Understanding the changes in the root system architecture of bread wheat under phosphorus (P)-limited conditions is critical for identifying specific traits contributing to improved P uptake. Phenotypic variability in root, biomass, and P index-related traits among 204 diverse wheat genotypes at the seedling stage was examined under low and optimum P treatments. Strong genotypic and phenotypic associations between P utilization efficiency (PUtE) and total root volume, dry weight of root and shoot, total P uptake, and total plant biomass were observed under optimum P. Under low P, strong positive correlations between PUtE and total root length, total root volume, total surface area, and total biomass were observed, while it was negatively correlated with average diameter. These traits exhibited medium to high heritability. Under low P, average root diameter, primary root length, root mass ratio, total root tips, and surface area showed high Shannon–Weaver diversity index (H’) values (>0.79). The agglomerative hierarchical clustering analysis grouped the genotypes into four distinct clusters. The best performing genotypes in Clusters I and II indicated their strong relationship with P use efficiency due to higher percent increases in total root length, total surface area, total root volume, total root tips, total biomass, P efficiency ratio, specific root length, and PUtE under low P as compared to optimum P conditions. The present study identified specific root system architectural traits and P use-efficient genotypes (SHANGHAI, Pavon F76, BWL 5233, SONALIKA, KHARCHIA LOCAL, WH 102, BWL 4425, HD 2888.2, CBW 12, MN75136/PGO, KRL 19, and WH 1022) associated with efficient P uptake and utilization. These identified genotypes and traits may be useful in wheat breeding programs to develop P-efficient varieties with better adaptations for sustainable agriculture.
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