Department of Biology, University of Saskatchewan, S7N OWO, Saskatoon, Sask, Canada.
Plant Cell Rep. 1994 Aug;13(11):601-6. doi: 10.1007/BF00232929.
This report describes a low-cost method for generating large numbers of high quality mature white spruce (Picea glauca [Moench.] Voss) somatic embryos which survived desiccation and grew to plantlets more vigorously than excised zygotic embryos cultured in vitro. Somatic embryos from suspension culture were supported within a culture chamber on a flat absorbent pad above the surface of a liquid culture medium containing 20-50 μM abscisic acid and 7.5 % polyethylene glycol. Throughout a 7 week culture period 3 L of fresh medium was pumped into one end of the chamber, while the spent medium exited by gravity from the opposite end. Over 6,300 cotyledonary stage white spruce somatic embryos were recovered after this time from a single culture chamber without manual manipulation. The somatic embryos were of excellent appearance with well developed cotyledons, and possessed high levels of storage lipids. They survived drying to about 8 % moisture content following treatment for 4 weeks at 63 % relative humidity, and following imbibition converted to normal plantlets at a frequency of 92 %, compared to 80 % for embryos grown in Petri dishes. Somatic embryos cultured within the bioreactor developed to plantlets that were 20 % longer than zygotic embryos excised from mature seed and grown in vitro, and were 38 % longer than somatic embryos cultured upon agar medium in Petri dishes.
本报告描述了一种低成本方法，可以大量生成高质量的成熟白云杉（Picea glauca [Moench.] Voss）体细胞胚，这些胚在干燥后存活下来，并比在体外培养的离体合子胚更旺盛地生长为小植株。悬浮培养的体细胞胚在培养室内的一个平面吸收垫上得到支撑，该吸收垫位于含有 20-50 μM 脱落酸和 7.5% 聚乙二醇的液体培养基表面上方。在 7 周的培养期间，新鲜培养基通过泵从一端输入到培养室内，而用过的培养基则通过重力从另一端流出。经过这段时间，从单个培养室内无需手动操作即可回收超过 6300 个子叶期白云杉体细胞胚。这些体细胞胚外观良好，子叶发育良好，并且具有高水平的储存脂质。经过 4 周在 63%相对湿度下处理后，它们可以干燥至约 8%的含水量而存活，并且经过吸胀后，转化为正常小植株的频率为 92%，而在 Petri 盘中生长的胚为 80%。在生物反应器中培养的体细胞胚发育成的小植株比从成熟种子中取出并在体外培养的合子胚长 20%，比在 Petri 盘中琼脂培养基上培养的体细胞胚长 38%。
