Novel polymorphic microsatellite loci and patterns of pollen-mediated gene flow in an ex situ population of <Emphasis Type="Italic">Eurycorymbus cavaleriei</Emphasis> (Sapindaceae) as revealed by categorical paternity analysis

Understanding the patterns of contemporary, pollen-mediated gene flow is of great importance for designing appropriate conservation strategies. In this study, ten novel polymorphic microsatellite loci were isolated for the rare dioecious tree, Eurycorymbus cavaleriei, and the patterns of pollen dispersal were investigated in an ex situ conserved population. A combination of microsatellite markers with high-collective exclusion power (0.932) was used to assign paternity to 240 seeds collected from eight maternal trees. The average effective pollen dispersal distance (δ) was 292.6 m and the frequency distribution of pollen movement suggested extensive pollen movement in the population. The effective pollen donors per maternal tree (N _ep) ranged from 5 to 10, and the most isolated maternal trees were observed with the largest number of N _ep = 10. Although a trend of near-neighbor mating was found in seven of eight maternal trees, no significant correlations were detected between the average effective pollen dispersal distance (δ) and the geographic distances (d1 and d2) between maternal and male trees. The increased average effective distance of pollen dispersal and number of N _ep for isolated maternal trees might be a compound consequence of low density and long-distance flight of pollinators of this species. The conservation implications of these results are discussed.     

Mating system and early viability resistance to habitat fragmentation in a bird-pollinated eucalypt

Habitat fragmentation has been shown to disrupt ecosystem processes such as plant-pollinator mutualisms. Consequently, mating patterns in remnant tree populations are expected to shift towards increased inbreeding and reduced pollen diversity, with fitness consequences for future generations. However, mating patterns and phenotypic assessments of open-pollinated progeny have rarely been combined in a single study. Here, we collected seeds from 37 Eucalyptus incrassata trees from contrasting stand densities following recent clearance in a single South Australian population (intact woodland=12.6 trees ha⁻¹; isolated pasture=1.7 trees ha⁻¹; population area=10 km²). 649 progeny from these trees were genotyped at eight microsatellite loci. We estimated genetic diversity, spatial genetic structure, indirect contemporary pollen flow and mating patterns for adults older than the clearance events and open-pollinated progeny sired post-clearance. A proxy of early stage progeny viability was assessed in a common garden experiment. Density had no impact on mating patterns, adult and progeny genetic diversity or progeny growth, but was associated with increased mean pollen dispersal. Weak spatial genetic structure among adults suggests high historical gene flow. We observed preliminary evidence for inbreeding depression related to stress caused by fungal infection, but which was not associated with density. Higher observed heterozygosities in adults compared with progeny may relate to weak selection on progeny and lifetime-accumulated mortality of inbred adults. E. incrassata appears to be resistant to the negative mating pattern and fitness changes expected within fragmented landscapes. This pattern is likely explained by strong outcrossing and regular long-distance pollen flow.     

Pollen dispersal and genetic structure of the tropical tree Dipteryx panamensis in a fragmented Costa Rican landscape

In the face of widespread deforestation, the conservation of rainforest trees relies increasingly on their ability to maintain reproductive processes in fragmented landscapes. Here, we analysed nine microsatellite loci for 218 adults and 325 progeny of the tree Dipteryx panamensis in Costa Rica. Pollen dispersal distances, genetic diversity, genetic structure and spatial autocorrelation were determined for populations in four habitats: continuous forest, forest fragments, pastures adjacent to fragments and isolated pastures. We predicted longer but less frequent pollen movements among increasingly isolated trees. This pattern would lead to lower outcrossing rates for pasture trees, as well as lower genetic diversity and increased structure and spatial autocorrelation among their progeny. Results generally followed these expectations, with the shortest pollen dispersal among continuous forest trees (240 m), moderate distances for fragment (343 m) and adjacent pasture (317 m) populations, and distances of up to 2.3 km in isolated pastures (mean: 557 m). Variance around pollen dispersal estimates also increased with fragmentation, suggesting altered pollination conditions. Outcrossing rates were lower for pasture trees and we found greater spatial autocorrelation and genetic structure among their progeny, as well as a trend towards lower heterozygosity. Paternal reproductive dominance, the pollen contributions from individual fathers, did not vary among habitats, but we did document asymmetric pollen flow between pasture and adjacent fragment populations. We conclude that long-distance pollen dispersal helps maintain gene flow for D. panamensis in this fragmented landscape, but pasture and isolated pasture populations are still at risk of long-term genetic erosion.     

