摘要
用B_5培养基酶解分离出百合花粉原生质体。原生质体经松胞素(5μg/ml)分别处理5、10、15、30、60 min,再用荧光标记的鬼笔碱染色,共焦激光扫描镜观察,跟踪了原生质体内的肌动蛋白微丝从一个组织复杂严密的网络转变为无数颗粒体的过程。松胞素处理过的原生质体移回至不含松胞素的培养基中后继续培养15 min,肌动蛋白颗粒快速地再延伸出微丝,重组成新的网络。存在于花粉原生质体中生殖细胞的微丝网络,在经松胞素处理后同样都形成为颗粒体。之后,如果原生质体再放入不含松胞素的培养基内继续培养,生殖细胞内的颗粒体同样会再延伸出微丝,重新组成网络。从原生质体胞质以及生殖细胞内所见到的微丝和颗粒相互转化的情况,可以断定,颗粒体不但具有凝聚微丝的功能,同时也具有重组微丝的功能。
Protoplasts were isolated from the mature pollen grains of Lily using cellulase 'Onozuka' R-10 and pectinase in B_5 medium. After isolation the protoplasts were treated with 5μg/ml cytochalasin B (CB) for 5, 10, 15, 30, and 60 min. The effects of the drug on the changes of the actin filamentous arrays in the pollen pro toplasts were monitored with TRITC-phalloidin staining without fixation. After the drug treatment, the filamentous arrays rapidly changed into granules. The sequence of events leading to the formation of the granules were as follows: Some of the filaments changed immediately into granules without apparently going through a transitional stage. Some, however, changed into thick, barbed wire-like bundles (as a result of coalescing of filaments). After about 10 min of CB treatment, the barbed wire-like bundles changed into short rods. After about 15 min of CB treatment all the short rods changed into either spherical granules or granules with a short rod attached. Upon the removal of the drug, the filamentous arrays rapidly returned and within about 15 min a well organized network re-appeared. The pattern of change of the actin filamentous arrays in the generative cells was quite similar to that seen in the vegetative cell cytoplasm. But the changes, from filaments to granules and then back to filaments, in the generative cell were extremely fast. Consequently, no transitional stages were observed. The granules formed after CB treatment served as actin nucleating centres and they did not undergo any further changes even after prolonged exposure to CB (up to 60 mins). Furthermore, the fact that new filaments could reform from these granules after the removal of the drug indicated that they might also play an active role in actin filament organization and/or initiation.
出处
《实验生物学报》
CSCD
1992年第4期377-389,共13页
Acta Biologiae Experimentalis Sinica
关键词
松胞素
百合
原生质体
微丝
Cytochalasin, Actin. Filaments. Pollen protoplasts. Lily.
