19 research outputs found

    The Arabidopsis minE mutation causes new plastid and FtsZ1 localization phenotypes in the leaf epidermis

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    Plastids in the leaf epidermal cells of plants are regarded as immature chloroplasts that, like mesophyll chloroplasts, undergo binary fission. While mesophyll chloroplasts have generally been used to study plastid division, recent studies have suggested the presence of tissue- or plastid type-dependent regulation of plastid division. Here, we report the detailed morphology of plastids and their stromules, and the intraplastidic localization of the chloroplast division-related protein AtFtsZ1-1, in the leaf epidermis of an Arabidopsis mutant that harbors a mutation in the chloroplast division site determinant gene AtMinE1. In atminE1, the size and shape of epidermal plastids varied widely, which contrasts with the plastid phenotype observed in atminE1 mesophyll cells. In particular, atminE1 epidermal plastids occasionally displayed grape-like morphology, a novel phenotype induced by a plastid division mutation. Observation of an atminE1 transgenic line harboring an AtMinE1 promoter::AtMinE1-yellow fluorescent protein fusion gene confirmed the expression and plastidic localization of AtMinE1 in the leaf epidermis. Further examination revealed that constriction of plastids and stromules mediated by the FtsZ1 ring contributed to the plastid pleomorphism in the atminE1 epidermis. These results illustrate that a single plastid division mutation can have dramatic consequences for epidermal plastid morphology, thereby implying that plastid division and morphogenesis are differentially regulated in epidermal and mesophyll plastids

    Differential responses to high- and low-dose ultraviolet-B stress in tobacco Bright Yellow-2 cells

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    Ultraviolet (UV)-B irradiation leads to DNA damage, cell cycle arrest, growth inhibition, and cell death. To evaluate the UV-B stressā€“induced changes in plant cells, we developed a model system based on tobacco Bright Yellow-2 (BY-2) cells. Both low-dose UV-B (low UV-B: 740 J māˆ’2) and high-dose UV-B (high UV-B: 2960 J māˆ’2) inhibited cell proliferation and induced cell death; these effects were more pronounced at high UV-B. Flow cytometry showed cell cycle arrest within 1 day after UV-B irradiation; neither low- nor high-UV-Bā€“irradiated cells entered mitosis within 12 h. Cell cycle progression was gradually restored in low-UV-Bā€“irradiated cells but not in high-UV-Bā€“irradiated cells. UV-A irradiation, which activates cyclobutane pyrimidine dimer (CPD) photolyase, reduced inhibition of cell proliferation by low but not high UV-B and suppressed high-UV-Bā€“induced cell death. UV-B induced CPD formation in a dose-dependent manner. The amounts of CPDs decreased gradually within 3 days in low-UV-Bā€“irradiated cells, but remained elevated after 3 days in high-UV-Bā€“irradiated cells. Low UV-B slightly increased the number of DNA single-strand breaks detected by the comet assay at 1 day after irradiation, and then decreased at 2 and 3 days after irradiation. High UV-B increased DNA fragmentation detected by the terminal deoxynucleotidyl transferase dUTP nick end labeling assay 1 and 3 days after irradiation. Caffeine, an inhibitor of ataxia telangiectasia mutated (ATM) and ataxia telangiectasia and Rad3-related (ATR) checkpoint kinases, reduced the rate of cell death in high-UV-Bā€“irradiated cells. Our data suggest that low-UV-Bā€“induced CPDs and/or DNA strand-breaks inhibit DNA replication and proliferation of BY-2 cells, whereas larger contents of high-UV-Bā€“induced CPDs and/or DNA strand-breaks lead to cell death

    IMACEL: A cloud-based bioimage analysis platform for morphological analysis and image classification.

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    Automated quantitative image analysis is essential for all fields of life science research. Although several software programs and algorithms have been developed for bioimage processing, an advanced knowledge of image processing techniques and high-performance computing resources are required to use them. Hence, we developed a cloud-based image analysis platform called IMACEL, which comprises morphological analysis and machine learning-based image classification. The unique click-based user interface of IMACEL's morphological analysis platform enables researchers with limited resources to evaluate particles rapidly and quantitatively without prior knowledge of image processing. Because all the image processing and machine learning algorithms are performed on high-performance virtual machines, users can access the same analytical environment from anywhere. A validation study of the morphological analysis and image classification of IMACEL was performed. The results indicate that this platform is an accessible and potentially powerful tool for the quantitative evaluation of bioimages that will lower the barriers to life science research

    Data from: The Arabidopsis arc5 and arc6 mutations differentially affect plastid morphology in pavement and guard cells in the leaf epidermis

