Fluorescent Labeling of the Cyst Nematode <i>Heterodera glycines</i> in Deep-Tissue Live Imaging

  • Ohtsu Mina
    Graduate School of Science, Nagoya University
  • Kurihara Daisuke
    Graduate School of Science, Nagoya University JST ERATO Higashiyama Live-Holonics Project, Nagoya University
  • Sato Yoshikatsu
    Institute of Transformative Bio-Molecules (ITbM), Nagoya University
  • Suzaki Takuya
    Graduate School of Life and Environmental Sciences, University of Tsukuba
  • Kawaguchi Masayoshi
    Division of Symbiotic Systems, National Institute for Basic Biology (NIBB)
  • Maruyama Daisuke
    Kihara Institute for Biological Research, Yokohama City University
  • Higashiyama Tetsuya
    Graduate School of Science, Nagoya University JST ERATO Higashiyama Live-Holonics Project, Nagoya University Institute of Transformative Bio-Molecules (ITbM), Nagoya University

Description

<p>Nematode infection of plant roots is a paradigm of host–parasite interactions. Although nematodes can be labeled with fluorescent dyes, migration of the worms into the deep regions of host roots makes them difficult to track. Here we report the use of two fluorescent dyes, FM4-64 and SYBR green I, to intensely label the soybean cyst nematode (SCN) Heterodera glycines for one week in host plants. Continuous monitoring of the labeled SCN juveniles was achieved with two-photon microscopy. Additionally, we developed a transient transformation system consisting of the non-model leguminous plant (fabaceous) roots, Astragalus sinicus and Agrobacterium rhizogenes to observe the cellular structures of the plant during SCN infection. By the combined use of fluorescent dyes and two-photon microscopy, clear images of infecting SCNs were obtained even in deep regions of A. sinicus roots. The fluorescent labeling described herein can also be used in detailed monitoring of the infection processes of other non-model nematodes, as well as the associated morphological changes in the host plant roots.</p>

Journal

  • CYTOLOGIA

    CYTOLOGIA 82 (3), 251-259, 2017

    Japan Mendel Society, International Society of Cytology

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