GmGASA6基因CRISPR-Cas9载体构建及大豆的遗传转化

Construction of GmGASA6 CRISPR-Cas9 vector and genetic transformation of soybean

通过对大豆GmGASA6基因的结构分析,选择了两个靶位点构建了GmGASA6 CRISPR-Cas9的基因编辑载体.利用农杆菌介导的大豆子叶节转化法进行了遗传转化,共转化外植体800个,获得了转基因阳性苗15株.通过测序证实部分T1代植株GmGASA6基因的靶位点发生了基因编辑.与野生型相比,种子萌发实验发现突变体的胚根伸长受到显著抑制,说明在种子的萌发过程中GmGASA6基因促进种子萌发和胚根的伸长.

Two target sites were selected to construct the GmGASA6 CRISPR-Cas9 gene editing vector by the structural analysis of soybean GmGASA6.Agrobacterium-mediated cotyledonary node transformation method was used for soybean genetic transformation and 800 explants were transformed,and 15 transgenic positive soybean seedlings were obtained.It was confirmed that the target sites of GmGASA6 in parts of T1 generation plants have been gene-edited by sequencing.The seed germination experiments found that the radicle elongation of the mutant was significantly inhibited than that of the wild type,indicating that the GmGASA6 gene promoted the seed germination and radicle elongation during the germination of soybean seeds.