Akula A, Dodd W A
Centre for Molecular Biotechnology, School of Life Science, Queensland University of Technology, GPO Box 2434, Brisbane, QLD 4001, Australia, , , , , , AU.
Plant Cell Rep. 1998 Jul;17(10):804-809. doi: 10.1007/s002990050487.
An efficient method for the in vitro propagation of a tea (Camellia sinensis (L) O. Kuntze) clone, `TRI-2025', from somatic embryos is described. This technique involves two phases; the induction of adventitious buds from nodal cuttings followed by the development of somatic embryos. Single nodal cuttings were excised from 1-year-old in vitro axenic cultures and inoculated on MS medium with different combinations of IBA/BAP/GA. Induction of multiple shoots from nodal explants occurred on MS medium with 0.5 mg l BAP, 0.1 mg l IBA and 0.0 mg l GA within 6 weeks of incubation. The cultures with multiple shoots were transferred to fresh medium, incubated for 120 days and transferred to MS medium with half-strength macro nutrients, full-strength micronutrients and vitamins and no growth regulator. The direct induction of somatic embryos without callus formation occurred on this medium at 60% frequency within 4 weeks. The production of embryos continued upon transfer of the cultures to fresh medium and a four- to eightfold multiplication rate was obtained during each 6-week culture cycle. The plantlets from these embryos were acclimatised with a 90% success rate. All plants were vigorous and hardy, with well-developed tap-root systems.
本文描述了一种从体细胞胚体外繁殖茶树(Camellia sinensis (L) O. Kuntze)无性系“TRI-2025”的有效方法。该技术包括两个阶段:从带节插穗诱导不定芽,然后发育成体细胞胚。从1年生体外无菌培养物中切下单个带节插穗,接种在含有不同组合的吲哚丁酸(IBA)/苄氨基嘌呤(BAP)/赤霉素(GA)的MS培养基上。在含有0.5毫克/升BAP、0.1毫克/升IBA和0.0毫克/升GA的MS培养基上,培养6周内带节外植体诱导出多个芽。将带有多个芽的培养物转移到新鲜培养基上,培养120天,然后转移到含有半量大量元素、全量微量元素和维生素且无生长调节剂的MS培养基上。在该培养基上,4周内60%的频率直接诱导出体细胞胚,无愈伤组织形成。将培养物转移到新鲜培养基上后,胚的产生持续进行,在每个6周的培养周期中获得了4至8倍的增殖率。这些胚发育成的植株驯化成功率为90%。所有植株生长旺盛、耐寒,主根系统发达。