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. 2011 Nov 23;6(11):e28114. doi: 10.1371/journal.pone.0028114

Figure 2. Generation of transgenic Arabidopsis thaliana lines with accumulation of siRNAs derived from transgene constructs.

Figure 2

(A) Schematic representation of the silencing constructs in the pART27 binary vector. The selected regions of GFP (612 bp) and PnPMA1 (262 bp) were cloned in an inverted-repeat configuration using the appropriate restriction enzymes into the sites spanned by 35 S promoter and OCS terminator regions in pKANNIBAL, respectively. The expression cassettes were subsequently transferred into the binary vector pART27, respectively. Accumulation of siRNAs in independent T2 A. thaliana lines transformed with (B) GFP dsRNA construct (lanes 1–8, T2 individuals from each of 8 independent T1 transgenic plants) (b1) or (C) PnPMA1 dsRNA (lanes 1–4, T2 individuals from each of 4 independent T1 transgenic plants) construct; WT, wild-type A. thaliana; M, RNA size marker, 21, 24 and 30 nt from bottom up, respectively. Lower panel indicates ethidium bromide staining of total RNAs to show equal loading of RNA samples.