Biology subjects

Matsubara, S.

Publications and source records attributed to Matsubara, S..

10 recordsLinked to original sources

bioRxiv2026

Heterogeneity-Resolved Ultrafast Transient Absorption Spectroscopy of Single Supramolecular Light-Harvesting Antennas

Photosynthetic light harvesting proceeds rapidly and robustly through precisely arranged pigment molecules. However, the molecular arrangements are not uniform because they exhibit structural heterogeneity among individual complexes and across spatial regions, and undergo dynamic fluctuations. Such static and dynamic disorder can substantially perturb excitation dynamics. Here, we report a highly sensitive transient absorption microscope that integrates single-objective absorption microscopy, balanced detection, and lock-in amplification, enabling quantitative analysis of heterogeneity and temporal fluctuations in excitation dynamics. By analyzing individual chlorophyll-derivative aggregates mimicking photosynthetic light-harvesting antennas, we demonstrated that two kinetic components with nearly identical time constants can be resolved based on differences in their time-constant distributions. We further quantified the photophysical properties of each component, including absorbance change, fluorescence intensity, fluorescence efficiency, and fluorescence peak intensity ratio. These results establish an analytical framework for excitation dynamics that leverages not only the mean values of time constants but also their distribution profiles.

biophysics↗

bioRxiv2025

Highly polygenic control of photosynthetic responses to nighttime temperature in Arabidopsis studied by genomic prediction

O_LIRising nighttime temperature (Tnight) can reduce crop yields, while low Tnight may restrict plant growth and development. Despite these quantifiable effects of Tnight, the genetic basis underlying plant responses to Tnight remains unclear. We investigated natural variation in long-term response of effective photosynthetic efficiency (Fq/Fm) to Tnight among Arabidopsis accessions. C_LIO_LIGenome-wide association study (GWAS) was conducted for Fq/Fm of the accessions grown under 15{degrees}C or 20{degrees}C Tnight. The associated single nucleotide polymorphisms (SNPs) were identified and incorporated in genomic prediction (GP) models to assess the improvement of prediction accuracy. The predictions were experimentally validated in an independent, genetically diverse population. C_LIO_LIGWAS revealed highly polygenic architecture of Fq/Fm, with associated SNPs varying across Tnight conditions and measurement days. Notably, 15{degrees}C Tnight stabilized the contributions of a subset of associated SNPs, whereas 20{degrees}C Tnight enhanced day-to-day variations in SNP-trait associations. The GWAS-derived SNPs significantly improved the prediction accuracy of GP models, indicating their collective influence. The validation experiment confirmed the identification of low-Fq/Fm accessions in 15{degrees}C Tnight. C_LIO_LIThe results uncover the genetic underpinnings of long-term Fq/Fm response to cool vs warm nights and establish a framework for leveraging GWAS and GP to explore complex traits, such as photosynthesis, toward breeding climate-resilient crops. C_LI

plant biology↗

bioRxiv2025

Gamete release in Ciona robusta: roles of gonadotropin-releasing hormone and the photoreception system

Gamete release, an essential event for animals, is regulated by various factors including environmental stimuli, neural circuit, and endocrine molecules. In this study, we investigated the mechanisms regulating gamete release in the ascidian Ciona robusta, part of the sister group of vertebrates. Ciona is a hermaphrodite, releasing sperm first from orange-pigmented organ (OPO) at the end of the spermiduct, followed by eggs from the oviduct beneath the spermiduct knob-like structure. In this study, behavioral and morphological analyses revealed that this sequential release occurs because the knob-like structure compresses the oviduct opening at the onset of gamete release. Observations of transgenic Ciona expressing Kaede under the gonadotropin-releasing hormone 2 (Gnrh2) promoter showed that Gnrh2-expressing neurons and fibers accumulate in the epithelium around the spermiduct openings. In contrast, Gnrh1-expressing neurons are localized in the cerebral ganglion and project toward the ovary, suggesting distinct roles of GnRH1 and GnRH2 in reproductive regulation. RNA-seq and real-time PCR analyses revealed that Opsin2, Opsin3, beta-carotene-15,15-monooxygenase (Bco), and several ion channel genes are specifically expressed in the OPO, along with Gnrh2. In situ hybridization showed that these genes are localized in the innermost OPO layer, suggesting that the OPO functions as a photoreceptive organ. Although, Gnrh2 expression has been considered low in adult Ciona, our study revealed strong OPO-specific expression with photoreceptive genes. Collectively, these findings suggest that GnRH2 plays a central role in gamete release regulation, potentially in coordination with the photoreceptive system, while GnRH1 may regulate ovarian functions. These results indicate that C. robusta employs distinct GnRH systems in a tissue-specific manner to regulate reproduction.

evolutionary biology↗

bioRxiv2025

Constitutive and regulatory responses of Arabidopsis thaliana to harmonically oscillating light

