Chen J -T., Chang W -C.
Institute of Botany, Academia Sinica, 115, Taipei, Taiwan, People's Republic of China
Plant Sci. 2000 Dec 7;160(1):87-93. doi: 10.1016/s0168-9452(00)00367-8.
An efficient method was established for high frequency somatic embryogenesis and plant regeneration from callus cultures of a hybrid of sympodial orchid (Oncidium 'Gower Ramsey'). Compact and yellow-white embryogenic calli formed from root tips and cut ends of stem and leaf segments on 1/2 MS [11] basal medium supplemented with 1-phenyl-3-(1,2,3-thiadiazol-5-yl)-urea (TDZ, 0.1-3 mg/l), 2,4-dichlorophenoxyacetic acid (2,4-D, 3-10 mg/l) and peptone (1 g/l) for 4-7 weeks. Embryogenic callus was maintained by subculture on the same medium for callus induction and proliferated 2-4 times (fresh weight) in 1 month. Initiation of somatic embryogenesis and development up to the protocorm-like-bodies (PLBs) from callus cultures was achieved on hormone-free basal medium. Regenerants were recovered from somatic embryos (SEs) after transfer to the same medium and showed normal development. The optimized protocol required about 12-14 weeks from the initiation of callus to the plantlet formation. Generally, the frequency of embryo formation of root-derived callus was higher than stem- and leaf-derived calli. Combinations of naphthaleneacetic acid (NAA) and TDZ significantly promoted embryo formation from callus cultures. The high-frequency (93.8%) somatic embryogenesis and an average of 29.1 SEs per callus (3x3 mm(2)) was found in root-derived callus on a basal medium supplemented with 0.1 mg/l NAA and 3 mg/l TDZ. Almost all the SEs converted and the plantlets grew well with an almost 100% survival rate when potted in sphagnum moss and acclimatized in the greenhouse.
建立了一种高效方法，用于从合轴兰花杂种（文心兰‘高尔·拉姆齐’）愈伤组织培养物中进行高频体细胞胚胎发生和植株再生。在添加了1-苯基-3-（1,2,3-噻二唑-5-基）脲（TDZ，0.1 - 3 mg/l）、2,4-二氯苯氧乙酸（2,4-D，3 - 10 mg/l）和蛋白胨（1 g/l）的1/2 MS [11]基本培养基上，根尖以及茎和叶切段的切口处形成紧密的黄白色胚性愈伤组织，培养4 - 7周。胚性愈伤组织通过在用于愈伤组织诱导的相同培养基上继代培养来维持，并在1个月内增殖2 - 4倍（鲜重）。在无激素基本培养基上实现了从愈伤组织培养物中起始体细胞胚胎发生并发育至原球茎状体（PLBs）。将体细胞胚（SEs）转移到相同培养基后获得再生植株，且植株发育正常。从愈伤组织起始到形成小植株，优化后的方案大约需要12 - 14周。一般来说，根来源愈伤组织的胚胎形成频率高于茎和叶来源的愈伤组织。萘乙酸（NAA）和TDZ的组合显著促进愈伤组织培养物的胚胎形成。在添加了0.1 mg/l NAA和3 mg/l TDZ的基本培养基上，根来源愈伤组织中发现了高频（93.8%）体细胞胚胎发生，每个愈伤组织（3×3 mm²）平均有29.1个SEs。当移栽到水苔中并在温室中驯化时，几乎所有的SEs都能转化，小植株生长良好，成活率几乎达到100%。
