Iron biofortification of rice using different transgenic approaches

Hiroshi Masuda, May Sann Aung, Naoko K Nishizawa

More than 2 billion people suffer from iron (Fe) deficiency, and developing crop cultivars with an increased concentration of micronutrients (biofortification) can address this problem. In this review, we describe seven transgenic approaches, and combinations thereof, that can be used to increase the concentration of Fe in rice seeds. The first approach is to enhance the Fe storage capacity of grains through expression of the Fe storage protein ferritin under the control of endosperm-specific promoters. Using this approach, the concentration of Fe in the seeds of transformants was increased by approximately 2-fold in polished seeds. The second approach is to enhance Fe translocation by overproducing the natural metal chelator nicotianamine; using this approach, the Fe concentration was increased by up to 3-fold in polished seeds. The third approach is to enhance Fe influx to the endosperm by expressing the Fe(II)-nicotianamine transporter gene OsYSL2 under the control of an endosperm-specific promoter and sucrose transporter promoter, which increased the Fe concentration by up to 4-fold in polished seeds. The fourth approach is introduction of the barley mugineic acid synthesis gene IDS3 to enhance Fe uptake and translocation within plants, which resulted in a 1.4-fold increase in the Fe concentration in polished seeds during field cultivation. In addition to the above approaches, Fe-biofortified rice was produced using a combination of the first, second, and third approaches. The Fe concentration in greenhouse-grown T2 polished seeds was 6-fold higher and that in paddy field-grown T3 polished seeds was 4.4-fold higher than in non-transgenic seeds without any reduction in yield. When the first and fourth approaches were combined, the Fe concentration was greater than that achieved by introducing only the ferritin gene, and Fe-deficiency tolerance was observed. With respect to Fe biofortification, the introduction of multiple Fe homeostasis genes is more effective than the introduction of individual genes. Moreover, three additional approaches, i.e., overexpression of the Fe transporter gene OsIRT1 or OsYSL15, overexpression of the Fe deficiency-inducible bHLH transcription factor OsIRO2, and knockdown of the vacuolar Fe transporter gene OsVIT1 or OsVIT2, may be useful to further increase the Fe concentration of seeds.