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.     

Jackknife resampling for precision measurement of direct gene flow estimates

Gene flow from outside pollen sources reduces the expected genetic gain of seed orchards, and has been quantified by genotyping seed samples from crop years to determine the proportion of seed sired by outside sources. Random bulk seed sampling and microsatellite genotyping allows for fast, unbiased identification of orchard contamination rates. Bulk seed sampling has been employed in multiple studies of seed orchard gene flow, but a method of determining precision of gene flow estimates from this sampling method has yet to be described. In this study, we used jackknife resampling to generate random samples of migrant and within-population genotypes obtained from an empirical sample of Douglas-fir (Pseudotsuga menziesii [Mirb.] Franco) genotypes of five microsatellite loci. We generated samples of 50, 100, 150, 200, and 250 genotyped individuals from a sample of 400 seeds, with genotypes corresponding to low (6%), moderate (14%), or high (56%) pollen contamination levels. The mean gene flow estimate and confidence interval were calculated for each sample size and gene flow scenario to determine the precision of gene flow estimates obtained from each sample size in each contaminant gene flow scenario. Genotyping 150 individuals was sufficient to produce gene flow estimates with calculated confidence interval values below ± 1%, indicating that 150 genotyped individuals were sufficient for precise gene flow estimation in this population. This resampling method will enable precise and unbiased estimation of gene flow into seed orchards for future monitoring of gene flow, to allow better management of seed orchard genetic resources.     

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.     

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.     

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.     

Swedish lodgepole pine seed orchard crops tested in north-west Russia

Abstract

Tree vitality and height of lodgepole pine (Pinus contorta var. latifolia) originating from the six Swedish seed orchards and native Scots pine (Pinus sylvestris L.) were estimated in three 8-yr-old test plantations established in the Komi Republic (north-west Russia). A randomized row-plot design with 9–13 replicates of each entry was used. In general, the Scots pine tree vitality was better than that of lodgepole pine but these differences were significant only for seed sources of southern origins Larslund, Rumhult, and Österby. On the contrary, the lodgepole pine height growth was faster (6–21%) except for seed sources of southern origins Rumhult and Österby. In comparison with the native Scots pine, lower mean heights (4–10%) were recorded for these seed sources. Among the lodgepole pine seed sources the northern origins had better vitality, while the effect of latitude on the tree height was insignificant.     

Background pollination in Pinus sylvestris seed orchards

The proportion of background pollen grains in the seed crops of two Pinus sylvestris seed orchards in Central Finland was estimated with the aid of multilocus allozyme markers. The orchards studied were more than 30 years old and in full pollen production. For the bulked seed crops of the seed orchard with southern clones the estimated average of background pollination over four years was 26%. There were statistically significant differences between years. No significant heterogeneity in the degree of background pollination between clones was found. Among single ramets there was significant heterogeneity in the estimated contamination rates, but the variation was not related to position in the orchard. For the seed orchard with northern clones the bulked seed crop was studied only for one year and the level of background pollination was found to be 33 %. These estimates are fairly high, but lower than for many other orchards. Background pollination at this level will cause losses in expected genetic gains. Part of the seeds from northern orchards will not be adapted to the intended area of use.     

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.     

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.     

Higher resistance of the offspring of Scots pine trees resulting from natural regeneration in old foci of Heterobasidion annosum root rot

Abstract

Heterobasidion annosum (Fr.) Bref. is a fungal pathogen causing annosum root rot – one of the most economically important diseases in coniferous stands. The major aim of this study was to compare the resistance of the offspring of Scots pine trees (Pinus sylvestris L.) from seed orchards and the offspring of trees that were naturally reproduced in old foci of the disease. In experiments conducted in vitro, we used 960 seedlings which were grown from seeds collected from 60 trees in 6 old foci of the disease and 640 seedlings grown from seeds collected in 4 seed orchards. The offspring of trees from seed orchards after inoculation with H. annosum had nearly twofold higher mortality rate than the seedlings developed from the seeds collected in old foci of the disease. This suggests that the offspring of self-sown trees in old foci of the disease has a greater, genetically conditioned resistance to annosum root rot.     

