The objective of this research is to explore the genetic diversity of Alfalfa germplasm resources by Phenotypic traits and Quality Characteristics provide basis for variety improvement and parent choice. 21 indexes of 75 alfalfa sativa accessions from different geographical origins were studied. The Phenotypic traits and Quality Characteristics were analyzed by principal component analysis and cluster analysis. The results showed that there were extensive variations in Phenotypic traits and Quality characteristics in different alfalfa accessions. The most significant variance was agronomic traits, followed by morphological characters and Quality Characteristics;Principal component analysis showed that The additive contributing rate of first eight principal components was 82.77%,four major discriminating characteristics were selected to reflect the comprehensive agronomic characters．Stem dry weight,leaf dry weight,plant fresh weight and plant dry weight were the main factors which produced morphological variation of 75 wild alfalfa accessions. Based on the 21 indexes, cluster analysis showed that all 75 accessions of alfalfa were divided into 5 groups.