Micropropagation by Somatic Embryogenesis of Coffea Species and Cryopreservation

用细胞分裂素单独诱导小粒咖啡和中粒咖啡成熟树幼叶的胚性愈伤组织和体细胞胚。细胞分裂素诱导的愈伤组织具有很高的胚性发生潜力，对无性繁殖非常有用。作为细胞分裂素，2-异戊烯基腺嘌呤（2 iP）效果最好，苄基腺嘌呤（BA）、玉米素和激动素对咖啡叶片体细胞胚胎发生也有一定的促进作用。附加生长素对体细胞胚胎发生有抑制作用。从小粒种和中杯种的胚性愈伤组织中制备原生质体，并将其培养在含有细胞分裂素的Gelrite培养基上。原生质体再生体细胞胚。这些简化的原生质体制备和培养方法对细胞融合和遗传工程具有重要意义。在暴露于一系列制备步骤之后，成功地冷冻保存了咖啡属的体细胞胚;在具有增加浓度的蔗糖的培养基上预培养，包封在藻酸盐珠中，脱水至30%并插入液氮中。

The embryogenic callus and somatic embryo of Coffea arabica and C.canephora can be induced by cytokinin alone from the section of young leaf of mature trees. The cytokinin induced callus has a high embryogenic potetial and is very useful for clonal propagation. As cytokinin, 2-isopentenyl-adenine(2iP)is the best, and benzyladenin(BA), zeatin and kinetin also are effective for somatic embryogenesis from coffee leaf slices. Additional auxins act inhibitory effect on somatic embryogenesis. Protoplast were prepared from embryogenic calluses from C.arabica and canephora and cultured on Gelrite medium with cytokinin as a plant regulator. Somatic embryos were regenerated from protoplasts. These simplified method for preparation and culture of protoplasts is very useful for cell fusion and genetic engeering. Somatic embryos of Coffea were successfully cryopreserved after being exposed to a series of preparative steps ; precultured on media with increasing concentration of sucrose, encapsulated in alginate beads, dehydrated up to 30% and pluged into liquid nitrogen.