Three hundred and twenty-eight individuals from 16 populations of the annual species, M. lupulina, and 447 individuals from 15 populations of the perennial species, M. ruthenica, were studied using 15 and 17 microsatellite loci, respectively. Conventional and Bayesian-clustering analyses were utilized to estimate population genetic structure, mating system and gene flow.
Genetic diversity of M. lupulina (mean H-E = 0.246) was lower than that of M. ruthenica (mean H-E = 0.677). Populations of M. lupulina were more highly differentiated (F-ST = 0.535) than those of M. ruthenica (F-ST = 0.130). For M. lupulina, 55.5 % of the genetic variation was partitioned among populations, whereas 76.6 % of the variation existed within populations of M. ruthenica. Based on the genetic data, the selfing rates of M. lupulina and M. ruthenica were estimated at 95.8 % and 29.5 %, respectively. The genetic differentiation among populations of both species was positively correlated with geographical distance.
The mating system differentiation estimated from the genetic data is consistent with floral morphology and observed pollinator visitation. There was a much higher historical gene flow in M. ruthenica than in M. lupulina, despite more effective seed dispersal mechanisms in M. lupulina. The population genetic structure and geographical distribution of the two Medicago species have been shaped by life form, mating systems and seed dispersal mechanisms.