- 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.
- 著者
- Endang Ayu Windari Mei Ando Yohei Mizoguchi Hiroto Shimada Keima Ohira Yasuaki Kagaya Tetsuya Higashiyama Seiji Takayama Masao Watanabe Keita Suwabe
- 出版者
- Japanese Society for Plant Biotechnology
- 雑誌
- Plant Biotechnology (ISSN:13424580)
- 巻号頁・発行日
- vol.38, no.1, pp.77-87, 2021-03-25 (Released:2021-03-25)
- 参考文献数
- 63
- 被引用文献数
- 18
Pollination is the crucial initial step that brings together the male and female gametophytes, and occurs at the surface of the stigmatic papilla cell in Arabidopsis thaliana. After pollen recognition, pollen hydration is initiated as a second critical step to activate desiccated mature pollen grains for germination, and thus water transport from pistil to pollen is essential for this process. In this study, we report a novel aquaporin-mediated water transport process in the papilla cell as a control mechanism for pollen hydration. Coupled with a time-series imaging analysis of pollination and a reverse genetic analysis using T-DNA insertion Arabidopsis mutants, we found that two aquaporins, the ER-bound SIP1;1 and the plasma membrane-bound PIP1;2, are key players in water transport from papilla cell to pollen during pollination. In wild type plant, hydration speed reached its maximal value within 5 min after pollination, remained high until 10–15 min. In contrast, sip1;1 and pip1;2 mutants showed no rapid increase of hydration speed, but instead a moderate increase during ∼25 min after pollination. Pollen of sip1;1 and pip1;2 mutants had normal viability without any functional defects for pollination, indicating that decelerated pollen hydration is due to a functional defect on the female side in sip1;1 and pip1;2 mutants. In addition, sip1;1 pip1;2 double knockout mutant showed a similar impairment of pollen hydration to individual single mutants, suggesting that their coordinated regulation is critical for proper water transport, in terms of speed and amount, in the pistil to accomplish successful pollen hydration.