Corredoira E, Ballester A, Vieitez A M
Instituto de Investigaciones Agrobiológicas de Galicia, CSIC, Apartado 122, 15080 Santiago de Compostela, Spain.
Ann Bot. 2003 Jul;92(1):129-36. doi: 10.1093/aob/mcg107. Epub 2003 May 21.
Experiments were performed to determine the influence of proliferation medium on the maintenance of embryogenic competence and on repetitive embryogenesis in Castanea sativa Mill. somatic embryos derived from leaf explants. Somatic embryo proliferation was carried out by both direct secondary embryogenesis and by the culture of nodular callus tissue originated from cotyledons of somatic embryos. Both systems led to the production of cotyledonary somatic embryos on Murashige and Skoog proliferation medium supplemented with 0.1 mg l-1 benzyladenine and 0.1 mg l-1 naphthaleneacetic acid. Carbon source and concentration had a marked influence on maturation and subsequent germination ability of chestnut somatic embryos. Plantlet conversion was achieved in embryos matured on media with 6 % sucrose, and on 3 or 6 % maltose, whereas mean shoot length, root length and leaf number of produced plants were not significantly affected by these maturation media. Overall, the best results were obtained with 3 % maltose-matured somatic embryos, giving rise to 6 % plant recovery in addition to 33 % of embryos exhibiting only shoot development. The application of a 2-month cold treatment at 4 degrees C to somatic embryos matured on medium with 3 % maltose was necessary for achieving plant conversion, while partial desiccation did not appear to influence this response. A total of 39 % of embryos eventually produced plants either through conversion to plantlets or indirectly through rooting of shoots. Shoots formed by somatic embryos could be excised, multiplied and rooted following the micropropagation procedures previously developed for chestnut.
进行了实验，以确定增殖培养基对欧洲栗体细胞胚胎胚性能力维持及重复胚胎发生的影响，这些体细胞胚胎源自叶片外植体。体细胞胚胎增殖通过直接次生胚胎发生以及源自体细胞胚胎子叶的结节状愈伤组织培养来进行。在添加了0.1 mg l-1苄基腺嘌呤和0.1 mg l-1萘乙酸的Murashige和Skoog增殖培养基上，这两种体系均能产生子叶体细胞胚胎。碳源和浓度对板栗体细胞胚胎的成熟及后续萌发能力有显著影响。在含有6%蔗糖、3%或6%麦芽糖的培养基上成熟的胚胎能够实现植株转化，然而这些成熟培养基对所产生植株的平均茎长、根长和叶片数量并无显著影响。总体而言，使用3%麦芽糖成熟的体细胞胚胎效果最佳，除了33%的胚胎仅表现出茎发育外，还能实现6%的植株再生。对在含有3%麦芽糖的培养基上成熟的体细胞胚胎进行2个月4℃的冷处理是实现植株转化所必需的，而部分干燥似乎并未影响这一反应。共有39%的胚胎最终通过转化为植株或间接通过茎生根产生了植株。体细胞胚胎形成的茎可以按照先前为板栗开发的微繁殖程序进行切割、增殖和生根。