Long-pollen Movement and Deviation of Random Mating in a Low-density Continuous Population of a Tropical Tree Hymenaea courbaril in the Brazilian Amazon

Mating system and pollen flow are two key elements to understand the genetic structure of tree species. Mating and pollen-dispersal patterns of a low-density population of bat pollinated Hymanea courbaril were examined before logging in a 546-ha plot in the Brazilian Amazon. The multilocus genotypes of nine microsatellite loci were determined for 130 adult-trees and 367 seeds collected from 20 seed-trees. Mating system analysis, using mixed-mating model and paternity analysis showed that the studied population is perfectly outcrossed ( t_m = 1.002), and probably self-incompatible. However, significant deviations from random mating were detected for mating among relatives ( t_m − t_s = 0.096, P < 0.05) and correlated matings ( r_p = 0.289, P < 0.05), indicating inbreeding in the population and that part of offspring are full-sibs (28.9%). Inbreeding was reflected in the positive and significant fixation index observed in adult trees ( F = 0.137, P < 0.05), although no significant inbreeding was detected in offspring ( F = 0.074, P > 0.05). The effective number of pollen donors mating with each seed-tree was determined to be low ( N_ep ≈ 4). The average of pollen flow distance was measured inside of the plot by both paternity (827 ± 429 m) and TwoGener analysis (115–363 m). However, this underestimated pollen dispersal distance, since the detected rate of pollen immigration inside of the plot was high (55%). The observed long-pollen dispersal distance is probably related to pollination by bats and the low density of reproductive trees in the site.     

Pollen Dispersal Between Isolated Trees in the Brazilian Savannah: A Case Study of the Neotropical Tree Hymenaea stigonocarpa

This case study examines the pollen dispersal distance, pollen dispersal patterns and intra‐family genetic structure for isolated trees in pastures of the bat‐pollinated Neotropical tree species Hymenaea stigonocarpa using six microsatellite loci and parentage analysis. The sampling included 28 grouped trees (referred to as the population) and six isolated trees in pastureland along a highway in Mato Grosso do Sul State, Brazil. From the population, we sampled 137 seeds from 12 seed‐trees, and from the isolated trees, we sampled 34 seeds from two seed‐trees. The results showed that pollen was dispersed over long distances (reaching 7353 m) and therefore the spatially isolated trees were not reproductively isolated. The pollen immigration rate in the population was also high (31%). Isolated trees presented a higher selfing rate (s=26%) than trees in the population (s=12%), suggesting that the spatial isolation of the trees increased selfing. However, selfing was responsible for only 30 percent of the inbreeding in offspring and mating among relatives was 70 percent. In the population, excluding selfing, ca 72 percent of the pollen was dispersed over distances <1000 m (average: 860 m). For the two isolated seed‐trees, excluding selfing, the average pollen dispersal distance was 5229 m. The results demonstrate that although pollen can be dispersed over long distances for H. stigonocarpa isolated trees, a high percentage of pollen comes from the same tree (selfing) and mating was correlated. Consequently, seeds must be collected from a large number of seed‐trees for conservation purposes.     

