Pollen contamination in a maritime pine polycross seed orchard and certification of improved seeds using chloroplast microsatellites

A new concept of seed orchard was developed by Institut National de la Recherche Agronomique for the maritime pine (Pinus pinaster Ait.) breeding program: the polycross seed orchard (PSO). The expected genetic gain of the PSO can only be reached if the fathers used in the pollen mix contribute equally to the next generation (i.e., to the base material of the PSO) and if pollen contamination from the surrounding stands is limited. Using chloroplast simple-sequence repeats (cpSSR), we showed that the chloroplast DNA was unipaternally inherited in maritime pine and verified that the chloroplast haplotype composition of the megagametophyte tissue corresponded to the chloroplast haplotype of the female parent. As a practical application, a statistical test based on cpSSR markers and simulation was established to verify the PSO origin of maritime pine seed lots. As a result of the cpSSR test, it was observed that (i) departure from even distribution of the fathers in the PSO was barely significant, (ii) the minimum pollen contamination rate in the PSO was 36%, and (iii) the contamination was not evenly distributed in the PSO. As a consequence, the expected genetic gain will range between 50 and 82% of what was initially foreseen.