The rate of net CO2 uptake is proportional to dim light and saturates when the light exceeds the plant assimilation capacity. This simple relationship between constant light and photosynthesis becomes intriguingly complex when the light oscillates. The rates of photosynthesis may differ between the descending and ascending phases of light oscillation. This hysteresis changes with the frequency and amplitude of the light and reports on the dynamics of the photosynthetic reactions and their regulation. Here, we investigated the chlorophyll fluorescence response of Arabidopsis thaliana to light oscillating with three different amplitudes: 100-200, 100-400, and 100-800 mol photons{middle dot}m-2{middle dot}s-1, each with periods ranging from 1 s to 8 min. The light amplitudes and periods were chosen to represent light patterns often appearing in nature. Three genotypes were compared: wild-type Col-0 and npq1 and npq4 mutants that are incapacitated in the rapidly reversible energy-dependent non-photochemical quenching (qE). The experiments identified two major dynamic patterns. One was found in oscillation periods shorter than 30 s, characterized by constitutive non-linearity and hysteresis. The other was mainly formed by regulatory non-linearity and hysteresis, occurring when the oscillation periods were longer than 30 s. The mathematical model simulating the chlorophyll fluorescence dynamics qualitatively reproduced the constitutive and regulatory dynamic patterns observed in the experiments. The model simulations illustrated the dynamics of non-photochemical quencher activation, plastoquinone pool reduction, and lumen pH that form the constitutive and regulatory non-linearities. The model simulations provided mechanistic insight into molecular processes forming the plant response to oscillating light.

plant biology↗

bioRxiv2023

Exploring natural genetic variation in photosynthesis-related traits of barley in the field

Optimizing photosynthesis is considered an important strategy for improving crop yields to ensure food security. To evaluate the potential of using photosynthesis-related parameters in crop breeding programs, we measured chlorophyll fluorescence along with growth-related and morphological traits of 23 barley inbreds across different developmental stages in field conditions. The photosynthesis-related parameters were highly variable, changing with light intensity and developmental progression of plants. Yet, the variations in photosystem II (PSII) quantum yield observed among the inbreds in the field largely reflected the variations in CO2 assimilation properties in controlled climate chamber conditions, confirming that the chlorophyll fluorescence-based technique can provide proxy parameters of photosynthesis to explore genetic variations under field conditions. Heritability (H2) of the photosynthesis-related parameters in the field ranged from 0.16 for the quantum yield of non-photochemical quenching to 0.78 for the fraction of open PSII center. Two parameters, the maximum PSII efficiency in light-adapted state ( H2 0.58) and the total non-photochemical quenching ( H2 0.53), showed significant positive and negative correlations, respectively, with yield-related traits (dry weight per plant and net straw weight) in the barley inbreds. These results indicate the possibility of improving crop yield through optimizing photosynthetic light use efficiency by conventional breeding programs.

plant biology↗

bioRxiv2023

A mathematical model of photoinhibition: exploring the impact of quenching processes

Plants are constantly exposed to changing environments, sometimes leading to extreme conditions and stress. For example, sudden exposure to high light leads to excess absorbed light energy, causing reactive oxygen species (ROS) formation. ROS damage the photosynthetic machinery, particularly the D1 protein in photosystem II (PSII), which therefore needs to be continuously repaired and replaced. The effect of the damage inflicted by high light is a prolonged decrease in photosynthetic efficiency. Hence, it is not surprising that photoinhibition has been subject to numerous experimental studies investigating its effects in the context of crop productivity. However, it has become apparent that classical measures of photoinhibition, i.e., changes in the chlorophyll fluorescence parameter Fv/Fm, are not only determined by the loss of PSII core function but also by processes such as energy transfer and quenching. Mathematical models can help dissect the influences on such fluorescence signals and quantify the contributions of various interacting mechanisms. We present a mathematical model with a dynamic description of the photosynthetic electron transport chain (PETC), non-photochemical quenching, and photoinhibition. With our model, we investigate the interconnection between quenching, photoprotection, and fluorescence using simulations and experimental data. We found that different energy-dissipating properties of intact and damaged PSIIs, as well as energy transfer between PSIIs, are critical components that need to be included in the model to ensure a satisfactory fit to the experimental data. We envisage that our model provides a framework for future investigations of photoinhibition dynamics and its importance for plant growth and yield.

plant biology↗

bioRxiv2022

The dynamics of non-photochemical quenching and cyclic electron transport in A.thaliana exposed to harmonically oscillating light

In natural environments, plants are exposed to rapidly changing light. Maintaining photosynthetic efficiency while avoiding photodamage requires equally rapid regulation of photoprotective mechanisms. We asked what the operation frequency range of regulation is in which plants can efficiently respond to varying light. Chlorophyll fluorescence, P700, plastocyanin, and ferredoxin responses of wild-type Arabidopsis thaliana were measured in oscillating light of various frequencies. We also investigated the npq1 mutant lacking violaxanthin de-epoxidase, the npq4 mutant lacking PsbS-protein, and the mutants crr2-2, and pgrl1ab impaired in different pathways of the cyclic electron transport. The fastest was the PsbS-regulation responding to oscillation periods longer than 10s. Processes involving violaxanthin de-epoxidase dampened changes of chlorophyll fluorescence in oscillation periods of 2min or longer. Knocking out the PGRL1-PGR5 pathway strongly reduced variations of all monitored parameters, probably due to congestion in the electron transport. Incapacitating the NDH-like pathway only slightly changed the photosynthetic dynamics. Our observations are consistent with the assumption that non-photochemical quenching in slow light oscillations involves violaxanthin de-epoxidase to produce, presumably, a stationary, non-oscillating level of zeaxanthin. We interpret the observed dynamics of Photosystem I components as being formed in slow light oscillations partially by thylakoid remodeling that modulates the redox rates.

