花粉介导的转Bt基因棉花田间基因流监测

采用花粉粒染色法对转Bt基因棉的花粉漂移距离和强度进行了观测,并应用PCR法检测转Bt基因棉的基因流频率.花粉粒染色法监测结果表明：同株异花间的花粉散布频率显著高于异株异花间(P<0.01);靠近转Bt基因棉花粉染色区1 m处的平均花粉散布频率,在东、南、西、北4个方向分别为44.8%、48.9%、57.1%和21.5%,但随着距转基因棉田距离的增大,4个方向的平均花粉散布频率都呈下降趋势.PCR结果的统计分析表明,在25m内,花粉散布距离和方向对基因流频率有极显著影响(P<0.01),随着距转基因棉田距离的增大,基因流频率呈下降趋势,最远距离为25 m时的最高基因流频率为2.0%.     

Long distance pollen-mediated gene flow at a landscape level: the weed beet as a case study

Gene flow is a crucial parameter that can affect the organization of genetic diversity in plant species. It has important implications in terms of conservation of genetic resources and of gene exchanges between crop to wild relatives and within crop species complex. In the Beta vulgaris complex, hybridization between crop and wild beets in seed production areas is well documented and the role of the ensuing hybrids, weed beets, as bridges towards wild forms in sugar beet production areas have been shown. Indeed, in contrast to cultivated beets that are bi-annual, weed beets can bolt, flower and reproduce in the same crop season. Nonetheless, the extent of pollen gene dispersal through weedy lineages remains unknown. In this study, the focus is directed towards weed-to-weed gene flow, and we report the results of a pollen-dispersal analysis within an agricultural landscape composed of five sugar beet fields with different levels of infestation by weed beets. Our results, based on paternity analysis of 3240 progenies from 135 maternal plants using 10 microsatellite loci, clearly demonstrate that even if weedy plants are mostly pollinated by individuals from the same field, some mating events occur between weed beets situated several kilometres apart (up to 9.6 km), with rates of interfield-detected paternities ranging from 11.3% to 17.5%. Moreover, we show that pollen flow appears to be more restricted when individuals are aggregated as most mating events occurred only for short-distance classes. The best-fit dispersal curves were fat-tailed geometric functions for populations exhibiting low densities of weed beets and thin-tailed Weibull function for fields with weed beet high densities. Thus, weed beet populations characterized by low density with geographically isolated individuals may be difficult to detect but are likely to act as pollen traps for pollen emitted by close and remote fields. Hence, it appears evident that interfield pollen-mediated gene flow between weed beets is almost unavoidable and could contribute to the diffusion of (trans)genes in the agricultural landscape.     

Extensive pollen dispersal in a bird-pollinated shrub, Calothamnus quadrifidus, in a fragmented landscape

Pollen dispersal was investigated in six populations of Calothamnus quadrifidus, a bird-pollinated shrub in the fragmented agricultural region of southern Western Australia. Paternity analysis using six microsatellite loci identified a pollen source within populations for 67% of seedlings, and the remainder were assumed to have arisen from pollen sources outside the populations. Outcrossing was variable, ranging from 5% to 82%, and long-distance pollen dispersal was observed in all populations with up to 43% of pollen sourced from outside the populations over distances of up to 5 km. This extensive pollen immigration was positively associated with population size but not isolation. Comparison of two populations of similar size but different density showed greater internal pollination and less selfing in the denser population, suggesting an influence of density on pollinator behaviour. The study revealed extensive long-distance pollen dispersal for C. quadrifidus within this fragmented agricultural landscape and highlighted the interaction between reserve populations and isolated road verge remnants in maintaining genetic connectivity at the landscape scale.     

Paternity analysis of the breeding structure of strangler fig populations: evidence for substantial long-distance wasp dispersal

Abstract. The mutualistic interaction of figs with their species-specific wasp pollinators and the role of figs as 'keystone' plant resources in tropical communities has received substantial attention from both plant and animal ecologists. Despite this focus on the reproductive biology of figs, the minute size of the wasps has effectively precluded our ability to monitor patterns of wasp dispersal and fig mating relationships in natural forest habitats. In this paper we use genetic markers and genealogy reconstruction techniques to examine the breeding structure of populations of four strangler fig species occurring in central Panama. The natural history of figs facilitates the genetic analysis of full-sib progeny arrays from which the genotypes of successful pollen donors can be reconstructed precisely. Paternity reconstruction in the four study species reveals that individual flowering trees may routinely receive pollen from numerous donors despite characteristically low densities of co-flowering individuals. These data indicate not only that breeding populations of these figs are larger than the minimum critical sizes predicted to be necessary to support populations of their species-specific pollinators, but are more extensive in size and area than has been described for any plant species yet examined. Further, the fig wasps are shown to be efficient agents of long-distance dispersal, routinely moving up to 10 km between flowering trees. In accord with the potential for substantial long-distance gene flow and large effective population sizes, ten of eleven species of Panamanian figs assayed were found to maintain exceptionally high levels of genetic variation within their populations. Combined with other reports of occasional long-distance dispersal, the results of this study suggest that fig wasps may be more effective at colonizing isolated fig populations than previously thought.     

