Phosphorus (P) deficiency is one of the main growth-limiting factors for plants. However, arbuscular mycorrhizal (AM) symbiosis can significantly promote P uptake. Generally, PHT1 transporters play key roles in plants’ P uptake, and thus, PHT1 genes have been investigated in some plants, but the regulation and functions of these genes in wheat (TaPHT1) during AM symbiosis have not been studied in depth. Therefore, a comprehensive analysis of TaPHT1 genes was performed, including sequence, phylogeny, cis-elements, expression, subcellular localization and functions, to elucidate their roles in AM-associated phosphate transport and immunity. In total, 35 TaPHT1s were identified in the latest high-quality bread wheat genome, 34 of which were unevenly distributed on 13 chromosomes, and divided into five groups. Sequence analysis indicated that there are 11 types of motif architectures and five types of exon-intron structures in the TaPHT1 family. Duplication mode analysis indicated that the TaPHT1 family has expanded mainly through segmental and tandem duplication events, and that all duplicated gene pairs have been under purifying selection. Transcription analysis of the 35 TaPHT1s revealed that not only known the mycorrhizal-specific genes TaPht-myc, TaPT15-4B (TaPT11) and TaPT19-4D (TaPT10), but also four novel mycorrhizal-specific/inducible genes (TaPT3-2D, TaPT11-4A, TaPT29-6A, and TaPT31-7A) are highly up-regulated in AM wheat roots. Furthermore, the mycorrhizal-specific/inducible genes are significantly induced in wheat roots at different stages of infection by colonizing fungi. Transient Agrobacterium tumefaciens-mediated transformation expression in onion epidermal cells showed that TaPT29-6A is a membrane-localized protein. In contrast to other AM-specific/inducible PHT1 genes, TaPT29-6A is apparently required for the symbiotic and direct Pi pathway. TaPT29-6A-silenced lines exhibited reduced levels of AM fungal colonization and arbuscules, but increased susceptibility to biotrophic, hemi-biotrophic and necrotrophic pathogens. In conclusion, TaPT29-6A was not only essential for the AM symbiosis, but also played vital roles in immunity.

Effector proteins secreted by plant pathogens play important roles in promoting colonization. Blumeria effector candidate (BEC) 1019, a highly conserved metalloprotease of Blumeria graminis f. sp. hordei (Bgh), is essential for fungal haustorium formation, and silencing BEC1019 significantly reduces Bgh virulence. In this study, we found that BEC1019 homologs in B. graminis f. sp. tritici (Bgt) and Gaeumannomyces graminis var. tritici (Ggt) have complete sequence identity with those in Bgh, prompting us to investigate their functions. Transcript levels of BEC1019 were abundantly induced concomitant with haustorium formation in Bgt and necrosis development in Ggt-infected plants. BEC1019 overexpression considerably increased wheat susceptibility to Bgt and Ggt, whereas silencing this gene using host-induced gene silencing significantly enhanced wheat resistance to Bgt and Ggt, which was associated with hydrogen peroxide accumulation, cell death, and pathogenesis-related gene expression. Additionally, we found that the full and partial sequences of BEC1019 can trigger cell death in Nicotiana benthamiana leaves. These results indicate that Bgt and Ggt can utilize BEC1019 as a virulence effector to promote plant colonization, and thus these genes represent promising new targets in breeding wheat cultivars with broad-spectrum resistance.

Potassium (K) plays important roles in plant growth and development processes, while low K (LK) stress inhibits plant growth by altering reactive oxygen species accumulation. Arbuscular mycorrhizal fungi (AMF) promote nutrient absorption and transport in plants. However, the roles of AMF in affecting K nutrition are less well studied than those of other nutrients, especially in wheat. In this study, the effects of AMF on four wheat varieties were evaluated; results showed that the inoculation with the AMF-Rhizophagus intraradices significantly increased mycorrhizal colonization, fresh and dry weights, ascorbic acid, and glutathione contents, while decreasing malondialdehyde contents under both normal and LK stress treatments. It is worth noting that the contents of K and several nutrient elements were more significantly increased in roots than in shoots, suggesting that AMF mainly affect the uptake of K and other nutrient elements in the roots. Moreover, the expression levels of K transporter genes were higher than those of nitrogen and phosphorus transporter genes, especially under AMF combined with LK stress treatments. These results indicate that AMF improves wheat growth and antioxidant activity by regulating K transporter gene expression and affecting K uptake and transport. Therefore, AMF could be used as a sustainable agricultural alternative in wheat under LK soils.

Drought stress is a major abiotic constraint that severely limits growth, yield formation, and grain quality in wheat. Potassium (K) application is known to alleviate drought stress in crops. However, the integrated effects of K fertilization on yield, grain nutritional and quality, and nutrient use efficiency under post-flowering drought stress remains poorly understood. In this study, two-year pot experiments were conducted with two wheat cultivars under well-watered (75% field capacity) or post-flowering drought (~40% field capacity) conditions, combined with K fertilization (120 kg K₂O ha⁻¹) or no K (0 kg K₂O ha⁻¹), using a randomized complete block design. The results showed that K application significantly improved wheat performance under post-flowering drought stress. Specifically, it increased plant biomass, thousand-grain weight, grain yield, K accumulation, and K uptake efficiency, while also elevating grain contents of calcium, magnesium, zinc, total starch, and wet gluten. In conclusion, K fertilization not only mitigates the adverse effects of post-flowering drought stress in wheat, but also improves yield, grain quality, and nutrient use efficiency. These findings offer valuable insights for high-efficiency nutrient management and wheat production under drought regions.