Pedigree and mating system analyses in a western larch (Larix occidentalis Nutt.) experimental population

?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.

     

Genetic variation, mating patterns and gene flow in a Pinus pinaster Aiton clonal seed orchard

? Relatedness among parents, variation in clonal fertility and background pollination deviate the realized genetic gain and the gene diversity of open pollinated seed orchard from expectation, in particular in wind pollinated species such as Pinus pinaster Aiton.

? This work investigates the genetic variation, the mating system and the pollen contamination in a P. pinaster clonal seed orchard (CSO), by screening the 60 clones from the CSO and the seeds collected from 21 mother-trees with three nuclear microsatellites.

? The expected diversity was similar, but the observed heterozygosity decreased 20% in the progenies compared with the parental trees. The outcrossing rate was 90.1%, the biparental inbreeding 21.7% computed through a multilocus approach, and the observed selfing 3.9%. The observed gene flow from outside the CSO was 52.4%.

? From the results we concluded that the observed gene flow and the biparental inbreeding were high, and care should be taken in the implementation and management of future CSO, in particular clones should be checked for relatedness and the ramet number could be directly proportional to their breeding value.

     

Pollen dispersal,mating patterns and pollen contamination in an insect-pollinated seed orchard of <Emphasis Type="Italic">Schima superba</Emphasis> Gardn. et Champ.

Pollen flow and mating patterns are the most important factors influencing the genetic structure of insect-pollinated forest trees and are essential parts of genetic management in seed orchards. We investigated pollen flow, the mating system and the level of pollen contamination in a clonal seed orchard of Schima superba Gardn. et Champ. In total, 328 open-pollination progenies coming from 11 mother trees were identified using 13 polymorphic microsatellite loci. A total of 203 full-sib families were identified and were nested among the 11 studied seed donors. The male reproductive success rate from 0.49 to 7.77% for most male parents, with an average of 2.44%. More than 80% of the crosses were found within a distance of 60 m, and the most frequent pollination distance between female parent and male parents was approximately 20 m. Mating system analysis showed that the outcrossing level was very high (t _m  = 1.000, outcrossing rate = 98.5%) in the seed orchard and that there was an average of 2.3 effective pollen donors (N _ep ) per female parent. In addition, the mating success of individual males within neighborhoods was moderately influenced by their fecundity and the direction of their location relative to mother trees. The pollen contamination from outside the seed orchard was high (7.01%). Our findings are valuable for the assessment of seed orchards, and it may be worthwhile to use pollen management strategies to decrease pollen contamination and increase the genetic quality of the seeds produced.     

Evaluating Supplemental Mass Pollination Efficacy in a Lodgepole Pine Orchard in British Columbia using Chloroplast DNA Markers

Supplemental mass pollination (SMP) success in a grafted lodgepole pine (Pinus contorta Dougl. ex Loud. var. latifolia Engelm.) seed orchard in southern British Columbia was studied by employing four chloroplast DNA (cpDNA) markers to determine fertilization success of 10 pollen parents. SMP was conducted operationally with a bulked pollen mix twice during peak receptivity in the seed orchard. Fertilization success of the 10 SMP parents averaged 16% greater than for wind-pollinated controls in a different section of the orchard. SMP also increased the uniformity of the male contribution in treated seeds.     

Success rate of supplemental mass pollination in a seed orchard of Pinus sylvestris L.

Two experiments (Expt 1, Expt 2) with supplemental mass pollination (SMP) were carried out in a seed orchard of Pinus sylvestris L. in order to assess the biological and operational prospects for SMP. The success of the SMP was studied by means of allozyme markers. In Expt 1 the pollen mix was applied very close to each female strobilus. The estimated success rates for treatments varied between 66% and 84%. When supplemental pollination was done at peak receptivity, one single pollination was enough to obtain high success rates. In Expt 2 the pollen mix was dusted over clusters of receptive strobili on whole‐trees. The estimated success rates for treatments varied between 10% and 23%. The success rates improved when the trees were pollinated on more than one occasion per day during the receptive period. The overall conclusion is that SMP is practical for P. sylvestris seed orchards.     

