穗茎注射法导入反义PLDγ基因小麦的分子检测

Molecular Detection of Wheat Transformed with Anti-PLD_γ Gene by Earstem Injecting

利用穗茎注射法将反义PLDγ(AntisensephospholipaseDγ)基因导入普通小麦兰考906,并用特异PCR标记和PCR-southern杂交技术对所获得的变异系进行了分子检测,旨在探讨该基因在小麦中的整合效果。结果表明:(1)经反义PLDγ基因的特异PCR标记,变异系03039、03048和03050扩增出与阳性质粒大小相同的400bp的目的片断,PCR-southern杂交结果与其一致。说明该基因已整合到小麦的基因组中,从而获得了转反义PLDγ基因的小麦。(2)转基因株系的成熟期比对照提前4～8d,后代中易产生雄性不育现象。

Antisense phospholipase D_γ (PLD_γ) gene was used to transform wheat Lankao 906 mediated by earstem injecting. Many transgenic spikes were detected by PCR and PCR-southern test in order to count the validity of integration. The results (indicated) that(1)A 400 bp fragment was amplified from three (individual) plants, 03039, 03048 and 03050, which was the same as that from the plasmid. Thus, we (primarily) (confirmed) that Anti-PLD_γ gene was integrated into wheat genomes and transgenic lines were (obtained).(2)The mature period of transgenic lines was 4 to 8 days shorter than that of receptor and Male sterility was (observed) in the following generations.