Southern pollen sired more seeds than northern pollen in southern seed orchards established with northern clones of Pinus sylvestris

Abstract

Contamination by southern pollen is a considerable problem in Scots pine (Pinus sylvestris L.) seed orchards established with northern clones. This study investigated whether the contamination was due to the competitive superiority of southern pollen by carrying out competition trials using mixtures of pollen from northern and southern populations of Scots pine. Trials were performed in a southerly seed orchard established with clones originating from northern populations. Seed paternity (siring) was determined through the analysis of allozyme variation. Southern genotypes sired significantly more seeds (76%) than their northern competitors and across all mixed-pollen crosses. Maternal genotype had no effect on seed siring success. The mean flower abortion rate was lower in southern pure-pollen crosses and mixed-pollen crosses than in northern pure-pollen crosses. The results show that local pollen may induce high levels of background pollination in southern seed orchards composed of northern genotype grafts. These results must be taken into account when aiming to produce suitable reforestation material for northern areas.     

Effects of crown-pruning on seed and pollen cone production in two lodgepole pine seed orchards in British Columbia

The effects of top pruning on subsequent seed and pollen cone production in lodgepole pine (Pinus contorta Dougl.) grafted clones in two seed orchards were evaluated. In the older orchard (14 years old), three pruning regimes were applied: an untreated control, a moderate pruning where 25% of the crown was removed and a severe pruning where 40% of the crown was removed. In the younger orchard (ten years old), only a control pruning and a severe pruning (50% of crown removed) were applied. Five-year seed cone production was lower in treated ramets compared to controls in both orchards. However, when cumulative seed cone yields were adjusted for reduced tree heights after pruning, differences were no longer significant in the older orchard. Strong clonal variation was observed in seed cone yields in both orchards. Pollen cone development was less variable among treatments in the older orchard. In the younger orchard, controls consistently produced heavier pollen crops. Pruning recommendations for similar orchards are made and results discussed for reducing the impacts of losses in seed cone yields.Deceased     

Levels of outcrossing and contamination in two Pinus sylvestris L. seed orchards in Northern Sweden

The joint levels of outcrossing and contamination in two young northern Swedish Scots pine (Pinus sylvestris L.) seed orchards were studied with the aid of allozyme markers. High levels of contamination (21–36%) were estimated and were attributed to the observed low within orchard pollen production. Outcrossing rate estimates were high (0.957–0.961), though they departed significantly from complete outcrossing (t= 1.0) in spite of the observed high contamination levels. These results confirmed reported relationships between outcrossing and contamination levels in open‐pollinated seed orchards. It was concluded that a crop management option relying on supplemental mass‐pollination would be effective in reducing both contamination and inbreeding levels.     

Allozyme frequencies,outcrossing rate and pollen contamination in Picea abies seed orchards

Allozymes of Swedish seed orchard clones of Norway spruce (Picea abies Karst.) were studied. There were differences between gene frequencies of origins from different regions. The gene pool of the fertilizating pollen of two different orchards was different. Pollen contamination and outcrossing rates were estimated in two seed orchards. The fraction of paternal gametes which did not originate from any seed orchard clone was 10% and 17%, respectively. Based on simulation it was estimated that only a quarter of the gametes with origin outside the orchard would be detected. Estimation of multilocus outcrossing rates were 0.95 and 0.98.     

Annual Variation in Pollen Contamination and Outcrossing in a Picea abies Seed Orchard

A mature Norway spruce [Picea abies (L.) Karst.] seed orchard, established in southern Finland with 67 clones from northern Finland, was analysed in three different years in order to estimate the pollen contamination ratios. Allozyme-based paternity analysis revealed that the contamination rate was high, 69-71%, and did not differ between the years studied. It appears that, in areas where spruce is the dominant tree species, the contamination rate will be very high even in mature seed orchards. However, the contamination rate in the thinned parts of the orchard was significantly lower than that in the unthinned parts in two of the three years studied. The outcrossing rate was also high, 96-100% in all years, even though the ramets of each clone were planted using a clonal-row design, and there were no significant differences in the outcrossing rates between the different parts of the orchard.     

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.     