Estimates of pollen contamination and selfing in a coastal Douglas-fir seed orchard

Seed orchards are the link from tree breeding to reforestation programs and are theoretically expected to function as closed, perfect populations, ensuring gain and diversity are consistently and predictively delivered as improved seed and seedlings. Seed orchard populations often deviate from panmixia due to fertility variation, reproductive asynchrony, and gene flow, leading to reduced seed crops genetic quality. Here, as a part of multiyear monitoring study, we used DNA fingerprinting (simple sequence repeat markers) to assess a Douglas-fir (Pseudotsuga menziesii) seed orchard's seed crop genetic quality (2009 seed crop). The studied seed crop was produced under ambient temperature (i.e. no reproductive phenology manipulation) and pollination was augmented by pollen from within orchard's pollen donors. DNA fingerprinting of the parental population (66 parents) along with 207 gametophyte (1n) – embryo (2n) pairs of random bulk sample of seed allowed parentage (maternal and paternal) assignment and the direct assessment of pollen contamination (0.18 ± 0.027) and selfing (0.17 ± 0.025) rates as well as parental (pollen, ovule and individual parent) gametic contribution was compared to a previous year's crop (2005). The extended reproductive phenology coupled with variable within-orchard pollen availability has created opportunities for both self and foreign pollen to be successful at various times resulting in the seemingly paradoxical scenario of high selfing and gene flow. These results (2005 and 2009) allowed comparison of seed orchard's crop management practices and are expected to provide scientific foundations to effective seed crops genetic quality improvement.     

Genetic parameters for grain angle in 28-year-old Norway spruce progeny trials and their parent seed orchard

? Sawn wood which acquires twist during drying is a substantial problem for the sawmill industry and is to a large extent caused by spiralled grain in the tree.

? In this study, the feasibility of using spiral grain angles as a selection trait in Norway spruce (Picea abies (L.) Karst) tree breeding and the extent of grain angle and its breeding values were investigated. Grain angles under bark and diameter were measured at breast height in three 28-year-old Norway spruce progeny trials and in their parent seed orchard.

? The mean grain angle value was 1.76° in the progeny trials and 1.84° in the seed orchard. Estimated genetic standard deviations for grain angle were almost 1°. Progeny trial heritabilities (H ² > 0.3) were larger than the seed orchard heritability (H ² = 0.24). Genotype by environment interaction among the progeny trials was negligible for grain angle. Progeny trial grain angles exhibited genetic correlation with seed orchard grain angles in the range 0.66 to 0.84.

? Thus, mature wood grain angles are also under considerable genetic control, raising the opportunity to reduce the occurence of left-grained trees, which are prone to producing twisted wood.

     

Measuring wood density by means of X-ray computer tomography

? Wood density is a characteristic of major interest. Usually, it is used as an indicator of wood quality; however, in the context of global change, it is increasingly used for biomass and carbon storage estimations. X-ray computer tomography is a method which enables quick estimates of wood density after applying a calibration procedure.

? A review of the literature is presented in this article. Most of the previous studies have been performed in the 80’s or at the beginning of the 90’s.

? In this study, the relationship between wood density and Hounsfield numbers was investigated using a recent medical scanner. A linear relationship was fitted using a calibration data set which consisted in tropical wood samples representing a large range of densities ranging between 133 and 1319 kg m^?3, and then validated using an independent data set (mainly temperate tree species). The fitted relationships were very strong (R ² > 0.999), whichever the tested scanner settings, with slight but significant effects of the current voltage and reconstruction filters. The RMSE values computed from the validation data set ranged between 5.4 and 7.7 kg m^?3 for densities ranging between 364 and 821 kg m^?3.

? In conclusion, this method of calibration enables the use of a medical scanner to obtain maps of wood density, in a fast and non destructive way, and with a very good accuracy. Very interesting perspectives are opened regarding biomass distribution within trees.

     

Using SSR markers to study the mechanism of 2n pollen formation in Populus × euramericana (Dode) Guinier and P. × popularis

? Using cytological methods and SSR DNA marker analysis, this study revealed the formation mechanisms and the genetic constitutions of the 2n pollen in Populus × euramericana (Dode) Guinier and P. × popularis.

? In P. × euramericana (Dode) Guinier, four abnormalities in microsporogenesis were observed: parallel spindle, fused spindle, tripolar spindle and premature cytokinesis. The first three can lead to first-division restitution (FDR) 2n pollen formation and the last one can form second-division restitution (SDR) 2n pollen. The SSR marker analysis results of parents and their tetraploidy filial generation confirmed that the genetic constitution of 2n pollen produced by P. × euramericana (Dode) Guinier was FDR.

? In P. × popularis, three of these abnormalities were observed: parallel spindle, fused spindle and premature cytokinesis. The SSR marker analysis results showed the genetic constitution of 2n pollen produced by P. × popularis was SDR. Natural 2n female gametes in P. × euramericana (Dode) Guinier are reported for the first time. SSR analysis indicated that natural 2n female gametes of P. × euramericana (Dode) Guinier did exist and were fertile, which could be FDR genetic constitution.

