园林生物技术.doc

1学院专业报名接收免试统考上线单考破格调剂招生数自费委培1564906041015169+1981043 000081303 城市规划与设计 317 1341 422 000090706 园林植物与观赏园艺※110 49 39 000560100 风景园林硕士 17+5563 免费考研网 园林植物与观赏园艺※120104748 33 免费考研网 Biotechnology of Horticultural Plant园艺植物生物技术园艺植物生物技术园艺植物生物技术园艺植物生物技术2（理论课教案）（理论课教案）华中农业大学园艺林学学院华中农业大学园艺林学学院2007-2008-13华华中中农农业业大大学学教教案案（（首首页页））授课时间 2007 年 9-11 月 教案编写时间 2007 年 8 月 课程编号 课程名称园艺植物生物技术 学 分 数2.5课型必修课（） 选修课（ ） 理论课（） 实验课（ ）任课教师郭文武、程运江职称教授、副教授（博士）总学时 40讲课： 30 学时 实验： 10 学时 实习： 学时授课对象2002（班）级 园艺 专业 1、2 班基本教材 或主要参 考书园艺植物生物技术，邓秀新、胡春根主编。

高等教育出版社，2005 H.S.Chawla 植物生物技术导论(许亦农 麻密 译) 化学工业出版社 肖尊安 植物生物技术 化学工业出版社教学目的 与要求通过本课程的学习，要求园艺专业学生掌握园艺植物生物技术相关基本理论和方法教学重点、 难点重点：生物技术的概念/应用现状、组织培养技术、病毒病脱除与无病毒繁殖 难点：原生质体操作技术、基因分离克隆、转基因技术教学进程第 次课12-56-78-910-1112-1314-15授课章节（30 学时）Theory part1) Introduction2) Tissue culture of hortic plant3) Protoplast culture shoots can be constantly divided * Stage III - Rooting * explant transferred to root media * Stage IV - Transfer to soil * explant returned to soil; hardened off二二二 外植体灭菌外植体灭菌外植体灭菌 1.过程 2.材料与前处理 3.灭菌强度 药剂种类与消毒处理时间三、三、三、组织培养过程中的变异现象组织培养过程中的变异现象组织培养过程中的变异现象 （一） 现象 Somaclonal Variation: variability in plants regenerated from tissue culture that is either induced or uncovered by a tissue culture process. Most somaclonal variation is negative, but if enough plants are examined, positive changes can usually be recovered. * The source for most breeding material begins with mutations, whether the mutation occurs in a modern cultivar, a landrace, a plant accession, a wild related species, or in an 12unrelated organism * Total sources of variation: * Mutation, Hybridization, Polyploidy * Somaclonal variation is a general phenomenon of all plant regeneration systems that involve a callus phase * There are two general types of Somaclonal Variation: * Heritable, genetic changes (alter the DNA) * Stable, but non-heritable changes (alter gene expression, AKA epigenetic) * Since utilizing somaclonal variation is a form of mutation breeding, we need to consider mutation breeding in more detail → InducingInducingInducing MutationsMutationsMutations * Physical Mutagens (irradiation) * Neutrons, Alpha rays *Densely ionizing (“Cannon balls”), mostly chromosome aberrations * Gamma, Beta, X-rays *Sparsely ionizing (“Bullets”), chromosome aberrations most mutations are recessive*Can avoid this constraint by not applying selection pressure in culture, but you loose the advantage of high through-put screening – have to grow out all regenerated plants, produce seed, and evaluate the M2 *Alternative: perform on haploid cell linesDiseaseDiseaseDisease ResistantResistantResistant SuccessSuccessSuccess usingusingusing SomaclonalSomaclonalSomaclonal VariationVariationVariation 第五节第五节第五节 几种组织培养技术在园艺中应用几种组织培养技术在园艺中应用几种组织培养技术在园艺中应用 一一一 胚培养（胚培养（胚培养（EmbryoEmbryoEmbryo CultureCultureCulture）））（一） Embryo Culture as a Source of Genetic Variation * Rescue F1 hybrid from a wide cross * Overcome seed dormancy, usually with addition of hormone to media (GA) * To overcome immaturity in seed * To speed generations in a breeding program * To rescue a cross or self (valuable genotype) from dead or dying plant * Hybridization * Can transfer mutant alleles between species * Can introduce new genetic combinations through interspecific crosses13* Polyploidy * Can combine embryo culture with chromosome doubling to create new polyploid species (allopolyploidy) * Embryo culture developed from the need to rescue embryos (embryo rescue) from wide crosses where fertilization occurred, but embryo development did not occur; * These techniques have been further developed for the production of plants from embryos developed by non-sexual methods (haploid production discussed later)（二）胚抢救（二）胚抢救（二）胚抢救 （（（EmbryoEmbryoEmbryo RescueRescueRescue ProcessProcessProcess））） * Make cross between two species * Dissect embryo (usually immature) * The younger the embryo, the more difficult to culture * Grow on culture medium using basic tissue culture techniques, use for breeding if fertile * Many times, resulting plants will be haploid because of lack of pairing between the chromosomes of the different species * This can be overcome by doubling the chromosomes, creating allotetraploids* Polyploids are another source of genetic variation → （三）幼胚离体培养的生长发育方式（三）幼胚离体培养的生长发育方式（三）幼胚离体培养的生长发育方式 1.胚性发育(embryonal development)： 2.早熟萌发(early mature sprouting)（四）（四）（四） 胚抢救应注意的几个因素胚抢救应注意的几个因素胚抢救应注意的几个因素 1 培养时期培养时期 自养（autotrophy） 异养（heterotrophy） 活体内胚>带胎座的胚珠>胚珠>退化或败育胚 2. 培养基和培养条件培养基和培养条件 3. 碳水化合物碳水化合物 4. 激素种类和水平激素种类和水平 5. 温度温度 两步培养法二二二 胚乳培养胚乳培养胚乳培养 （（（EndospermEndospermEndosperm culturecultureculture））） 一一一 概况概况： ： 胚乳培养始于胚乳培养始于1933年年， ，进进行玉米胚乳培养。

胚乳培养研究主要集中探行玉米胚乳培养胚乳培养研究主要集中探讨讨以下以下问题问题： ： 1、胚乳胚乳细细胞全能性：在离体培养条件下，胚乳胞全能性：在离体培养条件下，胚乳细细胞是否与其它体胞是否与其它体细细胞和花粉一胞和花粉一样样， ，具有全能性而再生植株具有全能性而再生植株 2、多数被子植物胚乳是三倍体，能否通、多数被子植物胚乳是三倍体，能否通过过胚乳培养胚乳培。