Patterns of pollen and seed dispersal in a small, fragmented population of the wind-pollinated tree Araucaria angustifolia in southern Brazil

Paternity analysis based on eight microsatellite loci was used to investigate pollen and seed dispersal patterns of the dioecious wind-pollinated tree, Araucaria angustifolia. The study sites were a 5.4 ha isolated forest fragment and a small tree group situated 1.7 km away, located in Paranalpha State, Brazil. In the forest fragment, 121 males, 99 females, 66 seedlings and 92 juveniles were mapped and genotyped, together with 210 seeds. In the tree group, nine male and two female adults were mapped and genotyped, together with 20 seeds. Paternity analysis within the forest fragment indicated that at least 4% of the seeds, 3% of the seedlings and 7% of the juveniles were fertilized by pollen from trees in the adjacent group, and 6% of the seeds were fertilized by pollen from trees outside these stands. The average pollination distance within the forest fragment was 83 m; when the tree group was included the pollination distance was 2006 m. The average number of effective pollen donors was estimated as 12.6. Mother-trees within the fragment could be assigned to all seedlings and juveniles, suggesting an absence of seed immigration. The distance of seedlings and juveniles from their assigned mother-trees ranged from 0.35 to 291 m (with an average of 83 m). Significant spatial genetic structure among adult trees, seedlings, and juveniles was detected up to 50 m, indicating seed dispersal over a short distance. The effective pollination neighborhood ranged from 0.4 to 3.3 ha. The results suggest that seed dispersal is restricted but that there is long-distance pollen dispersal between the forest fragment and the tree group; thus, the two stands of trees are not isolated.     

Low Interannual Variation of Mating System and Gene Flow of Symphonia globulifera in the Brazilian Amazon

Six microsatellite loci were used to compare the mating system and gene flow in two consecutive years of a natural, unlogged population of Symphonia globulifera in a 500 ha experimental plot in the Brazilian Amazon (Flona Tapajós). The species had a low density of reproductive trees per hectare (   d = 0.46  trees/ha). We analyzed 205 trees and 261 and 487 open-pollinated seeds from 26 and 30 mother-trees in the years 2002 and 2003, respectively. A significant spatial genetic structure was detected for the adult trees for distances up to 100 m. We observed only small interannual differences in multilocus outcrossing rate (     ,     ), biparental inbreeding (     ,     ), and paternity correlation (     ,     ). The number of pollen donors contributing to mating of each tree in both years was estimated to be low (     ). Using TwoGener analysis to calculate the density of reproductive trees and the distance of pollen dispersal for normal and exponential models, the lowest error was detected for exponential model. For this model, the estimated density of reproductive trees was lower in 2002 (     trees/ha) than 2003 (     trees/ha), resulting in a higher distance of pollen dispersal in 2002 (     m) than 2003 (     m), although these changes did not affect the outcrossing and correlated mating rates.     

Microsatellite analysis of maternal half-sib families of Quercus robur, pedunculate oak: II. inferring the number of pollen donors from the offspring

We present an approach to infer the number of pollen donors directly from genotype data of open- pollinated progeny of Quercus robur (pedunculate oak), a highly outcrossing tree species. The approach is based on closely linked, highly polymorphic codominant microsatellite markers. Initially the close linkages between three previously mapped microsatellite loci were confirmed by studies of linkage disequilibrium (LD). Then an approach to track the pollen donors contributing to maternal half-sib families (open-pollinated families) was developed by analysing haplotype arrays of closely linked microsatellite markers transmitted from the fathers to the progeny. Simulated data of five linked microsatellite loci segregating in eight open-pollinated families were used to study the relationship between the number of paternal chromosomes detected by this ”haplotype approach” and the number of diploid fathers contributing to the families. The results showed that the number of diploid pollen donors can be expressed as an exponential function of the number of paternal chromosomes inferred from the progeny. The 95% confidence interval of this regression function is used to determine the minimum number of fathers contributing to a genotyped open-pollinated family of Quercus robur. Finally this open-pollinated family is used to demonstrate the resolution obtained with the ”haplotype approach”. Six independent microsatellite loci were used to study relatedness among all pairs of pollen gametes that share a haplotype of three linked markers. The results suggest that the majority of such gametes are identical by descent from the same father. The ”haplotype approach” presented here can be used to monitor the number of contributing pollen donors in commercial seedlot samples from oak or any other outcrossing tree species for which closely linked, highly polymorphic, codominant genetic markers are available. Received: 5 June 1999 / Accepted: 30 July 1999     