? The results from this study showed a great potential for using 2n gametes to produce polyploid poplar clones, which can be used effectively for polyploid breeding for poplar species in the section Aigeiros.

     

Hybridization between Quercus robur and Q. petraea in a mixed oak stand in Denmark

? Hybridization and mating pattern between Quercus robur and Q. petraea was studied in a 5.8 ha mixed forest stand in Jutland, Denmark which comprises in total 135 Quercus robur and 230 Q. petraea trees. Classification of the oak trees into species was performed using canonical discriminant analysis of a range of leaf morphological traits. Adult trees (365) and offspring (582) were genotyped with eight microsatellite markers. Seedlings were sampled in 2003 and acorns were collected in 2004.

? Mating patterns of Q. robur and Q. petraea are expected to be different in the northern range of the distribution area and a larger hybridization rate is expected. It is further expected, that pollination from outside sources will be relatively less in small fragmented forest management systems compared to large scale oak forest. The conclusions should be verified through repeated year to year analysis of the mating pattern.

? Phenological studies revealed that there was no difference in flowering time between species. Data for the adult trees revealed no significant departures from Hardy-Weinberg proportions and there was weak, but significant spatial genetic structure, which supports the idea that the stand is of natural origin. Spatial genetic structure in the first distance class is stronger for Q. petraea. The genetic composition of the offspring was remarkably consistent from year to year. Paternity analysis revealed that, on average, 85% pollination came from fathers within the stand. The direction of the pollen flow varied from year to year. Inter-specific hybridization was high and ranged from 15–17% and from 48–55% for Q. petraea and Q. robur mothers respectively. Paternity analysis revealed that the population was basically outcrossing and only 3.7% of the analysed progeny were the product of selfing. Over the two years of study, approximately 200 trees contributed to the paternity of the next generations.

? The study confirms earlier studies with a greater tendency for Q. robur mothers to produce hybrid seeds than Q. petraea mothers. The rate of hybridization is higher in this Danish stand than in comparable studies elsewhere in Europe. Gene flow from outside sources are relatively low.

     

Forward selection in a maritime pine polycross progeny trial using pedigree reconstruction

Key message

Molecular markers were used for paternity recovery in a maritime pine ( Pinus pinaster Ait.) polycross trial, facilitating forward selection. Different breeding strategies for seed orchard establishment were evaluated by comparing genetic gains and diversity. This work opens up new perspectives in maritime pine breeding.

Context

Polycross mating designs are widely used in forest tree breeding to evaluate parental breeding values for backward selection. Alternatively, polycross progeny trials may be used to select the best trees on the basis of individual breeding values and molecular pedigree analysis.

Aims

This study aimed to test such a forward selection strategy for the maritime pine breeding program.

Methods

In a maritime pine polycross trial, progeny with higher breeding values for growth and stem straightness was first preselected with or without relatedness constraints. After paternity recovery, the preselected trees were ranked on the basis of their breeding values, estimated from the recovered full pedigree. Finally, the best candidates were selected with three different strategies (forward, backward, mixed) and three levels of coancestry constraints to establish a virtual clonal seed orchard.

Results

Complete pedigrees were successfully recovered for most of the preselected trees. There was no major difference in expected genetic gains between the two preselection strategies which differed for relatedness constraints. Genetic gains were slightly higher for forward selection than for classical backward selection.

Conclusion

This seminal study opens up new perspectives for using forward selection within the French maritime pine breeding program.

     

Genetic structure of Tunisian natural carob tree (Ceratonia siliqua L.) populations inferred from RAPD markers

? Seven RAPD markers were used to assess the genetic diversity and structure of ten Tunisian natural Ceratonia siliqua L. populations from different geographic and bioclimatic zones.

? The species maintain a high diversity within population as estimated by the percentage of polymorphic loci and Shannon’s index (P% = 76.31, $\bar H'_{pop} = 0.569$ ). The range of variation between populations was large. Populations from the upper semi-arid bioclimates, with more continuous distribution area showed the highest level of variation.

? A high genetic differentiation among populations (Φ_ST = 0.250 and $\bar G_{ST} = 0.347$ ), as a result of population isolation was revealed. Nevertheless, the genetic structure is in accordance with bioclimate indicating that ecological factors also should influence differentiation. Populations from the sub-humid, upper semi-arid and mean semi-arid zones clustered together and were distinct from those of the lower semi-arid ones.

? Conservation strategy should be made according to the level of polymorphism within population and bioclimate.

