Anthocyanins have high antioxidant activities, and engineering of anthocyanin biosynthesis in staple crops, such as rice （Oryza sativa L.）, could provide health-promoting foods for improving human health. However, e...Anthocyanins have high antioxidant activities, and engineering of anthocyanin biosynthesis in staple crops, such as rice （Oryza sativa L.）, could provide health-promoting foods for improving human health. However, engineering metabolic pathways for biofortification remains difficult, and previous attempts to engineer anthocyanin production in rice endosperm failed because of the sophisticated genetic regulatory network of its biosynthetic pathway. In this study, we developed a high-efficiency vector system for transgene stacking and used it to engineer anthocyanin biosynthesis in rice endosperm. We made a construct containing eight anthocyanin-related genes （two regulatory genes from maize and six structural genes from Coleus） driven by the endosperm-specific promoters,plus a selectable marker and a gene for marker excision. Transformation of rice with this construct generated a novel biofortified germplasm ＂Purple Endosperm Rice＂ （called ＂Zijingmi＂ in Chinese）, which has high anthocyanin contents and antioxidant activity in the endosperm. This anthocyanin production results from expression of the transgenes and the resulting activation （or enhancement） of expression of 13 endogenous anthocyanin biosynthesis genes that are silenced or expressed at low levels in wild-type rice endosperm. This study provides an efficient, versatile toolkit for transgene stacking and demonstrates its use for successful engineering of a sophisticated biological pathway, suggesting the potential utility of this toolkit for synthetic biology and improvement of agronomic traits in plants.展开更多
基金This work was supported by grants from National Natural Science Foundation of China （31000698）, the Ministry of Agriculture of China （2016ZX08010001 2016ZX08009002+1 种基金 2014ZX08010001）, and Guangdong Province Public Interest Research and Capacity Building Special Fund （2015B020201002 2016A020210084）.
文摘Anthocyanins have high antioxidant activities, and engineering of anthocyanin biosynthesis in staple crops, such as rice （Oryza sativa L.）, could provide health-promoting foods for improving human health. However, engineering metabolic pathways for biofortification remains difficult, and previous attempts to engineer anthocyanin production in rice endosperm failed because of the sophisticated genetic regulatory network of its biosynthetic pathway. In this study, we developed a high-efficiency vector system for transgene stacking and used it to engineer anthocyanin biosynthesis in rice endosperm. We made a construct containing eight anthocyanin-related genes （two regulatory genes from maize and six structural genes from Coleus） driven by the endosperm-specific promoters,plus a selectable marker and a gene for marker excision. Transformation of rice with this construct generated a novel biofortified germplasm ＂Purple Endosperm Rice＂ （called ＂Zijingmi＂ in Chinese）, which has high anthocyanin contents and antioxidant activity in the endosperm. This anthocyanin production results from expression of the transgenes and the resulting activation （or enhancement） of expression of 13 endogenous anthocyanin biosynthesis genes that are silenced or expressed at low levels in wild-type rice endosperm. This study provides an efficient, versatile toolkit for transgene stacking and demonstrates its use for successful engineering of a sophisticated biological pathway, suggesting the potential utility of this toolkit for synthetic biology and improvement of agronomic traits in plants.