Mating patterns and gene flow in the neotropical epiphytic orchid, Laelia rubescens

Understanding mating patterns and gene movement in plant populations occupying highly disturbed landscapes is essential for insights into their long-term survival. We used allozyme genetic markers to examine mating patterns and to directly measure pollen flow in the Central American epiphytic orchid, Laelia rubescens. Study populations were located in disturbed, seasonally dry tropical forest in Costa Rica. Every flowering individual within 15 populations and 12-18 seedlings from each maternal individual were genotyped over two reproductive seasons. Strict correlated mating by orchids produces full-sib progeny arrays from which the multilocus diploid genotype of the pollen parent can be inferred. These paternity analyses produced detailed quantitative estimates of pollen movement within and among populations of this species. Although our data illustrate that mating patterns vary spatially and temporally among trees, among pastures, and between years, overall patterns were surprisingly consistent. Thirty-four per cent of the capsules produced in both years resulted from gene flow events. Where pollen parents were identified, pollen moved mean distances of 279 m and 519 m in 1999 and 2000 respectively and a maximum documented distance of 1034 m. A substantially larger floral display in 2000 corresponded to a marked increase in pollen dispersal distances. Smaller populations, which more closely resembled those in undisturbed forest, had higher rates of gene flow than the large populations that characterize disturbed sites. We predict the occurrence of greater gene flow between low-density populations occupying undisturbed habitats.     

A comment on García et al. (2005, 2007) and related papers on mating patterns and gene dispersal in Prunus mahaleb

In two studies on mating patterns and spatial components of pollen and seed dispersal of Prunus mahaleb based on parentage analysis, García et al. (2005, 2007) depicted their 196 focal trees as a spatially isolated population where all reproductive trees had been genotyped. Additional distributional data for P. mahaleb trees in their study area, however, revealed that García and colleagues’ depiction of their study system bears little resemblance to reality. The trees these authors studied did not form a discrete, geographically isolated population. Around 300 ungenotyped reproductive trees occurred within the 1.5‐km distributional gap to the nearest population proclaimed by García and colleagues. Since exhaustive sampling of potential parental genotypes is essential in parentage analyses, the occurrence of a large number of ungenotyped trees in the immediate neighbourhood of focal trees can severely affect the main conclusions of García et al. (2005, 2007) as well as of several related publications on gene dispersal and mating patterns of P. mahaleb conducted on the same trees and relying on the same false premises of spatial isolation and exhaustive sampling.     

POLDISP: a software package for indirect estimation of contemporary pollen dispersal

poldisp 1.0 is a free software package to estimate the distribution of pollen dispersal distances from mother–offspring diploid genotypic data. It requires the spatial coordinates and genotypes of a sample of seed plants and their respective maternal progenies, providing estimates of the average, variance and kurtosis of the pollen dispersal curve. poldisp also estimates the effective reproductive density of pollen donors and the correlation of paternity within and among maternal sibships. poldisp is useful for characterizing the spatial scale of pollen dispersal, for assessing the variation in male fertility and for investigating biological factors affecting correlated paternity in plants.     

Viability and sunlight sensitivity of oak pollen and its implications for pollen-mediated gene flow

Pollen-mediated gene flow and the male reproductive success of wind-pollinated trees depend on the initial viability of the pollen and the changes that occur in its viability during transport in the atmosphere. The viability of Quercus robur pollen was determined before and during exposure to sunlight by in vitro germination and the fluorescein diacetate reaction (FCR) in 2002 and 2003, respectively. These experiments allowed us to calculate initial pollen viability and pollen sensitivity to sunlight. The germination test revealed a lower initial pollen viability (25–65%) than the FCR (53–92%). Following 9.5 h of irradiation the viability was reduced to 75–100% as determined by the in vitro germination test or to 40–70% as determined by the FCR. The actual values of initial pollen viability and pollen sensitivity to sunlight were used to define a range of values for modelling pollen dispersal using the mesoscale meteorological model METRAS. The deposition patterns of viable pollen varied by as much as a factor of 14 by changing the viability parameters in the range of the observed values. This suggests significant differences in male reproductive success. Variations in initial pollen viability have stronger effects on the gene-flow pattern than do variations in pollen sensitivity to sunlight. In particular, pollen distribution throughout the local environment is shaped by the initial pollen viability, while pollen sensitivity to sunlight mainly influences long-distance pollen dispersal.This revised version was published online in February 2005 with corrections to an incorrect sentence in the Results section. Under Pollen Sensitivity it should read: decreased faster when assessed by the germination test than by the FCR.     

