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In Vitro Propagation of Zingiberaceae Species with Medicinal Properties
Keng, Chan Lai,Hing, Thong Weng The Korean Society of Plant Biotechnology 2004 Plant molecular biology and biotechnology research Vol.6 No.3
Zingiber officinale buds from the rhizomes were used to produce in vitro shoots. These explants produced the largest number of multiple shoots, 9.8 shoots per explant, when were cultured on MS (Murashige and Skoog 1962) medium supplemented with 2.0 mg/L benzyladenine (BA) and 2.0 mg/L indole butyric acid (IBA). This medium was also found to be suitable for in vitro propagation of other Zingiberaceae species: Alpinia conchigera, Alpinia galanga, Curcuma domestica, C. zedoaria and Kaempferia galanga. Both C. domestica and C. zedoaria produced more multiple shoots when were cultured in the liquid proliferation medium, MS medium containing 2.0 mg/L BA and 2.0 mg/L IBA. To maintain the in vitro plantlets of Zingiberaceae species, they were required to subculture every four weeks. After executing proper acclimatization protocol, in vitro plantlets of Alpinia galanga, A. conchigera, Curcuma domestica, C. zedoaria, Kaempferia galanga and Zingiber officinale could be successfully planted in the field with high percentage of survival.
( Lai Keng Chan ),( Wan Yee Koh ),( Tengku S. Tengku Muhammad ) 한국식물학회 2005 Journal of Plant Biology Vol.48 No.1
An in vitro cytotoxicity screening of the Typhonium flagelliforme extracts indicated high cytotoxicity effect on human lung carcinoma NCI-H23 cells and human mammary gland carcinoma T-47D cells, but the extracts were not active on human liver carcinoma HepG2 cells. NCI-H23 cells were more susceptible to T. flagelliforme extracts than T -47D cells. ED50 values of the hexane fractions of the mature plant and the in vitro plantlet of T. flagelliforme on NCI-H23 cells were less than 2 ㎍/mL. Extract from the mature plant was relatively more cytotoxic than the one from in vitro plantlet except for the hexane fraction. The chloroform and butanol fraction of the mature plant had higher cytotoxicity effect than the fraction from in vitro plantlet on NCI-H23 cells. All the 3 fractions (hexane, chloroform, and butanol) of the mature plant exhibited higher cytotoxicity effects on human mammary gland carcinoma T -47D cells than the 3 fractions of in vitro plantlet. However, the human liver carcinoma cells were resistant to T. flagelliforme extracts except for higher concentration of hexane fractions of both the mature and the in vitro plants and the chloroform fraction of the mature plant. Micropropagated plantlets of T. flagelliforme could hence be used as herbal materials for the treatment of human lung and breast cancers.
Chan, Lai Keng,Dewi, Patra Rina,Boey, P.L. 한국식물학회 2005 Journal of Plant Biology Vol.48 No.1
Cymbopogon nardus L could be propagated via tissue culture using axillary buds as explants. The aseptic bud explants obtained using double sterilization methods produced stunted abnormal multiple shoots when they were cultured on Murashige and Skoog medium (MS) supplemented with 1.0 ㎎ L^(-1) or 2.0 ㎎ L^(-1) benzyladenine (BA). Stunted shoots that cultured on MS + 1.0 ㎎ L^(-1) BA + 1.0 ㎎ N^6-isopentenyl-adenine (2iP) could induce elongation of shoots from about 60% of the stunted shoots. Normal multiple shoots could be induced at the highest (19.7 shoots per bud) from the bud explants within six weeks when cultured on proliferation medium consisted of MS supplemented with 0.3 ㎎ L^(-1) BA + 1.0 ㎎ L^(-1) indole-3-butyric acid (IBA). The separated individual shoot produced roots when transferred to basic MS solid medium. The essential oils that were contained in the mature plants namely citronella), geraniol and citronellol were also found in the in vitro C. nardus plant-lets. Citronella) was the main essential oil component in the matured plants while geraniol was the main component in the in vitro plantlets.
( Chan Chee Weng ),( Chan Lai Keng ),( Boey Peng Lim ) 한국식물학회 2004 Journal of Plant Biology Vol.47 No.3
Juvenile hormone Ⅲ (JH Ⅲ) is one of the five closely related sesquiterpenoid compounds important in the regulation of an insect physiological processes in both the developing and the mature insect. JH Ⅲ and its biosynthetic intermediates, farnesol and methyl farnesoate were detected in the in vitro Cyperus aromaticus plantlets and the mature plants growing in the field. A higher amount of methyl farnesoate (52.7%) and farnesol (31.4%) were found to be present in the in vitro plantlets as compared to the field grown mother plants which contained lower methyl farnesoate (19.5%) and farnesol (14.8%). On the other hand, the mature mother plant (4 months old) contained a higher percentage of JH Ⅲ than the in vitro plantlets. The estimated content of JHⅢ analyzed by gas chromatography indicated that the amount of JHⅢ in the 10 weeks old in vitro plantlets and the four months old mature plants of C. aromaticus was 0.076 ㎍/g and 0.085 ㎍/g respectively. This study indicated that in vitro culture system could be a potential tool for the production of this natural derived bio-insecticide and could provide a material source for further investigations of the biosynthesis of JH Ⅲ in C. aromaticus.
Leng, Tan Chee,Haw, Ang Boon,Chan, Lai-Keng 한국식물학회 2004 Journal of Plant Biology Vol.47 No.1
Nodal explants of Spilanthes acmella produced normal multiple shoots when cultured vertically on Murashige and Skoog medium (1962) supplemented with 0.5 ㎎L^(-1) BA. The number of shoots formed from each explant was doubled after first 5-week subculture. The nodal explants placed vertically in Erlenmeyer flask (250,500, or lo00 ml) produced more multiple shoots than those cultured in 350 mL jam bottles and 500 mL tex-Z flask. Temperature above 28℃ caused abnormalities of in vitro plantlets. All in vitro plantlets survived after acclimatized and transferred to the outside environment. The survived plantlets did not show any morphological abnormalities in the field condition.
Siregar Luthfi Aziz Mahmud,Chan Lai-Keng,Boey Peng-Lim The Korean Society of Plant Biotechnology 2003 Plant molecular biology and biotechnology research Vol.5 No.2
Callus and cell suspension cultures of Eurycoma longifolia Jack were initiated from leaves of different trees. The leaf explant of tree Eu9 produced the most calli and also induced high cell biomass in the cell suspension culture. Optimum production of cell biomass could be initiated in proliferating culture medium with a pH of 5.75 prior to autoclaving. The effects of macronutrient inorganic salts of Murashige and Skoog (MS) liquid medium supplemented with X on production of cell biomass of Eurycoma longifolia were also investigated. The highest cell biomass was produced in MS medium containing macronutrients of $21\;mM\;NH_4NO_3,\;12.25\;mM\;KNO_3,\;3.00\;mM\;CaCl_2.2H_2O,\;0.575\;mM\;MgSO_4.7H_2O$, and $1.83\;mM\;KH_2PO_4$. A new medium labeled as TAM was formulated for the production of Eurycoma longifolia cell biomass in the cell suspension culture.