Popelka Juan Carlos, Altpeter Fredy
Institut für Pflanzengenetik und Kulturpflanzenforschung Gatersleben, Corrensstrasse 3, 06466 Gatersleben, Germany.
Theor Appl Genet. 2003 Aug;107(4):583-90. doi: 10.1007/s00122-003-1314-0. Epub 2003 Jun 26.
The efficient and reproducible production of stably expressing transgenic rye plants is described. Analysis of the genotype-specific callus culture-response of 21 rye inbred lines, single crosses and a population variety resulted in the identification of the highly responsive inbred line L22. Biolistic transformation experiments were performed using line L22 and the impact of different selection agents on the regeneration capacity was analyzed. Using the selectable marker gene nptII and corresponding paromomycin selection resulted in transformation efficiencies of up to 4.0% of the bombarded explants. A total of 17 independent transgenic rye plants were produced, and their stability and level of transgene expression was analyzed. The majority of these lines showed stable transgene expression. In contrast a few transgenic lines with multiple transgene inserts, provided evidence of transcriptional and post-transcriptional gene silencing.
本文描述了稳定表达转基因黑麦植株的高效且可重复的生产方法。对21个黑麦自交系、单交种和群体品种的基因型特异性愈伤组织培养反应进行分析，鉴定出了高反应性自交系L22。使用L22系进行了基因枪转化实验，并分析了不同选择剂对再生能力的影响。使用选择标记基因nptII和相应的巴龙霉素选择，转化效率高达轰击外植体的4.0%。共产生了17株独立的转基因黑麦植株，并分析了它们转基因表达的稳定性和水平。这些品系中的大多数表现出稳定的转基因表达。相比之下，一些具有多个转基因插入的转基因品系显示出转录和转录后基因沉默的证据。