Modelling pollen‐mediated gene flow in rice: risk assessment and management of transgene escape

Fast development and commercialization of genetically modified plants have aroused concerns of transgene escape and its environmental consequences. A model that can effectively predict pollen‐mediated gene flow (PMGF) is essential for assessing and managing risks from transgene escape. A pollen‐trap method was used to measure the wind‐borne pollen dispersal in cultivated rice and common wild rice, and effects of relative humidity, temperature and wind speed on pollen dispersal were estimated. A PMGF model was constructed based on the pollen dispersal pattern in rice, taking outcrossing rates of recipients and cross‐compatibility between rice and its wild relatives into consideration. Published rice gene flow data were used to validate the model. Pollen density decreased in a simple exponential pattern with distances to the rice field. High relative humidity reduced pollen dispersal distances. Model simulation showed an increased PMGF frequency with the increase of pollen source size (the area of a rice field), but this effect levelled off with a large pollen‐source size. Cross‐compatibility is essential when modelling PMGF from rice to its wild relatives. The model fits the data well, including PMGF from rice to its wild relatives. Therefore, it can be used to predict PMGF in rice under diverse conditions (e.g. different outcrossing rates and cross‐compatibilities), facilitating the determination of isolation distances to minimize transgene escape. The PMGF model may be extended to other wind‐pollinated plant species such as wheat and barley.     

A model-based method for estimating effective dispersal distance in tropical plant populations

Dispersal is a key mechanism to help populations propagate across space and thus is important in helping to understand spatial patterns. However, it is often difficult to quantify empirically as it requires intensive and detailed field study. Here we describe a method for estimating the effective dispersal distance of tropical plant populations. The method integrates a simple spatially explicit, individual-based dynamic model and spatial statistical analysis. The model is partly parameterized from spatial point pattern data as well as time series data from a 50 ha tropical forest plot in Barro Colorado Island (BCI) in Panama. Correlation between our estimated dispersal distances and those from inverse modeling based on field studies to date on BCI raises some questions about the match between our methods and those previously used. The method we propose can be generalized to any population for which spatial point pattern data are available. Additional field studies would be useful to further validate our method.     

Seasonal changes in pollinator activity influence pollen dispersal and seed production of the alpine shrub Rhododendron aureum (Ericaceae)

In alpine ecosystems, microscale variation in snowmelt timing often causes different flowering phenology of the same plant species and seasonal changes in pollinator activity. We compared the variations in insect visitation, pollen dispersal, mating patterns, and sexual reproduction of Rhododendron aureum early and late in the flowering season using five microsatellites. Insects visiting the flowers were rare early in the flowering season (mid-June), when major pollinators were bumblebee queens and flies. In contrast, frequent visitations by bumblebee workers were observed late in the season (late July). Two-generation analysis of pollen pool structure demonstrated that quality of pollen-mediated gene flow was more diverse late in the season in parallel with the high pollinator activity. The effective number of pollen donors per fruit (N(ep)) increased late in the season (N(ep) = 2.2-2.7 early, 3.4-4.4 late). However, both the outcrossing rate (t(m)) and seed-set ratio per fruit were smaller late in the season (t(m) = 0.89 and 0.71, seed-set ratio = 0.52 and 0.18, early and late in the season, respectively). In addition, biparental inbreeding occurred only late in the season. We conclude that R. aureum shows contrasting patterns of pollen movement and seed production between early and late season: in early season, seed production can be high but genetically less diverse and, during late season, be reduced, possibly due to higher inbreeding and inbreeding depression, but have greater genetic diversity. Thus, more pollinator activity does not always mean more pollen movement.     