     

Saprotrophic versus symbiotic strategy during truffle ascocarp development under holm oak. A response based on 13C and 15N natural abundance

? The development of truffles in the soil is not well understood. It is not known if a direct transfer of carbohydrates takes place between the host tree and the developing ascocarps through ectomycorrhizal structures or whether sporophores become independent from their hosts after several weeks or months and are able to use dead host tissues or soil organic matter as carbon (C) and nitrogen (N) sources.

? To study saprophytic or symbiotic capacities of truffle ascocarps the natural abundance of ¹⁵N and ¹³C in foliage, wood, fine roots, mycorrhizae, fungal sporophores and soil were determined in a truffle orchard.

? The processes of carbon and nitrogen allocation remained unchanged during the entire period of ascocarp development of Tuber melanosporum. From ¹³C and ¹⁵N natural abundance measurements, T. melanosporum, T. brumale and T. rufum did not exhibit saprotophic strategy during ascocarp development, which is contradictory to common statements found in handbooks regarding truffle cultivation.

     

Variation in the decay resistance of larch to fungi

? Decay resistance of larch (Larix sp.) to fungi was evaluated on heartwood samples belonging to 3 species (L. decidua, L. kaempferi and their hybrid), 3 races of European larch (polonica, sudetica and alpine), 13 wood lots (populations) and 313 trees.

? Larch wood appeared, on average, as moderately durable although a high variability was observed. At the sample level as well as at the mean individual tree level, durability ranged from class 1 to 5 according to EN 350-1 standard. At the population level, larch wood varied from ‘durable’ to ‘slightly durable’. Genetics played a major role in decay resistance at the species, provenance and tree levels. Environmental factors such as the position of heartwood samples and the age of trees were also identified as a source of variability.

? The most durable wood was not necessarily from old native alpine stands of European larch: some young larches from faster growing lowland origins also produced durable wood.

? Genetic improvement of larch wood durability appeared therefore likely by the selection of the best populations for decay resistance as well as from the selection of individuals.

     

Modulation of leaf physiology by age and in response to abiotic constraints in young cuttings of two Populus deltoides × P. nigra genotypes

? It is of importance, when comparing physiological responses of leaves to environmental constraints among different genotypes, to take into account any effect related to leaf position and age within the canopy that might interfere with the response to the constraints.

? To document such effects, photosynthetic capacity and tolerance to heat and to oxidation were measured on leaves from the top to the bottom of three-month-old single-stem rooted cuttings of Populus deltoides × P. nigra genotypes, ‘Dorskamp’ and ‘Luisa_Avanzo’, thus taking into account a gradient of ages from youngest and still expanding (top) to oldest and fully expanded (bottom) leaves.

? All recorded traits were tightly linked to the age of the leaves. Photosynthetic capacity gradually increased during leaf expansion, in parallel with chlorophyll content and relative nitrogen allocation to RuBisCO and to bioenergetics. On the contrary, dark respiration gradually decreased during leaf expansion until a minimum value was reached at maturity. Compared to expanding leaves, young mature leaves were characterized by a lower sensitivity to heat and a higher one to oxidations generated by methyl-viologen.

? Leaf characteristics appeared to vary along the stem to a larger extent than between the two genotypes that display largely different productivities in plantations.

     

Black pine (Pinus nigra Arn.) growth divergence along a latitudinal gradient in Western Mediterranean mountains

? Most studies of tree-growth and climate report positive responses to global warming in high latitudes and negative responses at lower ones.

? We analyzed tree-ring width of Pinus nigra Arn. along a 500 km latitudinal transect in the Iberian Peninsula to study the temporal trend and climate forcing in tree radial growth during the last century.

? Tree growth was enhanced by cool summers and moist cold seasons. Increased moisture stress has decreased tree growth rates. However, we present evidence of growth increases in some trees in all sampled populations after 1980’s. Climate change negatively (positively) affected between 72% (5%) of trees in the southern populations and 40% (25%) in the north Trees with positive growth trends were favored by winter temperatures and their abundance was inversely correlated with forest productivity.

? Our findings add evidences of tree growth divergence in the Mediterranean basin and show the gradual transition between forests where positive (temperate and boreal) and negative (Mediterranean) growth trends dominate.

     

Stimulation of tree defenses by Ophiostomatoid fungi can explain attack success of bark beetles on conifers

? Our aim is to present why the hypothesis, that Ophiostomatoid fungi play an important role in the establishment of most bark beetle species on living conifers, is valuable.

