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利用正义RNAi技术提高玉米直链淀粉含量效果的研究

DOI: 10.7668/hbnxb.2010.04.020, PP. 92-96

Keywords: 玉米,淀粉分支酶,正义RNAi,直链淀粉

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Abstract:

为了研究利用正义RNAi技术提高玉米直链淀粉含量效果,用基因枪法将构建的sbeⅡb正义RNAi表达载体pBAC06和pBAC08(筛选标记基因分别为3S启动子及Adh1-intron1增强驱动的epsps和bar)导入玉米(Zeamays)自交系幼胚愈伤组织,经过筛选、分化和再生获得了44株转基因当代植株,经PCR扩增、PCR-Southern检测有30株为阳性.选择9株健壮的阳性植株进行基因组Southern-blotting分析,结果表明7株T0植株整合了目的基因,其中4株整合了1个转基因拷贝,2株整合了2个转基因拷贝,1株整合了3个转基因拷贝.这7株中有2株结实,直链淀粉含量分别为23.22%和24.60%,有一定的小幅提高.下一步要进行较大规模的基因枪转化,得到比较大的转基因群体,以对更多转基因后代进行鉴定,期望筛选出外源基因多拷贝插入和直链淀粉含量大幅提高的株系.

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