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Research ArticlePLANT-MICROBE AND PLANT-INSECT INTERACTIONS
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Tobamovirus and Potyvirus Accumulation in Minor Veins of Inoculated Leaves from Representatives of the Solanaceae and Fabaceae

Xin Shun Ding, Shelly A. Carter, C. Michael Deom, Richard S. Nelson
Xin Shun Ding
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Shelly A. Carter
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C. Michael Deom
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Richard S. Nelson
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Published January 1998. DOI: https://doi.org/10.1104/pp.116.1.125

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    Fig. 1.

    Electron micrographs of typical category V veins (9–12 vein cells, not including BS cells) in sections from mature leaves of N. benthamiana (A and B) or C. annuum (C and D) treated with distilled water or 1.0 m sorbitol solution before fixation. A and C, Veins treated with distilled water before fixation. Cells are numbered to allow text discussion. Note in A that there are two cells at the position of cell 4. B and D, Veins treated with 1.0 m sorbitol before fixation. Note that the plasmalemma of cells 4, 6, and 8 are separated from their cell walls and thus plasmolyzed, whereas those of cells 5, 7, and 9 are not. Therefore, cells 4, 6, and 8 are considered to be VP cells, and cells 5, 7, and 9 are considered to be C cells. S, SE; X, xylem TE. A and B, bars = 3 μm; C and D, bars = 4 μm.

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    Fig. 2.

    Accumulation of genomic TMV RNA in cells of a category V vein (defined in Fig. 1 legend) from a mature inoculated leaf of Xanthi at 4 dpi. A, Section from an inoculated leaf analyzed by in situ hybridization using a biotinylated-RNA probe complementary to nucleotides 1 to 3332 of the TMV genome. The biotinylated-RNA probe was then detected using a streptavidin-gold conjugate followed by silver enhancement. The section was photographed under a light microscope. The labeling signal appears as white specks in cells in the dark-field image. Three VP cells and one C cell of the vein had accumulated viral RNA. The infected C cell is denoted with an arrow. B, Section from a mock-inoculated Xanthi leaf analyzed as for the vein in A. TMV RNA was not detected in vein cells from this section. Bars = 12 μm.

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    Fig. 3.

    Accumulation of TMV in cells of a typical category V vein (defined in Fig. 1) from a mature inoculated leaf ofC. annuum at 3 dpi. TMV accumulation was visualized by immunocytochemistry with primary antibody against TMV and secondary antibody conjugated with gold followed by silver enhancement. The sections were photographed under a light microscope. The labeling signal appears as white specks in cells in the dark-field image. A, Section from TMV-inoculated leaf; B, section from a mock-inoculated C. annuum leaf. Bar = 7.1 μm.

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    Fig. 4.

    Accumulation of SHMV in a typical category V vein (defined in Fig. 1) from a mature inoculated leaf of N. benthamiana at 4 dpi. SHMV accumulation was visualized as described in Figure 3 using antibody against SHMV. A, Section from leaf tissue inoculated with SHMV. Virus was detected in three VP cells but not in C cells of this vein. B, Section from a mock-inoculated leaf tissue analyzed as for the vein in A. Bars = 12 μm.

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    Fig. 5.

    Accumulation of SHMV in cells from a P. vulgaris leaf at 5 dpi. Accumulation of SHMV in cells was analyzed by immunocytochemistry using an antibody against SHMV and a goat anti-rabbit gold (20 nm) conjugate. Sections were examined and photographed under an electron microscope. A, Virion (V) aggregates labeled with immunogold in the cytoplasm of an infected mesophyll cell. Ch, Chloroplast; M, mitochondrion. Bar = 0.7 μm. B, Virion aggregates labeled with immunogold in the cytoplasm of a VP cell from a category V vein. No virion aggregates were detected in C cells of the vein. Inset, Magnification of several aggregates labeled with immunogold. S, SE; X, xylem TE. Bar = 2 μm.

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    Fig. 6.

    Accumulation of SHMV in minor vein cells ofP. sativum. Accumulation of SHMV in cells was analyzed as described in Figure 6. A, Accumulation of SHMV in a category V vein from a mature inoculated leaf at 5 dpi. SHMV was detected in a BS cell, a VP cell, but not in the adjacent T cells. Bar = 1.5 μm. B, Accumulation of SHMV in a category VI vein from a systemically infected leaf at 8 dpi. Viral aggregates of SHMV were detected in both VP and T cells of the vein. Bar = 2 μm. Arrows point to magnified images of boxed areas showing immunogold labeling. S, SE; X, xylem TE.

