Development of “Purple Endosperm Rice” by Engineering Anthocyanin Biosynthesis in the Endosperm with a High-Efficiency Transgene Stacking System

Abstract

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.

Overexpression of the caryophyllene synthase gene GhTPS1 in cotton negatively affects multiple pests while attracting parasitoids

“…Recently, the stacked genetic engineering of multiple genes has been applied to modify crops, such as the successful generation of 'Purple Endosperm Rice', in which a high-efficiency vector system was used to transfer eight anthocyanin-related genes. 57 Another multigene expression system (MES) combined wileyonlinelibrary.com/journal/ps the advantages of a bidirectional promoter and self-cleaving 2A peptides, was used to generate 'Purple Embryo Maize'. 58 These high-efficiency, multigene expression systems may also make it feasible to combine multiple strategies for efficient pest management in cotton.…”

Composição físico-químicas de cookies adicionados de resíduos do arroz vermelho

“…Segundo Zhu et al, (2017) o arroz apresenta uma grande diversidade genética na cor dos grãos e as antocianinas concentram-se principalmente no farelo de arroz descascado, onde para o resíduo foi quantificado um total de 2,23 mg/100g. Diferentes partes de um núcleo de arroz contêm diferentes quantidades de antocianinas .…”

From Golden Rice to aSTARice: Bioengineering Astaxanthin Biosynthesis in Rice Endosperm

“…The endosperm of rice (Oryza sativa) seeds is an ideal bioreactor for metabolic engineering and food biofortification (Paine et al, 2005;Blancquaert et al, 2015;Zhu et al, 2017). Engineering complex metabolic pathways often require introducing and expressing multiple related genes simultaneously in host plants.…”