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Figure 4 | BMC Plant Biology

Figure 4

From: Agrobacterium-derived cytokinin influences plastid morphology and starch accumulation in Nicotiana benthamiana during transient assays

Figure 4

LBA transformed with 4.3 kb tzs gene from pMP90 induces a GVR-like phenotype. Four different LBA/pLSU-ptzs-tzs transformants (LtZ1-4) were infiltrated into the lower epidermis of three different N. benthamiana plants. A Bar graph of average stromule frequency (SF) in non-infiltrated (NI) LBR-infiltrated (LBR), LtZ-infiltrated (LtZ1-4) and GVR-infiltrated (GVR) tissue. Rank sum (NI-LtZ1, 2, 3, 4):U=0, p<0.001; (LtZ1-LBR, LtZ2-LBR): U=7, p<0.001; (LtZ3-LBR): U=8, p<0.001; (LtZ4-LBR): U=4, p<0.001); (LtZ1-GVR): U=39, p=0.061; (LtZ2-GVR): U=40, p=0.069; (LtZ4-GVR): U=62, p=0.829; (LtZ3-GVR): U=32, p=0.039. Sample sizes: n(GV, NI, LtZ1, LtZ2)=12, n(LtZ3, LtZ4)=11. Raw data were arcsine transformed and bars represent back-transformed means. Error bars represent back-transformed 95% confidence intervals. B Box plot representing median stromule lengths in non-infiltrated (NI) tissue, LBR-infiltrated (LBR), LtZ-infiltrated (LtZ1-4) and GVR-infiltrated (GVR) tissues. Rank sum (LtZ1-GVR): U=54007, p<0.001; (LtZ2-GVR): U=51988, p<0.001; (LtZ3-GVR): U=53068.5, p=0.015; (LtZ4-GVR): U=50161, p<0.001; (LtZ1-LBR): U=38767, p<0.001; (LtZ2-LBR): U=39391.5, p<0.001; (LtZ3-LBR): U=33777, p<0.001; (LtZ4-LBR): U=35246.5, p<0.001; (LtZ1-NI): U=21007, p<0.001; (LtZ2-NI): U=21005, p<0.001; (LtZ3-NI): U=18181.5, p<0.001; (LtZ4-NI): U=19019.5, p<0.001. Sample sizes: n(NI)=235, n(LBR)=308, n(LtZ1)=360, n(LtZ2)=360, n(LtZ3)=330, n(LtZ4)=330, n(GVR)=360. Boxes contain 50% of data, the median is represented by black line. Error bars represent 90% confidence intervals. C ‘Stacked’ fluorescence image of extreme clustering of plastids around the nucleus three days post-infiltration with LtZ4. The cell is outlined in red and the nucleus is indicated by ‘n’. Image was converted to gray scale and inverted to better visualize plastids. Scale=20μm. D Iodine stained leaf three days after infiltration with GVR, LtZ strains 1–4, LBR and non-infiltrated control (NI). Infiltrated areas marked by a black dotted outline. E LC/MS/MS measurements of absolute trans-zeatin in fresh N. benthamiana leaf tissue 3 days post-infiltration with AIM buffer (AI), GVC7 (GVC7), LBR (LBR), GVR (GVR), and LtZ4 (LtZ4). Rank sum (GVR-NI, GVR-AI, GVR-GVC7, GVR-LBR, LtZ4-NI, LtZ4-AI, LtZ4-GVC7): U=0, p=0.029; (LtZ4-GVR): U=0, p=0.029. Sample size: n=4 for each. F LC/MS/MS measurements of absolute trans-zeatin-9-riboside in fresh N. benthamiana leaf tissue 3 days post-infiltration with AIM buffer (AI), GVC7 (GVC7), LBR (LBR), GVR (GVR), and LtZ4 (LtZ4). Rank sum (for all pair wise comparisons): U=0, p=0.029). Sample size: n=4 for each.

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