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).     

杉木初级种子园种用价值测定初步结果

本文报道了我国近50个已形成商品生产力的杉木种子园,种子遗传品质的大田测验幼林结果。试验表明:①各地种子园的种子与未经改良的种子一样,均有不同的遗传品质,说明并非所有种子园种子都有同等使用价值;②不少种子园的种子比当地一般种子的遗传品质要高,说明具有一定的改良效果,但比优良种源区的种子的种用价值又低了些。可见,不经实地试验,推广杉木种子园种子,应取慎重态度。为了提高初级种子园的遗传品质,极需要做种子园的去劣淘汰。     

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。     

Cost-benefit analysis of using orchard or stand seed in Scots pine sowing,the case of northern Finland

A cost-benefit analysis of orchard seed versus seed from existing natural stands is presented for the scenario of direct sowing of Scots pine (Pinus sylvestris L.). The differential benefits of orchard seed are seed quality and improved yield due to genetic gain. These benefits are expected to further reduce the total cost of orchard seed. Seedlings from seed orchard seed had 11% better survival rate (p < .001, the best clone over 17%, p < .010) than seedlings from local stand seed. This was observed in a case study of 15 young seeding trials located in Central and North Finland. Expected yield improvements for stands established with orchard seed were obtained from local progeny test results, and they were further simulated on tree growth using the MELA forest simulation program. The cost-benefit analysis showed that, with a 7% yield improvement and a 15% better seed quality, the net cost of orchard seed was less than that of stand seed (discount rate 3%).     

Magnitude and efficiency of genetic diversity captured from seed stands of <Emphasis Type="Italic">Pinus nigra</Emphasis> (Arnold) subsp. <Emphasis Type="Italic">pallasiana</Emphasis> in established seed orchards and plantations

To test how efficiently plantations and seed orchards captured genetic diversity from natural Anatolian black pine (Pinus nigra Arnold subspecies pallasiana Holmboe) seed stands, seed sources were chosen from 3 different categories (seed stands (SS), seed orchards (SO) and plantations (P)) comprising 4 different breeding zones of the species in Turkey. Twenty-five trees (mother trees) were selected from each SS, SO and P seed sources and were screened with 11 Random Amplified Polymorphic DNA (RAPD) markers. Estimated genetic diversity parameters were found to be generally high in all Anatolian black pine seed sources and the majority of genetic diversity is contained within seed sources (94%). No significant difference in genetic diversity parameters (numbers of effective alleles, % of polymorphic loci and heterozygosity) among seed source categories was found, except for a slight increase in observed heterozygosities in seed orchards. For all seed source categories, observed heterozygosity values were higher (H_o = 0.49 for SS, 0.55 for SO and 0.49 for P) than expected ones (H_e = 0.40 for SS, 0.39 for SO and 0.38 for P) indicating the excess of heterozygotes. In general, genetic diversity in seed stands has been transferred successfully into seed orchards and plantations. However, the use of seeds from seed orchards can increase the amount of genetic diversity in plantations further. The study also demonstrated that number of plus-tree clones (25–38) used in the establishment of seed orchards was adequate to capture the high level of diversity from natural stands.     

