Evolutionary divergence of the rye Pm17 and Pm8 resistance genes reveals ancient diversity.

KEY MESSAGE: We have isolated a novel powdery mildew resistance gene in wheat that was originally introgressed from rye. Further analysis revealed evolutionary divergent history of wheat and rye orthologous resistance genes. Wheat production is under constant threat from a number of fungal pathogens, among them is wheat powdery mildew (Blumeria graminis f. sp. tritici). Deployment of resistance genes is the most economical and sustainable method for mildew control. However, domestication and selective breeding have narrowed genetic diversity of modern wheat germplasm, and breeders have relied on wheat relatives for enriching its gene pool through introgression. Translocations where the 1RS chromosome arm was introgressed from rye to wheat have improved yield and resistance against various pathogens. Here, we isolated the Pm17 mildew resistance gene located on the 1RS introgression in wheat cultivar 'Amigo' and found that it is an allele or a close paralog of the Pm8 gene isolated earlier from 'Petkus' rye. Functional validation using transient and stable transformation confirmed the identity of Pm17. Analysis of Pm17 and Pm8 coding regions revealed an overall identity of 82.9% at the protein level, with the LRR domains being most divergent. Our analysis also showed that the two rye genes are much more diverse compared to the variants encoded by the Pm3 gene in wheat, which is orthologous to Pm17/Pm8 as concluded from highly conserved upstream sequences in all these genes. Thus, the evolutionary history of these orthologous loci differs in the cereal species rye and wheat and demonstrates that orthologous resistance genes can take different routes towards functionally active genes. These findings suggest that the isolation of Pm3/Pm8/Pm17 orthologs from other grass species, additional alleles from the rye germplasm as well as possibly synthetic variants will result in novel resistance genes useful in wheat breeding.

Single genetic locus improvement of iron, zinc and ß-carotene content in rice grains.

Nearly half of the world's population obtains its daily calories from rice grains, which lack or have insufficient levels of essential micronutrients. The deficiency of micronutrients vital for normal growth is a global health problem, and iron, zinc and vitamin A deficiencies are the most prevalent ones. We developed rice lines expressing Arabidopsis NICOTIANAMINE SYNTHASE 1 (AtNAS1), bean FERRITIN (PvFERRITIN), bacterial CAROTENE DESATURASE (CRTI) and maize PHYTOENE SYNTHASE (ZmPSY) in a single genetic locus in order to increase iron, zinc and ß-carotene content in the rice endosperm. NAS catalyzes the synthesis of nicotianamine (NA), which is a precursor of deoxymugeneic acid (DMA) iron and zinc chelators, and also chelate iron and zinc for long distance transport. FERRITIN provides efficient storage of up to 4500 iron ions. PSY catalyzes the conversion of GGDP to phytoene, and CRTI performs the function of desaturases required for the synthesis of ß-carotene from phytoene. All transgenic rice lines have significantly increased ß-carotene, iron, and zinc content in the polished rice grains. Our results establish a proof-of-concept for multi-nutrient enrichment of rice grains from a single genetic locus, thus offering a sustainable and effective approach to address different micronutrient deficiencies at once.

Rice NICOTIANAMINE SYNTHASE 2 expression improves dietary iron and zinc levels in wheat.

KEY MESSAGE: Iron and zinc deficiencies negatively impact human health worldwide. We developed wheat lines that meet or exceed recommended dietary target levels for iron and zinc in the grains. These lines represent useful germplasm for breeding new wheat varieties that can reduce iron and zinc deficiency-associated health burdens in the affected populations. Micronutrient deficiencies, including iron and zinc deficiencies, have negative impacts on human health globally. Iron-deficiency; anemia affects nearly two billion people worldwide and is the cause of reduced cognitive development, fatigue and overall low productivity. Similarly, zinc deficiency causes stunted growth, decreased immunity and increased risk of respiratory infections. Biofortification of staple crops is a sustainable and effective approach to reduce the burden of health problems associated with micronutrient deficiencies. Here, we developed wheat lines expressing rice NICOTIANAMINE SYNTHASE 2 (OsNAS2) and bean FERRITIN (PvFERRITIN) as single genes as well as in combination. NAS catalyzes the biosynthesis of nicotianamine (NA), which is a precursor of the iron chelator deoxymugeneic acid (DMA) required for long distance iron translocation. FERRITIN is important for iron storage in plants because it can store up to 4500 iron ions. We obtained significant increases of iron and zinc content in wheat grains of plants expressing either OsNAS2 or PvFERRTIN, or both genes. In particular, wheat lines expressing OsNAS2 greatly surpass the HarvestPlus recommended target level of 30 % dietary estimated average requirement (EAR) for iron, and 40 % of EAR for zinc, with lines containing 93.1 µg/g of iron and 140.6 µg/g of zinc in the grains. These wheat lines with dietary significant levels of iron and zinc represent useful germplasm for breeding new wheat varieties that can reduce micronutrient deficiencies in affected populations.