- 著者
- Rina Fujihara Naoyuki Uchida Toshiaki Tameshige Nozomi Kawamoto Yugo Hotokezaka Takumi Higaki Rüdiger Simon Keiko U Torii Masao Tasaka Mitsuhiro Aida
- 出版者
- Japanese Society for Plant Biotechnology
- 雑誌
- Plant Biotechnology (ISSN:13424580)
- 巻号頁・発行日
- vol.38, no.3, pp.317-322, 2021-09-25 (Released:2021-09-25)
- 参考文献数
- 27
- 被引用文献数
- 5
The shoot organ boundaries have important roles in plant growth and morphogenesis. It has been reported that a gene encoding a cysteine-rich secreted peptide of the EPIDERMAL PATTERNING FACTOR-LIKE (EPFL) family, EPFL2, is expressed in the boundary domain between the two cotyledon primordia of Arabidopsis thaliana embryo. However, its developmental functions remain unknown. This study aimed to analyze the role of EPFL2 during embryogenesis. We found that cotyledon growth was reduced in its loss-of-function mutants, and this phenotype was associated with the reduction of auxin response peaks at the tips of the primordia. The reduced cotyledon size of the mutant embryo recovered in germinating seedlings, indicating the presence of a factor that acted redundantly with EPFL2 to promote cotyledon growth in late embryogenesis. Our analysis suggests that the boundary domain between the cotyledon primordia acts as a signaling center that organizes auxin response peaks and promotes cotyledon growth.
- 著者
- Kotomi Kikukawa Ryota Sato Masaaki Iwamoto Takumi Higaki
- 出版者
- Japan Mendel Society, International Society of Cytology
- 雑誌
- CYTOLOGIA (ISSN:00114545)
- 巻号頁・発行日
- vol.86, no.3, pp.189-194, 2021-09-25 (Released:2021-09-25)
- 参考文献数
- 26
- 被引用文献数
- 2
Cell segmentation from microscopic images is conventionally used to investigate cell morphology. However, the time expense for manual segmentation becomes extreme with increasing numbers of cells to be analyzed. Recent progress in automated image analysis techniques can facilitate efficient and accurate cell segmentation in wide-range confocal images. Pavement cells, which mainly comprise the epidermal tissue of plant leaves, show jigsaw puzzle-like shapes and provide a model for elucidating the mechanisms underlying the complex morphology of plant cells. This mini-review demonstrates the effectiveness of using a confocal image processing pipeline for morphometric analysis and mechanical simulation using Arabidopsis thaliana cotyledon pavement cells as an example. We examined A. thaliana cotyledon surfaces using wide-range confocal images and used an image processing pipeline in ImageJ software to extract epidermal cell contours. We then used the segmented epidermal cell images to provide examples of how this information can be used for morphometry and mechanical simulation. The use of this high-throughput segmentation method is not limited to plant epidermal tissue and can be applied to various biological materials. Therefore, our approach to microscopic image analysis will hopefully contribute to the advancement of quantitative cell morphology research.
- 著者
- Takumi Higaki Hidenobu Mizuno
- 出版者
- Japanese Society for Plant Biotechnology
- 雑誌
- Plant Biotechnology (ISSN:13424580)
- 巻号頁・発行日
- pp.20.0605a, (Released:2020-09-05)
- 参考文献数
- 33
- 被引用文献数
- 5
In most dicotyledonous plants, leaf pavement cells exhibit complex jigsaw puzzle-like cell morphogenesis during leaf expansion. Although detailed molecular biological information and mathematical modeling of this jigsaw puzzle-like cell morphogenesis are now available, a full understanding of this process remains elusive. Recent reports have highlighted the importance of three-dimensional (3D) structures (i.e., anticlinal and periclinal cell wall) in understanding the mechanical models that describe this morphogenetic process. We believe that it is important to acquire 3D shapes of pavement cells over time, i.e., acquire and analyze four-dimensional (4D) information when studying the relationship between mechanical modeling and simulations and the actual cell shape. In this report, we have developed a framework to capture and analyze 4D morphological information of Arabidopsis thaliana cotyledon pavement cells by using both direct water immersion observations and computational image analyses, including segmentation, surface modeling, virtual reality and morphometry. The 4D cell models allowed us to perform time-lapse 3D morphometrical analysis, providing detailed quantitative information about changes in cell growth rate and shape, with cellular complexity observed to increase during cell growth. The framework should enable analysis of various phenotypes (e.g., mutants) in greater detail, especially in the 3D deformation of the cotyledon surface, and evaluation of theoretical models that describe pavement cell morphogenesis using computational simulations. Additionally, our accurate and high-throughput acquisition of growing cell structures should be suitable for use in generating in silico model cell structures.
