United AgriSeeds, Inc., P.O. Box 4011, 61820, Champaign, IL, USA.
Plant Cell Rep. 1989 Mar;7(8):673-6. doi: 10.1007/BF00272058.
Anthers from a highly androgenic genotype of maize (139/39-02), when cultured in a modified, liquid YP medium, dehisced within 2-7 days resulting in a stationary suspension of microspores. After 12-15 days, the microspore suspension was found to contain multicellular masses which went on to produce macroscopic embryo-like structures within 20-25 days of culture initiation. Embryogenic callus could be obtained by transferring microspore-derived embryos onto a modified N6 medium supplemented with 2.5 mg/l dicamba and 0.1 mg/l 2,4-D. Subculture onto hormone-free medium resulted in plant regeneration. Over 400 embryo-like structures per 100 anthers cultured have been obtained from liquid induction medium as compared to 55 embryos per 100 anthers cultured on an agar-solidified medium. Approximately 5-25% of these embryo-like structures went on to produce callus from which plants could be recovered. Mechanical isolation of microspores from anthers precultured for 0, 3, and 7 days also resulted in embryo production and plant regeneration. This represents the first report of plant recovery from isolated maize microspores. The use of a liquid induction medium applied to a highly androgenic genotype allows for the production of large numbers of microspore-derived plants and provides a single, haploid cell regeneration system for maize.
来自玉米高度雄性基因型（139/39-02）的花粉，在改良的液体 YP 培养基中培养时，在 2-7 天内开裂，导致微孢子的固定悬浮液。12-15 天后，发现微孢子悬浮液中含有多细胞团块，这些团块在培养起始后 20-25 天内继续产生宏观胚胎样结构。通过将微孢子衍生的胚胎转移到添加了 2.5 mg/l 二甲苯脒和 0.1 mg/l 2,4-D 的改良 N6 培养基上，可以获得胚性愈伤组织。继代到不含激素的培养基上导致植物再生。与在琼脂固体培养基上培养的每 100 个花粉中获得 55 个胚胎相比，从液体诱导培养基中培养的每 100 个花粉中获得了超过 400 个胚胎样结构。这些胚胎样结构中约有 5-25%继续产生愈伤组织，从中可以回收植物。对预培养 0、3 和 7 天的花粉进行机械分离也导致胚胎产生和植物再生。这代表了从分离的玉米小孢子中回收植物的首次报道。液体诱导培养基的使用应用于高度雄性基因型，可以产生大量的小孢子衍生植物，并为玉米提供了一个单一的、单倍体细胞再生系统。
