细胞工程-2-2014.ppt

Tuberization Flowering in vitro What is needed?Tissue culture has several critical requirements: Appropriate tissue (some tissues culture better than others) A suitable growth medium containing energy sources and inorganic salts to supply cell growth needs. Aseptic (sterile) conditions, as microorganisms grow much more quickly than plant and animal tissue and can over run a culture Growth regulators - both auxins cytokinins. Frequent subculturing to ensure adequate nutrition and to avoid the build up of waste metabolites * * * * * The most important naturally occurring auxin is IAA (indole-3-acetic acid), but its use in plant cell culture media is limited because it is unstable to both heat and light. It is more common to use stable chemical analogues of IAA as a source of auxin in plant cell culture media. 2,4-Dichlorophenoxyacetic acid (2,4-D) is the most commonly used auxin and is extremely effective in most circumstances. 胚胎发生的显著优点 1、数量多 2、速度快 3、结构完整 数量多: 一般在同一个外植体上诱导产生胚状体的数量要比诱导产生芽的数量高。这方面的优点在细胞悬浮培养中更为突出。如胡萝卜悬浮培养，在一个培养瓶中就可以产生10万个胚状体，这使人工种子的利用成为现实。 速度快: 因为胚状体是以单细胞直接分化产生小植株，所以比经过愈伤组织再分化的器官发生途径要快一些。例如，大麦小孢子培养，从接种花药到获得绿色花粉植株，胚状体途径大约需要25天，而器官发生途径大约需要50-55天。 结构完整: 由于胚状体具有同合子胚相类似的结构，容易萌发形成小植株。 Celery plant regeneration Embryogenic calluses Formation of somatic embryos Mature somatic embryos Plantlets Peanut somatic embryogenesis 三、影响形态发生的因素 Genotype Physiological state Medium Chemical Plant Growth Regulators (PGRs) Environment Genotype Different species show differences in amenability to tissue culture 如茄科中的烟草、矮牵牛，伞形花科中的胡萝卜，十字花科中的油菜很易再生成植株；而禾本科等单子叶植物则较难再生。 In many cases, different genotypes within a species will have variable responses to tissue culture Physiological state 1．外植体的类型 同一植物不同器官或组织的再生能力不同。 外植体的大小对再生也有影响，外植体太小很难存活生长。 2．个体发育年龄 Usually, the younger, less differentiated the explant, the better for tissue culture 愈伤组织的年龄也有明显的影响，年龄越老，当转移到分化培养基生长时，其苗分化频率越低。因此在组织培养中，要及时转移已形成的愈伤组织进行分化培养，可大大提高苗的分化频率。 ? Medium Macroelements Nitrogen (N)