- 著者
- Eri Kamon Chihiro Noda Takumi Higaki Taku Demura Misato Ohtani
- 出版者
- Japanese Society for Plant Biotechnology
- 雑誌
- Plant Biotechnology (ISSN:13424580)
- 巻号頁・発行日
- pp.21.0519a, (Released:2021-09-18)
- 参考文献数
- 52
- 被引用文献数
- 4
Secondary cell walls (SCWs) accumulate in specific cell types of vascular plants, notably xylem vessel cells. Previous work has shown that calcium ions (Ca2+) participate in xylem vessel cell differentiation, but whether they function in SCW deposition remains unclear. In this study, we examined the role of Ca2+ in SCW deposition during xylem vessel cell differentiation using Arabidopsis thaliana suspension-cultured cells carrying the VND7-inducible system, in which VND7 activity can be post-translationally upregulated to induce transdifferentiation into protoxylem-type vessel cells. We observed that extracellular Ca2+ concentration was a crucial determinant of differentiation, although it did not have consistent effects on the transcription of VND7-downstream genes as a whole. Increasing the Ca2+ concentration reduced differentiation but the cells could generate the spiral patterning of SCWs. Exposure to a calcium-channel inhibitor partly restored differentiation but resulted in abnormal branched and net-like SCW patterning. These data suggest that Ca2+ signaling participates in xylem vessel cell differentiation via post-transcriptional regulation of VND7-downstream events, such as patterning of SCW deposition.
- 著者
- Masahiro Hirota Shogo Higaki Takumi Higaki Seiichiro Hasezawa
- 出版者
- 日本放射線安全管理学会
- 雑誌
- Radiation Safety Management (ISSN:13471511)
- 巻号頁・発行日
- vol.12, no.2, pp.43-47, 2013 (Released:2013-12-19)
- 参考文献数
- 10
- 被引用文献数
- 2
The contamination of spinach collected immediately following the Fukushima Daiichi nuclear disaster was investigated. The radionuclides deposited in the spinach were 132Te, 131I, 132I, 134Cs, 136Cs and 137Cs. Only 40% of these radionuclides were removed when the spinach was washed with water or detergent. The two outside leaves of each spinach stump were contaminated with the radionuclides, but the three inside leaves were nearly uncontaminated. The most significant contamination was observed on the concavities, creases, veins and leaf or stem injuries of the spinach. Most of the radionuclides deposited on the surface, leading to a radioactivity concentration of the epidermal tissue 9 times that of the mesophyll tissue.
- 著者
- Manami Ichita Takumi Higaki
- 出版者
- Japan Mendel Society, International Society of Cytology
- 雑誌
- CYTOLOGIA (ISSN:00114545)
- 巻号頁・発行日
- vol.88, no.3, pp.169-173, 2023-09-25 (Released:2023-09-25)
- 参考文献数
- 43
The plasma membrane H+-ATPase in plant cells functions as a critical transporter, propelling protons against an electrochemical gradient. This action generates pH and potential differences across the plasma membrane, contributing to various plant physiological processes, including cell growth, nutrient absorption, and stomatal movements. Plant plasma membrane H+-ATPase activity is tightly regulated in response to a multitude of abiotic and biotic environmental cues. Recent studies have shed light on the multi-level regulation of H+-ATPase, extending beyond its proton transport activity and recognizing the importance of its abundance at the plasma membrane, achieved by a balance of endocytosis and exocytosis in response to environmental factors. This review provides an overview of the cell biological regulation of plasma membrane H+-ATPase by balancing its exocytosis and endocytosis. As key factors in regulation of the abundance of the plasma membrane H+-ATPase at the plasma membrane, the membrane trafficking proteins PATROL1 and SYP132 have been identified. The intracellular dynamics and functions are suggested to be tightly associated with the environmental response of plant cells. Despite substantial progress in deciphering the intracellular traffic regulation of the plasma membrane H+-ATPase, a comprehensive molecular understanding remains elusive. Elucidating the molecular pathways linking environmental factors with the regulation of membrane trafficking is pivotal for comprehending the cell biological regulation of environmental responses in plants.