Female fertility variation in mature Pinus sylvestris clonal seed orchards

Abstract

Female fertility is the basis for the output of seeds from clonal orchards and its variation is of major interest for the economics and biology of seed orchards, especially for the efficiency and diversity of seed orchard crops. Assessments of female fertility variation in 10 mature (>15 years old) seed orchards of Scots pine (Pinus sylvestris L.) were evaluated and compared. Depending on the individual orchard, fertility variation for each clone was assessed in slightly different ways, e.g. number of strobili, cones, seeds or litre of cones per ramet. Assessments in five of the orchards were made over consecutive years. The main result was that the clonal variation in mean female fertility per surviving ramet was lower than expected from the literature; the Kang–Lindgren sibling coefficient (Ψ) within individual years averaged 1.35. The variation between ramets within clones and years was of similar magnitude as that between clones. Clone by year interactions were noticeable, but were slightly lower than the variation between as well as that within clones in individual years. There was considerable variation in the variance components between years. The limited variation in female fertility indicated that it should not be a selection criterion when selecting clones for a seed orchard. Furthermore, it will not result in large differences between clones in seed set or large reductions in gene diversity in productive Scots pine seed orchards.     

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.     

Mating patterns in an indoor miniature <Emphasis Type="Italic">Cryptomeria japonica</Emphasis> seed orchard as revealed by microsatellite markers

To evaluate the practical application of an indoor seed orchard, we compared the seed production, final germination rates and mating patterns of potted Cryptomeria japonica D. Don clones in two miniature seed orchards: one in a glasshouse and the other outdoors. There was no statistically significant difference in total seed production between the orchards (P = 0.275, ANOVA). However, the final germination rate of seeds produced in the indoor orchard was significantly lower than that of seeds produced in the outdoor orchard (P < 0.01, Wilcoxon test). The average self-fertilization rate was higher in the indoor orchard (27.2%) than in the outdoor orchard (5.6%), and this parameter was strongly associated with the reduced final germination rates of the seeds harvested in the glasshouse. Pollen contamination was considerably lower in the indoor seed orchard (48.9% in the outdoor seed orchard and 4.4% in the indoor seed orchard). χ² tests indicated that the paternal contributions of each constituent clone differed significantly in both orchards (P < 0.001 in both cases). The strong variations in paternal contributions among indoor orchard clones was significantly correlated with their respective pollen production capacities (Spearman ρ = 0.693, P < 0.001). To reduce the high self-fertilization rates, we recommend the use of orchard clones with similar pollen production capacity in indoor seed orchards.     

Reduction of pollen contamination in Scots pine seed orchard crop by tent isolation

This study examined the differences in mating system between indoor versus outdoor Scots pine (Pinus sylvestris L.) seed orchard using DNA markers. By analyzing the paternities in seed crops from isolation tents, we found a significant reduction in contamination rate, from 23% in the outdoor open blocks to 0% inside the tents. Increase in selfing rate and a decrease in genetic diversity in the crops were also observed inside the tents. These undesirable effects can be mitigated to some extent by the supplementation of extra pollen genotypes, e.g. selfing rate decreased from 14.4% to 6.6% and the effective number of fathers increased from 6 to 11.4 in the tents without and with supplemental mass pollination, respectively. Our study showed that tent isolation may become an effective rescuer for seed orchards where pollen contamination is severe.     

Flowering and Airborne Pollen Occurrence in a Pinus sylvestris Seed Orchard Consisting of Northern Clones

To explain the high background pollination rate detected in earlier studies, this study investigated the flowering phenology of a Scots pine seed orchard with respect to airborne pollen occurrence during 1992 and 1993. The receptive period of the female strobili and the shedding period of the male strobili were observed visually. Airborne pollen was monitored with Rotorod traps located inside and outside the seed orchard. There was no time gap in the onset of flowering between the earliest female and male strobili in 1992. Female flowering began 3 days earlier than male flowering in 1993, and the earliest strobili were probably pollinated by pollen transported long distance from more southerly areas. The almost simultaneous peak in pollen occurrence inside the seed orchard and in surrounding pine stands indicates that the seed orchard clones are not temporally isolated from sources of contamination. However, pollen concentrations inside the seed orchard were higher than those in the surrounding stands. Therefore, flowering phenology cannot sufficiently explain the observed pollen contamination rates in this seed orchard.     

Autumn frost hardiness of Pinus sylvestris offspring from seed orchard grafts of different ages

Autumn frost hardiness of offspring from clones (originating from 66.0–68.8° N) in six southern (62° N) and one northern (66.5° N) Pinus sylvestris seed orchards in Finland, was evaluated. One‐year‐old seedlings derived from open pollinated grafts of young, intermediate, and fairly mature age, and natural stand seedlings, were artificially freeze tested. The northerly located orchard produced the least injured offspring. There were no significant systematic differences between age classes. Neither were there any indications of decreased freezing injury with later age classes. The family variance was significant in four orchards. The estimated injury levels of orchard offspring were, in general, closer to those of natural stands in the neighbourhood of the orchard than natural stands at the place of clonal origin. It is suggested that not only differences in mother clone hardiness but also clonal differences in pollen contamination rate caused the observed family variation.     

