Comparative immunocytochemistry of the cytoskeleton in filamentous fungi with dikaryotic and multinucieate hyphae
Comparative immunocytochemistry of the cytoskeleton in filamentous fungi with dikaryotic and multinucieate hyphae
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双核和多核菌丝丝状真菌细胞骨架的比较免疫细胞化学
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发表时间：
1989
期刊：
影响因子：
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通讯作者：
M. Raudaskoski
中科院分区：
文献类型：
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作者：
V. Salo;S. Niini;I. Virtanen;M. Raudaskoski
对代表腐生和菌根物种的五种丝状真菌、一种子囊菌、Gyromitra esculents 和四种同担子菌、Amanita regalis、Heterobasidion annosum、Paxillus involutus 和 Suillus bovinus 的微管进行间接免疫荧光 (HF) 显微镜检查，表明微管细胞骨架的总体结构是可比的，尽管它们是多核和多核的。双核细胞条件。在每个物种的间期顶端细胞中，纵向微管轨迹从横壁延伸到顶端，并且在菌丝的顶端部分出现严格的纵向取向。免疫印迹表明，双核物种 A. regalis、P. involutus 和 S. bovinus 的 α- 和 β-微管蛋白迁移率是相同的，而同担子菌 H. annosum 和子囊菌 G. esculenta 的微管蛋白迁移率模式均具有多核菌丝，彼此之间以及与双核物种的微管蛋白迁移率略有不同。使用超滤显微镜可以检查真菌菌丝长顶端细胞中细胞质微管和纺锤体形成之间的关系。在核分裂过程中，细胞质微管在分裂核/核周围延伸 40-60 μm 的区域中分解。在含有靠近顶端的分裂核的多核菌丝和具有短顶端细胞的双核菌丝中，细胞质微管的解体延伸到顶端，当考虑顶端细胞中细胞质微管的功能时，这可能是一个重要的观察结果。细胞质微管的分解首先与纺锤体极体（SPB）尺寸的增加有关，然后与纺锤体从SPB的发育有关。这表明 SPB 在通过细胞质微管分解形成的微管蛋白亚基池中形成纺锤体微管的核心作用中发挥着核心作用。在 A. regalis 的大核和 H. annosum 的顶端细胞中，可以清楚地看到核分裂过程中 SPB 结构的变化，这些细胞含有一排非同步分裂的核，每个核都有一个处于不同阶段的 SPB。在同担子菌的双核和多核菌丝中，纺锤体的发育和结构相似，而在子囊菌 G. esculenta 中记录了某些异常特征。讨论了后期观察到的纺锤体紧密定向对于维持同担子菌菌丝细胞中异核核条件的重要性。在菌丝顶端区域以及隔膜的发育阶段检测到肌动蛋白，并首次通过免疫印迹分析在同担子菌中鉴定出肌动蛋白。与快速生长的菌种相比，缓慢生长的 P. involutus 和 S. bovinus 的菌丝尖端被 NBD-phallacidin 强烈染色。通过单克隆抗肌动蛋白抗体检测所有物种中的肌动蛋白，表明缓慢生长的菌丝顶端丝状肌动蛋白的稳定性高于快速生长的菌丝。
Indirect immunofluorescence (HF) microscopy of microtubules of five filamentous fungi, one ascomycete, Gyromitra esculents, and four homobasidiomycetes, Amanita regalis, Heterobasidion annosum, Paxillus involutus and Suillus bovinus , representing saprophytic and mycorrhizal species, indicated that the gross structure of the microtubule cytoskeleton was comparable in spite of the multinucleate and dikaryotic cell conditions. In the apical cells of each species at interphase, longitudinally oriented microtubule tracks extended from the cross wall to the apex with strictly longitudinal orientation occurring in the apical part of the hypha. Immunoblots indicated that α- and β-tubulin mobilities of the dikaryotic species A. regalis, P. involutus and S. bovinus were identical, while the tubulin mobility patterns of the homobasidiomycete H. annosum and the ascomycete G. esculenta , both with multinucieate hyphae, differed slightly from each other and from those of the dikaryotic species. The use of UF microscopy made it possible to examine the relationship between the cytoplasmic microtubules and spindle formation in the long apical cells of the fungal hyphae. During nuclear division cytoplasmic microtubules disassembled in a region extending 40–60 μm around the dividing nucleus/nuclei. In multinucieate hyphae containing dividing nuclei close to the apex and in dikaryotic hyphae with short apical cells, disassembly of the cytoplasmic microtubules extended up to the apex, which may be an important observation when the function of cytoplasmic microtubules in the apical cells is considered. The disassembly of the cytoplasmic microtubules was first associated with an increase in the size of the spindle pole body (SPB), and then with the development of the spindle from the SPB. This suggested that the SPB had a central role in nucleating spindle microtubules from the pool of tubulin subunits built up through the disassembly of the cytoplasmic microtubules. The changes in structure of the SPB during nuclear division were clearly seen in the large nuclei of A. regalis and in the apical cells of H. annosum containing a row of non-synchronously dividing nuclei, each with an SPB at a different phase. In the dikaryotic and multinucieate hyphae of homobasidiomycetes the development and structure of the spindle was similar, while in the ascomycete G. esculenta certain deviant features were recorded. The significance of the close orientation of the spindles, observed at anaphase, for the maintenance of the heterokaryotic nuclear condition in the hyphal cells of the homobasidiomycetes is discussed. Actin was detected in the apical region of the hyphae as well as at the developmental phase of the septum and was identified by immunoblot analyses for the first time in the homobasidiomycetes. The tips of the slow-growing hyphae of P. involutus and S. bovinus stained intensely with NBD-phallacidin as compared to the weak reaction seen in the fast-growing species. The detection of actin in all the species by a monoclonal anti-actin antibody, suggested that the stability of filamentous actin was greater at the hyphal apex in slowgrowing than in fast-growing hyphae.