- 著者
- Yuuki Sakai Takumi Higaki Kimitsune Ishizaki Ryuichi Nishihama Takayuki Kohchi Seiichiro Hasezawa
- 出版者
- Japanese Society for Plant Biotechnology
- 雑誌
- Plant Biotechnology (ISSN:13424580)
- 巻号頁・発行日
- vol.39, no.1, pp.5-12, 2022-03-25 (Released:2022-03-25)
- 参考文献数
- 32
- 被引用文献数
- 1 2
The development of the plant body starts with spore germination in bryophytes. In many cases, the first division of the spore occurs after germination and cell elongation of the spore. In Marchantia polymorpha, asymmetric division occurs upon spore germination to generate two daughter cells: the larger one retains the ability to divide and develops into the thallus via sporeling or protonema, while the smaller one maintains tip growth and differentiates into the first rhizoid, providing a scaffold for initial development. Although spore germination of M. polymorpha was described in the 19th century, the intracellular processes of the first asymmetric division of the spore have not been well characterized. In this study, we used live-cell imaging analyses to elucidate microtubule dynamics during the first asymmetric division concomitantly with germination. In particular, we demonstrated that the preprophase band was not formed in the spore and that the bipolar prospindle, which is a microtubule structure surrounding the nucleus during prophase, migrated from the center to the periphery in the spore, suggesting that it was the earliest visible sign of cell polarity. We also showed that the occurrence of asymmetric division depended on actin filaments. Our findings regarding the first division of the spore in M. polymorpha will lead to a better model for cell-autonomous asymmetric division in plants.
- 著者
- Eri Kamon Chihiro Noda Takumi Higaki Taku Demura Misato Ohtani
- 出版者
- Japanese Society for Plant Biotechnology
- 雑誌
- Plant Biotechnology (ISSN:13424580)
- 巻号頁・発行日
- vol.38, no.3, pp.331-337, 2021-09-25 (Released:2021-09-25)
- 参考文献数
- 52
- 被引用文献数
- 4
Secondary cell walls (SCWs) accumulate in specific cell types of vascular plants, notably xylem vessel cells. Previous work has shown that calcium ions (Ca2+) participate in xylem vessel cell differentiation, but whether they function in SCW deposition remains unclear. In this study, we examined the role of Ca2+ in SCW deposition during xylem vessel cell differentiation using Arabidopsis thaliana suspension-cultured cells carrying the VND7-inducible system, in which VND7 activity can be post-translationally upregulated to induce transdifferentiation into protoxylem-type vessel cells. We observed that extracellular Ca2+ concentration was a crucial determinant of differentiation, although it did not have consistent effects on the transcription of VND7-downstream genes as a whole. Increasing the Ca2+ concentration reduced differentiation but the cells could generate the spiral patterning of SCWs. Exposure to a calcium-channel inhibitor partly restored differentiation but resulted in abnormal branched and net-like SCW patterning. These data suggest that Ca2+ signaling participates in xylem vessel cell differentiation via post-transcriptional regulation of VND7-downstream events, such as patterning of SCW deposition.
