Summary Bread wheat ( Triticum aestivum L.) is cultivated on more land than any other crop and produces a fifth of the calories consumed by humans. Wheat endosperm is rich in starch yet contains low concentrations of dietary iron (Fe) and zinc (Zn). Biofortification is a micronutrient intervention aimed at increasing the density and bioavailability of essential vitamins and minerals in staple crops; Fe biofortification of wheat has proved challenging. In this study we employed constitutive expression ( CE ) of the rice ( Oryza sativa L.) nicotianamine synthase 2 ( Os NAS 2 ) gene in bread wheat to up‐regulate biosynthesis of two low molecular weight metal chelators – nicotianamine ( NA ) and 2′‐deoxymugineic acid ( DMA ) – that play key roles in metal transport and nutrition. The CE ‐ Os NAS 2 plants accumulated higher concentrations of grain Fe, Zn, NA and DMA and synchrotron X‐ray fluorescence microscopy ( XFM ) revealed enhanced localization of Fe and Zn in endosperm and crease tissues, respectively. Iron bioavailability was increased in white flour milled from field‐grown CE ‐ Os NAS 2 grain and positively correlated with NA and DMA concentrations.

Biofortification of rice (Oryza sativa L.) with micronutrients is widely recognized as a sustainable strategy to alleviate human iron (Fe) and zinc (Zn) deficiencies in developing countries where rice is the staple food. Constitutive overexpression of the rice nicotianamine synthase (OsNAS) genes has been successfully implemented to increase Fe and Zn concentrations in unpolished and polished rice grain. Intensive research is now needed to couple this high-micronutrient trait with high grain yields. We investigated associations of increased grain Fe and Zn concentrations with agro-morphological traits of backcross twice second filial (BC2F2) transgenic progeny carrying OsNAS1 or OsNAS2 overexpression constructs under indica/japonica and japonica/japonica genetic backgrounds. Thirteen agro-morphological traits were evaluated in BC2F2 transgenic progeny grown under hydroponic conditions. Concentrations of eight mineral nutrients (Fe, Zn, copper, manganese, calcium, magnesium, potassium, and phosphorus) in roots, stems/sheaths, non-flag leaves, flag leaves, panicles, and grain were also determined. A distance-based linear model (DistLM) was utilized to extract plant tissue nutrient predictors accounting for the largest variation in agro-morphological traits differing between transgenic and non-transgenic progeny. Overall, the BC2F2 transgenic progeny contained up to 148% higher Fe and 336% higher Zn concentrations in unpolished grain compared to non-transgenic progeny. However, unpolished grain concentrations surpassing 23 μg Fe g-1 and 40 μg Zn g-1 in BC2F2 indica/japonica progeny, and 36 μg Fe g-1 and 56 μg Zn g1 in BC2F2 japonica/japonica progeny, were associated with significant reductions in grain yield. DistLM analyses identified grain-Zn and panicle-magnesium as the primary nutrient predictors associated with grain yield reductions in the indica/japonica and japonica/japonica background, respectively. We subsequently produced polished grain from high-yield BC2F2 transgenic progeny carrying either the OsNAS1 or OsNAS2 overexpression constructs. The OsNAS2 overexpressing progeny had higher percentages of Fe and Zn in polished rice grain compared to the OsNAS1 overexpressing progeny. Results from this study demonstrate that genetic background has a major effect on the development of Fe and Zn biofortified rice. Moreover, our study shows that high-yielding rice lines with Fe and Zn biofortified polished grain can be developed by OsNAS2 overexpression and monitoring for Zn overaccumulation in the grain.

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We tested the effect of the addition of biosolids combined with a native mycorrhizal inoculum (Arbuscular Mycorrhizal Fungi [AMF]) on growth of a native Australian grass, and on trace element stabilization of sulphidic gold mine tailings. A glasshouse trial was established on four substrates: tailings (T); tailings with a layer of 5 cm topsoil (TS); tailings amended with 100 dry t ha −1 biosolids (LB), and tailings amended with 500 dry t ha −1 biosolids (HB). Pots of 1.2 L of capacity were established; some were inoculated with a mixture of Glomus sp. (WUM51-9227), Scutelospora aurigloba (WUM51-53), and Acaulospora levis (WUM46) culture mix, and others were uninoculated controls. Seeds of the native Australian grass, Bothriochloa macra were sown in the pots. Root infection, plant biomass production, nutrients and trace element concentrations in shoots were investigated.Addition of biosolids significantly increased AMF infection of roots compared to unamended substrates. No clear qualitative differences in colonization were detected. Addition of biosolids and AMF together clearly improved the establishment and growth of the native grass. Similar trends in nutritional status were shown for biosolids and inoculation with AMF treatments. Mycorrhizal inoculation increased plant biomass production and the effectiveness of nutrient uptake. The combined use of biosolids and mycorrhizal inoculation could be a reliable method for phytostabilization purposes in polluted substrates.

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