Ole K. Hansen

Chloroplast microsatellite variation in Abies nordmanniana and simulation of causes for low differentiation among populations

Fifteen populations of Abies nordmanniana, originating from all main parts of its distributional area in the Caucasian region, were genotyped for three chloroplast microsatellites as well as one mitochondrial marker. The chloroplast microsatellites were highly variable, resulting in a total of 111 haplotypes in 361 analysed individuals, while the mitochondrial marker showed no variation. Analysis of molecular variance attributed 2.1% of the variation in the microsatellites to be among populations, and no correlation between geographic distribution and genetic distances among populations could be observed. A simulation study was conducted to investigate to what extent the low genetic differentiation among populations could be a result of size homoplasy in the applied microsatellites. However, the simulations indicated that the low differentiation more likely is caused by high gene flow among populations.

Establishment of a quasi-field trial in Abies nordmanniana—test of a new approach to forest tree breeding

This study used DNA markers to establish a quasi-field trial within a production Christmas tree stand produced from seed collected in an open-pollinated clonal seed orchard (CSO). A total of 660 offspring from the CSO, which comprised 99 clones of Abies nordmanniana, were genotyped with 12 microsatellites. Parentage was assigned successfully to 93% and 98% of the progeny at 95% and 80% confidence, respectively. The assignment rate declined only to 90% when the number of markers was reduced to 10. The distribution of parentage to the offspring among the CSO clones was highly skewed. The most successful clone was assigned as parent in 7% of the cases, and only 92 of the 119 potential parental genotypes were assigned as parents. The obtained pedigree was used to estimate breeding values for the CSO clones for five characters relevant for Christmas tree breeding. For high-heritability traits, such as flushing, accurate breeding values could be estimated for a considerable proportion of the clones. To estimate breeding values for low-heritability traits, such as Christmas tree quality score, more genotyped offspring will be required. The largest drawback of the method is the highly skewed distribution of parentage among the parents in the seed orchards, making it difficult to calculate breeding values for all clones. The approach seems well suited for tree breeding that puts more emphasis on pure selection of parental genotypes and less on estimating quantitative genetic parameters.

Microsatellites used to establish full pedigree in a half-sib trial and correlation between number of male strobili and paternal success

Abstract• Paternity was established in a field trial of Abies nordmanniana with open-pollinated (OP) offspring from a clonal seed orchard (CSO) comprising 23 clones.• The purposes were to: (1) investigate the violation of the assumption of true half-sibs in OP progeny trials; (2) assess the value of male strobili scorings to predict paternal contributions in CSOs; and (3) study the bias in breeding values and heritabilities obtained in breeding with OP family trials due to unknown paternity.• The paternal contribution to the offspring varied a lot among the clones, but the resulting violation of the assumption of true half-sibs in the progeny trial was only modest.• On average 84% of the relationships among the offspring were true half-sib, resulting in an average genetic correlation of 0.29 (range 0.26 to 0.33).• Male strobili scorings from the seed harvest year did well in forecasting the paternal contribution to the offspring. The linear regression of sired progeny on the estimated clonal proportion of male strobili explained 76% of the variation.• The large variation in siring success and existence of other types of relationships than true half-sibs among the offspring only gave minor bias in estimated genetic parameters.

Nordic provenance trials with Abies lasiocarpa and Abies lasiocarpa var. arizonica: three-year results

In three Nordic field trials, 26 provenances of Abies lasiocarpa (Hook.) Nutt. and Abies lasiocarpa var. arizonica (Merriam) Lemmon were evaluated 3 yrs after establishment. The focus was on 16 traits important for adaptation, establishment and production of Christmas trees. For all three sites in Denmark, Norway and Iceland, survival ranged from 89 to 96%. Provenances showed statistically significant differences for all but two traits. For all traits there was significant interaction between provenance and site. Analysis of ecovalens showed that for most traits, only a limited number of provenances contributed substantially to the interaction. Therefore, a large proportion of the provenances showed a fairly consistent ranking across sites. The frequency of potential Christmas trees in the provenances ranged from 10 to 49%. Southern provenances from New Mexico and Arizona seemed to have the best potential for producing high-quality Christmas trees. The southern provenances added substantially to the interaction between provenance and site for budset, lammas growth and colour. When using multivariate statistical methods on all measured traits, the provenance clusters fitted neatly into six distinct geographical regions.