- 著者
- Takumi Higaki Fumiya Sato Koh Iba
- 出版者
- Japan Mendel Society, International Society of Cytology
- 雑誌
- CYTOLOGIA (ISSN:00114545)
- 巻号頁・発行日
- vol.86, no.2, pp.101-102, 2021-06-25 (Released:2021-06-25)
- 参考文献数
- 4
- 被引用文献数
- 2
- 著者
- Yumi Hirakawa Seiichiro Hasezawa Takumi Higaki
- 出版者
- Japan Mendel Society, International Society of Cytology
- 雑誌
- CYTOLOGIA (ISSN:00114545)
- 巻号頁・発行日
- vol.83, no.3, pp.289-293, 2018-09-25 (Released:2018-10-12)
- 参考文献数
- 29
- 被引用文献数
- 1
We previously established an experimental system for efficient pathogenic signal induced-cell death using tobacco BY-2 cells and culture filtrate of a plant pathogenic bacterium, Erwinia carotovora. Using this experimental system, cytoskeletal and vacuolar changes during the process of filtrate-induced cell death were characterized in detail. However, the initial events induced by the filtrate were largely unknown. In this study, reactive oxygen species (ROS) production and endocytosis stimulation were examined in BY-2 cells treated with the filtrate of E. carotovora. Transient ROS production within 2 min was observed after the filtrate treatment. Treatment with the Ca2+ chelator BAPTA or the protein kinase inhibitor K252a significantly inhibited ROS production in a dose-dependent manner, suggesting that ROS production depends on Ca2+ influx or protein phosphorylation. In addition, internalization of the endocytic marker FM4-64 was promoted by filtrate treatment within 30 min, suggesting that the filtrate treatment stimulated endocytosis. These results showed that ROS production and subsequent endocytosis stimulation are the initial events induced by culture filtrate of E. carotovora in BY-2 cells.
- 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
- Rina Fujihara Naoyuki Uchida Toshiaki Tameshige Nozomi Kawamoto Yugo Hotokezaka Takumi Higaki Rüdiger Simon Keiko U Torii Masao Tasaka Mitsuhiro Aida
- 出版者
- Japanese Society for Plant Biotechnology
- 雑誌
- Plant Biotechnology (ISSN:13424580)
- 巻号頁・発行日
- pp.21.0508a, (Released:2021-09-18)
- 参考文献数
- 27
- 被引用文献数
- 5
The shoot organ boundaries have important roles in plant growth and morphogenesis. It has been reported that a gene encoding a cysteine-rich secreted peptide of the EPIDERMAL PATTERNING FACTOR-LIKE (EPFL) family, EPFL2, is expressed in the boundary domain between the two cotyledon primordia of Arabidopsis thaliana embryo. However, its developmental functions remain unknown. This study aimed to analyze the role of EPFL2 during embryogenesis. We found that cotyledon growth was reduced in its loss-of-function mutants, and this phenotype was associated with the reduction of auxin response peaks at the tips of the primordia. The reduced cotyledon size of the mutant embryo recovered in germinating seedlings, indicating the presence of a factor that acted redundantly with EPFL2 to promote cotyledon growth in late embryogenesis. Our analysis suggests that the boundary domain between the cotyledon primordia acts as a signaling center that organizes auxin response peaks and promotes cotyledon growth.
- 著者
- Fumiya Sato Koh Iba Takumi Higaki
- 出版者
- Japan Mendel Society, International Society of Cytology
- 雑誌
- CYTOLOGIA (ISSN:00114545)
- 巻号頁・発行日
- vol.86, no.2, pp.119-126, 2021-06-25 (Released:2021-06-25)
- 参考文献数
- 16
- 被引用文献数
- 4
The Arabidopsis thaliana stomatal complex contains a pair of guard cells surrounded by subsidiary cells, which assist in turgor-driven stomatal movement and receive water and ions. This transport, driven by environmental signals, involves a translocation factor of the plasma membrane proton pump H+-ATPase AHA1, PATROL1. In this study, we investigated the responses of PATROL1 to salinity and hyperosmotic stresses. Specifically, we analyzed the effects of 125 mM NaCl or 231 mM mannitol on the cotyledon pavement cell cortexes in transgenic A. thaliana seedlings expressing green fluorescent protein (GFP)-tagged PATROL1. Cells treated with NaCl had few GFP-PATROL1-labeled dot-like structures but contained unusual labeled large bodies and rod-like structures. Cells treated with mannitol had similar large bodies, but not rods, indicating that the rod-like structures form specifically under salinity stress conditions. Dual observations of GFP-PATROL1 and red fluorescent protein (RFP)-tagged AHA1 in stress-treated cells revealed that the latter did not accumulate in the stress-induced GFP-PATROL1 structures, suggesting that the stress-induced GFP-PATROL1 structures are not involved in RFP-AHA1 localization. Additionally, the primary root growth of the patrol1 mutant was more sensitive to NaCl treatment than was that of wild type. Thus, PATROL1 appears to contribute to salinity stress tolerance, possibly by regulating membrane trafficking.