plant biology↗

bioRxiv2021

Comparative analysis of transcriptomic profiles among ascidians, zebrafish, and mice: insights from tissue-specific gene expression

Tissue/organ-specific genes (TSGs) are important not only for understanding organ development and function, but also for investigating the evolutionary lineages of organs in animals. Here, we investigate the TSGs of 9 adult tissues of an ascidian, Ciona intestinalis Type A (Ciona robusta), which lies in the important position of being the sister group of vertebrates. RNA-seq and qRT-PCR identified the Ciona TSGs in each tissue, and BLAST searches identified their homologs in zebrafish and mice. Tissue distributions of the vertebrate homologs were analyzed and clustered using public RNA-seq data for 12 zebrafish and 30 mouse tissues. Among the vertebrate homologs of the Ciona TSGs in the neural complex, 48% and 63% showed high expression in the zebrafish and mouse brain, respectively, suggesting that the central nervous system is evolutionarily conserved in chordates. In contrast, vertebrate homologs of Ciona TSGs in the ovary, pharynx, and intestine were not consistently highly expressed in the corresponding tissues of vertebrates, suggesting that these organs have evolved in Ciona-specific lineages. Intriguingly, more TSG homologs of the Ciona stomach were highly expressed in the vertebrate liver (17-29%) and intestine (22-33%) than in the mouse stomach (5%). Expression profiles for these gene suggest that the biological roles of the Ciona stomach are distinct from those of their vertebrate counterparts. Collectively, Ciona tissues were categorized into 3 groups: i) high similarity to the corresponding vertebrate tissues (neural complex and heart), ii) low similarity to the corresponding vertebrate tissues (ovary, pharynx, and intestine), and iii) low similarity to the corresponding vertebrate tissues, but high similarity to other vertebrate tissues (stomach, endostyle, and siphons). The present study provides transcriptomic catalogs of adult ascidian tissues and significant insights into the evolutionary lineages of the brain, heart, and digestive tract of chordates.

evolutionary biology↗

bioRxiv2021

Chlorophyll fluorescence: How the quality of information about PAM instrument parameters may affect our research

Chlorophyll a fluorescence is a powerful indicator of photosynthetic energy conversion in plants and photosynthetic microorganisms. One of the most widely used measurement techniques is Pulse Amplitude Modulation (PAM) fluorometry. Unfortunately, parameter settings of PAM instruments are often not completely described in scientific articles although their variations, however small these may seem, can influence measurements. We show the effects of parameter settings on PAM measurements. We first simulated fluorescence signals using a previously published computational model of photosynthesis. Then, we validated our findings experimentally. Our analysis demonstrates how the kinetics of non-photochemical quenching (NPQ) induction and relaxation are affected by different settings of PAM instrument parameters. Neglecting these parameters may mislead data interpretation and derived hypotheses, hamper independent validation of the results, and cause problems for mathematical formulation of underlying processes. Given the uncertainties inflicted by this neglect, we urge PAM users to provide detailed documentation of measurement protocols. Moreover, to ensure accessibility to the required information, we advocate minimum information standards that can serve both experimental and computational biologists in our efforts to advance system-wide understanding of biological processes. Such specification will enable launching a standardized database for plant and data science communities. HighlightPAM fluorometry measurement is sensitive to instrument settings and protocols. Yet, protocols are published incompletely. We urge to reach an agreement on minimal protocol information of PAM experiments to be shared publicly.

plant biology↗

bioRxiv2020

Removal of repressive histone marks creates epigenetic memory of recurring heat in Arabidopsis

Acclimation to high temperature increases plants tolerance of subsequent lethal high temperatures1-3. Although epigenetic regulation of plant gene expression is well studied, how plants maintain a memory of environmental changes over time remains unclear. Here, we show that JUMONJI (JMJ) proteins4-8, demethylases involved in histone H3 lysine 27 trimethylation (H3K27me3), are necessary for Arabidopsis thaliana heat acclimation. Acclimation induces sustained H3K27me3 demethylation at key HEAT SHOCK PROTEIN (HSP) loci by JMJs, poising the HSP genes for subsequent activation. Upon sensing heat after a 3-day interval, JMJs directly reactivate HSP genes. Finally, jmj mutants fail to maintain heat memory under fluctuating field temperature conditions. Our findings of an epigenetic memory mechanism involving histone demethylases may have implications for environmental adaptation of field plants.

plant biology↗