Biology subjects

Karahara, I.

Publications and source records attributed to Karahara, I..

6 recordsLinked to original sources

bioRxiv2026

Effects of microgravity on the three-dimensional morphology of rhizoids in Physcomitrium patens

Rooting systems of plants perceive environmental stimuli and flexibly regulate their growth. Therefore, understanding stimulus perception and response mechanisms is essential for optimizing cultivation. During the transition from aquatic to terrestrial environments, land plants have acquired mechanisms to adapt to gravitational force on land. Thus, elucidating gravity responses of rhizoids in bryophytes, early diverging land plants, provides important insights into how gravity-response mechanisms were established during land plant evolution. Analyzing rhizoid morphology under microgravity, where gravitational effects are largely eliminated, provides an effective approach to examine the gravity-response mechanisms that evolved after terrestrialization. In this study, to elucidate microgravity effects on rhizoid growth of Physcomitrium patens, we analyzed 3D datasets obtained by refraction-contrast micro-CT using synchrotron radiation after fixation and embedding of samples from the Space Moss experiment conducted on the International Space Station. Because each CT volume contains numerous rhizoids, we optimized a WEKA-based machine-learning segmentation approach by improving preprocessing, training, and postprocessing steps, resulting in a significantly improved segmentation accuracy. Comparison of 3D morphological indices between manually segmented rhizoids and predicted results supported the validity of the proposed method for morphological analysis. Morphological analyses revealed that, compared with both ground and artificial 1 x g conditions, rhizoid elongation and gravitropic responses were suppressed under microgravity, leading to reduced vertical growth. These findings indicate that gravity plays a fundamental role in rhizoid morphogenesis, and their absence affects growth orientation and elongation. This study provides foundational data for research on the rooting systems of bryophytes in space.

plant biology↗

bioRxiv2024

Prolonged exposure to hypergravity increases biomass and alters biomass allocation in Arabidopsis thaliana (L.) Heynh. with no apparent impact on element content in the shoot system

Previous studies have already shown that plants can complete their life cycle under microgravity. However, the effects of long-term exposure to altered gravity conditions, including microgravity, on most of the biological processes of a plants life cycle remain largely unexplored. Given the limited opportunities for space experiments, it is crucial to conduct ground-based experiments, such as hypergravity experiments. To investigate the longer-term effects of hypergravity, we have developed and utilized a custom-built hypergravity cultivation system using a centrifuge equipped with lighting, enabling the continuous growth of seed plants under hypergravity conditions. In this study, we examined the effects of 10 g hypergravity on the biomass of the shoot system (stems and rosette leaves) and the root system of vascular plants for the first time, covering the entire cultivation period from germination to 40 days. Our results showed that the dry mass of the stem per unit length was significantly higher under 10 g compared to the 1 g control, indicating a typical gravity resistance response of the stem. Moreover, the total dry mass of the stems, rosette leaves, and roots was higher under 10 g hypergravity compared to the 1 g control, suggesting an increase in biomass at the individual plant level. We also observed that the leaf mass per area of the rosette leaf was higher under hypergravity compared to the 1 g control, indicating enhanced photosynthesis rates in Arabidopsis and resulting in increased biomass of individual plants. In terms of biomass allocation, both root-shoot ratio and root mass fraction were significantly higher under hypergravity conditions compared to the 1 g control. Furthermore, we measured the content of mineral elements (Ca, Co, Cu, Fe, K, Mg, Mn, Mo, P, Zn) in the roots and rosette leaves using inductively coupled plasma optical emission spectrometry. Despite the increase in dry mass of the root system, we found no significant differences in the content of any of the ions analyzed between 10 g and 1 g conditions, indicating that mineral nutrient uptake homeostasis is maintained even under hypergravity conditions.

plant biology↗

bioRxiv2023

Prolonged exposure to hypergravity increases number and size of cells and enhances lignin deposition in the stem of Arabidopsis thaliana (L.) Heynh.