Long-distance pollen flow and tolerance to selfing in a neotropical tree species

Outcrossing rates, pollen dispersal and male mating success were assessed in Dicorynia guianensis Amshoff, a neotropical tree endemic to the Guiana shield. All adult trees within a continuous area of 40 ha (n = 157) were mapped, and were genotyped with six microsatellite loci. In addition, progenies were genotyped from 22 mature trees. At the population level, the species was mostly outcrossing (tm = 0.89) but there was marked variation among individuals. One tree exhibited mixed mating, confirming earlier results obtained with isozymes that D. guianensis can tolerate selfing. A Bayesian extension of the fractional paternity method was used for paternity analysis, and was compared with the neighbourhood method used widely for forest trees. Both methods indicated that pollen dispersal was only weakly related to distance between trees within the study area, and that the majority (62%) of pollen came from outside the study stand. Using maximum likelihood, male potential population size was estimated to be 1119, corresponding to a neighbourhood size of 560 hectares. Male mating success was, however, related to the diameter of the stem and to flowering intensity assessed visually. The mating behaviour of D. guianensis is a combination of long-distance pollen flow and occasional selfing. The species can still reproduce when it is extremely rare, either by selfing or by dispersing pollen at long distances. These results, together with the observation that male mating success was correlated with the size of the trees, could be implemented in management procedures aiming at regenerating the species.     

Historical and contemporary mating patterns in remnant populations of the forest tree Fraxinus excelsior L

Genetic variation at microsatellite markers was used to quantify genetic structure and mating behavior in a severely fragmented population of the wind-pollinated, wind-dispersed temperate tree Fraxinus excelsior in a deforested catchment in Scotland. Remnants maintain high levels of genetic diversity, comparable with those reported for continuous populations in southeastern Europe, and show low interpopulation differentiation (E = 0.080), indicating that historical gene exchange has not been limited (Nm = 3.48). We estimated from seeds collected from all trees producing fruits in three of five remnants that F. excelsior is predominantly outcrossing (t(m) = 0.971 +/- 0.028). Use of a neighborhood model approach to describe the relative contribution of local and long-distance pollen dispersal indicates that pollen gene flow into each of the three remnants is extensive (46-95%) and pollen dispersal has two components. The first is very localized and restricted to tens of meters around the mother trees. The second is a long-distance component with dispersal occurring over several kilometers. Effective dispersal distances, accounting for the distance and directionality to mother trees of sampled pollen donors, average 328 m and are greater than values reported for a continuous population. These results suggest that the opening of the landscape facilitates airborne pollen movement and may alleviate the expected detrimental genetic effects of fragmentation.     

Remnant Pachira quinata pasture trees have greater opportunities to self and suffer reduced reproductive success due to inbreeding depression

Habitat fragmentation is extensive throughout the world, converting natural ecosystems into fragments of varying size, density and connectivity. The potential value of remnant trees in agricultural landscapes as seed sources and in connecting fragments has formed a fertile area of debate. This study contrasted the mating patterns of bat-pollinated Pachira quinata trees in a continuous forest to those in pasture through microsatellite-based paternity analysis of progeny. The breeding system was determined by analysis of pollen tube growth and seed production from controlled pollinations. Fitness of selfed and outcrossed seed was compared by germination and seedling growth. There was more inbreeding within pasture trees (outcrossing=0.828±0.015) compared with forest trees (0.926±0.005). Pasture trees had fewer sires contributing to mating events, but pollen dispersal distances were greater than those in the forest. Paternity analysis showed variation in outcrossing rates among pasture trees with high proportions of external and self pollen sources detected. A leaky self-incompatibility system was found, with self pollen having reduced germination on stigmas and slower growth rate through the style. Controlled pollinations also showed a varied ability to self among trees, which was reflected in the selfing rates among pasture trees shown by the paternity analysis (0–80% selfing). Self pollination resulted in lower seed set, germination and seedling growth compared with outcrossing. While remnant trees in agricultural landscapes are involved in broader mating patterns, they show increased but varied levels of inbreeding, which result in reduced fitness.     

