Additional file 1 of Cucumber mosaic virus-induced gene and microRNA silencing in water dropwort (Oenanthe javanica (Blume) DC)
Creators: name: He, Zhen
name: Sheng, Shuangyu
name: Wang, Lingqi
name: Dong, Tingting
name: Zhang, Kun
name: Li, Liangjun
Publisher: name: figshare
Year: 2024
Subjects: subject: Genetics
subject: FOS: Biological sciences
subject: Plant Biology

description: Additional file 1: Figure S1. Construction of pCB301-CMV-RNA2△2b. (A) Map of pCB301-RNA2 and pCB301-RNA2△2b. (B) Inverse-PCR detection of CMV-RNA2. Figure S2. The CMV-Fny 2b deletion mutant infected N. benthamiana by agro-infection. (A) Symptoms of the CMV-Fny 2b deletion mutant in N. benthamiana. Scale bars are 2 cm. (B) Detection of CMV-Fny 2b deletion mutant-infected N. benthamiana by agro-infection (8 dpi). The vector plasmid with the CMV RNA3 insert was amplified as the positive (lane P) control. Figure S3. Silencing of the NbPDS gene in N. benthamiana using the CMV VIGS vector. (A) Construct diagram of infectious clones of pCB301CMV-NbPDS. (B) RT‒PCR detection of the PDSC of N. benthamiana. (C) Phenotypes of pCB301CMV-NbPDS in N. benthamiana (14 dpi). Scale bars are 2 cm. (D) Detection of pCB301CMV-NbPDS-infected N. benthamiana by agro-infection (14 dpi). (E) RT‒qPCR analysis of NbPDS expression levels in CMV△2b- or CMV-NbPDS-infected plants. Figure S4. Identification of PDS gene in O. javanica. (A) Phylogenetic analysis of PDS of O. javanica. (B) Map of PDS gene of O. javanica. (C) Silencing Sequence of PDS gene in O. javanica. Figure S5. Identification of GID1 gene in O. javanica. (A) Phylogenetic analysis of GID1 of O. javanica. (B) Map of GID1 gene of O. javanica. (C) Silencing Sequence of GID1 gene in O. javanica. Figure S6. Silencing of the OjGID1 gene in O. javanica using the CMV VIGS vector. (A) Phenotypes of pCB301CMV-OjGID1 in O. javanica (30 dpi). Scale bars are 2 cm. (B) Systematic leaf morphology analysis of pCB301CMV-OjGID1 in ‘Fq1’ (30 dpi). (C) Detection of pCB301CMV-OjGID1-infected ‘Fq1’ by agro-infection (30 dpi). (D) RT‒qPCR analysis of OjGID1 expression levels in CMV△2b- or CMV-OjGID1-infected plants. (E) RT‒qPCR analysis of GA-related genes expression in CMV△2b- or CMV-OjGID1-infected plants. Figure S7. RT‒qPCR analysis of 6 OjGID1 genes expression in CMV△2b- or CMV-OjGID1-infected O. javanica. (A) RT‒qPCR analysis of 6 OjGID1 genes expression in CMV△2b- or CMV-OjGID1-infected ‘Yzcbq’. (B) RT‒qPCR analysis of 6 OjGID1 genes expression in CMV△2b- or CMV-OjGID1-infected ‘Fq1’. Figure S8. Silencing of the NbGID1 gene in N. benthamiana using the CMV VIGS vector. (A) Phenotypes of pCB301CMV-NbGID1 in N. benthamiana (17 dpi). Scale bars are 2 cm. (B) Systematic leaf morphology analysis of pCB301CMV-NbGID1 in N. benthamiana (17 dpi). (C) Detection of pCB301CMV-NbGID1-infected N. benthamiana by agro-infection. (D) RT‒qPCR analysis of NbGID1 expression levels in CMV△2b- or CMV-NbGID1-infected plants. (E) RT‒qPCR analysis of NbGID1 expression levels in CMV△2b- or CMV-NbGID1-infected plants. Figure S9. Silencing of miR319 in O. javanica using CMV-based VbMS vector. (A) Phenotypes of plants inoculated with CMV-STTM319 at 30 dpi. Scale bars are 2 cm. (B) Systematic leaf morphology analysis of pCB301CMV-STTM319 in ‘Yzcbq’. (C) Detection of pCB301CMV-STTM319-infected ‘Yzcbq’ by agro-infection. (D) Stem‒loop RT‒qPCR analysis of miR319 expression levels in CMV△2b- or CMV-STTM319-infected plants. (E) Detection of the relative expression levels of the miR319 target genes TCP2 and TCP4. Figure S10. Silencing of miR319 in N. benthamiana using CMV-based VbMS vector. (A) Phenotypes of plants inoculated with CMV-STTM319 at 15 dpi. Scale bars are 2 cm. (B) Systematic leaf morphology analysis of pCB301CMV-STTM319 in N. benthamiana. (C) Detection of pCB301CMV-STTM319-infected N. benthamiana by agro-infection. (D) Stem‒loop RT‒qPCR analysis of miR319 expression levels in CMV△2b- or CMV-STTM319-infected plants. (E) Detection of the relative expression levels of the miR319 target genes TCP2 and TCP4. Figure S11. Silencing of miR396 in O. javanica using CMV-based VbMS vector. (A) Phenotypes of plants inoculated with CMV-STTM396 at 30 dpi. Scale bars are 2 cm. (B) Systematic leaf morphology analysis of pCB301CMV-STTM396 in ‘Yzcbq’. (C) Detection of pCB301CMV-STTM396-infected ‘Yzcbq’ by agro-infection. (D) Stem‒loop RT‒qPCR analysis of miR396 expression levels in CMV△2b- or CMV-STTM396-infected plants. (E) Detection of the relative expression level of the miR396 target gene Growth-Regulating Factor 9-like. Figure S12. Silencing of miR396 in N. benthamiana using CMV-based VbMS vector. (A) Phenotypes of plants inoculated with CMV-STTM396 at 15 dpi. Scale bars are 2 cm. (B) Systematic leaf morphology analysis of pCB301CMV-STTM396 in N. benthamiana. (C) Detection of pCB301CMV-STTM396-infected N. benthamiana by agro-infection. (D) Stem‒loop RT‒qPCR analysis of miR396 expression levels in CMV△2b- or CMV-STTM396-infected plants. (E) Detection of the relative expression level of the miR396 target gene polyamine oxidase 1. Figure S13. Naturally infected of water dropwort by CMV. (A) Symptoms of CMV in O. javanica. (B) RT-PCR and western blot detection of the water dropwort samples.