Genetic resources of teak (Tectona grandis Linn. f.)—strong genetic structure among natural populations

Twenty-nine provenances of teak (Tectona grandis Linn. f.) representing the full natural distribution range of the species were genotyped with microsatellite DNA markers to analyse genetic diversity and population genetic structure. Provenances originating from the semi-moist east coast of India had the highest genetic diversity while provenances from Laos showed the lowest. In the eastern part of the natural distribution area, comprising Myanmar, Thailand and Laos, there was a strong clinal decrease in genetic diversity the further east the provenance was located. Overall, the pattern of genetic diversity supports the hypothesis that teak has its centre of origin in India, from where it spread eastwards. The analysis of molecular variance (AMOVA) gave an overall highly significant Fst value of 0.227—population pairwise Fst values were in the range 0.01–0.48. Applying the G″st differentiation parameter, the estimated overall differentiation was 0.632, implying a strong genetic structure among populations. A neighbour-joining (NJ) tree, using the pairwise population matrix of G″st values as input, contained three distinct groups: (1) the eight provenances from Thailand and Laos, (2) the Indian provenances from the dry interior and the moist west coast and (3) the provenances from northern Myanmar. The provenances from southern Myanmar were placed close to the root of the tree together with the three provenances from the semi-moist east coast of India. A Bayesian cluster analysis using the STRUCTURE software gave very similar results, with three main clusters, each containing two sub-clusters, while Bayesian cluster analysis in the Geneland software, exploiting the spatial coordinates of the provenances, resulted in five clusters in accordance with the former results. The implications of the findings for conservation and use of genetic resources of the species are discussed.

Species variation in susceptibility to the fungus Neonectria neomacrospora in the genus Abies

ABSTRACT The fungus Neonectria neomacrospora has recently caused an epidemic outbreak in conifer species within the genus Abies in Denmark and Norway. Christmas tree producers in Europe and North America rely, to a large extent, on Abies species. The damage caused by N. neomacrospora, including dead shoot tips, red flagging of branches and potentially dead trees, have therefore caused concern about reduced quality and loss of trees, and thereby of revenue. Field observations of natural infection of 39 taxa, from 32 species, within the genus Abies in the Hørsholm Arboretum, Denmark, were evaluated; significant differences were seen between taxa, that is, species, and between some species and their subspecies. The Greek fir, Abies cephalonica, was the only species without damage. An inoculation experiment on detached twigs with mycelium plugs from a N. neomacrospora culture showed that all species could be infected. The damage observed in the inoculation experiment could explain 30% of the variation in the field observations based on species mean values. The epidemic outbreak and the high number of species susceptible to this fungus indicate that N. neomacrospora requires attention in the cultivation and conservation of Abies species.

Response to selfing in seed set, seedling establishment and nursery growth based on controlled crosses of Abies nordmanniana clones.

Abstract Nordmann fir (Abies nordmanniana) is used for production of high value Christmas trees in short rotation. Potentially all trees are intended to be sold - no thinnings are normally applied. A high proportion of saleable trees per ha is the main key to secure economic return to the growers. Consumers demand a symmetric and rather uniform tree and, dependent on local traditions, it should be more or less dense and narrow. In Denmark, breeding of Nordmann fir and establishment of seed orchards focus on seed supply and improved Christmas tree quality - aiming at adaption and a rather uniform ideotype Christmas tree. Benefits from employing rather few clones, to maximise gain and uniformity, may potentially be counteracted by increased selfing. The objective of this study was to quantify the effect of selfing on percentage of filled seed and nursery establishment of seedlings. Inbreeding depression was seen for filled seeds (40%), growth traits (5-17%), plus mortality and axial damage (5-12 percent units). Many selfed seedlings survive and develop into marketable seedlings, although with a depression in numbers of 23-37%, or 9-12 percent units. Nursery sorting procedures can only partially reduce the number of inbred seedlings in Nordmann fir. The large variation among clones in the response to selfing indicates that knowledge of the behaviour of selfed progeny from specific clones in ‘small number seed orchards’ is of practical interest. Trade-offs between increased gain by selection of few clones and a penalty paid for increased inbreeding need further studies during a full Christmas tree rotation.

