小麦编码高分子量谷蛋白亚基基因的转化

Transformation of Genes Encoding High Molecular Weight Glutenin Subunit (HMW-GS)into Common Wheat (Triticum aestivum L. )

以小麦的幼穗和幼胚作为转化受体,首次用抗除草剂草甘膦的EPSPs基因作为选择标记,通过基因枪法共转化,将小麦编码高分子量谷蛋白亚基的基因1Dx5和1Dy10转移到普通小麦京花1号中,获得33株再生植株,经PCR初步检测有12株同时扩增到了3个处在不同质粒上的外源基因EP-SPs、1Dx5和1Dy10的目标片段。将部分PCR检测为阳性的转化体进行Southern分析,发现其中有1株再生植株基因组中整合了3个外源基因,有2株的基因组中同时整合了EPSPs和1Dx5基因,1株的基因组中整合了EPSPs和1Dy10基因。此外,在转化过程中发现,幼穗是比幼胚更为合适的转化受体,并且转化受体具有合适的转化时期。

Two separate plasmids, harboring 1Dx5 gene encoding wheat high molecular weight glutenin subunit (HMW-GS) and 1Dy10 gene emcoding wheat HMW-GS 1Dy10 respectively, were first co-transformed with the EPSPs gene as selectable marker via particle bombardment. Tweleve of 33 regenerated plants obtained were amplified target fragments of three separate plasmids, harboring EPSPs, 1Dx5 and 1Dy10 genes respectively, by PCR analysis. Part transfor-mants integrated three foreign genes scored by PCR were further comfirmed by Southern blotting. One of these transformants was integrated into its genome(s) with three foreign genes, two with EPSPs and 1Dx5 genes, one with EPSPs and 1Dy10 genes, Furthermore, immature spikes were found more suitable for foreign gene transformation to direct DNA delivery. The results also indicated the target materials have their own suitable transformation periods.