caa a22 4500 467900132 CHVBK 20180406152815.0 cr unu---uuuuu 170328e20060901xx s 000 0 eng 10.1007/s00299-006-0153-2 doi (NATIONALLICENCE)springer-10.1007/s00299-006-0153-2 Stability of transgene integration and expression in subsequent generations of doubled haploid oilseed rape transformed with chitinase and β-1,3-glucanase genes in a double-gene construct [Elektronische Daten] [Margareta Melander, Iréne Kamnert, Ingrid Happstadius, Erland Liljeroth, Tomas Bryngelsson] A double-gene construct with one chitinase and one β-1,3-glucanase gene from barley, both driven by enhanced 35S promoters, was transformed into oilseed rape. From six primary transformants expressing both transgenes 10 doubled haploid lines were produced and studied for five generations. The number of inserted copies for both the genes was determined by Southern blotting and real-time PCR with full agreement between the two methods. When copy numbers were analysed in different generations, discrepancies were found, indicating that at least part of the inserted sequences were lost in one of the alleles of some doubled haploids. Chitinase and β-1,3-glucanase expression was analysed by Western blotting in all five doubled haploid generations. Despite that both the genes were present on the same T-DNA and directed by the same promoter their expression pattern between generations was different. The β-1,3-glucanase was expressed at high and stable levels in all generations, while the chitinase displayed lower expression that varied between generations. The transgenic plants did not show any major impact on fungal resistance when assayed in greenhouse, although purified β-1,3-glucanase and chitinase caused retardment of fungal growth in vitro. Springer-Verlag, 2006 Brassica napus nationallicence Doubled haploids nationallicence Expression stability nationallicence Gene silencing nationallicence Transgene deletion nationallicence Melander Margareta Svalöf Weibull AB, 268 81, Svalöv, Sweden aut Kamnert Iréne Svalöf Weibull AB, 268 81, Svalöv, Sweden aut Happstadius Ingrid Svalöf Weibull AB, 268 81, Svalöv, Sweden aut Liljeroth Erland Department of Crop Science, Swedish University of Agricultural Sciences, P.O. Box 44, 230 53, Alnarp, Sweden aut Bryngelsson Tomas Department of Crop Science, Swedish University of Agricultural Sciences, P.O. Box 44, 230 53, Alnarp, Sweden aut Plant Cell Reports Springer-Verlag 25/9(2006-09-01), 942-952 0721-7714 25:9<942 2006 25 299 https://doi.org/10.1007/s00299-006-0153-2 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00299-006-0153-2 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Melander Margareta Svalöf Weibull AB, 268 81, Svalöv, Sweden aut NATIONALLICENCE 700 1- Kamnert Iréne Svalöf Weibull AB, 268 81, Svalöv, Sweden aut NATIONALLICENCE 700 1- Happstadius Ingrid Svalöf Weibull AB, 268 81, Svalöv, Sweden aut NATIONALLICENCE 700 1- Liljeroth Erland Department of Crop Science, Swedish University of Agricultural Sciences, P.O. Box 44, 230 53, Alnarp, Sweden aut NATIONALLICENCE 700 1- Bryngelsson Tomas Department of Crop Science, Swedish University of Agricultural Sciences, P.O. Box 44, 230 53, Alnarp, Sweden aut NATIONALLICENCE 773 0- Plant Cell Reports Springer-Verlag 25/9(2006-09-01), 942-952 0721-7714 25:9<942 2006 25 299 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer