[工学]原生质体培养与融合28h.ppt

* * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * 分析所鉴定植株是否具有融合亲本的遗传物质 再生植株同时具有亲本的带； 再生植株在一种情况下具有一个亲本的带，在另一种情况下具有另一个亲本的带。 RAPD带型 RAPD带型 Protoplast fusion - a new way in hybridisation Protoplast fusion provides a unique tool to combine valuable traits beyond the feasibility of normal sexual crossing. Protoplasts are single cells without a cell wall. In potato and hops they are isolated from leaves using cell wall degrading enzymes. Triggered by electric pulses protoplasts of two selected breeding lines can be fused in an electric field. This fusion (= somatic hybridisation) encompasses the combination of all chromosomes from both fusion partners. Therefore the starting material should only possess half the number of chromosomes which can be achieved by specific crosses or through anther culture.   ???  Benefits of protoplast fusion No loss of genetic information during the formation of gametes Specific addition of the genomes of two plants Combination of complex traits without loosing any gene Elimination of crossing barriers Protoplast fusion in potato Application of protoplast culture Exploitation and incorporation of new genes (wild species) Construction of basis breeding populations and cultivars Specific breeding for quality and resistance Since 1990 protoplast fusion is well established in potatoes. Each year approximately 400 somatic hybrids derived from protoplast fusion are tested in the fields. At current this technique is being established in hops. Regeneration of the first protoplast derived hop plants has been achieved.  A mesophyll protoplast-derived hop plant A两个彼此独立的原生质体 B两个原生质体间发生凝聚作用 C、D在局部位置上膜融合 E、F形成1个异核体 原生质体融合过程示意图 A B C D E F A两个彼此独立的原生质体 B两个原生质体间发生凝聚作用 C、D在局部位置上膜融合 E、F形成1个异核体 原生质体融合过程示意图 植物A细胞 植物B细胞 去壁 原生质体A 原生质体B 原生质体融合 融合体 再生壁 杂种细胞 细胞分裂 愈伤组织 杂种植株 去壁的常用方法： 酶解法（纤维素酶、果胶酶等） 原生质体融合方法： 物理法：离心、振动、电刺激等 化学法：聚乙二醇（PEG） 植物体细胞杂交过程示意