Additional file 1 of Non-transgenic, PAMAM co-delivery DNA of interactive proteins NbCRVP and NbCalB endows Nicotiana benthamiana with a stronger antiviral effect to RNA viruses
Creators: name: Song, Liyun
name: Zhang, Daoshun
name: Liu, Tianbo
name: Jiang, Changqing
name: Li, Bin
name: Li, Changquan
name: Shen, Lili
name: Li, Ying
name: Wang, Fenglong
name: Jiao, Yubing
name: Yang, Jinguang
Publisher: name: figshare
Year: 2024
Subjects: subject: Biophysics
subject: Biochemistry
subject: Medicine
subject: Microbiology
subject: FOS: Biological sciences
subject: Genetics
subject: Molecular Biology
subject: Biotechnology
subject: Chemical Sciences not elsewhere classified
subject: Cancer
subject: Infectious Diseases
subject: FOS: Health sciences
subject: Plant Biology
subject: Virology

description: Additional file 1: Figure S1. The expression pattern of NbCRVP. (a) Expression of NbCRVP under the stress of viral infection at 1, 3, 5, 7 dpi. The data were analyzed by Duncan’s multiple range tests in the ANOVA program of SPSS, different letters indicate that values of the four treatments were significantly different (P &lt; 0.05), which were the same as b, c, d. (b) The NbCRVP expression trends in flower, stem, leaf and root. (c) The NbCRVP expression trends after spraying with 0.5 mM SA, 0.05 mM ethephon, or 0.1 mM Me-JA, water with 0.02% Tween 20 was used as a control. (d) The expression of NbCRVP after silencing the key signaling genes, NPR1, COI1 and EIN2. (e) Subcellular distribution of NbCRVP observed in healthy N. benthamiana and virus infected N. benthamiana. Figure S2. Subcellular distribution of NbCRVP using DAPI as a nuclear localization marker. Figure S3. The silencing efficiency of NbCRVP. (a) Phenotype after 14 days of silencing NbCRVP. (b) The silencing efficiency was detected after 14 days of treatment. Figure S4 Overexpression NbCRVP on multi-infection of TMV, PVY and CMV. (a) The detection of NbCRVP after overexpression NbCRVP. The data were analyzed with independent sample t test, * indicated that values of the two treatments were significantly different (P &lt; 0.05), the same as b. (b) Effect of overexpression NbCRVP on multi-infection of TMV, PVY and CMV detected by RT-qPCR at 1, 2, 3, 4 dpi. (c) Differences in biological symptoms of overexpression NbCRVP on multi-infection of TMV, PVY and CMV. Figure S5 Screening of interactive proteins for NbCRVP in yeast two-hybrid assay. (a) Screening of interacting proteins by dot culture on QDO/X/A. (b) PCR amplification, gel electrophoresis, and sequencing to identify interacting proteins. Figure S6 SEM and TEM images of PAMAM@CRVP nanocomposites. Figure S7 Detect the expression of PAMAM@CRVP:CalB in the Solanum lycopersicum and Capsicum annuum protoplast. (a) Detect the expression of PAMAM@CRVP:CalB in Solanum lycopersicum. (b) Detect the expression of PAMAM@CRVP:CalB in Capsicum annuum. Figure S8 In vitro shelf life and delivery expression of PAMAM@pDNA nanocomplexes in plant cells. (a) Monitoring the delivery and expression of PAMAM@pDNA nanocomplex in plant cells under laser confocal. (b) Monitor the stability of PAMAM@pDNA nanocomplexes in vitro. Figure S9 Characterization of PAMAM@CRVP:CalB nanocomposites. (a) Average particle size distribution of PAMAM@CRVP:CalB nanocomposites. (b) Intensity distribution of Zeta potential of PAMAM@CRVP:CalB nanocomposites. Figure S10 Toxicity assay of PAMAM@CRVP:CalB nanocomposites. (a) Dry cell weight determination treated with PAMAM@CRVP:CalB nanocomposites. (b) cell mortality rate determination treated with PAMAM@CRVP:CalB nanocomposites. Figure S11 Validation that PAMAM-mediated resistance protein delivery was non-transgenic. Lane 1 was PAMAM@CRVP:CalB-treated wild-type N. benthamiana seeds, Lane 2 was PAMAM@CRVP:CalB-treated NbCRVP-OE seeds.