Nitrogen deficiency-induced protein changes in immature and mature leaves of creeping bentgrass

Abstract

Nitrogen (N) deficiency inhibits plant growth and induces leaf senescence through regulating various metabolic processes. The objectives of this study were to examine protein changes in response to N deficiency in immature and mature leaves of a perennial grass species and determine major metabolic processes affected by N deficiency through proteomic profiling. Creeping bentgrass (Agrostis stolonifera cv. Penncross) plants were originally fertilized with a diluted 36N-2.6P-5K fertilizer. After 14 days acclimation in a growth chamber, plants were grown in a nutrient solution containing 6 mM nitrate (control) or without N (N deficiency). Immature leaves (upper first and second not yet fully expanded leaves) and mature leaves (lower fully expanded leaves) were separated at 28 days of treatment for protein analysis. Two-dimensional electrophoresis and mass spectrometry analysis were used to identify protein changes in immature and mature leaves in response to N deficiency. The abundance of many proteins in both immature and mature leaves decreased with N deficiency, including those involved in photosynthesis, photorespiration, and amino acid metabolism (hydroxypyruvate reductase, serine hydroxymethyltransferase, alanine aminotransferase, glycine decarboxylase complex, glycolate oxidase), protein protection [heat shock protein (HSP)/HSP 70, chaperonin 60 and FtsH-like protein], and RNA stability (RNA binding protein). The reduction in protein abundance under N deficiency was greater in mature leaves than in immature leaves. The abundance of small HSP and metalloendopeptidase increased under N deficiency only in immature leaves. These results suggest that N deficiency accelerated protein degradation in immature and mature leaves of creeping bentgrass, particularly those proteins associated with energy and metabolism, but to a lesser extent in immature leaves. Immature leaves were also able to accumulate proteins with chaperone functions and for N reutilization, which could protect leaves from senescence under N deficiency.