Arbuscular mycorrhizal fungi and rhizobia accelerate plant growth and N accumulation and contribution to soil total N in white clover by difficultly extractable glomalin-related soil protein

Arbuscular mycorrhizal fungi and rhizobia are soil symbiotic microorganisms involved in plant nitrogen (N) acquisition, whereas it is unclear how single or combined inoculation of both contributes to soil total N in legume crops. This study analyzed the effects of single or combined inoculation with arbuscular mycorrhizal fungus (Paraglomus occultum) and rhizobium (Rhizobium leguminosarum bv. trifolii) on growth performance, root soluble protein, leghemoglobin, NH4+-N, NO3−-N, and N concentrations, nitrogenase activities, soil total N contents, N contents in glomalin-related soil protein (GRSP), as well as the contribution of N in GRSP to soil total N in white clover. Twelve weeks after inoculation, P. occultum promoted nodule formation, and R. leguminosarum bv. trifolii accelerated root mycorrhizal colonization. Single or combined inoculation with P. occultum and R. leguminosarum bv. trifolii improved biomass, root total length, surface area and volume, root soluble protein concentrations, nodule leghemoglobin concentrations, and nitrogenase activities to varying degrees, with the trend of improvement being more pronounced with combined inoculation. R. leguminosarum bv. trifolii and/or P. occultum significantly increased NO3−-N, NH4+-N, and total N concentrations in nodules and roots, as well as soil total N concentrations. The N in easily extractable GRSP (EE-GRSP) and difficultly extractable GRSP (DE-GRSP) was 4.37–5.64 mg/g and 6.69–7.83 mg/g, respectively, contributing 2.68–3.26 % and 2.28–3.10 % of soil total N. P. occultum and combined inoculation accelerated the production of EE-GRSP and DE-GRSP and N in EE-GRSP. R. leguminosarum bv. trifolii not only significantly increased DE-GRSP production but also reduced N in DE-GRSP. R. leguminosarum bv. trifolii or P. occultum significantly increased the contribution of N in DE-GRSP, not EE-GRSP, to soil total N. It was concluded that arbuscular mycorrhizal fungus and rhizobium accelerated N accumulation and contribution to soil total N in white clover by DE-GRSP.