PHOSPHATE OVERACCUMULATOR 2 (PHO2) is a negative regulator of arbuscular mycorrhizal symbiosis

Arbuscular mycorrhizal (AM) symbiosis is an ancient relationship formed between most plants and Glomeromycotina fungi, typically in response to phosphate (Pi) limitation in soils. By hosting these fungi in their roots, plants extend their access to essential mineral nutrients and water beyond the rhizosphere, while providing the fungus with carbon in return. This mutualistic symbiosis presents a promising tool for enhancing sustainability in agriculture, as it not only supports plant nutrition but also immunity and wider soil health. However, achieving high crop yields currently relies on supplementing plants with excess Pi, which suppresses AM symbiosis. We found that this suppression is mediated by a key negative regulator of the Pi starvation response (PSR) in rice ( Oryza sativa ), Phosphate overaccumulator 2 (PHO2). PHO2 encodes an E2 ubiquitin-conjugating enzyme which targets various proteins involved in the PSR in Pi-sufficient conditions for protein degradation. Here we report that pho2 mutants of rice and Nicotiana benthamiana retained high AM fungal colonisation even in high Pi conditions. Our transcriptomic analysis of uninoculated rice roots revealed that pho2 mutants are less sensitive to Pi treatment and retain susceptibility to AM symbiosis by maintaining expression of a core set of AM-related genes gating the early stage AM fungal entry, such as genes involved in strigolactone biosynthesis, LysM-containing plant receptors for fungal molecules, and components of the common symbiosis signalling pathway (CSSP). Furthermore, isotope tracing using ³³ P and phosphate transporter ( PHT1 ) gene expression patterns collectively suggest enhanced direct and symbiotic Pi overaccumulation in pho2 mutant leaves. Together, our data reveal a new role for PHO2, as a negative regulator of AM colonisation and symbiotic Pi accumulation in shoots.