Generation and identification of amylose-only barley. (a) Chimeric RNAi hairpin construct simultaneously targeting the three different SBE genes SBEI, SBEIIa and SBEIIb. Expression was driven by the maize Ubiquitin promoter. Promoter and intron are not drawn to scale. The actual length of the intron is 1290 bp. The amplification product of the primer set Hairpin Fw and Hairpin Rev, which specifically recognizes the hairpin construct is indicated (b) Relative gene expression levels of the three SBEs isoforms (SBEI, SBEIIa and SBEIIb) assessed by RT qPCR in three individual grains, A, B and C at (20DAP) each of control and transgenic T1 lines (three technical replicates each). SE bars are indicated. (c) SBE enzyme activity in developing endosperm of SBE RNAi 4.1 and control grains, based on the average value of 3 experiments. Bars indicate standard error (d) Size exclusion chromatography (SEC) profile of starch from control and SBE RNAi4.1 lines. Black lines show the elution profile determined measuring the total sugar content of each fraction. λ-max absorbance of the α-glucan-iodine complex in each fraction is indicated by grey dots.
Carciofi et al. BMC Plant Biology 2012 12:223 doi:10.1186/1471-2229-12-223