著者
Mina Ohtsu Daisuke Kurihara Yoshikatsu Sato Takuya Suzaki Masayoshi Kawaguchi Daisuke Maruyama Tetsuya Higashiyama
出版者
日本メンデル協会、国際細胞学会
雑誌
CYTOLOGIA (ISSN:00114545)
巻号頁・発行日
vol.82, no.3, pp.251-259, 2017-06-25 (Released:2017-09-06)
参考文献数
35

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.
著者
Junko Hasegawa Takumi Higaki Yuki Hamamura Daisuke Kurihara Natsumaro Kutsuna Tetsuya Higashiyama Seiichiro Hasezawa Sachihiro Matsunaga
出版者
日本メンデル協会、国際細胞学会
雑誌
CYTOLOGIA (ISSN:00114545)
巻号頁・発行日
vol.79, no.4, pp.467-474, 2014-12-25 (Released:2015-01-16)
参考文献数
31
被引用文献数
1 7

Vacuoles occupy 80–90% of a mature plant cell and mainly contribute to all types of cell expansion. Zeocin, an inducer of DNA double-strand breaks, causes cell expansion with endoreduplication. The vacuolar structure after zeocin treatment was examined in tobacco (Nicotiana tabacum) cultured cells expressing GFP fused to a vacuole membrane protein. We found that the genotoxic stress induced the cell expansion with subdivision of the vacuolar lumen by cytoplasmic strands. When a femtosecond laser was used to cut off the cytoplasmic strand, mitochondrial transport along the strand stopped. This suggested that in the elongated cells under the genotoxic stress, the transport of subcellular materials was activated for DNA repair within the damaged cell nucleus by the construction of a network of cytoplasmic strands in the vacuolar lumen.