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植物細胞與組織培養 Plant Cell and Tissue Culture,精緻農業學系 鍾仁彬 助理教授,課程內容: 植物細胞與組織培養發展至今，技術已經相當純熟，試管內培養成功的植物種類也愈來愈多，廣泛應用於植物學各方面研究、植物繁殖、作物良等，更是生物技術發展的最重要基礎。 本課程期望修課同學瞭解基本原理、技術與應用。包括器官、組織、細胞及原生質體培養的基本技術、器官與胚發生的原理，及組織培養於作物試管內繁殖、無病毒植株培育、單被體培育、誘變育種、體細胞雜種培育、二次代謝物生產、遺傳工程之應用等。,Ch. 1. Introduction 1-1. Definitions 1-2. Advantages of Plant Tissue Culture 1-3. History of Plant Tissue Culture 1-4. Applications,1-1. Definitions,Plant Tissue Culture Plant Cell, Tissue and Organ Culture In vitro Culture Aseptic Culture,1-2. Advantages of Plant Tissue Culture,1. For propagation,2. Genotype modification (i.e., plant breeding),3. Biomass production of biochemical secondary products,4. Plant pathology,5. Germplasm preservation,6. Scientific investigations,植物生物技術生產應用關係圖,器官培養 (Organ culture),種苗生產 (Seedling production),育 種 (Breeding),生物產量分離與精製 (Bio-mass for separation and purification),醫葯品等有用物質生產 Production for medicaments,細胞培養 (Cell culture),植物無性繁殖 (Asexal propagation),細胞固定 (Cell fixation),種源保存 (Germplasm preservation),遺傳修飾 (Genetic modification),必要技術,應用技術,Q : Key points for successful plant tissue culture,1-3. History of Plant Tissue Culture,1861Knop : Knops solution, inorganic salt nutrition,Haberlandt (Austria) : Cultivate isolated plant cells in vitro on an artificial medium -,Concept of “ Totipotency ”,1922Kotte (student of Haberlandt): Improve medium : sugar, Organic N-compounds,Father of Plant Tissue Culture,Plants can be separated into their component parts ( organs, tissues, or cells ), which can be manipulated in vitro and then grown back to complete plants., Concept of “ Totipotency ”,植物可取其部分器官、組織或細胞進行體外操作，然後生長為一個完整植株。,每個細胞具有全套基因組，可再生成為完整個體的能力,1922 Robbins (USA) : Root tip culture - Pisum (豌豆屬), Zea and Gossypium (棉屬) First success of culturing plant tissue in vitro 1923 Robbins & Maneval : Maintained maize roots for 20 weeks with the aid of subcultures 1926 Went : Discovery of auxin 1933 Kogl, haagen-Smit & Erxleben : Isolation of IAA (indo-3-acetic acid),Gautheret : I IRoot tip & root fragment culture I 1934 Cambium culture I ( woody plant ) I I maintain several months I I 1937IAA promotes root growth I I 1939Carrot explants undifferentiated mass ( callus ) repeated subculture,1932White (in Rodkefeller Institute) : RRoot tip culture - Triticum 1934 Indefinite culture of isolated roots tomato - subculture / 7 days 1936 160 passages Importance of vitamin B complex in root culture ( use yeast extract ) 1939Procambial tissue culture - callus ( Nicotiana ) - small leaf and shoot 1942Tumor culture - callus without plant hormone,1950Steward : carrot culture : quantitative analysis of culture growth Discovered embryogenesis in vitro,1955Steward & Shantz (in Cornell) : Carrot root phloem in liquid medium single cell suspension 1958 whole plant, Prove the concept of totipotency,1952 Morel & Martin : First virus-free plant through shoot tips culture ( Dahlia ),1960Shoot tip culture of Cymbidium, Propagation many plants from one plant,1954Muir, Hildebrandt & Riker (in Wisconsin) : Suspension culture by reciprocal shaker ( Nicotiana ),Single cell clone by nurse cell culture method,1955Miller, Skoog, etc. Discovery of kinetin (the first cytokinin) Tobacco culture - bud formation,1957Skoog & Miller : auxin / cytokinin balance in root or shoot formation high kinetin - shoot high auxin - root,Auxin / Cytokinin balance,1956Nickell (Chas, Pfizer & Co. N.Y.) : Single cell suspension culture of Phaseolus vulgaris,1960Bergmenn : Plating culture Nicotiana tobacum & Phaseolus vulgaris Single cell suspension culture - plating in 0.6% agar medium, thin layer in petri dish - cell divided to form colony, Idea of producing plant alkaloids from tissue culture,1957Tulecke : Pollen culture of Ginkgo biloba,Cocking : (UK) Enzymatic isolation and culture of protoplast,1962 Murashige & Skoog : Tobacco tissue culture Development of MS medium,Guha & Maheshwari : (India) Anther culture - haploid plant ( Datura ),Niizeki & Oono : (Japan) Haploid plant of rice Started for plant breeding, One pollen grain - one plant,Nagata & Takebe : (Japan) Plant regeneration from tobacco protoplast,Carlson : Somatic hybrid plant from protoplst fusion Nicotiana glauca + N. langsdorffii ( 2n = 24 ) I( 2n = 18 ) I somatic hybrid ( 2n = 42 ),1986Hauptmann : Transgenic tobacco,A. Basic research 1. Morphogenesis 2. Biochemical : higher plant 3. Genetics : somatic cell genetics, mutation, polyploidization 4. Host-pathogen studies : obligate parasites, plant tumors,1-4. Applications,B. Economic application 1. Production of secondary substances 2. Rapid clonal progagation 3. Artificial seeds 4. Pathogen free plants 5. Plant breeding & Hybridization a. Embryo culture b. Pollen tube pollination c. Mutation induction & isolation d. Haploids e. Preservation & International movement of germplasm f. Somatic hybridization g. Genetic engineering,1-4. Applications,
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