Chloroplast movement in leaves has been shown to help minimize photoinhibition and increase growth under certain conditions. Much can be learned about chloroplast movement by studying the chloroplast positioning in leaves using e.g., confocal fluorescence microscopy, but access to this type of microscopy is limited. This protocol describes a method that uses the changes in leaf transmission as a proxy for chloroplast movement. If chloroplasts are spread out in order to maximize light interception, the transmission will be low. If chloroplasts move towards the anticlinal cell walls to avoid light, the transmission will be higher. This protocol describes how to use a straightforward, home-built instrument to expose leaves to different blue light intensities and quantify the dynamic changes in leaf transmission. This approach allows researchers to quantitatively describe chloroplast movement in different species and mutants, study the effects of chemicals and environmental factors on it, or screen for novel mutants e.g., to identify missing components in the process that leads from light perception to the movement of chloroplasts.
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http://dx.doi.org/10.3791/62881 | DOI Listing |
Plant Mol Biol
January 2025
Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Chuo-Ku, Kumamoto, 860-8555, Japan.
The applicability of a deep learning model for the virtual staining of plant cell structures using bright-field microscopy was investigated. The training dataset consisted of microscopy images of tobacco BY-2 cells with the plasma membrane stained with the fluorescent dye PlasMem Bright Green and the cell nucleus labeled with Histone-red fluorescent protein. The trained models successfully detected the expansion of cell nuclei upon aphidicolin treatment and a decrease in the cell aspect ratio upon propyzamide treatment, demonstrating its utility in cell morphometry.
View Article and Find Full Text PDFTree Physiol
January 2025
Institute of Soil and Water Conservation, Northwest A&F University, Yangling 712100, China.
Modulation of stomatal development and movement is a promising approach for creating water-conserving plants. Here, we identified and characterized the PagHCF106 gene of poplar (Populus alba × Populus glandulosa). The PagHCF106 protein localized predominantly to the chloroplast, and the PagHCF106 gene exhibited tissue-specific expression pattern.
View Article and Find Full Text PDFPlants (Basel)
December 2024
Henan Key Laboratory of Fruit and Cucurbit Biology, Zhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Zhengzhou 450009, China.
Potyviral P3 is involved in viral replication, movement, and pathogenicity; however, its biochemical function is unknown. In this study, the P3 of the zucchini yellow mosaic virus (ZYMV) interacted with ClBBD, a protein with high ortholog bifunctional nuclease activity, in watermelon. The binding site was shown via yeast two-hybrid screening and BiFC assay to be located at the N-terminus of P3 rather than P3N-PIPO.
View Article and Find Full Text PDFJ Microsc
December 2024
Biology Department and the Molecular and Environmental Plant Sciences Program, Texas A&M University, College Station, Texas, USA.
The endoplasmic reticulum (ER) forms contact sites with the chloroplast. Exposing contact sites that contain both the chloroplast and the ER to localised high-fluence, wavelength specific, 405 nm violet light, hereinafter referred to as photostimulation, induces multiple, potentially interacting intra- and intercellular responses. The responses vary depending on the tissue type of the cell and the chloroplast.
View Article and Find Full Text PDFPlant J
January 2025
Center for Bioscience Research and Education, Utsunomiya University, Tochigi, 321-8505, Japan.
In the accumulation response, chloroplasts move toward weak blue light (BL) to maximize photosynthetic efficiency; in the avoidance response, they move away from strong BL to reduce photodamage. The BL receptor kinase phototropin (phot) mediates these chloroplast relocation responses, and the chloroplast relocation response requires phot kinase activity. Upon receiving BL, phot undergoes autophosphorylation; however, the molecular mechanisms that regulate chloroplast relocation through phot autophosphorylation remain unclear.
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