? After summarizing knowledge about the relationships of bark beetles with conifers and fungi, we conclude that controversy results from misinterpretations when using fungal pathogenicity to demonstrate the role of Ophiostomatoid fungi in beetle establishment on host trees.

? We demonstrate that fungal pathogenicity is not the right parameter to appreciate the role of fungus in beetle establishment on host trees. We argue that artificial low density inoculations that allow the appreciation of fungus ability to stimulate tree defenses and thus to help beetles in overcoming tree resistance must be used in complement to mass inoculations. In both cases, results must be expressed in terms of tree defense stimulation rather than in terms of tree killing.

1. Fungal species stimulating tree defenses are generally not those that grow the best in the sapwood.
2. We argue that beetle development in the phloem, fungal invasion of the sapwood and phloem, and tree death, occur after tree defenses are exhausted, and that any fungus present in the beetle gallery could thus potentially invade the sapwood after defense exhaustion.

? We conclude that stimulation of the tree defense reactions in both the phloem and the superficial sapwood is a real benefit brought by fungi to the beetles during the first phase of establishment (overcoming tree resistance).

? Considering the origin of the bark beetle fungus associations attacking living trees and their general functioning based on stimulation of tree defenses, we develop three hypotheses:

1. any beetle species would be helped in its establishment in a given tree species by developing an association, even loosely, with a fungus species belonging to the Ophiostomatoid flora of that tree species;
2. the necessity of a considerably low level of tree resistance for fungus extension into the tree is the selection pressure that has led fungi to develop their intrinsic ability to stimulate tree defenses, through their ability to grow into the phloem. This association can be completed by antagonistic fungal species controlling extension of the previous fungal species in the tree tissues;
3. Beetle species using the strategy of overcoming tree resistance are associated with a fungal complex, of which species could assume three roles regarding relationships between beetles and trees: 1- to stimulate tree defenses in the phloem and superficial sapwood, 2- to grow into the sapwood after tree resistance is overcome, and 3- to control phloem extension of the first other two categories. Bringing nutrients to the beetle progeny can be a fourth role.

? We propose that bark beetle — Ophiostomatoid associations can be categorized, based on associations’ frequency and complexity while taking into account beetle aggressiveness. We show that a close correspondence exists between beetles’ aggressiveness and the ability of their main associated fungal species to stimulate the defenses of their host tree.

? We conclude with suggesting that most sapwood invading fungi might be “cheaters” which have taken advantage of the efficiency of the relationship between beetles and fungi that stimulate tree defenses.

     

Diminished vessel diameter as a possible factor in the decline of European ash (Fraxinus excelsior L.)

? The aim of this work was to examine the anatomy and functioning of secondary xylem in stems in relation to the decline of European ash (Fraxinus excelsior L.).

? We tested the hypothesis that declining trees show changes in the structure of wood, which result in impaired water transport.

? Anatomical analyses were carried out on wood samples (comprising all annual rings formed during the 30 years life of the analysed trees) collected at breast height from the main stem of healthy, weakened and dead ash trees. The width of annual wood increments, the diameter and density of earlywood vessels were measured and the theoretical hydraulic conductivity index through the secondary xylem calculated by application of Hagen-Poisseuille’s formula.

? Anatomical characteristics changed both with the age of trees and in response to unfavorable factors. The largest vessels were observed in healthy trees, which implied that they had the highest hydraulic conductivity index, whereas trees considered to be in decline produce smaller vessels and hence had reduced conductivity.

     

Integrating functional diversity into tropical forest plantation designs to study ecosystem processes

? The elucidation of relationships between biodiversity and ecosystem processes has been limited by the definition of metrics of biodiversity and their integration into experimental design. Functional trait screening can strengthen the performance of these designs.

? We suggest the use of Rao’s quadratic entropy to measure both functional diversity and phylogenetic diversity of species mixtures proposed for an experimental design, and demonstrate how they can provide complementary information.

? We also present an index assessing the statistical performance of these independent variables in different experimental designs. Measurement of independent variables as continuous vs. discrete variables reduces statistical performance, but improves the model by quantifying species differences masked by group assignments.

? To illustrate these advances, we present an example from a tropical forest tree community in which we screened 38 species for nine functional traits. The proposed TropiDEP design is based on the relative orthogonality of two multivariate trait axes defined using principal component analysis.

? We propose that independent variables describing functional diversity might be grouped to calculate independent variables describing suites of different traits with potentially different effects on particular ecosystem processes. In other systems these axes may differ from those reported here, yet the methods of analysis integrating functional and phylogenetic diversity into experimental design could be universal.