Low Inbreeding and High Pollen Dispersal Distances in Populations of Two Amazonian Forest Tree Species

Recent studies suggest that tropical tree species exhibit low inbreeding and high gene dispersal levels despite the typically low density of conspecifics in tropical forests. To examine this, we undertook a study of pollen gene dispersal and mating system of two Amazonian tree species. We analyzed 341 seeds from 33 trees at four microsatellite loci in a Carapa guianensis population from Brazil, and 212 seeds from 22 trees at four microsatellite loci in a Sextonia rubra population from French Guiana. Differentiation of allele frequencies among the pollen pool of individual trees was Φ_FT= 0.053 (95% CI: 0.027–0.074) for C. guianensis and Φ_FT= 0.064 (95% CI: 0.017–0.088) for S. rubra. The mean pollen dispersal distances were estimated at 69–355 m for C. guianensis , and 86–303 m for S. rubra , depending on the pollen dispersal model and the estimate of reproductive tree density used. The multi-locus outcrossing rate was estimated at 0.918 and 0.945, and the correlation of paternity at 0.089 and 0.096, for C. guianensis and S. rubra , respectively, while no significant levels of biparental inbreeding were detected. Comparing trees with high and low local density of conspecifics, we found no evidence for differences in inbreeding levels. The results are discussed within the framework of the emerging picture of the reproductive biology of tropical forest trees.     

Does long‐distance pollen dispersal preclude inbreeding in tropical trees? Fragmentation genetics of Dysoxylum malabaricum in an agro‐forest landscape

Tropical trees often display long‐distance pollen dispersal, even in highly fragmented landscapes. Understanding how patterns of spatial isolation influence pollen dispersal and interact with background patterns of fine‐scale spatial genetic structure (FSGS) is critical for evaluating the genetic consequences of habitat fragmentation. In the endangered tropical timber tree Dysoxylum malabaricum (Meliaceae), we apply eleven microsatellite markers with paternity and parentage analysis to directly estimate historic gene flow and contemporary pollen dispersal across a large area (216 km²) in a highly fragmented agro‐forest landscape. A comparison of genetic diversity and genetic structure in adult and juvenile life stages indicates an increase in differentiation and FSGS over time. Paternity analysis and parentage analysis demonstrate high genetic connectivity across the landscape by pollen dispersal. A comparison between mother trees in forest patches with low and high densities of adult trees shows that the frequency of short‐distance mating increases, as does average kinship among mates in low‐density stands. This indicates that there are potentially negative genetic consequences of low population density associated with forest fragmentation. Single isolated trees, in contrast, frequently receive heterogeneous pollen from distances exceeding 5 km. We discuss the processes leading to the observed patterns of pollen dispersal and the implications of this for conservation management of D. malabaricum and tropical trees more generally.     

Pollen mediated gene flow in a native population of Malus sylvestris and its implications for contemporary gene conservation management

Pollen mediated gene flow was studied in a Danish population of Malus sylvestris with the objective to support the Danish genetic conservation and management activities. A total of 50 mature trees (potential pollen donors) along with 180 seedlings (originating from 12 of the mature trees) were genotyped at 10 nuclear microsatellite loci. Paternity could be established for 46 seedlings, and a pollination distribution curve based on these observations was developed. Further, two indirect methods, Kindist and Twogener were applied for estimation of gene flow parameters. Pollinations were mostly between nearby trees with a median of observed pollination distances of approximately 23 m. However, a few long distance pollinations were observed and this increased the average pollination distance to approximately 60 m. The Kindist analysis of the data seemed to underestimate the average pollination distance as the fat-tailed distribution of the pollen dispersal distribution was not taken into account. Application of the results in a conservation and domestication context is discussed.     

