Evidence of spontaneous selfing and disomic inheritance in Geranium robertianum

Knowing species' breeding system and mating processes occurring in populations is important not only for understanding population dynamics, gene flow processes, and species' response to climate change, but also for designing control plans of invasive species. Geranium robertianum, a widespread biennial herbaceous species showing high morphological variation and wide ecological amplitude, can become invasive outside its distribution range. A mixed-mating system may be expected given the species' floral traits. However, autonomous selfing is considered as a common feature. Genetic variation and structure, and so population mating processes, have not been investigated in wild populations. We developed 15 polymorphic microsatellite markers to quantify genetic variation and structure in G. robertianum. To investigate whether selfing might be the main mating process in natural conditions, we sampled three generations of plants (adult, F1, and F2) for populations from the UK, Spain, Belgium, Germany, and Sweden, and compared open-pollinated with outcrossed hand-pollinated F2 progeny. The highly positive Wright's inbreeding coefficient (F _IS) values in adults, F1, and open-pollinated F2 progeny and the low F _IS values in outcross F2 progeny supported autonomous selfing as the main mating process for G. robertianum in wild conditions, despite the presence of attractive signals for insect pollination. Genetic differentiation among samples was found, showing some western-eastern longitudinal trend. Long-distance seed dispersal might have contributed to the low geographic structure. Local genetic differentiation may have resulted not only from genetic drift effects favored by spontaneous selfing, but also from ecological adaptation. The presence of duplicate loci with disomic inheritance is consistent with the hypothesis of allotetraploid origin of G. robertianum. The fact that most microsatellite markers behave as diploid loci with no evidence of duplication supports the hypothesis of ancient polyploidization. The differences in locus duplication and the relatively high genetic diversity across G. robertianum range despite spontaneous autonomous selfing suggest multiple events of polyploidization.