Conifers normally go through a long juvenile period, for Norway spruce (Picea abies) around 20 to 25 years, before developing male and female cones. We have grown plants from inbred crosses of a naturally occurring spruce mutant (acrocona). One-fourth of the segregating acrocona plants initiate cones already in their second growth cycle, suggesting control by a single locus. The early cone-setting properties of the acrocona mutant were utilized to identify candidate genes involved in vegetative-toreproductive phase change in Norway spruce. Poly(A + ) RNA samples from apical and basal shoots of cone-setting and noncone-setting plants were subjected to high-throughput sequencing (RNA-seq). We assembled and investigated 33,383 expressed putative protein-coding acrocona transcripts. Eight transcripts were differentially expressed between selected sample pairs. One of these (Acr42124_1) was significantly up-regulated in apical shoot samples from cone-setting acrocona plants, and the encoded protein belongs to the MADS box gene family of transcription factors. Using quantitative real-time polymerase chain reaction with independently derived plant material, we confirmed that the MADS box gene is up-regulated in both needles and buds of cone-inducing shoots when reproductive identity is determined. Our results constitute important steps for the development of a rapid cycling model system that can be used to study gene function in conifers. In addition, our data suggest the involvement of a MADS box transcription factor in the vegetative-to-reproductive phase change in Norway spruce.The two most commonly grown and economically important conifers in Sweden, Scots pine (Pinus sylvestris) and Norway spruce (Picea abies), both go through a long vegetative growth phase before they begin to produce cones; the vegetative growth period is 20 to 25 years and 8 to 20 years for Norway spruce and Scots pine, respectively. Mature Scots pine trees produce cones every year, while mature Norway spruce trees produce cones only every 3rd to 5th year. The timing of cone establishment is largely synchronized within the various spruce populations and is determined by a combination of genetic and environmental factors (Lindgren et al., 1977;Högberg and Eriksson, 1994). The long generation time and the irregular cone setting between different years pose major obstacles for breeding of Norway spruce both with respect to genetic gain and retained diversity. To guarantee a sufficient seed supply, forest companies have to plant and maintain large seed orchards and in addition store large quantities of seeds to cover the demand during years of little or no seed production. Both conifer breeding programs and the production of improved seed for forest regeneration would benefit from methods to control the length of the vegetative period before the trees start to produce cones as well as the possibilities to control cone setting and cone production itself. Despite that, our knowledge of the genetic mechanisms that regulate the transition from vegetati...

Seed orchards are forest tree production populations for supplying the forest industry with consistent and abundant seed crops of superior genetic quality. However, genetic quality can be severely affected by non-random mating among parents and the occurrence of background pollination. This study analyzed mating structure and background pollination in six large isolation tents established in a clonal Scots pine seed orchard in northern Sweden. The isolation tents were intended to form a physical barrier against background pollen and induce earlier flowering relative to the surrounding trees. We scored flowering phenology inside and outside the tents and tracked airborne pollen density inside and outside the seed orchard in three consecutive pollination seasons. We genotyped 5683 offspring collected from the tents and open controls using nine microsatellite loci, and assigned paternity using simple exclusion method. We found that tent trees shed pollen and exhibited maximum female receptivity approximately 1 week earlier than trees in open control. The majority of matings in tents (78.3 %) occurred at distances within two trees apart (about 5 m). Selffertilization was relatively high (average 21.8 %) in tents without supplemental pollination (SP), but it was substantially reduced in tents with SP (average 7.7 %). Pollen contamination was low in open controls (4.8-7.1 %), and all tents remained entirely free of foreign pollen. Our study demonstrates that tent isolation is effective in blocking pollen immigration and in manipulating flowering phenology. When complimented with supplemental pollination, it could become a useful seed orchard management practice to optimize the gain and diversity of seed orchard crops.

A total of 172 clones of Eucalyptus camaldulensis were tested in three clonal tests in northern, north-central and southern Vietnam, with 32 of them planted across all three sites. At age 3-5 years, the clonal repeatabilities were 0.18-0.42 for growth traits, 0.71-0.78 for wood basic density and 0.56-0.66 for pilodyn penetration. Genotypic correlations between growth and density at the three sites were from -0.24 to 0.17, and did not differ significantly from zero. Genotypic correlations between sites were 0.32-0.56 for growth traits at age 3 years, and 0.72-0.88 for density and pilodyn penetration. Selection gains for breast height diameter at individual sites at a selection proportion of 5% were 22-32%, with minor effects on density. Selection for diameter at one site gave indirect responses in diameter at the other two sites that were only 40-60% of the gains obtainable from direct selection at those sites. This study shows that fast-growing E. camaldulensis clones can be selected in Vietnam with only minor effects on density. Selection for growth should be regionally based to maximize selection gain whereas clonal rankings for density will change little across regions.

