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Figure legends
Figure 1. Formation of wheat anther cell layers. The anther primordium only contains L1, L2 and L3 layers. Some cells from L2 become archesporial (AR) cells < 150 µm, which differentiate into epidermis (EN), secondary parietal layer (SPL) and pollen mother cells (PMC) > 150 µm anthers. Then SPL divides to form middle layer (ML) and tapetum (TA) almost at 400 µm. Subsequently, PMC form the MP at 700 µm, which differentiate to pollen grain (PG) almost at 1300 µm.
Figure 2. Confocal images of anther developmental cells.(A) The EPI cells have obvious shape and AR cells began to form at 100 µm. (B) At 310 µm AR cells differentiate into EN, SPL and PMC cells after mitosis. (C) EN cells clearly defined, which have oval shape at 580 µm. (D) SPL are round at 380 µm.(E) PMC cells have round shape and almost have clear structure at 370 µm. (F, G) Tds formed after meiosis in PMC cells at 660 and 680 µm, respectively. PMC cells totally disappear, and ML cell began to crush (H) Tds differentiate into MP at 700 µm and ML become disappear at almost 1100 µm and MP grow large create space from TA cells(I) PG become round at 1630 µm and gametes develop within pollen grains and ML, TA, MP disappear after PG development. Scale bar: a, c, d, f, g = 25 µm, b, e, I = 50 µm, h= 75 µm.
Figure 3. The cell numbers and cell sizes of control wheat anther was measured using velocity software. (A) Cell count of anther layers along X-axis. (B) Cell count of anther layers along Y-axis. (C) Total cells/locule (X*Y-tips) of anther layers. (D) Percentage of locule cell count of anther layers.(E) Cell width of anther layers along X-axis. (F) Cell length of anther layers along Y-axis. (G) Cell depth of anther layers along Z-axis. (H) Cell volume of anther layers was derived by multiplying the width, length and depth (I) Y/X value of anther layers were measured by dividing the length and width of anther layers (J) Percentage of locule volume of anther layers. The cell counts, and sizes were quantified with at least 60 anthers per stage.
Figure 4. T. laevis location on anther developmental cells of wheat anther. (A-C) Hyphae on EPI cells (800 µm);(D-F) Hyphae on EN cells (1220 µm); (G-I) Hyphae on TA cells (1300 µm) (J-L) Hyphae on mature PG (1520 µm). Scale bars: a-c = 50 µm; d-f = 25 µm; g-i = 10 µm; j-l = 25 µm.
Figure 5. The cell numbers and cell sizes of T. laevisinfected wheat anther was measured using velocity software. (A)Cell count of anther layers along X-axis. (B) Cell count of anther layers along Y-axis. (C) Total cells/locule (X*Y-tips) of anther layers. (D) Percentage of locule cell count of anther layers. (E) Cell width of anther layers along X-axis.(F) Cell length of anther layers along Y-axis. (G)Cell depth of anther layers along Z-axis. (H) Cell volume of anther layers was derived by multiplying the width, length and depth(I) Y/X value of anther layers were measured by dividing the length and width of anther layers (J) Percentage of locule volume of anther layers. The cell counts, and sizes were quantified with at least 60 anthers per stage.
Figure 6. T. controversa location on anther developmental cells of wheat anther. (A-C) Hyphae on EPI cells 1000 (µm); (D-F) Hyphae on EN cells (1300 µm); (G-I)Hyphae on TA cells (2100 µm) (J-L) Hyphae on mature PG (2100 µm). Scale bars: a-c = 50 µm; d-l= 25 µm.
Figure 7. The cell numbers and cell sizes of T. controversa infected wheat anther was measured using velocity software.(A) Cell count of anther layers along X-axis. (B) Cell count of anther layers along Y-axis. (C) Total cells/locule (X*Y-tips) of anther layers. (D) Percentage of locule cell count of anther layers. (E) Cell width of anther layers along X-axis. (F) Cell length of anther layers along Y-axis.(G) Cell depth of anther layers along Z-axis. (H) Cell volume of anther layers was derived by multiplying the width, length and depth (I) Y/X value of anther layers were measured by dividing the length and width of anther layers (J) Percentage of locule volume of anther layers. The cell counts, and sizes were quantified with at least 60 anthers per stage.
Figure 8 . The DEGs between T. controversa and T. laevis samples. (A ) Numbers of DEGs compared between T. laevis and control samples. (B ) Numbers of DEGs compared between T. controversa and control samples. DEGs are shown in dark (up-regulated) and light white (down-regulated). Control_1_vs_ infected 1_G = 0~150 μm, control_2_vs_ infected 2_G= 150~400 μm, control_3_vs_ infected 3_G = 400~700 μm, control_4_vs_ infected 4_G =700~1100 μm and control_5_vs_ infected 5_G=1100~1300 μm.
Figure 9. Analysis of differentially expressed genes in different anther length stages. (a ) Venn diagram analysis of the DEGs in different anther length stages after inoculation withT. laevis . (b ) Hierarchical clustering heatmap of expression profiles of selected DEGs in response to T. laevisinfection. (c ) Venn diagram analysis of the DEGs in different anther length stages after inoculation with T. controversa . (d ) Hierarchical clustering heatmap of expression profiles of selected DEGs in response to T. controversa infection. Each column shows a library, and each row shows DEG expression. The colors blue, white and red indicate low, medium, and high expression patterns of genes, respectively.
Figure 10. TaEC increase wheat resistance to bunt pathogens. (A) a : The wheat leaves inoculated with BSMV: TaEC at 13 days post inoculation (dpi). b: The wheat leaves inoculated with BSMV: γ at 13 dpi. c : The wheat leaves inoculated with BSMV:TaEC at 30 (dpi). d : The wheat leaves inoculated with BSMV:γ at 30 (dpi). (B ) e: The kernels inoculated with BSMV:γ (left) and BSMV:TaEC (right) after T. controversa inoculation f : The kernels inoculated with BSMV:γ (left) and BSMV:TaEC (right) after T. laevis inoculation.g : The kernels only infected with BSMV:γ(left) and BSMV:TaEC (right) without inoculation of T. controversa and T. laevis . (C) h : Lateral arrangement of wheat grains infected with BSMV:γ and BSMV:TaEC. i : Longitudinal arrangement of wheat grains infected with BSMV:γ and BSMV:TaEC. (D ) The relative expression level of TaEC after T. controversa inoculation. (E ) The relative expression level of TaEC after T. laevis inoculation.