The BnamiR827–BnaA09.NLA1–BnaPHT1 module regulates phosphate homeostasis, pollen viability, and seed yield in <i>Brassica napus</i>

Phosphorus (P) is an essential macronutrient for the growth and yield of crops. However, there is limited understanding of the regulatory mechanisms of phosphate (Pi) homeostasis, and its impact on growth, development, and yield-related traits in Brassica napus. Here, we identified four NITROGEN LIMITATION ADAPTATION1 (BnaNLA1) genes in B. napus; their expression was predominant in roots and suppressed by Pi starvation-induced BnamiR827. All the BnaNLA1 proteins have similar sequences, subcellular localizations, and abilities to rescue the growth defects of the atnla1 mutant. One of the genes, BnaA09.NLA1, is expressed abundantly in roots, and also in old leaves, anthers, and pollen. Knocking out BnaNLA1 genes or overexpressing BnamiR827 resulted in increased concentrations of Pi in leaves and stamens and reduced pollen viability, thereby negatively impacting seed yield. Bimolecular fluorescence complementation (BiFC) and split-ubiquitin yeast two-hybrid (Y2H) analyses demonstrated that BnaA09.NLA1 interacted with seven Pi transporters highly expressed in roots and/or anthers (i.e. BnaPT8/10/11/27/35/37/42) to regulate Pi uptake and Pi allocation in anthers. Taken together, this study demonstrates that the BnamiR827-BnaA09.NLA1-BnaPHT1 module is involved in the regulation of Pi uptake and Pi allocation in floral organs, which is vital for the growth, pollen viability, and seed yield of B. napus.