Mating System and Pollen Contamination in a Pinus brutia Seed Orchard

Mating system parameters and pollen contamination were estimated in an 11-year-old Pinus brutia Ten. clonal seed orchard, located in Antalya, Turkey, with the aid of isozymes. Isozyme analysis was performed on both maternal (megagametophyte) and embryo tissues of seeds collected from the seed orchard and the surrounding natural stand. Fourteen loci encoding nine enzyme systems were assayed. Based on a multilocus estimator, the proportion of viable seeds originating from outcrossing (tˆ_m) in the seed orchard was found to be 0.947; the remainder (0.053) was due to selfing. We estimated that 85.7% of the orchard seed resulted from pollen contamination by trees in surrounding stands. This may be related to the young age and subsequent low pollen production of the orchard. Only 9% [i.e., 1.0−0.857 (contamination)−0.053 (selfing) = 0.090] of the orchard offspring was due to cross-pollination among clones within the orchard. Due to the high level of pollen contamination, it is estimated that genetic gain in this seed crop is less than 57% of that expected if all offspring resulted from random mating among orchard clones (i.e., no contamination).     

Are Phenological Observations Sufficient to Estimate the Quality of Seed Crops from a <Emphasis Type="Italic">Eucalyptus grandis</Emphasis> Open-pollinated Seed Orchard? Consequences for Seed Collections

Reproductive phenology was observed over three years in a Eucalyptus grandis seed orchard in Madagascar to determine the impact of geographic differences of parental selections on the expected genetic composition of seed crops. Pollen flow (observed pollen cloud) was deduced from a paternity assignment based on sampling from one year’s seed production. The two approaches were used to verify whether knowledge of phenology is sufficient to predict the genetic quality of the seeds collected. Despite the high flowering level, with a constant cycle over three years, the results demonstrated wide reproductive phenological differences associated with the parents’ origin, suggesting putative pollination disequilibrium. From both observed and expected pollen clouds, the results showed preferential mating among different provenances, which has consequences for seed crop composition. Phenological observations, which can be made inexpensively, can be used to promote methods of effective seed orchard management in order to improve the genetic quality of seeds.     

林木种子园花粉污染研究进展

林木种子园花粉污染是影响种子园了遗传品质的重要因子，本文综述了国外林木种子园花粉污染研究的概况、污染水平、影响污染水平的因子、减少污染的措施等。并对我国种子园花粉污染研究和花粉管理提出了建议。     

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.     

Parental participation in progeny and effective population sizes in experimental seed orchards of wild cherry Prunus avium L. (Batsch)

Parents and progenies genetic diversity, and male and female contributions to the seed crop were assessed in three experimental Prunus avium seed orchards. Collected data were used to compare different effective population sizes, based on phenological, seed crop and paternity analysis. Our results did not show any difference of genetic diversity between parents and progenies. A limited pollen pollution was detected. We showed that distance and coflowering among clones had a significant effect on effective pollination, and a significant effect of the production of flowers was revealed in one of the seed orchards. Our study also revealed a quite low number of effective size of fathers per mother, but high effective sizes of mothers, fathers and parents at the level of the seed orchard. Finally, the calculation of effective size of mothers, fathers and parents was not highly modified when having the complete information based on the paternity analysis.     

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.     

Pollination dynamics in a Douglas-fir seed orchard as revealed by pedigree reconstruction

• ? Pollination dynamics was studied in a Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) seed orchard using 8 nuclear microsatellite markers and pedigree reconstruction.
• ? The seed orchard consisted of 49 parents (clones). Cone-crop management included bloom delay and supplemental mass pollination (SMP) using 12 internal and 4 external pollen donors.
• ? A random sample of 801 bulk seeds was genotyped for both haploid megagametophyte and corresponding diploid embryo.
• ? Using the parental population’s multilocus genotypes, full pedigree reconstruction generated all the information needed to estimate the maternal, paternal, and parental reproductive success, selfing, pollen contamination, and pollination success of the 4 external pollen donors.
• ? Maternal, paternal, and parental reproductive success varied with 80% of gametes being produced by 23, 45, and 37% of the orchard’s parents, respectively, resulting in a drastically reduced effective population size as compared to the census number (14 vs. 53).
• ? Selfing, pollen contamination, and aggregate SMP success (internal and external) were estimated to be 15.2, 10.4, and 15.0%, respectively.
• ? Full pedigree reconstruction was effective in unraveling the orchard’s pollination dynamics and both female and male reproductive success.

     

Low rates of pollen contamination in a Scots pine seed orchard in Sweden: the exception or the norm?

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.