W. T. Adams

Complex patterns of mating revealed in a Eucalyptus regnans seed orchard using allozyme markers and the neighbourhood model.

The neighbourhood model apportions offspring of individual mother plants to self‐fertilization, outcrossing to males within a circumscribed area around the mother plant (the neighbourhood), and outcrossing to males outside the neighbourhood. Formerly the model was applied only to haploid pollen gametes in the offspring of conifers, but is extended so that it can be used with genotypic data from diploid offspring of both angiosperms and gymnosperms. In addition, it is shown that the mating parameters can be estimated without independent estimates of allele frequencies in the pollen pools outside the neighbourhood; thus the model might be applied effectively to natural populations exposed to unknown external pollen sources. Parameters of the neighbourhood mating model were estimated for a 10‐year‐old seed orchard population of the insect‐pollinated tree, Eucalyptus regnans, in southeast Australia, which contained a mixture of two geographical provenances (Victoria and Tasmania). The mating patterns revealed were complex. Crosses between trees of the same provenance occurred three times more often than crosses between trees of different provenances. Levels of self‐fertilization and patterns of mating within neighbourhoods were influenced by provenance origin, crop fecundity and orchard position (central vs. edge) of mother trees. Gene dispersal, however, was extensive, with approximately 50% of effective pollen gametes coming from males more than 40 m away from mother trees (average distance between neighbouring trees was 7.4 m). Thus, insect pollinators are efficient promoters of cross‐fertilization in this orchard, with the result that the effective number of males mating with each female is large.

Mating patterns and pollen dispersal in a natural knobcone pine (Pinus attenuata Lemmon.) stand

Mating system and effective pollen dispersal were studied in a natural stand of knobcone pine (Pinus attenuata Lemmon.) using 11 isozyme loci. Analyses were performed by fitting neighbourhood and mixed-mating models to multilocus genotypic arrays of offspring from four mother trees. Neighbourhoods consisted of all potential outcross males within 11 m of each mother tree (44, on average). Average outcrossing rates of the mother trees were estimated to be 0.97 and 0.96 for the respective models, whereas the population-wide outcrossing rate based on the mixed-mating model and a broader sample of mother trees was 0.92. The estimated proportion of offspring sired by males outside the neighbourhood of each mother tree was 0.56. Thus about 41 per cent of matings resulted from outcrossing with nearby males (within neighbourhoods). Distance and direction of individual males from mother trees and the size (tree height) of males played significant roles in determining outcross mating patterns within neighbourhoods. Male mating success increased with both proximity and tree size, although males east of mother trees accounted for more offspring than males in other directions. Despite the role of proximity, directionality and tree size in determining mating success within neighbourhoods, the effective number of males mating with each female seems to be large.

Gene dispersal within forest tree populations

Patterns of gene dispersal by seeds and pollen greatly influence the genetic structure of plant populations and their effective size. This paper reviews methods of measuring gene dispersal and current information on patterns of dispersal within local populations of forest trees. Recently, a number of statistical procedures for investigating gene movement based on the use of large numbers of isozyme loci have been described. These procedures include various forms of parentage analysis and the fitting of mating models to genotypic arrays of offspring from individual maternal plants. With the levels of genetic discrimination currently possible in forest trees, the model approach appears to be the most reliable means of estimating gene dispersal parameters. Too little data are available to draw general conclusions about patterns of gene movement within natural populations. Nevertheless, reports to date indicate that dispersal by both pollen and seed can be considerable. For any one mother tree, the bulk of effective pollen in conifers may come from distant males in the same stand or from surrounding stands (gene flow). In insect-pollinated angiosperms, gene flow may also be substantial, but cross-fertilization within stands may primarily be between nearest flowering trees.

Estimation of gene flow into two seed orchards of loblolly pine (Pinus taeda L.)

SummaryMultilocus allozyme markers, which were present in background stands but not in two seed orchards, enabled estimation of gene flow into two seed orchards of loblolly pine. Estimates averaged 36% in two orchards over three years. When pollen pool frequencies in the background stands were used rather than gene frequencies in the non-orchard trees, the estimates averaged 60%. In one year, trees in the row on the edge of one orchard received significantly more pollen from background stands. Estimates of gene flow between the two 2-ha orchards average 10%. Implications of the observed levels of gene flow are discussed in terms of population genetics and seed orchard management.

