Consecutive five-year analysis of paternal and maternal gene flow and contributions of gametic heterogeneities to overall genetic composition of dispersed seeds of Pinus densiflora (Pinaceae).

PREMISE OF THE STUDY: Genetic variability in monoecious woody plant populations results from the assemblage of individuals issued from asymmetrical male and female reproductive functions, produced during spatially and temporarily heterogeneous reproductive and dispersal events. Here we investigated the dispersal patterns and levels of genetic diversity and differentiation of both paternal and maternal gametes in a natural population of Pinus densiflora at the multiple-year scale as long as five consecutive years. • METHODS: We analyzed the paternity and maternity for 1576 seeds and 454 candidate adult trees using nuclear DNA polymorphisms of diploid biparental embryos and haploid maternal megagametophytes at eight microsatellite loci. • KEY RESULTS: Despite the low levels of genetic differentiation among gamete groups, a two-way AMOVA analysis showed that the parental origin (paternal vs. maternal gametes), the year of gamete production and their interaction had significant effects on the genetic composition of the seeds. While maternal gamete groups showed a significant FST value across the 5 years, this was not true for their paternal counterparts. Within the population, we found that the relative reproductive contributions of the paternal vs. the maternal parent differed among adult trees, the maternal contributions showing a larger year-to-year fluctuation. • CONCLUSIONS: The overall genetic variability of dispersed seeds appeared to result from two sources of heterogeneity: the difference between paternal and maternal patterns of reproduction and gamete dispersal and year-to-year heterogeneity of reproduction of adult trees, especially in their maternal reproduction.

Effects of pollen supply and quality on seed formation and maturation in Pinus densiflora.

To understand the detailed mechanisms underlying variations in seed productivity per cone, it is important to examine simultaneously the effects of two pollination mode components (pollen supply and quality) on two seed production processes (seed formation and maturation). We conducted artificial pollination experiments with four pollination treatments (selfing, polycross, no-pollination and open-pollination treatments) in each of two vertical crown layers (upper and lower) for 19 Pinus densiflora ramets. We measured formed seeds as a proportion of ovules (P(Form)), and filled seeds as a proportion of formed seeds (P(Fill)) per cone in each treatment and layer, and inferred the relative influences of pollination mode and resource availability on seed productivity. In the no-pollination treatment, no seeds were formed in any cones of all five ramets. The Generalized Linear Model showed that there were no significant differences in P(Form) both between selfing and polycross treatments and upper and lower layers. The mean P(Fill) values in the selfing treatment were significantly lower than those in the polycross treatment in both layers. The mean P(Fill)s of the two layers did not differ significantly in the selfing treatment, but did in the open-pollination and polycross treatments. The results show that pollen supply affects mainly seed formation, whereas pollen quality affects mainly seed maturation. Resource availability also affects mainly seed maturation, if pollen quality is higher than a certain threshold.

Simultaneous evaluation of paternal and maternal immigrant gene flow and the implications for the overall genetic composition of Pinus densiflora dispersed seeds.

When considering the genetic implications of immigrant gene flow, it is important to evaluate both the proportions of immigrant gametes and their genetic composition. We simultaneously investigated paternal and maternal gene flow in dispersed seeds in a natural population of Pinus densiflora located along a ridge. The paternity and maternity of a total of 454 dispersed seeds (in 2004 and 2005) were accurately and separately assigned to 454 candidate adult trees, by analyzing the nuclear DNA of both diploid biparentally derived embryos and haploid maternally derived megagametophytes of the seeds. The relative genetic diversities and differences between within-population and immigrant groups of both paternally and maternally derived gametes (4 groups) that formed the genotypes of the seeds were evaluated. Using 8 microsatellite markers, we found that 64.0-72.6% of paternally derived gametes, and 17.8-20.2% of maternally derived gametes, were from other populations. Principal coordinate analysis showed that the 4 gamete groups tended to be plotted at different locations on the scattergram, indicating that they each have different genetic compositions. Substantial paternal and maternal immigrant gene flow occurred in this population, and therefore, the overall genetic variation of dispersed seeds is enhanced by both paternally and maternally derived immigrant gametes.