Transformational restoration: novel ecosystems in Denmark

Abstract Restoring the estimated 1 billion hectares of degraded forests must consider future climate accompanied by novel ecosystems. Transformational restoration can play a key role in adaptation to climate change but it is conceptually the most divergent from contemporary approaches favoring native species and natural disturbance regimes. Here, we review concepts of novelty in ecosystems with examples of emergent/neo-native and designed novel ecosystems, with application to transformational restoration. Danish forests have a high degree of novelty and provide a realistic context for discussing assisted migration, one method of transformational adaptation. Deforestation and impacts of past land use created a highly degraded landscape dominated by heathland in western Denmark. Restoration with non-native species began 150 years ago because the native broadleaves could not establish on the heathlands. Danish forestry continues to rely extensively on non-native species. Preparing for transformational adaptation requires risky research today to prepare for events in the future and refugia from the last glaciation may provide genetic material better adapted to future climate. A new project will test whether species and provenances from the Caspian forests in Iran possess greater genetic diversity and superior resistance (physiological adaptability) and resilience (evolutionary adaptability) and possibly a gene pool for future adaptation.

Worldwide translocation of teak—origin of landraces and present genetic base

Teak (Tectona grandis Linn. f.) is one of the major plantation timbers of the world. The species is native to India, Myanmar, Thailand and Laos in South East Asia but was translocated to several countries in Africa and Central and South America during the past century. Today, large areas of plantations are grown outside the species native range. It is speculated that genetic bottlenecks and founder effects combined with new selection pressures under new growing conditions have led to the formation of distinct landraces; this hypothesis is supported by results from international provenance tests. In the present study, we apply genetic markers to identify the likely origin of teak grown outside its native range and examine if the landraces show signs of reduced genetic diversity. We find large variation in the level of diversity among landraces, although not larger than that observed among native populations. We conclude that variation in the studied teak landraces probably reflects their areas of genetic origin rather than severe founder effects created during their introduction. The genetic data suggests that the studied landraces originated from either the semi-moist east coast of India, southern Myanmar or western Thailand. These results indicate that translocation of teak has mainly come from a certain part of the native distribution and that this did not include the widespread natural teak areas of southern, dry interior or western India or northern Myanmar.

Ad hoc breeding of Abies bornmülleriana for Christmas tree production using a combination of DNA markers and quantitative genetics—a case study

Quasi-field trials (QFTs) using offspring from first-generation plus trees of Abies bornmülleriana were established on two sites in Denmark via DNA markers and parentage analysis. The aim of this study was to demonstrate an example of ad hoc breeding to make better genetic material available for Christmas tree production. Assessments of several important Christmas tree quality-associated phenotypic traits were made for all offspring individuals in delineated areas of the production stands, and moreover the best individuals in the whole production stands were identified. Postharvest needle retention was evaluated in the parent population in both 2016 and 2017. The results showed that by combining available methodology and pre-selected material, substantial genetic gain in relation to Christmas tree production in A. bornmülleriana could be achieved. Backward selection seems tempting due to its shorter time horizon for delivering improved genetic material. Postharvest needle retention evaluation can be a cheap but important last step in modifying the backward selection. We propose to combine the results based on QFTs with selective seed harvest and paternity tests to give the best possible starting point for production of clonal A. bornmülleriana plants using somatic embryogenesis (SE).