Photoautotrophic growth of soybean cells in suspension culture: I. Establishment of photoautotrophic cultures.
Photoautotrophic growth of soybean cells in suspension culture: I. Establishment of photoautotrophic cultures.
复制标题
悬浮培养中大豆细胞的光自养生长：I.光自养培养物的建立。
作者：
Michael E. Horn;J. Sherrard;J. Widholm
从大豆（Glycine max (L.)）愈伤组织中目视筛选出叶绿素含量高的光嗜养愈伤组织。稳定。子叶。从愈伤组织开始的悬浮培养在5% CO(2)（平衡空气）的连续光照下成为光自养的。在2周的传代期内，干重增加了1000 ~ 1400%。细胞内Chl含量为4.4 ~ 5.9微克/毫克干重，约为同等光照（300微爱因斯坦/平方米/秒）下大豆叶片Chl含量的75 ~ 90%。在黑暗中，在缺乏蔗糖的培养基中或在0.5微摩尔的3-（3,4-二氯苯基）-1,1-二甲基脲的存在下，不能观察到生长，这种浓度不会抑制异养生长（在蔗糖上）。光自养生长绝对需要升高的CO(2)浓度（约1%）。在14天的传代期内，生长（鲜重和干重）呈对数关系。光合作用在第4天后迅速增加，达到每小时每毫克Chl吸收83微摩尔CO(2)的峰值，而黑暗呼吸从第2天到第6天减少90%。生长培养基的pH从7.0迅速下降到4.5，在第14天缓慢上升到5.0。在这个pH值范围和光强（200-300微爱因斯坦/平方米每秒）下，虽然在高pH值和光强下记录到了O(2)的演化，但没有检测到O(2)的演化。
Highly chlorophyllous photomixotrophic callus was visually selected from callus originating from soybean (Glycine max (L.) Merr. var. Corsoy) cotyledon. Suspension cultures initiated from this callus became photoautotrophic under continuous light with an atmosphere of 5% CO(2) (balance air). Dry weight increases of 1000 to 1400% in the 2-week subculture period have been observed. The cellular Chl content ranged from 4.4 to 5.9 micrograms per milligram dry weight which is about 75 to 90% of the Chl content in soybean leaves under equivalent illumination (300 micro-Einsteins per square meter per second).No growth can be observed in the dark in sucrose-lacking medium or in the presence of 0.5 micromolar 3-(3,4-dichlorophenyl)-1,1-dimethylurea, a concentration which does not inhibit heterotrophic growth (on sucrose). Photoautotrophic growth has an absolute requirement for elevated CO(2) concentrations (>1%). During the 14-day subculture period, growth (fresh weight and dry weight) is logarithmic. Photosynthesis quickly increases after day 4, reaching a peak of 83 micromoles CO(2) incorporated per milligram Chl per hour while dark respiration decreases 90% from day 2 to day 6. The pH of the growth medium quickly drops from 7.0 to 4.5 before slowly increasing to 5.0 by day 14. At this pH range and light intensity (200-300 microEinsteins per square meter per second), no O(2) evolution could be detected although at high pH and light intensity O(2) evolution was recorded.