The objective of the study was to find the explant resources, the type and concentration of plant growth regulators (PGRs), as well as the in vitro culture conditions suitable for different morphogenesis processes - callus induction, direct embryogenesis, shoot and root formation, etc. The growth and development of in vitro Ngoc Linh ginseng derived from thin cell layer (TCL) was examined in Quang Nam to assess the adaptability of in vitro Ngoc Linh plantlets in its natural territory, compared to those growing in Bidoup - Nui Ba National Park, Lam Dong.
MINISTRY OF EDUCATION AND TRAINING VIETNAM ACADEMY OF SCIENCE AND TECHNOLOGY GRADUATE UNIVERSITY OF SCIENCE AND TECHNOLGY - VU THI HIEN REGENERATION AND MICROPROPAGATION OF Panax vietnamensis Ha et Grushv USING THIN CELL LAYER TECHNOLOGY Major: Plant Physiology Code: 9.42.01.12 SUMMARY OF PHILOSOPHY DOCTORAL DISSERTATION ON BIOLOGY Ho Chi Minh City - 2018 The work was realized in Graduate University of Science and Technology, Vietnam Academy of Science and Technology Advisor 1: Prof Duong Tan Nhut, Ph.D Advisor 2: Thai Xuan Du, Ph.D Reviewer 1: Reviewer 2: Reviewer 3: The thesis will be evaluated by doctoral committee at Graduate University of Science and Technology, Vietnam Academy of Science and Technology on ……… 2018 The thesis is available at: - Library of Graduate University of Science and Technology - National Library of Vietnam INTRODUCTION The necessity of the dissertation Ngoc Linh ginseng is a Vietnamese endemic ginseng with the scientific name Panax vietnamensis Ha et Grushv Since discovered in 1973, it can be said that Ngoc Linh ginseng is one of the most important medicinal plants Many reports indicated that Ngoc Linh ginseng has not only the pharmacological characteristics of a Ginseng, but also the individual characteristics such as anti-stress, decrease of depression and anxiety, stimulation of the immune system, resistance to cytotoxic toxins, antioxidant in vitro and in vivo, etc The success of propagation of Ngoc Linh ginseng is still limited because this species is only grown on Ngoc Linh mountain To harvest gingseng roots, the propagation period lasts to years to store enough bioactivities Our thesis entitled "Regeneration and micropropagation of Ngoc Linh ginseng (Panax vietnamensis Ha et Grushv.) using thin cell layer technique" has been carried out The aim of this study is to obtain a number of vigorous plantlets with and high quality roots and tubers, especially, they are well adapted to the natural conditions, thereby contributing to preserving this precious medicinal plant Objective The objective of the study was to find the explant resources, the type and concentration of plant growth regulators (PGRs), as well as the in vitro culture conditions suitable for different morphogenesis processes - callus induction, direct embryogenesis, shoot and root formation, etc The growth and development of in vitro Ngoc Linh ginseng derived from thin cell layer (TCL) was examined in Quang Nam to assess the adaptability of in vitro Ngoc Linh plantlets in its natural territory, compared to those growing in Bidoup - Nui Ba National Park, Lam Dong The contents of the thesis 3.1 Research on the morphogenesis from different explant resources 3.2 Research on growth and subsequent development of plantlets in vitro in different ecological conditions 3.3 Qualitative and quantitative saponin in plant in vitro and at nursery CHAPTER I OVERVIEW The thesis has consulted 34 Vietnamese documents and 94 English documents and internet documents; (1) Introduction to ginseng; (2) Ngoc Linh ginseng (Panax vietnamensis Ha et Grushv.); (3) cell culture technology; (4) factors influencing morphogenesis; (5) plant growth regulators; (6) the role of light in the regeneration, growth and development of plants; (7) The plant regeneration CHAPTER II RESEARCH OBJECTIVE AND METHODS 2.1 Materials 2.1.1 Plant materials Explant source for morphogenesis: including leaf, petiole and rhizome of 3-month old Ngoc Linh ginseng in vitro plants; Explants grew under different ex vitro conditions: Ngoc Linh ginseng intact plants with rhizomes and leaves, about cm in height; explant sources for determining bioactive agents: in vitro plantlets, 6-month old, 1-year old and 2-years old plants 2.1.2 Equipment - tools, standard chemicals and solvents Light intensity meter LI-250A Light meter; microscope; Equipment used in HPLC analysis (High Performance Liquid Chromatography, Rg1, Rb1, MR2) Chloroform: methanol: water (65:35:10) 2.2 Research Methods 2.2.1 Plant morphogenesis method 2.2.2 Plant morphology and microscope observation method 2.2.4 Saponin content analysis method 2.2.4.1 Thin layer chromatography method 2.2.4.2 High Performance Liquid Chromatography (HPLC) 2.3 Research establishment methods 2.3.1 Content 1: Researching the morphogenesis from different explant sources 2.3.1.1 Evaluating the effect of single PGRs on morphogenesis of leaf explant tTCL_L under light and dark conditions 2.3.1.2 Evaluating the effect of single PGRs on morphogenesis of petiole explant tTCL_L under light and dark conditions 2.3.1.3 Evaluating the effect of single PGRs on morphogenesis of lTCL_C petiole explant under light and dark conditions 2.3.1.4 Evaluating the effect of single PGRs on morphogenesis of TTCL_R rhizome explant under light and dark conditions 2.3.1.5 Evaluating the effect of the combination of auxin and cytokinin on the morphogenesis of the leaf explant tTCL_L under light and dark conditions 2.3.1.6 Evaluating the effect of the combination of auxin and cytokinin on the morphogenesis of the tTCL_C petiole explant under light and dark conditions 2.3.1.7 Evaluating the effect of the combination of auxin and cytokinin on the morphogenesis of the lTCL_C petiole explant under light and dark conditions 2.3.1.8 Evaluating the effect of the combination of auxin and cytokinin on the morphogenesis of the rhizome explant tTCL_R under light and dark conditions 2.3.1.9 Morphological anatomy 2.3.1.10 Develop Ngoc Linh plantlets from somatic embryos 2.3.2 Research on growth and subsequent development of in vitro plantlets under different ecological conditions 2.3.2.1 Research on the growth and development of Ngoc Linh ginseng cultured in vitro grown in Quang Nam 2.3.2.2 Research on the growth and development of Ngoc Linh ginseng cultured in vitro grown in the Heaven Gate of Bidoup - Nui Ba National Park (Lam Dong) 2.3.3 Content 3: Qualitative and quantitative saponin in ginseng in vitro and in complete ginseng at nursery stage 2.3.3.1 Determine the content of saponin in Ngoc Linh ginseng in vitro, month ginseng, year and year old seedlings planted in Quang Nam 2.3.3.2 Quantification of saponins in Ngoc Linh ginseng in vitro, month ginseng, year and year old trees were planted in Quang Nam 2.4 Statistics The experiment was completely randomized (CDR) The mean of the follow-up indices among the treatment formulas was analyzed by ANOVA method, then compared with the Ducan test at confidence level P