Scanning electron micrographs of the conidia germination and infection of <i>C</i>. <i>pseudoreteaudii</i> on <i>Eucalyptus</i> leaf.

<p>a-b, conidia germinated from both ends or septum at 6 hpi; c-d, each germ tube could produce multiple branches at 8 hpi; e-h, the ends of germ tubes expanded and invaded the leaves from the stoma at 12 hpi.</p

Transcriptomic Analysis of <i>Calonectria pseudoreteaudii</i> during Various Stages of <i>Eucalyptus</i> Infection

<div><p><i>Eucalyptus</i> leaf blight caused by <i>Calonectria</i> spp. is a serious disease in <i>Eucalyptus</i> seedling and plantations. However, the molecular mechanisms of the infection process and pathogenesis of <i>Calonectria</i> to <i>Eucalyptus</i> is not well-studied. In this study, we analyzed the transcriptomes of <i>C</i>. <i>pseudoreteaudii</i> at three stages of <i>Eucalyptus</i> leaf infection, and in mycelium grown in potato dextrose broth using Illumina RNA-Seq technology. We identified 161 differentially expressed genes between <i>C</i>. <i>pseudoreteaudii</i> from leaf and mycelium grown in potato dextrose broth. GO and KEGG enrichment analyses of these genes suggested that they were mainly involved in oxidoreductase activity, hydrolase activity, and transmembrane transporter activity. Most of the differentially expressed genes at the early infection stage were upregulated. These upregulated genes were mainly involved in cell wall hydrolysis and toxin synthesis, suggesting a role for toxin and cell wall hydrolases in the establishment of <i>Calonectria</i> leaf blight. Genes related to detoxification of phytoalexins were continually upregulated during infection. The candidate effectors and putative pathogenicity determinants identified in this study will help in the functional analysis of <i>C</i>. <i>pseudoreteaudii</i> virulence and pathogenicity.</p></div

Mode of hypha action of <i>C</i>. <i>pseudoreteaudii</i> on host cell.

<p>The fungal plasma membrane is shown in black, the fungal cytoplasm is shown in pink, the plant plasma membrane is shown in light green, and the plant cell wall is shown in dark green. The interface between the fungal hypha and the plant plasma membrane is shown as a light blue area. Fungal effectors and targeted plant substrate are shown in various colors. CWDE, cell wall degrading enzyme; PDE, phytoalexin degrading enzyme; ROS, reactive oxygen species; POD, peroxidase; MFS, major facilitator superfamily; SIT, siderophore iron transporter; SAH, salicylate hydroxylase; SA, salicylic acid.</p

Differentially expressed genes (DEG) of <i>C</i>. <i>pseudoreteaudii</i>.

<p>A, venn diagram showed common DEGs at three infection stages. B, number of DEGs at different colonization stages of <i>C</i>. <i>pseudoreteaudii</i>.</p

The 20 most enriched GO terms.

<p>Bar chart of DEGs enriched in GO term; it can directly reflect the number of DEGs distributing into different GO terms. CC, cellular component; MF, molecular function; BP, biological process.</p

List of the 20 most up-regulated genes of <i>C</i>. <i>pseudoreteaudii</i> during infection.

<p>List of the 20 most up-regulated genes of <i>C</i>. <i>pseudoreteaudii</i> during infection.</p

Summary of mapping results.

<p>Summary of mapping results.</p

Additional file 1 of Boosting cisplatin chemotherapy by nanomotor-enhanced tumor penetration and DNA adducts formation

Additional file 1: Figure S1. DLS analysis of F68@TA prepared by different mass ratio of F68 and TA. (n=3). Figure S2. TEM images of F68@TA@Ag NPs prepared with different content of AgNO3. Figure S3. (A) Size distributions and (B) zeta potentials of F68@TA, F68@TA/CDDP, and AINR solutions. Figure S4. Tyndall Effects of F68@TA/CDDP solution at different pHs for 1 h. Figure S5. DLS analysis of F68@TA/CDDP treated with PBS at different pHs for 48 h. Figure S6. HPLC chromatograms of different solvents and corresponsive CDDP (40 μg/mL) solution for 2 d. Figure S7. (A) Graphs and (B) UV-vis spectrum of F68@TA@Ag solutions with different concentration of H2O2. Figure S8. TBE-PAGE gel electrophoregram of DNA. 1: unmethylated single strand DNA; 2: unmethylated single strand DNA cut by restriction enzyme DnpII; 3: m1A methylated DNA; 4: m1A DNA sheared by DnpII. Figure S9. A) The tracking paths of F68@TA@Ag over 20 s in 0, 2.5, 5, 10, 25 and 50 mM H2O2. B) Average MSD versus time interval (Δt) analyzed from tracking trajectories. C) Corresponding diffusion coefficient values of F68@TA@Ag at various concentration of H2O2. (n=30). Figure S10. Fluorescence emission spectrums of FITC and RB dissolved in PBS at different pHs. Figure S11. A) Flow cytometry analysis and B) Fluorescence semi-quantitative analysis of 4T1 cells treated with F68@TA/RB for different time. Figure S12. A) CLSM images and B) Fluorescence semi-quantitative analysis of 4T1 cancer cells after treatments with F68@TA/RB (RB: 10 μg/mL) for different time. Scale bar: 25 μm. Figure S13. Fluorescence semi-quantitative analysis of H2O2 in 4T1 cells after treatment with different concentration of CDDP. Figure S14. Fluorescence semi-quantitative analysis of Cl- in 4T1 cells after treatment with CDDP, F68@TA, F68@TA@Ag and AINR. Figure S15. Fluorescence semi-quantitative analysis of Fe2+ in 4T1 cells after treatment with CDDP, F68@TA, F68@TA@Ag and AINR. Figure S16. Flow cytometry analysis of the fluorescence intensity in A) Hs578Bst cells and B) 4T1 cells treated with F68@TA/RB (RB: 5 μg/mL) for different time. C) Fluorescence semi-quantitative analysis of 4T1 cells and Hs578Bst cells. Figure S17. Fluorescence semi-quantitative analysis of Cl- in tumor tissues. Figure S18. Fluorescence semi-quantitative analysis of Fe2+ in tumor tissues. Figure S19. Body weight changes of different groups. (n = 5). Figure S20. H&E staining of tissues exfoliated from different groups. Figure S21. Serum biochemical index. A) Liver function and B) Kidney function index of mice treated with different formulation. 1: Control, 2: F68@TA, 3: F68@TA@Ag, 4: CDDP, 5: F68@TA/CDDP, 6: AINR, and concentration of CDDP was set as 1 μg/mL. Figure S22. The changes of tumor volume of 4T1 cell bearing mice with the treatment of AINR (CDDP: 2 mg/kg), 2 mg/kg CDDP, and 3 mg/kg CDDP. (n=5). Table S1. The DNA sequences of unmethylated DNA and methylated DNA