Chemical analysis of cell wall regeneration and reversion of protoplasts from Schizophyllum commune
Chemical analysis of cell wall regeneration and reversion of protoplasts from Schizophyllum commune
复制标题
裂褶菌细胞壁再生和原生质体回复的化学分析
DOI：
10.1007/bf00428371
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发表时间：
1975
影响因子：
2.8
通讯作者：
J. Wessels
中科院分区：
文献类型：
--
作者：
O. M. H. Vries;J. Wessels
在MgSO_4作为渗透压稳定剂的存在下，裂褶菌有核原生质体发育成大的液泡，并可根据其低浮力密度进行分离。所有这些原生质体都能够壁再生，并且约50%在液体培养基中恢复到菌丝生长模式。从再生开始研究了三种主要细胞壁组分S-葡聚糖（α-1，3-葡聚糖）、R-葡聚糖（β-1，3，β-1，6-葡聚糖）和几丁质的形成动力学。S-葡聚糖和几丁质的积累以及RNA和蛋白质的合成在短暂的滞后后同时开始，但R-葡聚糖的形成被延迟。只有在R-葡聚糖快速合成几个小时后，才开始回复到菌丝管。环己酰亚胺（0.5 μg/ml）抑制蛋白质合成98%，抑制R-葡聚糖的形成和菌丝生长的逆转，但几丁质和S-葡聚糖的形成确实开始，并持续几个小时，似乎没有受到影响。这表明负责合成S-葡聚糖和几丁质的酶在原生质体制备期间保持完整。多霉素D抑制几丁质和R-葡聚糖的合成，也抑制菌丝生长的逆转。然而，S-葡聚糖的合成没有受到抑制。这些抑制剂研究以及正常再生过程中R-葡聚糖形成的动力学表明，R-葡聚糖的合成是菌丝形态发生起始所必需的。
In the presence of MgSO4 as osmotic stabilizer, nucleated protoplasts of Schizophyllum commune developed a large vacuole and could be isolated on the basis of their low buoyant density. All these protoplasts were capable of wall regeneration and about 50 percent reverted to the hyphal mode of growth in liquid medium. The kinetics of the formation of three main cell-wall components, S-glucan (α-1,3-glucan), R-glucan (β-1,3, β-1,6-glucan) and chitin were studied from the onset of regeneration. S-glucan and chitin accumulation as well as RNA and protein synthesis started simultaneously after a short lag, but R-glucan formation was delayed. The reversion to hyphal tubes only began after several hours of rapid R-glucan synthesis. Cycloheximide (0.5 μg/ml), inhibiting protein synthesis by 98% inhibited the formation of R-glucan and the reversion to hyphal growth but the formation of chitin and S-glucan did start and continued seemingly unimpaired for several hours. This indicates that the enzymes responsible for the synthesis of S-glucan and chitin remained intact during protoplast preparation. Polyoxin D inhibited both the synthesis of chitin and R-glucan and also the reversion to hyphal growth. However, the synthesis of S-glucan was not suppressed. These inhibitor studies as well as the kinetics of R-glucan formation during normal regeneration suggest that the synthesis of R-glucan is required for the initiation of hyphal morphogenesis.