RNAi 靶向沉默opaque2基因对高粱醇溶蛋白含量的影响

doi:10.3969/j.issn.1673-1689.2019.10.011

首页 > 过刊浏览>2019年第38卷第10期 >72-78. DOI:10.3969/j.issn.1673-1689.2019.10.011

RNAi 靶向沉默opaque2基因对高粱醇溶蛋白含量的影响
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                        10.3969/j.issn.1673-1689.2019.10.011
                    
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Effects of RNAi Targeting opaque2 Gene Expression on the Kafirin Content in Sorghum

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摘要:

为研究opaque2基因沉默对高粱醇溶蛋白含量的影响，采用RNAi技术构建opaque2基因抑制表达载体，利用农杆菌介导转化高粱幼胚，测定转基因植株籽粒中醇溶蛋白含量，SDS-PAGE电泳检测醇溶蛋白带型，并使用透射电镜观察胚乳细胞中蛋白质体变化。结果表明，与野生型相比，5株转基因植株中4株籽粒的醇溶蛋白含量显著降低（P<0.05）；野生型和各转基因植株的醇溶蛋白SDS-PAGE电泳带型相同；野生型胚乳细胞中淀粉粒排列紧密，部分大淀粉粒融合成大块状，蛋白质体数量较多且有一些体积较大；而转基因植株淀粉粒排列疏松，细胞中存在多处孔隙，蛋白质体数量较少，体积较小。opaque2基因沉默引起胚乳细胞蛋白质体数量变少，体积变小，进而影响胚乳细胞内部结构和籽粒醇溶蛋白含量。

Abstract:

In order to explore the effect of silencing opaque2 gene on the kafirin content in sorghum，the suppressed expression vector of opaque2 gene is constructed by RNAi technology，and then transformed to young embryos by Agrobacterium tumefaciens-mediated transformation system. The kafirin content in these transgenic seeds is determined and indentified by SDS-PAGE，and also the changes of protein bodies in these endosperm cells are observed by transmission electron microscope. The results show that the kafirin content of four fifths of transgenic plants is significantly lower than that of the wild type（P<0.05）. Those of the protein bands between the wild type and all transgenic plants are similar. The starch granules in the endosperm cells of the wild type arranged closely and most of them are fused into large blocks，and large numbers of protein bodies are indicated with large volumes. While the starch granules in the transgenic plants arrayed loosely and indicated many gaps in the endosperm cells，and while those numbers and volumes of the protein bodies are less and smaller than the wild type，respectively. In conclusion，opaque2 gene silencing induced changes in number and volume of protein bodies in endosperm cells，which additionally affects the internal structure of endosperm cells and the kafirin content in sorghum.

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蔡欣月,李志江,牛江帅,包丹,苏晨,孙艺墨,戴凌燕. RNAi 靶向沉默opaque2基因对高粱醇溶蛋白含量的影响[J].食品与生物技术学报,2019,38(10):72-78.

CAI Xinyue, LI Zhijiang, NIU Jiangshuai, BAO Dan, SU Chen, SUN Yimo, DAI Lingyan. Effects of RNAi Targeting opaque2 Gene Expression on the Kafirin Content in Sorghum[J]. Journal of Food Science and Biotechnology,2019,38(10):72-78.

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