Figure 1.
Schematic representation of Datura stramonium as a differential host to Iris yellow spot virus (IYSV) and Tomato spotted wilt virus (TSWV).
(A) IYSV infection is localized in inoculated leaves; (B) TSWV causes systemic infection; (C) Co-inoculation of IYSV and TSWV results in severe systemic infection.
Figure 2.
Datura stramonium is a restrictive host to infection by Iris yellow spot virus (IYSV).
(A) Inoculated leaves with local lesions as a result of virus infection; (B) younger, uninoculated leaves of the same plant remain symptomless and virus-free; (C) Detection of IYSV genes in inoculated leaves of D. stramonium using reverse transcription-polymerase chain reaction. Total RNA from TSWV inoculated leaf (1) and healthy plant leaf as negative control (2) was used as a template for amplifying the genes using specific primers as described in Table 1. M: 1 kb Marker; N: IYSV nucleocapsid (N) protein gene; NSs: IYSV nonstructural (NSs) protein gene; NSm: IYSV nonstructural (NSm) protein gene; GN/GC: IYSV glycoprotein precursor (GN/GC); RdRp: IYSV RNA dependent RNA polymerase (RdRp).
Table 1.
Detection of Iris yellow spot virus (IYSV) and Tomato spotted wilt virus (TSWV) in Datura stramonium plants.
Figure 3.
Datura stramonium is a permissive host for Tomato spotted wilt virus (TSWV).
D. stramonium infected with TSWV (A) Inoculated leaves with local lesions as a result of virus infection; (B) Younger, uninoculated leaves of the same plant shows severe necrotic symptoms; (C) Detection of TSWV genes in inoculated leaves of D. stramonium using reverse transcription-polymerase chain reaction. Total RNA from TSWV inoculated leaf (1) and healthy plant leaf as negative control (2) was used as a template for amplifying the genes using specific primers as described in Table 1. M: 1 kb Marker; N: IYSV nucleocapsid (N) protein gene; NSs: IYSV nonstructural (NSs) protein gene; NSm: IYSV nonstructural (NSm) protein gene; GN/GC: IYSV glycoprotein precursor (GN/GC); RdRp: IYSV RNA dependent RNA polymerase (RdRp).
Figure 4.
Datura stramonium plants co-inoculated with Iris yellow spot virus (IYSV) and Tomato spotted wilt virus (TSWV).
(A) Leaves inoculated with both IYSV and TSWV Dual infection results in more severe systemic symptoms, necrotic spots and concentric rings on inoculated leaves; (B) systemic leaves from dual inoculated plants showing symptoms including leaf curling, severe venial chlorosis, and yellowing on younger un-inoculated leaves; (C) Detection of IYSV genes in systemic leaves of D. stramonium co-inoculated with TSWV and IYSV using reverse transcription-polymerase chain reaction., (D) Detection of TSWV genes in systemic leaves of D. stramonium co-inoculated with TSWV and IYSV using reverse transcription-polymerase chain reaction; Lane M: Marker; N: nucleocapsid (N) protein gene; NSs: nonstructural (NSs) protein gene; NSm: nonstructural (NSm) protein gene; GN/GC: glycoprotein precursor (GN/GC); RdRp: RNA dependent RNA polymerase (RdRp);.
Table 2.
List of primers used in reverse transcription-polymerase chain reaction.
Figure 5.
Detection of nucleocapsid (N) gene and non-structural (NSs) genes in inoculated and uninoculated systemic leaves of Datura stramonium plants infected with Iris yellow spot virus (IYSV) and Tomato spotted wilt virus (TSWV) in plants inoculated with either IYSV, TSWV or both.
Figure 6.
Expression of N and NSs genes of Iris yellow spot virus (IYSV) and Tomato spotted wilt virus (TSWV) in single or dually infected Datura stramonium plants using gene-specific cDNA probes.
Leaf samples were harvested 15–20 days post inoculation from inoculated and systemic leaves. Lanes: 1-Healthy, uninfected control; 2-IYSV only inoculated leaf; 3-IYSV only systemic leaf; 4-TSWV only inoculated leaf; 5-TSWV only systemic leaf; 6-IYSV+ TSWV-inoculated leaf; 7-IYSV+ TSWV systemic leaf.
Figure 7.
Small interfering (si) RNAs of N and NSs genes of Tomato spotted wilt virus (TSWV) in single or dually infected Datura stramonium plants.
The leaf samples were harvested 25–30 days after inoculation from inoculated and systemic leaves A: TSWV N gene specific small interfering (Si) RNAs B: TSWV NSs gene-specific siRNAs, U6: Ubiquitin as loading control for RNA. Lanes: 1-Healthy, uninfected control; 2- IYSV-inoculated leaf; 3- IYSV systemic leaf; 4-Blank; 5-IYSV+ TSWV-inoculated leaf; 6-IYSV+ TSWV systemic leaf; 7-Blank; 8-TSWV-inoculated leaf; 9-TSWV systemic leaf.