Modulation of protein expression in alfalfa (Medicago sativa L.) root and leaf tissues by Fusarium proliferatum

Alfalfa (Medicago sativa L.) is an important forage crop and is also a target of many fungal diseases including Fusarium spp. As of today, very little information is available about molecular mechanisms that contribute to pathogenesis and defense responses in alfalfa against Fusarium spp. and specifically against Fusarium proliferatum, the causal agent of alfalfa root rot. In this study, we used a proteomic approach to identify inducible proteins in alfalfa during a compatible interaction with F. proliferatum strain YQC-L1. Samples used for the two-dimensional gel electrophoresis (2-DE) and MALDI-TOF/TOF mass spectrometry were from roots and leaves of alfalfa cultivar AmeriGraze 401+Z and WL656HQ. Plants were grown in hydroponic conditions and at 4 days post inoculation with YQC-L1. Our disease symptom assays indicated that AmeriGraze 401+Z was tolerant to YQC-L1 infection while WL656HQ was highly susceptible. Analysis of differentially expressed proteins found in the 2-DE was further characterized using the MASCOT MS/MS ion search software and associated databases to identify multiple proteins that might be involved in F. proliferatum resistance. A total of 66 and 27 differentially expressed proteins were found in the roots and leaves of the plants inoculated with YQC-L1, respectively. These identified proteins were placed in various categories including defense and stress response related metabolism, photosynthesis and protein synthesis. Thirteen identified proteins were validated for their expressions by quantitative reverse transcription (qRT)-PCR. Our results suggested that some of the identified proteins might play important roles in alfalfa resistance against Fusarium spp. These finding could facilitate further dissections of molecular mechanisms controlling root rot disease in alfalfa and potentially other legume crops.