Advanced generation seed orchards’ turnover as affected by breeding advance,time to sexual maturity and costs,with special reference to Pinus sylvestris in Sweden

Abstract

The effect of various biological, genetic, economic and management factors relevant to advanced generation seed orchard establishment was investigated using numerical estimates for Scots pine (Pinus sylvestris L.) in Sweden. Factors considered were planting density, rate of genetic advance in the breeding population, timing of first seed collection, seed value, seed production cost (stratified to establishment, annual management, cone harvest and seed extraction), orchard rotation age and contamination level. The developed model demonstrated its utility in studying and evaluating various economic and biological options associated with advanced generation seed orchard establishment/turnover. Sensitivity analyses demonstrated the robustness of the developed model through various arbitrarily changes in genetic gain advances, establishment, management and seed production, particularly those associated with cone harvests from upper crown and costs. The Swedish Scots pine case study produced results supporting faster turnover of seed orchard generations (30 vs 40 years) with shorter orchard lifespan (early start of seed cone after 8 years vs 15 years) delivering higher gain through minimizing the genetic gain differential between the breeding and production populations and allowing the capture of this gain for inclusion in the seed orchards. Orchard planting densities of 400 and 600 grafts per hectare produced similar results with marginal differences, and the latter was recommended for future orchard establishment.     

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.     

The influence of pine twist rust (Melampsora pinitorqua) on growth and development of Scots pine (Pinus sylvestris)

The influence of pine twist must (Alelampsora pinitorqsat) on growth and development of Scots pine was studied during three consecutive years. The experimental material included three provenances from natural stands and one open pollinated seed orchrd progeny. Trees severely affected by pine twist must reached an average of 38% lower height than did not-affected trees. while mildly affected trees grew somewhat better, on the average, than did non-affected trees. Pine twist must caused tha greatest loss of height increment in the seed orchard progeny.     

Predicting individual breeding values and making forward selections from open-pollinated progeny test trials for seed orchard establishment of interior Lodgepole Pine (<Emphasis Type="Italic">Pinus contorta</Emphasis> ssp. <Emphasis Type="Italic">latifolia</Emphasis>) in British Columbia

Ten-year height data from three open-pollinated lodgepole pine progeny test trials were analysed in order to select individuals for seed orchard establishment in the low and high elevation seed-planning units (TOLO and TOHI) of the new Thompson Okanagan seed planning-zone in interior British Columbia, Canada. The total test population had about 35,000 individuals from 362 families of 9 provenances and was divided into two portions, each serving as the base population for its appropriate seed-planning unit. For each population, various genetic parameters were estimated and individual breeding values were predicted. Combined-site estimates of individual, family, and within-family heritability ranged from 0.23 to 0.38, 0.77 to 0.84, and 0.17 to 0.29, respectively. Estimated coefficients of type-b genetic correlation were greater than 0.81, indicating little G × E interaction in this region. Provenance effects were large. Larch Hills and Inonoaklin River were the two superior provenances with an expected gain of 13% and 10% in stem volume at rotation age of 60 years in TOLO. Individual breeding values was not uniformly distributed among provenances and the top 100 individuals originated from 20 families in TOLO and 32 families in TOHI. Co-ancestry among these individuals should not be a serious concern in seed orchard establishment as the predicted genetic loss due to excessive inbreeding is only 1.06% in TOLO and 0.6% in TOHI. The status numbers of the two prospective seed orchards are 32 and 44, respectively, and both are well above the required minimum of 10 in British Columbia. Using unrelated clones for seed orchard establishment may result in reduced optimum genetic gain and thus is not a favourable option.     

山地条件下油松花粉的传播及其对该地形中种子园的花粉侵染

1988～1989年在卢氏油松无性系种子园和人工油松林内外设立花粉采样点,观测花粉在不同山地类型的传播特点及对下风方向种子园侵染的危险性。结果表明:油松花粉传播受地形、散粉日期、风速、大气湿度等因素的影响。风速大、空气干燥,天气晴朗的日子,散粉速度快,传粉距离远,在开阔地、抬升地、对坡地形中,距离花粉源数百米甚至数千米地点的花粉浓度常高于花粉源的花粉浓度,对坡地形尤为显著。因而,在营建山地种子园时,至少应保持1000 m的隔离距离。本文还结合实验结果,对山地种子园的建立和管理提出了一些看法。