农杆菌介导法将反义蜡质基因转入杂交稻亲本

Antisense waxy Gene Introduced into the Calli in Parents of Hybrid Rice by Agrobacterium tumefaciens

通过农杆菌介导法将反义蜡质基因导入特青(OryzasativaL.subsp.indica)和广粳1号(OryzasativaL.subsp.japonica)这两个杂交稻亲本的愈伤组织中,获得了经PCR检测证明外源基因已整合进植物基因组中的再生植株,其中含潮霉素抗性基因的植株有6株,但仅有2株能够扩增出反义蜡质基因的特异条带,其原因可能是在转基因植株中出现了基因丢失或沉默现象。试验表明,粳型稻广粳1号的不同外植体(成熟胚、幼胚、幼穗、花药)的愈伤诱导率比特青的高,且随着继代时间的延长,不同基因型水稻诱导的愈伤之间的转化率存在着极显著性差异(P<0.01);经过转染的水稻愈伤在一定的筛选压力下会出现“逃逸”现象,抗性筛选后成活愈伤PCR检测发现广粳1号和特青的愈伤转化率只有44.4%和29.4%;不同凝固剂对愈伤诱导的质量有着明显的影响,植物凝胶作凝固剂比琼脂作凝固剂时的愈伤褐化率低。

In order to improve the quality of the rice, we had tansferred the anti-sense waxy gene into calli in parents of super hybrid flee by Agrobacterium tumefaciens. Regeneration plants were obtained.The exogenous gene had been integrated into the genome of the regeneration plants by PCR analysis. We found that the frequence of transformation were significant in calli from different genotype rice. Meanwhile some factors affecting transformation efficiency of calli was studied in this paper. The ratio of all of explants including mature embryos, young embryos, young panicle and anther from japonica flee are higher than that from indica flee. The calli＇ s rate of brown which are culture on phytagel was lower than that on agar.