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Cellular organization in germ tube tips of Gigaspora and its phylogenetic implications

机译：长臂猿生殖管尖端的细胞组织及其系统发育意义

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摘要

Comparative morphology of the fine structure of fungal hyphal tips often is phylogenetically informative. In particular, morphology of the Spitzenkorper varies among higher taxa. To date no one has thoroughly characterized the hyphal tips of members of the phylum Glomeromycota to compare them with other fungi. This is partly due to difficulty growing and manipulating living hyphae of these obligate symbionts. We observed growing germ tubes of Gigaspora gigantea, G. margarita and G. rosea with a combination of light microscopy (LM) and transmission electron microscopy (TEM). For TEM, we used both traditional chemical fixation and cryo-fixation methods. Germ tubes of all species were extremely sensitive to manipulation. Healthy germ tubes often showed rapid bidirectional cytoplasmic streaming, whereas germ tubes that had been disturbed showed reduced or no cytoplasmic movement. Actively growing germ tubes contain a cluster of 10-20 spherical bodies approximately 3-8 mu m behind the apex. The bodies, which we hypothesize are lipid bodies, move rapidly in healthy germ tubes. These bodies disappear immediately after any cellular perturbation. Cells prepared with cryo-techniques had superior preservation compared to those that had been processed with traditional chemical protocols. For example, cryo-prepared samples displayed two cell-wall layers, at least three vesicle types near the tip and three distinct cytoplasmic zones were noted. We did not detect a Spitzenkorper with either LM or TEM techniques and the tip organization of Gigaspora germ tubes appeared to be similar to hyphae in zygomycetous fungi. This observation was supported by a phylogenetic analysis of microscopic characters of hyphal tips from members of five fungal phyla. Our work emphasizes the sensitive nature of cellular organization, and the need for as little manipulation as possible to observe germ tube structure accurately.
机译：真菌菌丝尖端细微结构的比较形态学通常在系统发育上提供信息。尤其是Spitzenkorper的形态在较高的分类单元中会有所不同。迄今为止，还没有人对球菌门菌门的菌丝尖端进行充分表征，以将它们与其他真菌进行比较。部分原因是由于这些专性共生体难以生长和操纵菌丝。我们观察到的Gigaspora gigantea，G。margarita和G. rosea的生殖管通过光学显微镜（LM）和透射电子显微镜（TEM）的组合而生长。对于TEM，我们同时使用了传统的化学固定和冷冻固定方法。所有种类的胚芽管对操纵极为敏感。健康的生殖管通常表现出快速的双向细胞质流，而受到干扰的生殖管显示出细胞质运动减少或没有。活跃生长的胚芽管在顶点后约3-8微米处包含10-20个球形体簇。我们假设的身体是脂质体，可以在健康的生殖管中快速移动。这些身体在任何细胞扰动后立即消失。与采用传统化学方法处理过的细胞相比，采用冷冻技术制备的细胞具有更好的保存性。例如，冷冻制备的样品显示出两个细胞壁层，在尖端附近至少有三种囊泡类型，并注意到了三个不同的胞质区。我们没有使用LM或TEM技术检测到Spitzenkorper，并且Gigaspora胚芽管的尖端组织似乎与合子真菌的菌丝相似。该观察得到了来自五个真菌门的菌丝尖端的微观特征的系统发育分析的支持。我们的工作强调细胞组织的敏感性，需要尽可能少的操作以准确地观察胚管结构。

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