The mating system in natural and shelterwood stands of Douglas-fir

SummaryMating systems in two pairs of old-growth uncut and adjacent shelterwood stands of Douglas-fir (Pseudotsuga menziesii var. ‘menziesii’) were compared by estimating the proportions of viable progenies due to outcrossing (t) with both single-locus and multilocus techniques. Single-locus population estimates (ťs), ranging from 0.41 to 1.16, were significantly (P<0.05) heterogeneous among loci in three of four stands; mean single-locus estimates for shelterwoods were not significantly different from those for uncut stands. Multilocus population estimates (ťm) ranged from 0.94 to 1.00; again, estimates for shelterwoods were not significantly different from those for uncut stands. Multilocus estimates were slightly higher than mean single-locus estimates for uncut stands but were nearly equivalent for shelterwoods, suggesting that related matings other than selfs may be associated with uncut stands, but not shelterwoods. Individual-tree outcrossing rates (ťmi), estimated for six trees in each shelterwood, ranged from 0.90 to 1.10 and were significantly heterogeneous among trees at one of the two shelterwoods. Outcrossing was high (> 0.90) in both uncult and shelterwood stands, and no evidence indicated that low parent-tree density had affected stand outcrossing rates.

Estimating pollen flow using SSR markers and paternity exclusion: accounting for mistyping

Highly informative genetic markers, such as simple sequence repeats (SSRs), can be used to directly measure pollen flow by parentage analysis. However, mistyping (i.e. false inference of genotypes caused by the occurrence of null alleles, mutations, and detection errors) can lead to substantial biases in the estimates obtained. Using computer simulations, we evaluated a direct method for estimating pollen immigration using SSR markers and a paternity exclusion approach. This method accounts for mistyping and does not rely on assumptions about the distribution of male reproductive success. If ignored, even minor rates of mistyping (1.5%) resulted in overestimating pollen immigration by up to 150%. When we required at least two mismatching loci before excluding candidate fathers from paternity, the resulting pollen immigration estimates had small biases for rates of mistyping up to 4.5%. Requiring at least three mismatches for exclusion was needed to minimize the upward biases of pollen immigration caused by rates of mistyping up to 10.5%. The minimum number of highly variable SSR loci needed to minimize cryptic gene flow and obtain reliable estimates of pollen immigration varied from five to seven for a sampling scheme applicable to most conifers (i.e. when paternal haplotypes can be unambiguously determined). Between five and nine highly variable SSR loci were needed for a more general sampling scheme that is applicable to all diploid seed plants. With moderately variable SSR markers, consistently accurate estimates of pollen immigration could be obtained only for rates of mistyping up to 4.5%. We developed the pollen flow (pfl) computer program which can be used to obtain unbiased and precise estimates of pollen immigration under a wide range of conditions, including population sizes as large as 600 parents and mistyping rates as high as 10.5%.

Mapping of quantitative trait loci controlling adaptive traits in coastal Douglas-fir. II. Spring and fall cold-hardiness

Abstract Quantitative trait loci (QTLs) affecting fall and spring cold-hardiness were identified in a three-generation outbred pedigree of coastal Douglas-fir [Pseudotsuga meniziesii (Mirb.) Franco var. menziesii]. Eleven QTLs controlling fall cold-hardiness were detected on four linkage groups, and 15 QTLs controlling spring cold-hardiness were detected on four linkage groups. Only one linkage group contained QTLs for both spring and fall cold-hardiness, and these QTLs tended to map in close proximity to one another. Several QTLs were associated with hardiness in all three shoot tissues assayed in the spring, supporting previous reports that there is synchronization of plant tissues during de-acclimatization. For fall cold-hardiness, co-location of QTLs was not observed for the different tissues assayed, which is consistent with previous reports of less synchronization of hardening in the fall. In several cases, QTLs for spring or fall cold-hardiness mapped to the same location as QTLs controlling spring bud flush. QTL estimations, relative magnitudes of heritabilities, and genetic correlations based on clonal data in this single full-sib family, supports conclusions about the genetic control and relationships among cold-hardiness traits observed in population samples of Douglas-fir in previous studies.