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    Fig. 7.

    Accumulation of PVY in cells of a typical category V vein from a mature inoculated leaf of Xanthi at 4 dpi. Analysis for PVY accumulation was done as described in Figure 3 using an antibody against PVY. A, Section from leaf tissue inoculated with PVY. The virus was detected in three VP cells but not in C cells of this vein. B, Section from a mock-inoculated leaf analyzed identically to tissue shown in A. Bars = 10 μm.

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    Table I.

    Symptom ontogeny in leaves from various hosts after virus infection

    Virus and HostLeaf
    InoculatedYoung systemic
    TMV
     N. benthamiana c.l.-a3 dpiv.c.-b5 dpi
     N. tabacum c.l. 3 dpiv.c. 5 dpi
     C. annuum c.l. 3 dpiv.c. 5 dpi
     L. esculentum n.s.-c v.c. and l.r.-d5 dpi
    SHMV
     N. benthamiana n.s.v.c. 5 dpi
     P. vulgaris c.l. 4 dpim.m.-e6 dpi
     P. sativum n.s.v.c. 6 dpi
    PVY
     N. tabacum n.s.v.c. 5 dpi
     N. benthamiana n.s.m.m. 5 dpi
    PStV
     N. benthamiana n.s.s.c.-f5 dpi
    • ↵F0-a c.l., Chlorotic lesion.

    • ↵F0-b v.c., Vein clearing.

    • ↵F0-c n.s., No symptoms.

    • ↵F0-d l.r., Leaf rolling.

    • ↵F0-e m.m., Mild mosaic.

    • ↵F0-f s.c., Systemic chlorosis.

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    Table II.

    Analysis of minor veins for virus accumulation in VP cells, C/T cells, or both

    PlantdpiVirusNo. of Veins ExaminedVeins with Only VP Cells InfectedVeins with Only C or T Cells InfectedVeins with Both VP and C or T Cells Infected
    %
    N. benthamiana 3TMV43 (10)1-a 62.8037.2
    C. annuum 337  (21)45.9054.1
    L. esculentum 34  (8)75.0025.0
    N. benthamiana 4SHMV40  (9)62.507.5
    P. vulgaris 522  (8)31.100
    P. sativum 519  (8)94.700
    Xanthi4PVY18  (5)38.8061.2
    N. benthamiana 417 (10)41.2058.8
    N. benthamiana 4PStV23  (5)30.4056.5
    • ↵F1-a Numbers in parentheses indicate the number of sections analyzed.

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    Table III.

    Percentage of infected VP and C/T cells within minor veins and their ratio

    PlantdpiVirusNo. of Veins ExaminedCell TypeVP:C/T2-b
    VPC/T2-a
    %
    N. benthamiana 3TMV4373.77.011:1
    C. annuum 33778.228.83:1
    L. esculentum 34100.08.312:1
    N. benthamiana 4SHMV4061.52.525:1
    P. vulgaris 52231.10.0>31:1
    P. sativum 51990.30.0>90:1
    Xanthi4PVY1837.09.34:1
    N. benthamiana 41734.05.96:1
    N. benthamiana 4PStV2358.729.62:1
    • ↵F2-a All species analyzed had smooth-walled C cells except P. sativum, which had T cells.

    • ↵F2-b Ratio of VP to C/T cells infected.

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Tobamovirus and Potyvirus Accumulation in Minor Veins of Inoculated Leaves from Representatives of the Solanaceae and Fabaceae
Xin Shun Ding, Shelly A. Carter, C. Michael Deom, Richard S. Nelson
Plant Physiology Jan 1998, 116 (1) 125-136; DOI: 10.1104/pp.116.1.125

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Tobamovirus and Potyvirus Accumulation in Minor Veins of Inoculated Leaves from Representatives of the Solanaceae and Fabaceae
Xin Shun Ding, Shelly A. Carter, C. Michael Deom, Richard S. Nelson
Plant Physiology Jan 1998, 116 (1) 125-136; DOI: 10.1104/pp.116.1.125
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Plant Physiology: 116 (1)
Plant Physiology
Vol. 116, Issue 1
Jan 1998
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