1、植物細胞與組織培養(yǎng)植物細胞與組織培養(yǎng) Plant Cell and Tissue Culture 精緻農(nóng)業(yè)學系精緻農(nóng)業(yè)學系 鍾仁彬鍾仁彬 助理教授助理教授課程內(nèi)容課程內(nèi)容: 植物細胞與組織培養(yǎng)發(fā)展至今，技術(shù)已經(jīng)相當純熟，試管內(nèi)培養(yǎng)植物細胞與組織培養(yǎng)發(fā)展至今，技術(shù)已經(jīng)相當純熟，試管內(nèi)培養(yǎng)成功的植物種類也愈來愈多，廣泛應(yīng)用於成功的植物種類也愈來愈多，廣泛應(yīng)用於植物學各方面研究植物學各方面研究、植物繁植物繁殖殖、作物良作物良等，更是等，更是生物技術(shù)生物技術(shù)發(fā)展的最重要基礎(chǔ)。發(fā)展的最重要基礎(chǔ)。 本課程期望修課同學瞭解本課程期望修課同學瞭解基本原理、技術(shù)與應(yīng)用基本原理、技術(shù)與應(yīng)用。包括器官、組。包括器官

2、、組織、細胞及原生質(zhì)體培養(yǎng)的基本技術(shù)、器官與胚發(fā)生的原理，及組織織、細胞及原生質(zhì)體培養(yǎng)的基本技術(shù)、器官與胚發(fā)生的原理，及組織培養(yǎng)於作物試管內(nèi)繁殖、無病毒植株培育、單被體培育、誘變育種、培養(yǎng)於作物試管內(nèi)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(chǎn)、遺傳工程之應(yīng)用等。體細胞雜種培育、二次代謝物生產(chǎn)、遺傳工程之應(yīng)用等。Ch. 1. Introduction 1-1. Definitions 1-2. Advantages of Plant Tissue Culture 1-3. History of Plant Tissue Culture 1. For propaga

3、tion 2. Genotype modification (i.e., plant breeding)3. Biomass production of biochemical secondary products4. Plant pathology5. Germplasm preservation6. Scientific investigations植物生物技術(shù)生產(chǎn)應(yīng)用關(guān)係圖植物生物技術(shù)生產(chǎn)應(yīng)用關(guān)係圖器官培養(yǎng)器官培養(yǎng)(Organ culture)種苗生產(chǎn)種苗生產(chǎn)(Seedling production)育育 種種(Breeding)生物產(chǎn)量分離與精製生物產(chǎn)量分離與精製(Bio-mass f

4、or separation and purification)醫(yī)葯品等有用物質(zhì)生產(chǎn)醫(yī)葯品等有用物質(zhì)生產(chǎn)Production for medicaments 細胞培養(yǎng)細胞培養(yǎng)(Cell culture)植物無性繁殖植物無性繁殖(Asexal propagation)細胞固定細胞固定(Cell fixation)種源保存種源保存(Germplasm preservation)遺傳修飾遺傳修飾(Genetic modification)必要技術(shù)應(yīng)用技術(shù) Q : Key points for successful plant tissue culture 1861 Knop : Knops solut

5、ion, inorganic salt nutrition1902 Haberlandt (Austria) : Cultivate isolated plant cells in vitro on an artificial medium -Concept of “ Totipotency ”1922 Kotte (student of Haberlandt): Improve medium : sugar, Organic N-compounds Father of Plant Tissue CulturePlants can be separated into their compone

6、nt 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，可再每個細胞具有全套基因組，可再生成為完整個體的能力生成為完整個體的能力1922 Robbins (USA) : Root tip culture - Pisum (豌豆屬豌豆屬), Zea and Gossypium (棉屬棉

7、屬) First success of culturing plant tissue in vitro1923 Robbins & Maneval : Maintained maize roots for 20 weeks with the aid of subcultures 1926 Went : Discovery of auxin1933 Kogl, haagen-Smit & Erxleben : Isolation of IAA (indo-3-acetic acid) 1932Gautheret : I IRoot tip & root fragment

8、culture I 1934 Cambium culture I ( woody plant ) I I maintain several months I I 1937IAA promotes root growth I I1939Carrot explants undifferentiated mass ( callus ) repeated subculture 1932White (in Rodkefeller Institute) : RRoot tip culture - Triticum 1934 Indefinite culture of isolated roots toma

9、to - 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 cultur

10、e 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

11、 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 format

12、ion 1957Skoog & Miller : auxin / cytokinin balance in root or shoot formation high kinetin - shoot high auxin - root Auxin / Cytokinin balance1956Nickell (Chas, Pfizer & Co. N.Y.) : Single cell suspension culture of Phaseolus vulgaris 1960Bergmenn : Plating culture Nicotiana tobacum & Ph

13、aseolus 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 culture19571957TuleckeTulecke : : Pollen culture of Pollen culture of Ginkgo bilobaGinkgo biloba 1960 Cocking : (UK)Enzy

14、matic isolation and culture of protoplast 1962 Murashige & Skoog : Tobacco tissue culture Development of MS medium 1964 Guha & Maheshwari : (India)Anther culture - haploid plant ( Datura ) 1968 Niizeki & Oono : (Japan)Haploid plant of rice Started for plant breeding One pollen grain - on

15、e plant 1971 Nagata & Takebe : (Japan)Plant regeneration from tobacco protoplast 1972 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 research1. Morphogenesis2. Biochemical : higher plant3. Genetics : somatic cell genetics, mutation, polyploidization4. Host-pathogen studies : obligate parasites, plant tumors B. Economic application1. Production of secondary substances2. Rapid clonal progagation 3. Artificial seeds4. Pathoge