Highly variable SSR markers in Douglas-fir: Mendelian inheritance and map locations

Twenty-two highly variable SSR markers were developed in Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco] from five SSR-enriched genomic libraries. Fifteen PCR primer pairs amplified a single codominant locus, while seven primer pairs occasionally amplified two loci. The Mendelian inheritance of all 22 SSRs was confirmed via segregation analyses in several Douglas-fir families. The mean observed heterozygosity and the mean number of alleles per locus were 0.855 (SE=0.020) and 23 (SE=1.6), respectively. Twenty markers were used in genetic linkage analysis and mapped to ten known linkage groups. Because of their high polymorphism and unambiguous phenotypes, 15 single-locus markers were selected as the most suitable for DNA fingerprinting and parentage analysis. Only three SSRs were sufficient to achieve an average probability of exclusion from paternity of 0.998 in a Douglas-fir seed orchard block consisting of 59 parents.

Family variation for fall cold hardiness in two Washington populations of coastal Douglas-fir (pseudotsuga menziesii var. menziesii (Mirb.) Franco)

Abstract In order to assess the genetics of fall cold hardiness in coastal Douglas-fir (Pseudotsuga menziesii var. menziesii (Mirb.) Franco), shoot cuttings were collected in October from saplings (9-year-old trees) of open-pollinated families in two progeny tests in each of two breeding zones in Washington, one in the Coast range (80 families) and one on the west slope of the Cascade Mountains (89 families). Samples from over 5500 trees were subjected to artificial freezing and visually evaluated for needle, stem and bud tissue injury. The extent to which cold injury is genetically related to tree height and shoot phenology (timing of bud burst and bud set) was also evaluated. Significant family variation was found for all cold hardiness traits; however, individual heritability estimates were relatively low (ranging from 0.09 to 0.22). Significant family-by-test site interaction was detected for needle injury in the Cascade breeding zone, but not in the coastal zone. Genetic correlations (rA) among needle, stem and bud tissues for cold damage were weak (0.16 ≤ rA ≤ 0.58) indicating that genes controlling fall hardening are somewhat different for different tissues. Timing of bud burst and bud set were only weakly correlated with cold injury (rA ≤ 0.49). Thus, bud phenology is a poor predictor of fall cold hardiness in this species. There was no consistent relationship between tree height and cold injury in the coastal zone. In the Cascade zone, taller trees appeared to be more susceptible to cold injury, but the association was weak (mean rA = 0.38, range 0.20–0.72).

Population substructure in continuous and fragmented stands of Populus trichocarpa.

Population substructure has important implications for both basic and applied genetic research. We used 10 microsatellite markers to characterize population substructure in two ecologically and demographically contrasting populations of the model tree Populus trichocarpa. The Marchel site was a continuous stand growing in a mesic habitat in western Oregon, whereas the Vinson site consisted of three disjunct and isolated stands in the high desert of eastern Oregon. A previous study revealed that pollen-mediated gene flow is extensive in both populations. Surprisingly, model-based clustering, principal components analysis and analyses of molecular variance provided overwhelming support for the existence of at least two intermingled sub-populations within the continuous Marchel population (FST=0.026, P<0.001), which occupied an area with a radius of only about 250 m. Genets in these two sub-populations appeared to have different relative clone ages and phenologies, leading us to hypothesize that they correspond to different seedling cohorts, each established from seeds produced by relatively few mothers. As expected, substructure was stronger in the fragmented Vinson population (FST=0.071, P=0.001), and this difference appeared to result from the more extensive family structure in this population. Using group-likelihood methods, we reconstructed multiple interconnected half-sib families in the Vinson population, with some genets having as many as eight putative siblings. Researchers involved in ongoing and future association studies in P. trichocarpa should account for the likely presence of subtle but practically significant substructure in populations throughout the range of this species.