb have been used for the treatment of inflammation, anorexia, indigestion and gastric infections. Iridoids and secoiridoids are the main bioactive compounds which attribute to the pharmacological properties of this plant. The species is difficult to mass propagate by seed due to the low percentage of germination and limited dormancy period. Wild populations in some locations are considered to be in the endangered category due to over exploitation. Results In the present study, we report an efficient micropropagation system. Shoot apices of six weeks old in vitro grown G. scabra plants were used as explants for the in vitro propagation. Induction of multiple shoots (9.1/explant) was achieved on the culture of shoot apices on half strength Murashige and Skoog’s basal medium (MSBM) containing 2.0 mg/L−1 6-ben...
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