TERI-Deakin Nanobiotechnology Centre, TERI Gram, The Energy and Resources Institute (TERI), Gwal Pahari, Gurugram, Haryana, India.
J Basic Microbiol. 2019 Aug;59(8):767-774. doi: 10.1002/jobm.201900138. Epub 2019 Jun 18.
Detailed information on structural changes that occur during ontogenesis of Rhizophagus irregularis in axenically developed coculture is limited. Our study aims to investigate the series of events that occur during mycorrhizal ontogenesis under axenic condition through basic and advanced microscopic techniques followed by comparison among these to identify the suitable technique for rapid and detailed analysis of mycorrhizal structures. Three stages were identified in mycorrhizal ontogenesis from initiation (preinfection stage of hyphae; its branching, infection and appressoria formation; epidermal opening; and hyphal entry), progression (arbuscular development; hyphal coils and vesicles) to maturity (extraradical spores). Scanning electron microscopy was found to be an efficient tool for studying spatial three-dimensional progression. Adding to the advantages of advanced microscopy, potential of autofluorescence to explore the stages of symbiosis nondestructively was also established. We also report imaging of ultrathin sections by bright field microscopy to provide finer details at subcellular interface. Owing to the merits of nondestructive sampling, ease of sample preparation, autofluorescence (no dye required), no use of toxic chemicals, rapid analysis and in depth characterization confocal laser scanning microscopy was identified as the most preferred technique. The method thus developed can be used for detailed structural inquisition of mycorrhizal symbiosis both in in planta and in an in vitro system.
关于在无菌培养的共培养物中Rhizophagus irregularis 个体发生过程中发生的结构变化的详细信息有限。我们的研究旨在通过基本和先进的显微镜技术来研究在无菌条件下发生的菌根个体发生过程中的一系列事件，然后对这些事件进行比较，以确定适合快速和详细分析菌根结构的技术。在菌根个体发生过程中，从起始阶段（菌丝的侵染前阶段；其分支、侵染和附着胞形成；表皮开口；和菌丝进入），进展阶段（丛枝发育；菌丝线圈和液泡）到成熟阶段（外生孢子），共确定了三个阶段。扫描电子显微镜被发现是研究空间三维进展的有效工具。除了先进显微镜的优势外，还确定了自动荧光在非破坏性探索共生阶段的潜力。我们还报告了通过明场显微镜对超薄切片进行成像，以提供亚细胞界面的更精细细节。由于具有非破坏性采样、易于制备样品、自动荧光（无需染色）、不使用有毒化学物质、快速分析和深入表征的优点，共聚焦激光扫描显微镜被确定为最优选的技术。所开发的方法可用于在植物体内和体外系统中对菌根共生进行详细的结构探究。