Contemporary pollen flow,mating patterns and effective population size inferred from paternity analysis in a small fragmented population of the Neotropical tree <Emphasis Type="Italic">Copaifera langsdorffii</Emphasis> Desf. (Leguminosae-Caesalpinioideae)

Pollen flow and dispersal patterns were investigated in a small, isolated forest fragment of the Neotropical insect pollinated tree Copaifera langsdorffii, using paternity analysis and eight microsatellite loci. We also investigated the coancestry and effective population size of progeny arrays for conservation and environmental restoration purposes. Open-pollinated seeds were collected from 15 seed trees within the forest fragment, in which all adult trees were mapped, measured and genotyped. Twenty seeds were also collected from the nearest neighbor tree located 1.2 km from the forest fragment. Our results show that levels of genetic diversity were significantly higher in adults than offspring and significant levels of inbreeding were detected in offspring (F = 0.226). From paternity analysis, we observed low levels of selfing (s = 8%) and pollen immigration (m = 8%) in the fragment, but very high levels were detected for the isolated tree (s = 20%; m = 75%), indicating that the population and the tree are not reproductively isolated and are connected by patterns of long distance pollen dispersal (maximum detected 1,420 m). Within the forest fragment, the pattern of pollen dispersal was a near neighbor pattern with 49% of the pollen being dispersed within 50 m. The effective population size of the progeny array was low, indicating the need to collect seeds from a large number of seed trees (at least 76) for conservation purposes. The results show that the spatial isolation of the population and isolated tree due to forest fragmentation has not disrupted genetic connectivity; however, spatial isolation does seem to increase selfing and correlated mating.     

Short-distance pollen dispersal in a protogynous Annonaceae tree species from the Brazilian Cerrado

Annona crassiflora (Annonaceae) is a protogynous beetle-pollinated savannah tree species, widely distributed in the savannahs of the Cerrado biome. Studies on the mating system and pollen dispersal of protogynous species are very scarce. Here, we used six microsatellite loci to assess the mating system and pollen dispersal of A. crassiflora in a savannah remnant in Central Brazil. We mapped and sampled leaves of 112 adult trees and collected 74 fruits from 20 mother trees (1–4 fruits per plant) to obtain the seeds used (460) for mating system and parentage analyses. Annona crassiflora has predominantly allogamous mating systems, with a high multilocus outcrossing rate (t_m?=?0.974, SE?=?0.011) that did not differ among mother trees (F?=?1.32, p?=?0.165). However, t_m–t_s was variable among seed trees, indicating that some seeds were produced by mating among relatives. Our results also showed multiple paternity within fruits. Multilocus correlation of outcrossed paternity was high (r_p?=?0.302, SE?=?0.045), indicating that for each mother tree, the probability that the same pollen donor sired two random sibs was 30.2%, and the mean number of pollen donors per mother tree was high (6.3). We detected a maximum pollen dispersal distance of 360.7 m and an average of 124.3 m (SD?=?80 m), but most pollination events (73%) occurred at shorter distances (<?160 m), indicating short-distance pollen dispersal, most likely due to the pollinator behaviour.     

Pollen competition as a reproductive isolation barrier represses transgene flow between compatible and co-flowering citrus genotypes

Background/Objective

Despite potential benefits granted by genetically modified (GM) fruit trees, their release and commercialization raises concerns about their potential environmental impact, and the transfer via pollen of transgenes to cross-compatible cultivars is deemed to be the greatest source for environmental exposure. Information compiled from field trials on GM trees is essential to propose measures to minimize the transgene dispersal. We have conducted a field trial of seven consecutive years to investigate the maximum frequency of pollen-mediated crop-to-crop transgene flow in a citrus orchard, and its relation to the genetic, phenological and environmental factors involved.

