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. 2006 Feb;140(2):681–692. doi: 10.1104/pp.105.074906

Figure 4.

Figure 4.

PVS3 promoter is controlled by both SIPK and WIPK. A, Scheme of reporter and effector plasmids used in transient assays. The reporter plasmid PVS3 promoter fragment (positions −2,334 to +30) was translationally fused to the GUS gene containing intron (GUSint). The effector plasmid estradiol-inducible promoter was fused to the StMEK1DD gene. B, Half leaf of gene-silenced N. benthamiana was coinfiltrated with a mixture of Agrobacterium harboring PVS3::GUSint (reporter) or pER8::StMEK1DD (effector). Estradiol (10 μm) was injected 48 h after agroinfiltration to activate the effector. GUS activities driven by PVS3 promoter in response to StMEK1DD were measured in gene-silenced N. benthamiana by PVX (PVX-control [PVX], WIPK [PVX:W], SIPK [PVX:S], SIPK/WIPK [PVX:S/W]) 24 h after estradiol injection. Results are presented as relative values calibrated by GUS activity, which were measured by fluorometric quantitation of 4-MU, under the control of 35S promoter in another half leaf. The PVX-control with StMEK1DD was arbitrarily assigned as 100% value against which all other values were plotted. C, Expression profiles of EAS in the gene-silenced N. benthamiana. Total RNA was isolated from gene-silenced N. benthamiana leaves after infiltration with Agrobacterium harboring 35S::StMEK1DD, and transcript levels of WIPK, SIPK, and EAS were determined by RNA gel-blot hybridization.