Paternity of Eschscholzia californica plants introduced to habitats comprising different floral cover

All open flowers were removed from the E. californica plants prior to their placement in pre-marked locations across the landscape. One bud on each plant was covered in a fine muslin to represent a 'pollinator excluded' flower. Plants remained in the field for 16 days to ensure full anthesis of new flowers and to allow for multiple pollination events. After this period, all fruit were tagged to ensure that only fruit development arising from the period of the field experiment were included in analyses. Plants were then collected and stored under controlled glasshouse conditions until fruit maturation. Ten progeny per plant from each of the field exposed plants were genotyped using 50mg of fresh leaf material and following standardised protocols. The incidence of self-fertilisation in plants from each habitat was calculated manually by individually comparing each successfully amplified progeny against their maternal plant. If, at each of the seven loci, the progeny was a complete match for the maternal genotype, or was homozygote for one of the maternal plants alleles, it was scored as selfed. Alternatively, if any novel alleles were observed in the progeny that were not present in the maternal plant, the progeny was classified as outcrossed. Paternity was determined using Cervus 3.0.7, where each progeny sample was listed detailing alleles at the seven microsatellite loci, specifying the known maternal sample as well as the potential paternal samples.