Tsw gene-based resistance is triggered by a functional RNA silencing suppressor protein of the Tomato spotted wilt virus - PubMed (original) (raw)

Enzyme‐linked immunosorbent assay (

ELISA

) measuring virus titres of different Tomato spotted wilt virus (

TSWV

) isolates and their ability to suppress green fluorescence protein (

GFP

) silencing. (a)

NSs

titres of

TSWV

resistance‐inducing (

RI

) and resistance‐breaking (

RB

) isolates were measured by antigen‐coated plate (

ACP)‐ELISA

and are shown relative to the virus titres measured by double antibody sandwich (

DAS)‐ELISA

. The average of four repetitions is shown, demonstrating that there is no significant difference between these isolates. (b) Western immunoblot detection of

NSs

in Nicotiana benthamiana, transiently expressed from p

EAQ

‐

HT

or after virus infection. Transiently expressed

NSs

shows a difference in amount between the

RI

(from

V

ir129) and

RB

isolates, whereas, in the virus setting, similar amounts can be observed. In addition, in both the transient setting and viral setting, a size difference between the

RI

and the

RB

isolates can be observed.

ATTA

, Agrobacterium tumefaciens transient transformation assay. (c) Local transient

GFP

expression was superimposed with a mechanical

TSWV

inoculation on the same leaf. Photographs were taken at 5 days post‐inoculation (dpi), and show that all isolates are able to suppress local

GFP

silencing. As a negative control, Potato virus X (

PVX

) was used, which does not show local suppression of

GFP

silencing.