Methodology/Principal Findings

Three different citrus genotypes carrying the uidA (GUS) tracer marker gene (pollen donors) and a non-GM self-incompatible contiguous citrus genotype (recipient) were used in conditions allowing natural entomophilous pollination to occur. The examination of 603 to 2990 seeds per year showed unexpectedly low frequencies (0.17–2.86%) of transgene flow. Paternity analyses of the progeny of subsets of recipient plants using 10 microsatellite (SSR) loci demonstrated a higher mating competence of trees from another non-GM pollen source population that greatly limited the mating chance of the contiguous cross-compatible and flowering-synchronized transgenic pollen source. This mating superiority could be explained by a much higher pollen competition capacity of the non-GM genotypes, as was confirmed through mixed-hand pollinations.

Conclusions/Significance

Pollen competition strongly contributed to transgene confinement. Based on this finding, suitable isolation measures are proposed for the first time to prevent transgene outflow between contiguous plantings of citrus types that may be extendible to other entomophilous transgenic fruit tree species.     

Mating dynamics of Scots pine in isolation tents

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). Self-fertilization 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.     

Mating patterns and pollen dispersal in a heterodichogamous tree, Juglans mandshurica (Juglandaceae)

Mating patterns in heterodichogamous species are generally considered to be disassortative between flowering morphs, but this hypothesis has hitherto not been vigorously tested. Here, mating patterns and pollen dispersal were studied in Juglans mandshurica, a heterodichogamous wind-pollinated species that is widely distributed in northern and north-eastern China. Paternity analyses carried out on 11 microsatellite loci were used to estimate morph-specific rates of outcrossing and disassortative mating. Pollen dispersal and genetic structure were also investigated in the population under study. The mating pattern of J. mandshurica was highly outcrossing and disassortative. Pairwise values of intramorph relatedness were much higher than those of intermorph relatedness, and a low level of biparental inbreeding was detected. There was no significant difference in outcrossing and disassortative mating rates between the two morphs. The effective pollen dispersal distribution showed an excess of near-neighbor matings, and most offspring of individual trees were sired by one or two nearby trees. These results corroborate the previous suggestion that mating in heterodichogamous plant species is mainly disassortative between morphs, which not only prevents selfing but also effectively reduces intramorph inbreeding.     

Morph-specific mating patterns in a low-density population of a heterodichogamous tree, Juglans ailantifolia

Heterodichogamy, including protandrous (PA) and protogynous (PG) morphs, is considered a mechanism to avoid selfing and promote disassortative mating. Although morphotypes are usually present in a population at a 1:1 ratio, this ratio may be biased in a low-density population by demographic stochasticity, resulting in a deficiency of mating partners in a neighbourhood dominated by a single morph. In this study, we determined morph ratio of the heterodichogamous tree species, Juglans ailantifolia by observing flowering in a low-density population during 2?years. The morph ratio (PG: PA) of 2.56:1 deviated significantly deviated from 1:1. We genotyped 59 reproductive trees and 405 offspring derived from eight PG-mother and three PA-mother trees with 11 microsatellite markers. Paternity analysis was conducted to clarify the effects of mother morph on the proportion of intra-morph mating. Then, we applied the Bayesian mixed effect mating model (MEMM) to clarify mating system, pollen dispersal, and individual fecundity of PG- and PA-mother trees. We found that the selfing rate and the distance of pollen dispersal were not clearly different between PG- and PA-mother trees. In contrast, the proportion of intra-morph mating was higher in the majority-morph (PG) mother trees than in the minor-morph (PA) mother trees. The MEMM indicated that mean dispersal distance of PG-mother trees was larger than that of PA-mother trees with large variance. Furthermore, we observed individuals with unusually high intra-morph fecundity for majority-morph (PG) trees. These findings indicate that intra-morph mating may occur when majority-morph mothers suffer a deficiency of potential inter-morph mates.