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    Chloroplasts, or photosynthetic plastids, multiply by binary fission, forming a homogeneous population in plant cells. In Arabidopsis thaliana, the division apparatus (or division ring) of mesophyll chloroplasts includes an inner envelope transmembrane protein ARC6, a cytoplasmic dynamin-related protein ARC5 (DRP5B), and members of the FtsZ1 and FtsZ2 families of proteins, which co-assemble in the stromal mid-plastid division ring (FtsZ ring). FtsZ ring placement is controlled by several proteins, including a stromal factor MinE (AtMinE1). During leaf mesophyll development, ARC6 and AtMinE1 are necessary for FtsZ ring formation and thus plastid division initiation, while ARC5 is essential for a later stage of plastid division. Here, we examined plastid morphology in leaf epidermal pavement cells (PCs) and stomatal guard cells (GCs) in the arc5 and arc6 mutants using stroma-targeted fluorescent proteins. The arc5 PC plastids were generally a bit larger than those of the wild type, but most had normal shapes and were division-competent, unlike mutant mesophyll chloroplasts. The arc6 PC plastids were heterogeneous in size and shape, including the formation of giant and mini-plastids, plastids with highly developed stromules, and grape-like plastid clusters, which varied on a cell-by-cell basis. Moreover, unique plastid phenotypes for stomatal GCs were observed in both mutants. The arc5 GCs rarely lacked chlorophyll-bearing plastids (chloroplasts), while they accumulated minute chlorophyll-less plastids, whereas most GCs developed wild type-like chloroplasts. The arc6 GCs produced large chloroplasts and/or chlorophyll-less plastids, as previously observed, but unexpectedly, their chloroplasts/plastids exhibited marked morphological variations. We quantitatively analyzed plastid morphology and partitioning in paired GCs from wild-type, arc5, arc6, and atminE1 plants. Collectively, our results support the notion that ARC5 is dispensable in the process of equal division of epidermal plastids, and indicate that dysfunctions in ARC5 and ARC6 differentially affect plastid replication among mesophyll cells, PCs, and GCs within a single leaf

    Plastid phenotypes in leaf PCs of <i>Arabidopsis</i> WT, <i>arc5</i>, <i>arc6</i>, and <i>atminE1</i>.

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    <p>(Aā€“J) Images of plastid-targeted YFP in the third-fourth leaf petioles of 4-week-old WT (A), <i>arc5</i> (B, E, H), <i>arc6</i> (C, F, I), and <i>atminE1</i> (D, G, J) seedlings. CLSM images of maximal intensity projection are shown. Giant plastids (arrows), putative dividing plastids (arrowheads), stomata (double arrowheads), and mini-plastids (asterisks) are indicated. Bar = 10 Ī¼m.</p

    The <i>Arabidopsis arc5</i> and <i>arc6</i> mutations differentially affect plastid morphology in pavement and guard cells in the leaf epidermis

    No full text
    <div><p>Chloroplasts, or photosynthetic plastids, multiply by binary fission, forming a homogeneous population in plant cells. In <i>Arabidopsis thaliana</i>, the division apparatus (or division ring) of mesophyll chloroplasts includes an inner envelope transmembrane protein ARC6, a cytoplasmic dynamin-related protein ARC5 (DRP5B), and members of the FtsZ1 and FtsZ2 families of proteins, which co-assemble in the stromal mid-plastid division ring (FtsZ ring). FtsZ ring placement is controlled by several proteins, including a stromal factor MinE (AtMinE1). During leaf mesophyll development, <i>ARC6</i> and <i>AtMinE1</i> are necessary for FtsZ ring formation and thus plastid division initiation, while <i>ARC5</i> is essential for a later stage of plastid division. Here, we examined plastid morphology in leaf epidermal pavement cells (PCs) and stomatal guard cells (GCs) in the <i>arc5</i> and <i>arc6</i> mutants using stroma-targeted fluorescent proteins. The <i>arc5</i> PC plastids were generally a bit larger than those of the wild type, but most had normal shapes and were division-competent, unlike mutant mesophyll chloroplasts. The <i>arc6</i> PC plastids were heterogeneous in size and shape, including the formation of giant and mini-plastids, plastids with highly developed stromules, and grape-like plastid clusters, which varied on a cell-by-cell basis. Moreover, unique plastid phenotypes for stomatal GCs were observed in both mutants. The <i>arc5</i> GCs rarely lacked chlorophyll-bearing plastids (chloroplasts), while they accumulated minute chlorophyll-less plastids, whereas most GCs developed wild type-like chloroplasts. The <i>arc6</i> GCs produced large chloroplasts and/or chlorophyll-less plastids, as previously observed, but unexpectedly, their chloroplasts/plastids exhibited marked morphological variations. We quantitatively analyzed plastid morphology and partitioning in paired GCs from wild-type, <i>arc5</i>, <i>arc6</i>, and <i>atminE1</i> plants. Collectively, our results support the notion that ARC5 is dispensable in the process of equal division of epidermal plastids, and indicate that dysfunctions in ARC5 and ARC6 differentially affect plastid replication among mesophyll cells, PCs, and GCs within a single leaf.</p></div
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