Insects feeding in conifer cones are difficult to control with nonsystemic insecticides. Newly developed systemic insecticides that can be injected into tree trunks may be a possible way of reducing both insect damage and negative side-effects to the surrounding environment, compared with conventional spraying. Several insecticides that could be injected into tree stems were tested on Picea abies (L.) Karst. In one experiment, insecticides (bifenthrin, deltamethrin, abamectin, and imidacloprid) were injected during flowering; in a second experiment two of these insecticides (abamectin and imidacloprid) were injected 1 yr before the expected flowering. In the second experiment insecticide treatment was also combined with treatments with the flower stimulating hormone, gibberellin (GA(4/7)). The only insecticide that reduced damage was abamectin, both after injection during flowering and after injection 1 yr before the expected flowering. Injections with GA(4/7) increased flowering and were as efficient as the conventional application method of drilling but abamectin was not effective in combination with the drilling method. There was no negative effect of the insecticide injections on seed quality. The injections were ineffective against the seed chalcid Megastigmus strobilobius (Ratzeburg), which was found to have an unexpected, negative effect on seed quality. Our results suggest that it may be possible to reduce damage from certain insect species, and to increase flowering by injecting abamectin and GA(4/7) in the year before a cone crop.

The effects of the application of wood ash and of fertilizer regimes including phosphorus (P) and potassium (K), with and without simultaneous addition of nitrogen (N), were investigated on a stand of Scots pine (Pinus sylvestris L.) saplings growing on a drained oligotrophic peatland site in southern Sweden. A randomized block design was used. Tree growth and concentrations of various elements in the needles were measured. The addition of similar doses of P (approx. 40 kg P ha-1) from different sources resulted in similar growth responses, amounting to 1.6-1.9 m 3 ha-1 yr-1 of stem wood over the 26-year study. The P source was either wood ash (2500 kg d.w. ha-1) or PK-fertilizer (raw phosphate and potassium chloride). In response to several treatments there were both increased numbers of trees and increased growth of individual trees. The high PK-dose (40 kg P ha-1 and 80 kg K ha-1) appeared to result in a larger growth increase than the low dose (20 kg P ha-1 and 40 kg K ha-1). The N treatment had no additional effect on growth. In the control plots, tree growth was more or less negligible (0.04 m 3 ha-1 yr-1). After almost 26 years, concentrations of P and K in the needles of treated plants were still higher than in the untreated control plants. Nevertheless, in spite of the elevated P concentration, P appears to limit the growth of Scots pine. In conclusion, after sufficient drainage of this type of peatland site, it is possible for a forest stand to develop to the pole stage if wood ash or PK-fertilizer is applied.

In Fennoscandia, both Scots pine (Pinus sylvestris L.) and Norway spruce (Picea abies (L.) Karst.) often fail to produce mature seed, especially in the northern parts of their range. Therefore, cone and seed crop predictions are of major strategic importance for maintaining sustainable multipurpose forestry. We present functions for predicting germination capacity of Pinus sylvestris and Picea abies seed over a wide geographic area. The functions are based on germination analyses for 1297 Pinus sylvestris and 597 Picea abies natural stands in Sweden during 1971-1994. Meteorological data from 71 weather stations were used to calculate heat sums with threshold values from 4 to 10°C and two durations of growing season (ending August 31 or September 30). Logistic regression was utilised for parameter estimates. Accumulated heat sum (threshold 5°C) from start of growing season until August 31 in combination with number of days from estimated time of fertilisation until approximate time for embryo growth cessation gave the best function. The function shows that Picea abies has lower temperature requirements for producing mature seed than Pinus sylvestris. A germination capacity of 95% is reached at a heat sum of 875 degree-days for Picea abies and at 975 degree-days for Pinus sylvestris.

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We investigated mating structure and gene flow in a clonal seed orchard of Scots pine (Pinus sylvestris L.) over three consecutive pollination seasons (2010-2012) with nine nuclear microsatellite markers. The paternity of 1991 offspring from four maternal parents was assigned to 28 candidate fathers using an exclusion procedure and a likelihood-based method implemented in the program CERVUS. Relative reproductive success was highly variable among pollen parents but consistent across years and ranged from 0.1% to 18.3%. Consequently, the seed crops' effective number of fathers was reduced to 52.9%, 48.8%, and 45.7% of the census in the three seasons, respectively. Self-fertilization fluctuated around the orchard's expected value of 5.1%, reaching 4.05%, 7.71%, and 6.61%, respectively. Pollen contamination was estimated to be 5.64%, 7.29%, and 4.89%, respectively, after correction for cryptic gene flow. CERVUS provided similar results as the exclusion method, but estimates greatly varied depending on the input parameters, mainly the proportion of fathers sampled. These results indicate the studied seed orchard is a well-functioning production population with only minor negative effects of self-fertilization and pollen contamination on the quality of seed crops. Genotyping issues associated with microsatellites as a potential source of false paternity assignment and exclusion are discussed.