- 著者
- Takumi Higaki Hidenobu Mizuno
- 出版者
- Japanese Society for Plant Biotechnology
- 雑誌
- Plant Biotechnology (ISSN:13424580)
- 巻号頁・発行日
- vol.37, no.4, pp.429-435, 2020-12-25 (Released:2020-12-25)
- 参考文献数
- 33
- 被引用文献数
- 1 5
In most dicotyledonous plants, leaf pavement cells exhibit complex jigsaw puzzle-like cell morphogenesis during leaf expansion. Although detailed molecular biological information and mathematical modeling of this jigsaw puzzle-like cell morphogenesis are now available, a full understanding of this process remains elusive. Recent reports have highlighted the importance of three-dimensional (3D) structures (i.e., anticlinal and periclinal cell wall) in understanding the mechanical models that describe this morphogenetic process. We believe that it is important to acquire 3D shapes of pavement cells over time, i.e., acquire and analyze four-dimensional (4D) information when studying the relationship between mechanical modeling and simulations and the actual cell shape. In this report, we have developed a framework to capture and analyze 4D morphological information of Arabidopsis thaliana cotyledon pavement cells by using both direct water immersion observations and computational image analyses, including segmentation, surface modeling, virtual reality and morphometry. The 4D cell models allowed us to perform time-lapse 3D morphometrical analysis, providing detailed quantitative information about changes in cell growth rate and shape, with cellular complexity observed to increase during cell growth. The framework should enable analysis of various phenotypes (e.g., mutants) in greater detail, especially in the 3D deformation of the cotyledon surface, and evaluation of theoretical models that describe pavement cell morphogenesis using computational simulations. Additionally, our accurate and high-throughput acquisition of growing cell structures should be suitable for use in generating in silico model cell structures.
- 著者
- Takumi Higaki Aya Kato Fumiyoshi Myouga Natsumaro Kutsuna Seiichiro Hasezawa Noriko Nagata
- 出版者
- 日本バイオイメージング学会
- 雑誌
- bioimages (ISSN:09192719)
- 巻号頁・発行日
- vol.22, pp.1-7, 2014 (Released:2015-06-30)
- 参考文献数
- 12
Recent advances in bioimaging equipment have allowed for the significant and rapid acquisition of biological images. Consequently, new methods are energetically being developed to quantitatively and objectively evaluate the biological features of many bioimages. Recently, we described the application of the CARTA (Clustering-Aided Rapid Training Agent) image classification framework, which can be used to obtain highly accurate classifications of a wide variety of biomedical images. However, the efficacy of this technology in association with transmission electron microscopy (TEM) images has not been evaluated. We herein report the automatic classification of chloroplast TEM images of chloroplasts of wild-type Arabidopsis thaliana and four kinds of mutants impaired in chloroplast-targeted proteins. CARTA successfully classified the TEM images based on their genetic backgrounds. As previously reported with fluorescent microscopic and magnetic resonance images, CARTA reduced the annotation cost while maintaining accuracy. Furthermore, using images obtained with different accelerating voltages resulted in biologically meaningless classifications, suggesting that TEM settings are critical for chloroplast morphological classification. CARTA is expected to be useful for future computational inspections of the ultrastructure visualized with TEM.