We have performed a lab-based hypergravity cultivation experiment using a centrifuge equipped with a lighting system and examined long-term effects of hypergravity on the development of the main axis (stem) of the Arabidopsis (Arabidopsis thaliana (L.) Heynh.) primary inflorescence. Plants grown under 1 x g (gravitational acceleration on Earth) conditions for 20-23 days and having the first visible flower bud were exposed to hypergravity at 8 x g for 10 days. We analyzed the effect of prolonged hypergravity conditions on growth, lignin deposition, and tissue anatomy of the main axis. As a result, the length of the main axis decreased and cross-sectional area, dry mass per unit length, cell number, lignin content of the main axis significantly increased under hypergravity. Lignin content in the rosette leaves also increased when they were exposed to hypergravity during their development. Except for interfascicular fibers, cross-sectional areas of the tissues composing the internode significantly increased under hypergravity in most type of the tissues in the basal part than the apical part of the main axis, indicating that the effect of hypergravity is more pronounced in the basal part than the apical part. The number of cells in fascicular cambium and xylem significantly increased under hypergravity both in the apical and basal internodes of the main axis, indicating a possibility that hypergravity stimulates procambium activity to produce xylem element more than phloem element. The main axis was suggested to be strengthened through changes in its morphological characteristics as well as lignin deposition under prolonged hypergravity conditions.

plant biology↗

bioRxiv2022

Three-dimensional visualization of moss rhizoid system by refraction-contrast X-ray micro-computed tomography

Land plants have two types of shoot-supporting systems, root system and rhizoid system, in vascular plants and bryophytes. However, since the evolutionary origin of the systems are different, how much they exploit common systems or distinct systems to architect their structures are largely unknown. To understand the regulatory mechanism how bryophytes architect rhizoid system responding to an environmental factor, such as gravity, and compare it with the root system of vascular plants, we have developed the methodology to visualize and quantitatively analyze the rhizoid system of the moss, Physcomitrium patens in 3D. The rhizoids having the diameter of 21.3 m on the average were visualized by refraction-contrast X-ray micro-CT using coherent X-ray optics available at synchrotron radiation facility SPring-8. Three types of shape (ring-shape, line, black circle) observed in tomographic slices of specimens embedded in paraffin were confirmed to be the rhizoids by optical and electron microscopy. Comprehensive automatic segmentation of the rhizoids which appeared in different three form types in tomograms was tested by a method using Canny edge detector or machine learning. Accuracy of output images was evaluated by comparing with the manually-segmented ground truth images using measures such as F1 score and IoU, revealing that the automatic segmentation using the machine learning was more effective than that using Canny edge detector. Thus, machine learning-based skeletonized 3D model revealed quite dense distribution of rhizoids, which was similar to root system architecture in vascular plants. We successfully visualized the moss rhizoid system in 3D for the first time.

plant biology↗

bioRxiv2021

Visualization of Arabidopsis root system architecture in 3D by refraction-contrast X-ray micro-computed tomography

Plant roots change their morphological traits in order to adapt themselves to different environmental conditions, resulting in alteration of the root system architecture. To understand this mechanism, it is essential to visualize morphology of the entire root system. To reveal effects of long-term alteration of gravity environment on root system development, we have performed an experiment in the International Space Station using Arabidopsis (Arabidopsis thaliana (L.) Heynh.) plants and obtained dried root systems grown in rockwool slabs (mineral wool substrate). X-ray computer tomography (CT) technique using an industrial X-ray scanner has been introduced for the purpose to visualize root system architecture of crop species grown in soil in 3D non-invasively. In the case of the present study, however, root system of Arabidopsis is composed of finer roots compared with typical crop plants and rockwool is also composed of fibers having similar dimension to that of the roots. A higher spatial resolution imaging method is required for distinguishing roots from rockwool. Therefore, in the present study, we tested refraction-contrast X-ray micro-CT using coherent X-ray optics available at the beamline BL20B2 of the synchrotron radiation facility SPring-8. Using this technique, both the primary and the secondary roots were successfully identified in the tomographic slices, clearly distinguished from the individual rockwool fibers and resulting in successful tracing of these roots from their basal regions. This newly-developed technique should contribute to elucidate the effect of microgravity on Arabidopsis root system architecture in space.

plant biology↗

bioRxiv2020

Life cycle of arabidopsis in the international space station - Growth direction of the inflorescence stems in the presence of light under microgravity

In the "Space Seed" experiment performed in Kibo module of the International Space Station, growth direction of the inflorescence stem of arabidopsis was examined under space 1 G, G, and ground 1 G conditions in the presence of light. The stems grew almost upright (vertical to the surface of seedbed) under ground 1 G. Although the stems were primarily upright both under space 1 G and G, they tilted slightly. The tilting of the stems under space 1 G was indicated to be due to tilting of the artificial gravitational acceleration vectors produced on the centrifuge. The tilting of the stems under G was suggested to be due to the pressure of directional airflow produced by ventilation.

plant biology↗