Iron and zinc biofortification of rice by synergistic expression of OsNAS2 gene with monocot (Pennisetum glaucum) and dicot (Phaseolus vulgaris) ferritins

Abstract

Iron and zinc deficiencies are the most prevalent cause of global hidden hunger. Rice, being one of the most consumed crops worldwide, is suitable to target for Fe and Zn biofortification. In present study, we generated rice transgenic lines to meet the recommended dietary requirement of iron and zinc through endosperm specific expression of dicot (kidney bean) and monocot (pearl millet) Ferritins along with constitutive expression of rice nicotianamine synthase 2 (OsNAS2) gene. Visualization through perls’ prussian staining and quantification by ICP-MS showed significant improvement in grain iron content in all the transgenic lines. The transgenic lines expressing any of the three selected gene combinations (PvFerrtin-OsNAS2, feedPgFerrtin-OsNAS2 and foodPgFerritin-OsNAS2), showed the potential to surpass the 30% of the estimated average requirement (13 μg/g Fe and 28 μg/g Zn) proposed for rice in HarvestPlus breeding program. Though the expression of PvFerritin along with OsNAS2 gene in IET10364 (indica) variety showed the best result, providing up to 4.2- and 3.5-fold increase in iron (30.56 μg/g) and zinc (60.1 μg/g) content, respectively; in polished grains compared to non-transgenic control. Thus, the lines developed in our study can be used for further breeding purpose to enhance the iron and zinc content in commercial rice varieties.