More than a billion people suffer from iron or zinc deficiencies globally. Rice (Oryza sativa L.) iron and zinc biofortification; i.e., intrinsic iron and zinc enrichment of rice grains, is considered the most effective way to tackle these deficiencies. However, rice iron biofortification, by means of conventional breeding, proves difficult due to lack of sufficient genetic variation. Meanwhile, genetic engineering has led to a significant increase in the iron concentration along with zinc concentration in rice grains. The design of impactful genetic engineering biofortification strategies relies upon vast scientific knowledge of precise functions of different genes involved in iron and zinc uptake, translocation and storage. In this review, we present an overview of molecular processes controlling iron and zinc homeostasis in rice. Further, the genetic engineering approaches adopted so far to increase the iron and zinc concentrations in polished rice grains are discussed in detail, highlighting the limitations and/or success of individual strategies. Recent insight suggests that a few genetic engineering strategies are commonly utilized for elevating iron and zinc concentrations in different genetic backgrounds, and thus, it is of great importance to accumulate scientific evidence for diverse genetic engineering strategies to expand the pool of options for biofortifying farmer‐preferred cultivars.

Iron deficiency leads to severe chlorosis in crop plants, including wheat, thereby reducing total yield and quality. Furthermore, grains of most bread wheat varieties are poor source of iron, which is vital for human nutrition. Despite the significance, iron uptake and translocation mechanisms in bread wheat have not been studied in detail, particularly under iron limited growth conditions. In this study, bread wheat plants were grown under iron deficiency stress until maturity. Data were collected at three distinct developmental time points during grain-filling. The plants experiencing low iron availability exhibited significantly lower chlorophyll content as well as low iron concentration in leaves and grains. The expression levels of bread wheat genes homologous to iron deficiency responsive genes of rice, barley, and Arabidopsis were significantly changed under iron deficiency stress. The wheat homologs of genes involved in phytosiderophore (PS) synthesis and transport were significantly up-regulated in the iron-deficient roots through all development stages, confirming an important role of deoxymugineic acid (DMA) in iron acquisition. The up-regulation of NICOTIANAMINE SYNTHASE (NAS) and DEOXYMUGINEIC ACID SYNTHASE (DMAS) in flag leaves and grains suggested the involvement of nicotianamine (NA) and DMA in iron chelation and translocation in wheat, particularly at the commencement of grain-filling. In line with this, the homolog of gene encoding TRANSPORTER OF MUGINEIC ACID (TOM) was up-regulated in the wheat roots under iron deficiency. Additionally, genes encoding long-distance iron transporter YELLOW STRIPE-LIKE (YSL), the vacuolar transporter NATURAL RESISTANCE ASSOCIATED MACROPHAGE PROTEIN (NRAMP), and the transcription factor BASIC HELIX-LOOP-HELIX (bHLH), were also up-regulated upon iron starvation. A tissue specific and growth stage specific gene expression differences in response to iron deficiency stress were observed, providing new insights into iron translocation, storage and regulation in bread wheat.

Human iron (Fe) deficiency is a widespread problem worldwide. Rice (Oryza sativa) Fe biofortification is a promising approach to address human Fe deficiency. Since its conceptualization, various biofortification strategies have been developed, some of which have resulted in significant gain in grain Fe concentration. On the other hand, there are still many aspects which are yet to be addressed in the studies to date. In this review, we first overview the important rice Fe biofortification strategies reported to date and the complication associated to them. Subsequently, we highlight the key outstanding questions and hypotheses related to rice Fe biofortification. Finally, we propose the direction of future rice biofortification studies.

Intrinsic improvement of Fe concentration in the rice grains, called rice Fe biofortification, is a promising countermeasure against widespread human Fe deficiency. In this study, two novel rice Fe biofortification approaches are reported. The first approach (Y approach) involved the expression of maize YELLOW STRIPE 1 controlled by the HEAVY METAL ATPASE 2 promoter. Y approach increased the polished grain Fe concentrations up to 4.8-fold compared to the non-transgenic (NT) line. The second approach (T approach) involved the expression of rice TRANSPORTER OF MUGINEIC ACID 1 controlled by the FERRIC REDUCTASE DEFECTIVE LIKE 1 promoter. T approach increased the polished grain Fe concentrations by up to 3.2-fold. No synergistic increases in the polished grain Fe concentrations were observed when Y and T approaches were combined (YT approach). However, the polished grain Fe concentrations further increased by 5.1- to 9.3-fold compared to the NT line, when YT approach was combined with the endosperm-specific FERRITIN expression (YTF approach) or when YTF approach was combined with the constitutive NICOTIANAMINE SYNTHASE expression (YTFN approach). Total grain weight per plant in most Y, T, YT and YTFN lines was comparable to that in the NT line, while it was significantly decreased in most YTF lines.