National Institute of Agricultural Science, Rural Development Administration, Jeonju 54874, Korea.
Department of Biotechnology, College of Agriculture and Life Sciences, Chonnam National University, Gwangju 61186, Korea.
Int J Mol Sci. 2017 Nov 18;18(11):2458. doi: 10.3390/ijms18112458.
Rice ( L.) is a primary global food cereal. However, when compared to wheat, rice has poor food processing qualities. Dough that is made from rice flour has low viscoelasticity because rice seed lacks storage proteins that are comparable to gluten protein from wheat. Thus, current research efforts aim to improve rice flour processing qualities through the transgenic expression of viscoelastic proteins in rice seeds. In this study, we characterized the transgenic expression of wheat glutenin subunits in rice seeds. The two genes and , which both encode wheat high-molecular-weight glutenin subunits that confer high dough elasticity, were cloned from Korean wheat cultivars KeumKang and JoKyung, respectively. These genes were inserted into binary vectors under the control of the rice endosperm-specific promoter and were expressed in the high-amylose Korean rice cultivar Koami ( L.). Individual expression of both glutenin subunits was confirmed by SDS-PAGE and immunoblot analyses performed using T₃ generation of transgenic rice seeds. The subcellular localization of 1Dx5_KK and 1Dy10_JK in the rice seed endosperm was confirmed by immunofluorescence analysis, indicating that the wheat glutenin subunits accumulate in protein body-II and novel protein body types in the rice seed. These results contribute to our understanding of engineered seed storage proteins in rice.
水稻（L.）是一种主要的全球粮食谷物。然而，与小麦相比，水稻的食品加工品质较差。由米粉制成的面团的黏弹性低，因为水稻种子缺乏与小麦面筋蛋白相当的储存蛋白。因此，目前的研究工作旨在通过在水稻种子中转基因表达黏弹性蛋白来改善米粉的加工品质。在这项研究中，我们研究了小麦谷蛋白亚基在水稻种子中的转基因表达情况。从韩国小麦品种 Kumkang 和 JoKyung 中克隆了编码小麦高分子量谷蛋白亚基的两个基因 和 ，这两个亚基赋予面团高弹性。这些基因被插入到含有水稻胚乳特异性 启动子的二元载体中，并在高直链淀粉韩国水稻品种 Koami（L.）中表达。通过 SDS-PAGE 和免疫印迹分析，从 T₃ 代转基因水稻种子中证实了这两个谷蛋白亚基的单独表达。通过免疫荧光分析证实了 1Dx5_KK 和 1Dy10_JK 在水稻胚乳中的亚细胞定位，表明小麦谷蛋白亚基在水稻种子的蛋白体-II 和新型蛋白体类型中积累。这些结果有助于我们了解水稻中工程化的种子储存蛋白。
