Plant Biotechnology Institute, National Research Council of Canada, S7N OW9, Saskatoon, Saskatchewan, Canda.
Plant Cell Rep. 1991 Mar;9(11):595-601. doi: 10.1007/BF00231796.
With barley a large variation in frequency of plant formation from microspores of spikes from the same plant has been observed. The highest frequency of plant formation was obtained when culturing anthers in the dark on a high Ficoll medium containing 2,4-D and kinetin to induce proembryo (or callus) formation. Subsequently the proembryos or calli were cultured in dim light on a high Ficoll-high sugar medium containing IBA and kinetin. Finally the embryos were transferred to a starch agar medium. A maximum of 13 green plants were obtained from microspores of a single anther.The ratios of green to albino microspore derived plants varied from 9∶1 to 1∶9 depending on culture conditions. Under anaerobic conditions, lactic acid and other organic acids may have damaged the organelles in the cells resulting in the formation of albino plants. Thus, direct embryogenesis by using a well-buffered, high Ficoll-high sugar medium and proper aeration are essential for obtaining high frequency of green plants from microspores.
从小麦花粉粒中观察到同一植株小穗的小孢子形成植株的频率有很大的变化。在含有 2,4-D 和激动素的高浓度 Ficoll 培养基上暗培养花药，以诱导原胚（或愈伤组织）形成时，可获得最高的植株形成频率。随后，将原胚或愈伤组织在含有 IBA 和激动素的高浓度 Ficoll-高糖培养基中在弱光下培养。最后，将胚转移到淀粉琼脂培养基上。从单个花药的花粉粒中最多可获得 13 株绿色植株。绿色和白化小孢子衍生植株的比例因培养条件而异，从 9∶1 到 1∶9 不等。在厌氧条件下，乳酸和其他有机酸可能会破坏细胞中的细胞器，导致白化植物的形成。因此，使用缓冲良好、高浓度 Ficoll-高糖培养基和适当通气的直接胚胎发生对于从小孢子中获得高频率的绿色植株是必不可少的。
