Microsatellites, or simple sequence repeats (SSRs), are usually regarded as the "markers of choice" in population genetics research because they exhibit high variability. The development cost of these markers is usually high. In addition, microsatellite primers developed for one species often do not cross-amplify in related species, requiring separate development for each species. However, microsatellites found in expressed sequence tags (ESTs) might better cross-amplify as they reside in or near conserved coding DNA. In this study, we identified 14 Pinus taeda (loblolly pine) EST-SSRs from public EST databases and tested for their cross-species transferability to P. contorta ssp. latifolia, P. ponderosa, and P. sylvestris. As part of our development of a P. contorta microsatellite set, we also compared their transferability to that of 99 traditional microsatellite markers developed in P. taeda and tested on P. contorta ssp. latifolia. Compared to traditional microsatellites, EST-SSRs had higher transfer rates across pine species; however, the level of polymorphism of microsatellites derived from ESTs was lower. Sequence analyses revealed that the frequencies of insertions/deletions and base substitutions were lower in EST-SSRs than in other types of microsatellites, confirming that EST-SSRs are more conserved than traditional SSRs. Our results also provide a battery of 23 polymorphic, robust microsatellite primer pairs for lodgepole pine.

Complete pedigree information is a prerequisite for modern breeding and the ranking of parents and offspring for selection and deployment decisions. DNA fingerprinting and pedigree reconstruction can substitute for artificial matings, by allowing parentage delineation of naturally produced offspring. Here, we report on the efficacy of a breeding concept called “Breeding without Breeding” (BwB) that circumvents artificial matings, focusing instead on a subset of randomly sampled, maternally known but paternally unknown offspring to delineate their paternal parentage. We then generate the information needed to rank those offspring and their paternal parents, using a combination of complete (full-sib: FS) and incomplete (half-sib: HS) analyses of the constructed pedigrees. Using a random sample of wind-pollinated offspring from 15 females (seed donors), growing in a 41-parent western larch population, BwB is evaluated and compared to two commonly used testing methods that rely on either incomplete (maternal half-sib, open-pollinated: OP) or complete (FS) pedigree designs. BwB produced results superior to those from the incomplete design and virtually identical to those from the complete pedigree methods. The combined use of complete and incomplete pedigree information permitted evaluating all parents, both maternal and paternal, as well as all offspring, a result that could not have been accomplished with either the OP or FS methods alone. We also discuss the optimum experimental setting, in terms of the proportion of fingerprinted offspring, the size of the assembled maternal and paternal half-sib families, the role of external gene flow, and selfing, as well as the number of parents that could be realistically tested with BwB.

. Pedigree and mating system analyses in a western larch (Larix occidentalis Nutt.) experimental population. Annals of Forest Science, Springer Verlag/EDP Sciences, 2008, 65 (7), pp.1. Ann. For. Sci. 65 (2008) Abstract•The mating pattern and gene flow in a western larch (Larix occidentalis Nutt.) experimental population was studied with the aid of microsatellite markers and a combination of paternity-mating system analysis. The commonly difficult to assess, male gametic contribution was determined with 95% confidence and its impact on genetic gain and diversity was determined.• Male fertility success rate ranged between 0 and 11%. Male reproductive output parental imbalance was observed with 50% of the pollen being produced by the top 5% of males while the lower 39% males only produced 10% of the pollen.• A significant difference was observed between male effective population size (genetic diversity) estimates from paternity assignment compared to those based on population's census number (21 vs. 41); however, this difference did not affect estimates of genetic gain.• A total of 221 full-fib families were identified (sample size range: 1-8) and were nested among the studied 14 seed-donors.• A combination of paternity-mating system analysis is recommended to provide a better insight into seed orchards' mating dynamics. While pollen flow tends to inflate mating system's outcrossing rate, the paternity analysis effectively determined the rate and magnitude of contamination across receptive females. • Le taux de succès reproductif mâle était compris entre 0 et 11 %. Un déséquilibre dans la contribution des parents mâles a été observé avec la production de 50 % du pollen par 5 % des pères alors que 39 % d'entre eux ne contribuaient que pour seulement 10 % du pollen. Mots-clés• Une différence significative a été observée entre la taille efficace de la population mâle (diversité génétique) estimée par la recherche de paternité et celle basée sur les effectifs recensés de la population (21 vs. 41) ; cependant, cette différence n'affecte pas l'estimation du gain génétique.• 221 familles de plein-frères ont été identifiées (effectifs entre 1 et 8), regroupées parmi les 14 arbresmères étudiés.• La combinaison d'une analyse de paternité et du système de reproduction est recommandée pour étudier de manière approfondie la dynamique de croisement en vergers à graines. Tandis que les flux de pollen tendent à augmenter le taux d'inter-croisements, l'analyse de paternité détermine de manière effective le taux et l'amplitude de contamination des arbres-mères.

Seed orchards’ parental reproductive success is often assessed through surveys of seed- and pollen-cone production (reproductive investment). Although the reliability of this approach has been questioned, its speed and ease made it the method of choice for providing approximate estimates for the genetic quality and quantity of seed orchard crops. In this study, reproductive investment data from various female and male phenotypic assessment methods from three seed orchards (lodgepole pine ( Pinus contorta Dougl. ex. Loud. var. latifolia Engelm.), Douglas-fir ( Pseudotsuga menziesii (Mirb.) Franco), and western larch ( Larix occidentalis Nutt.)) were compared with reproductive success estimates based on DNA fingerprinting and parentage analyses. The use of reproductive investment as a proxy to reproductive success on estimating seed crop’s volume genetic worth and diversity (measured by effective number of parents) was evaluated. Regression analyses indicated that the phenotypic assessment methods reflect the true parental reproductive success and that estimates of seed- and pollen-cone volume production appear to be the most appropriate indicators of female and male reproductive success, respectively. Although the best combination of female–male survey methods produced inflated effective number of parents estimates, the genetic worth estimates were similar to those based on DNA, highlighting their robustness to fertility variation assessment.