Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Rozvojová 263, CZ-16502 Prague 6, Czech Republic.
Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Rozvojová 263, CZ-16502 Prague 6, Czech Republic.
Plant Sci. 2014 May;221-222:90-9. doi: 10.1016/j.plantsci.2014.02.006. Epub 2014 Feb 26.
The role of the actin cytoskeleton in somatic embryo development was investigated using latrunculin B and cytochalasin D. Brief treatments (1h) with either drug at the start of maturation fragmented the actin in suspensor cells and/or depolymerized actin filaments in meristematic cells. The drugs targeted different cells: latB primarily affected the suspensor cells, but cchD damaged both suspensor and meristematic cells. Lethal damage to the meristematic and suspensor cells was observed when the drugs were applied throughout the maturation period, although the severity of this effect depended on their concentrations. The drugs' effects on the yield of mature somatic embryos were investigated by applying them to embryo cultures throughout the maturation period or for one week at three different points in the maturation process: immediately prior to the start of maturation, during the first week of maturation, and during the fourth week of maturation. The strongest effects were observed when the drugs were applied at the start of maturation. Under these conditions, latB destroyed the suspensors, eliminating the underdeveloped embryos that depend on them. This accelerated the development of embryos that were capable of separating from the suspensors. Thus, while the total number of embryos at the end of the maturation period was lower than in untreated control cultures, the surviving mature embryos were of high quality. cchD treatment at the start of maturation strongly inhibited embryo development. Drug treatment at the end of the maturation period did not significantly affect embryo development: latB caused no change in the yield of somatic embryos, but cchD treatment increased the number of malformed embryos compared to untreated controls.
使用拉普丁素 B 和细胞松弛素 D 研究了肌动蛋白细胞骨架在体细胞胚胎发育中的作用。在成熟开始时用药物短暂处理 1 小时，会使悬浮细胞中的肌动蛋白断裂，并使分生细胞中的肌动蛋白丝解聚。这些药物针对不同的细胞：拉普丁素 B 主要影响悬浮细胞，但细胞松弛素 D 会损伤悬浮细胞和分生细胞。当药物在整个成熟过程中使用时，会观察到分生细胞和悬浮细胞的致命损伤，尽管这种效应的严重程度取决于其浓度。通过在整个成熟过程中或在成熟过程中的三个不同时间点（即成熟开始前、第一周成熟期间和第四周成熟期间）将药物应用于胚胎培养，研究了药物对成熟体细胞胚胎产量的影响。当药物在成熟开始时应用时，观察到最强的效果。在这些条件下，拉普丁素 B 破坏了悬浮细胞，消除了依赖于悬浮细胞的发育不良的胚胎。这加速了能够从悬浮细胞中分离的胚胎的发育。因此，虽然在成熟期末的胚胎总数低于未处理的对照培养物，但存活的成熟胚胎质量很高。在成熟开始时用细胞松弛素 D 处理强烈抑制胚胎发育。在成熟期末进行药物处理不会显著影响胚胎发育：拉普丁素 B 对体细胞胚胎的产量没有变化，但与未处理的对照相比，细胞松弛素 D 处理增加了畸形胚胎的数量。
