caa a22 4500 46789387X CHVBK 20180406152756.0 cr unu---uuuuu 170328e20060301xx s 000 0 eng 10.1007/s11103-005-5280-6 doi (NATIONALLICENCE)springer-10.1007/s11103-005-5280-6 Post-transcriptional Gene Silencing of GBSSI in Potato: Effects of Size and Sequence of the Inverted Repeats [Elektronische Daten] [H. Heilersig, A. Loonen, M. Bergervoet, A. Wolters, R. Visser] In the past, silencing of granule-bound starch synthase (GBSSI) in potato was achieved by antisense technology, where it was observed that inclusion of the 3′ end of the GBSSI coding region increased silencing efficiency. Since higher silencing efficiencies were desired, GBSSI inverted repeat constructs were designed and tested in potato. First, large inverted repeats comprising the 5′ and the 3′ half of the GBSSI cDNA were tested. The 5′ IR construct gave a significantly higher silencing efficiency than the 3′ IR construct. Since it was not known whether the observed difference was due to the sequence or the orientation of the inverted repeat, the GBSSI cDNA was divided into three regions, after which each region was tested in small inverted repeats in two orientations. To this end large numbers of independent transformants were produced for each construct. The results suggested that there was no effect of inverted repeat orientation on silencing efficiency. The percentage of transformants showing strong inhibition varied from 48% for a 3′-derived construct to 87% for a 5′ as well as a middle region-derived construct. Similar to the large inverted repeats, the 3′ sequences induced the least efficient silencing implying that the observed differences in silencing efficiency are caused by sequence differences. The small inverted repeat constructs with a repeat size of 500-600bp and a spacer of about 150bp were more efficient silencing inducers than the large inverted repeat constructs where the size of the repeat was 1.1 or 1.3kb whilst the size of spacer was 1.3 or 1.1kb. The results presented here show that size and sequence of the inverted repeat influenced silencing efficiency. Springer, 2006 dsRNA nationallicence granule-bound starch synthase nationallicence inverted repeat nationallicence post-transcriptional gene silencing nationallicence potato nationallicence dsRNA : double-stranded RNA nationallicence GBSSI : granule-bound starch synthase I nationallicence IR : inverted repeat nationallicence PTGS : post-transcriptional gene silencing nationallicence RISC : RNA-induced silencing complex nationallicence siRNA : small interfering RNA nationallicence UTR : untranslated region nationallicence VIGS : virus-induced gene silencing nationallicence Heilersig H. Graduate School Experimental Plant Sciences, Laboratory of Plant Breeding, Department of Plant Sciences, Wageningen University, P.O. Box 386, 6700, Wageningen, AJ, The Netherlands aut Loonen A. Graduate School Experimental Plant Sciences, Laboratory of Plant Breeding, Department of Plant Sciences, Wageningen University, P.O. Box 386, 6700, Wageningen, AJ, The Netherlands aut Bergervoet M. Graduate School Experimental Plant Sciences, Laboratory of Plant Breeding, Department of Plant Sciences, Wageningen University, P.O. Box 386, 6700, Wageningen, AJ, The Netherlands aut Wolters A. Graduate School Experimental Plant Sciences, Laboratory of Plant Breeding, Department of Plant Sciences, Wageningen University, P.O. Box 386, 6700, Wageningen, AJ, The Netherlands aut Visser R. Graduate School Experimental Plant Sciences, Laboratory of Plant Breeding, Department of Plant Sciences, Wageningen University, P.O. Box 386, 6700, Wageningen, AJ, The Netherlands aut Plant Molecular Biology Kluwer Academic Publishers 60/5(2006-03-01), 647-662 0167-4412 60:5<647 2006 60 11103 https://doi.org/10.1007/s11103-005-5280-6 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11103-005-5280-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Heilersig H. Graduate School Experimental Plant Sciences, Laboratory of Plant Breeding, Department of Plant Sciences, Wageningen University, P.O. Box 386, 6700, Wageningen, AJ, The Netherlands aut NATIONALLICENCE 700 1- Loonen A. Graduate School Experimental Plant Sciences, Laboratory of Plant Breeding, Department of Plant Sciences, Wageningen University, P.O. Box 386, 6700, Wageningen, AJ, The Netherlands aut NATIONALLICENCE 700 1- Bergervoet M. Graduate School Experimental Plant Sciences, Laboratory of Plant Breeding, Department of Plant Sciences, Wageningen University, P.O. Box 386, 6700, Wageningen, AJ, The Netherlands aut NATIONALLICENCE 700 1- Wolters A. Graduate School Experimental Plant Sciences, Laboratory of Plant Breeding, Department of Plant Sciences, Wageningen University, P.O. Box 386, 6700, Wageningen, AJ, The Netherlands aut NATIONALLICENCE 700 1- Visser R. Graduate School Experimental Plant Sciences, Laboratory of Plant Breeding, Department of Plant Sciences, Wageningen University, P.O. Box 386, 6700, Wageningen, AJ, The Netherlands aut NATIONALLICENCE 773 0- Plant Molecular Biology Kluwer Academic Publishers 60/5(2006-03-01), 647-662 0167-4412 60:5<647 2006 60 11103 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer