AetSRG1 contributes to the inhibition of wheat Cd accumulation by stabilizing phenylalanine ammonia lyase.

Cadmium (Cd) is a toxic heavy metal that poses a serious threat to crop safety, productivity, and human health. Aegilops tauschii is the D genome donor of common wheat and shows abundant genetic variation. However, the tolerance of Ae. tauschii toward Cd at the molecular level is poorly understood. In this study, key factors involved in the Cd stress response of Ae. tauschii were investigated by RNA sequencing. Differentially expressed genes (DEGs) under Cd stress were identified in Ae. tauschii roots and shoots. A Fe(II)/2-oxoglutarate dependent dioxygenase (designated as AetSRG1), with an unknown function in Cd stress, was of particular interest. The open reading frame of AetSRG1 was cloned and overexpressed in wheat, which resulted in reduced Cd accumulation along with a lower Cd2+ flux, decreased electrolyte leakage, and higher reactive oxygen species production. The protein of AetSRG1 interacted with phenylalanine ammonia lyase (PAL). Finally, we found that AetSRG1 stabilizes PAL and promotes the synthesis of endogenous salicylic acid. This study provides novel insights into the molecular mechanisms underlying the response of Ae. tauschii toward Cd stress. The key genes identified in this work serve as potential targets for developing low cadmium wheat.

TmNAS3 from Triticum monococum directly regulated by TmbHLH47 increases Fe content of wheat grain.

Biofortification is an effective way to enhance wheat grain Fe content. However, Fe overload inhibits the growth and development of wheat. In this work, the impact of Triticum monococcum nicotianamine synthase 3 (TmNAS3) on Fe accumulation in wheat grain was analyzed. Transgenic wheat expressing TmNAS3 was obtained via Agrobacterium-mediated transformation. The concentrations of Fe in the grains of two transgenic wheat lines were 62.42 µg/g and 68.75 µg/g, while that in the non-transgenic line (NT) was only 29.51 µg/g. Exogenous Fe application induced the expression of natural resistance-associated macrophage protein 3 (NRAMP3), NRAMP6, yellow stripe-like protein 3 (YSL3), YSL6, and vacuolar iron transporter 2 in transgenic wheat. The transcription factor that bound to the TmNAS3 promoter was identified, and the findings suggested that TmbHLH47 directly interacted and promoted the transcription of TmNAS3. Moreover, TmbHLH47 was observed to bind directly to the G-box in TmNAS3 promoter and regulated the transcriptional level of TmNAS3. Our findings contribute a TmbHLH47/TmNAS3 transcriptional pathway and thereby provide a potential strategy for improving the Fe concentration of wheat through genetic engineering.

Low-molecular-weight glutenin subunit LMW-N13 improves dough quality of transgenic wheat.

In our previous study, a novel LMW-GS designated as LMW-N13 with a unique molecular structure was identified from Aegilops uniaristata. LMW-N13 has been characterized as the largest LMW-GS, so far, and possesses an extra cysteine residue compared with typical LMW-GS. In order to analyze the contribution of LMW-N13 to dough quality, in this work, three transgenic wheat lines overexpressing LMW-N13 were generated. Compared with non-transformation (NT) lines, transgenic (TG) lines demonstrated superior dough properties. These superior dough properties were accompanied by the higher contents of glutenin macropolymer (GMP) and total protein. The microstructure of the dough was further investigated by scanning electron microscopy; starch granules in NT lines were smaller than those in transgenic lines. The protein matrix in NT lines was relatively loose and discontinuous. Conversely, the protein matrix in transgenic lines was more continuous and tight. The application of LMW-N13 in wheat breeding is also discussed.