Park S-U, Facchini P J
Department of Biological Sciences, University of Calgary, Calgary, Alberta, T2N 1N4, Canada e-mail:
Plant Cell Rep. 2000 Mar;19(4):421-426. doi: 10.1007/s002990050750.
The development of a rapid protocol for high-efficiency somatic embryogenesis and plant regeneration from seed-derived embryogenic callus cultures of California poppy (Eschscholzia californica Cham.) is reported. The optimized procedure required less than 13 weeks from the initiation of seed cultures to the recovery of plantlets and involved the sequential transfer of cultures onto solid Murashige and Skoog basal medium containing three different combinations of growth regulators. All steps were performed at 25 °C. Friable primary callus was induced from seeds of E. californica cultured on medium supplemented with 1.0 mg l 2,4-dichlorophenoxyacetic acid. The primary callus was transferred to medium containing 1.0 mg l 1-naphthaleneacetic acid and 0.5 mg l 6-benzylaminopurine to establish embryogenic callus and promote somatic embryogenesis. Regenerated plantlets were recovered after the conversion of somatic embryos on medium containing 0.05 mg l 6-benzylaminopurine and showed normal development. Embryogenic callus was induced at a frequency of 85%, an average of 45 somatic embryos were produced per callus, 90% of the somatic embryos converted, and about 70% of the plantlets were recovered in soil. The growth rate of somatic embryo-derived shoots could be increased by gibberellic acid treatment, but the resulting plantlets were hyperhydritic.
报道了一种从加州罂粟（Eschscholzia californica Cham.）种子来源的胚性愈伤组织培养物中高效体细胞胚胎发生和植株再生的快速方案。从种子培养开始到植株再生，优化后的程序所需时间不到13周，包括将培养物依次转移到含有三种不同生长调节剂组合的固体Murashige和Skoog基础培养基上。所有步骤均在25°C下进行。在添加了1.0 mg/L 2,4-二氯苯氧乙酸的培养基上培养加州罂粟种子，诱导出易碎的初级愈伤组织。将初级愈伤组织转移到含有1.0 mg/L萘乙酸和0.5 mg/L 6-苄基腺嘌呤的培养基上，以建立胚性愈伤组织并促进体细胞胚胎发生。在含有0.05 mg/L 6-苄基腺嘌呤的培养基上，体细胞胚转化后获得再生植株，且发育正常。胚性愈伤组织的诱导频率为85%，每个愈伤组织平均产生45个体细胞胚，90%的体细胞胚转化，约70%的植株在土壤中得以再生。赤霉素处理可提高体细胞胚来源芽的生长速率，但所得植株会出现玻璃化现象。
