Department of Botany, University of British Columbia, 6270 University Boulevard, Vancouver, British Columbia, V6T-1Z4, Canada.
Mycologia. 2019 Nov-Dec;111(6):904-918. doi: 10.1080/00275514.2019.1669999. Epub 2019 Oct 30.
The advantage of filamentous growth to the fungal lifestyle is so great that it arose multiple times. Most zoosporic fungi from phylum Chytridiomycota exhibit a monocentric thallus form consisting of anucleate filamentous rhizoids that anchor reproductive sporangia to substrata and absorb nutrients. Actin function during polarized growth and cytokinesis is well documented across eukaryotes, but its role in sculpting nonhyphal, nonyeast fungal cells is unknown. We sought to provide a basis for comparing actin organization among major fungal lineages and to investigate the effects of actin disruption on morphogenesis in a monocentric thallus. Using fluorescence microscopy, we observed fixed, rhodamine phalloidin-stained actin in chemically fixed , exemplifying monocentric thallus development within the diverse, zoosporic phylum Chytridiomycota. We also compared rhizoid lengths and rhizoid branching of thalli incubated with the actin inhibitor latrunculin B to determine the effects of actin disruption on morphology. Actin was concentrated at the tips of growing rhizoids. Actin cables typically formed cortical, parallel arrays in hyphae, but in mature sporangia they were concentrated in a funnel-shaped array in the central region. Thalli treated with latrunculin B had shorter rhizoids with fewer branches than controls. In both hyphae and monocentric thalli, actin localization coincides with active, polarized growth and cytokinesis. Specific actin localization patterns are largely shared between monocentric species but differ significantly from patterns observed in hyphae. Actin integrity is critical for sustaining filamentous growth in all fungi.
丝状生长对真菌生活方式的优势如此之大,以至于它多次出现。大多数来自接合菌门的游动孢子真菌表现出单核的营养体形式,由无核丝状根状菌丝组成,根状菌丝将生殖孢子囊锚定在基质上并吸收营养。肌动蛋白在极化生长和胞质分裂过程中的功能在真核生物中得到了很好的证明,但它在塑造非丝状、非酵母真菌细胞中的作用尚不清楚。我们试图为比较主要真菌谱系中的肌动蛋白组织提供基础,并研究肌动蛋白破坏对单核营养体形态发生的影响。我们使用荧光显微镜观察固定的、吖啶橙鬼笔环肽染色的肌动蛋白,以化学固定的方式举例说明在多样化的游动孢子门接合菌中单核营养体的发育。我们还比较了用肌动蛋白抑制剂拉曲布宁 B 孵育的根状菌丝的长度和分支,以确定肌动蛋白破坏对形态的影响。肌动蛋白集中在生长根状菌丝的顶端。肌动蛋白电缆通常在菌丝中形成皮质、平行排列,但在成熟的孢子囊中,它们集中在中央区域的漏斗形排列中。用拉曲布宁 B 处理的营养体的根状菌丝比对照的短,分支也少。在菌丝和单核营养体中,肌动蛋白的定位都与活跃的、极化生长和胞质分裂相一致。单核种之间的特定肌动蛋白定位模式在很大程度上是共享的,但与在菌丝中观察到的模式有很大的不同。肌动蛋白的完整性对于维持所有真菌的丝状生长是至关重要的。