Influence of cryptic population structure on observed mating patterns in the wild progenitor of maize (Zea mays ssp. parviglumis)

Indirect two-generation analysis of pollen flow has proven to be an effective alternative to exhaustive paternity analysis in numerous plant populations. In this investigation, the method is extended to an annual wild maize species, Zea mays ssp. parviglumis (Poaceae). Our analysis of mating system in parviglumis revealed high levels of outcrossing and higher biparental inbreeding than typically observed in grasses. Pollen dispersal analysis suggested low levels of long-distance dispersal. Given previous evidence for intrapopulation genetic structure in parviglumis populations, we explored the impact of cryptic population structure on estimates of mating system and pollen flow. Subpopulations inferred through spatially explicit Bayesian assignment showed markedly different values for both mating system parameters and pollen flow than the entire population. Finally, a novel method of pollen haplotype assignment revealed nonrandom mating consistent with intrapopulation structure. These results indicate parviglumis could be particularly susceptible to habitat fragmentation currently occurring throughout Mexico due to recent changes in land use.     

Long-distance seed dispersal in plant populations

Long-distance seed dispersal influences many key aspects of the biology of plants, including spread of invasive species, metapopulation dynamics, and diversity and dynamics in plant communities. However, because long-distance seed dispersal is inherently hard to measure, there are few data sets that characterize the tails of seed dispersal curves. This paper is structured around two lines of argument. First, we argue that long-distance seed dispersal is of critical importance and, hence, that we must collect better data from the tails of seed dispersal curves. To make the case for the importance of long-distance seed dispersal, we review existing data and models of long-distance seed dispersal, focusing on situations in which seeds that travel long distances have a critical impact (colonization of islands, Holocene migrations, response to global change, metapopulation biology). Second, we argue that genetic methods provide a broadly applicable way to monitor long-distance seed dispersal; to place this argument in context, we review genetic estimates of plant migration rates. At present, several promising genetic approaches for estimating long-distance seed dispersal are under active development, including assignment methods, likelihood methods, genealogical methods, and genealogical/demographic methods. We close the paper by discussing important but as yet largely unexplored areas for future research.     

Influence of pollinator grooming on pollen‐mediated gene dispersal in Mimulus ringens (Phrymaceae)

Pollinator foraging patterns and the dynamics of pollen transport influence the quality and diversity of flowering plant mating opportunities. For species pollinated by grooming pollinators, such as bees, the amount of pollen carried between a donor flower and potential recipient flowers depends on how grooming influences pollen transfer. To investigate the relationship between grooming and pollen‐mediated gene dispersal, we studied bumblebee (Bombus fervidus) foraging behavior and resulting gene dispersal in linear arrays of Mimulus ringens. Each of the 14 plants in an array had a unique multilocus genotype, facilitating unambiguous assignment of paternity to 1050 progeny. Each plant was trimmed to a single flower so that pollinator movements could be linked directly to resulting gene dispersal patterns. Pollen‐mediated gene dispersal was very limited. More than 95% of the seeds sired by a donor flower were distributed over the first three recipient flowers in the visitation sequence. However, seeds were occasionally sired on flowers visited later in the pollinator's floral visitation sequence. Intensive grooming immediately following pollen removal from a donor flower significantly increased the decay rate of the donor flower's gene dispersal curve. These results suggest that the frequency and relative intensity of grooming can have significant effects on patterns of pollen‐mediated gene dispersal from individual pollen donors.     

Cautions on direct gene flow estimation in plant populations

Through simulations we have investigated the statistical properties of two of the main approaches for directly estimating pollen gene flow (m) in plant populations: genotypic exclusion and mating models. When the assumptions about accurately known background pollen pool allelic frequencies are met, both methods provide unbiased results with comparable variances across a range of true m values. However, when presumed allelic frequencies differ from actual ones, which is more likely in research practice, both estimators are biased. We demonstrate that the extent and direction of bias largely depend on the difference (measured as genetic distance) between the presumed and actual pollen pools, and on the degree of genetic differentiation between the local population and the actual background pollen sources. However, one feature of the mating model is its ability to estimate pollen gene flow simultaneously with background pollen pool allelic frequencies. We have found that this approach gives nearly unbiased pollen gene flow estimates, and is practical because it eliminates the necessity of providing independent estimates of background pollen pool allelic frequencies. Violations of the mating model assumptions of random mating within local population affect the precision of the estimates only to a limited degree.     

Immigration history and gene dispersal: allozyme variation in Nordic populations of the red campion, Silene dioica (Caryophyllaceae)

Most of the Nordic region was ice-covered during the last (Weichselian) glaciation. During the postglacial period, plant and animal species recolonized the region from several directions and the geographic structuring of genetic variation within Nordic species may still contain a historic component that reflects patterns of postglacial immigration. The present investigation of 69 populations of Silene dioica represents the first large-scale allozyme study of a widespread herbaceous plant in the Nordic region. Although the frequencies of individual alleles showed a range of different geographic patterns, mapping of the axis scores from an ordination of variation at eight polymorphic loci revealed a division into two main geographic groups of populations. The broadly south-western and north-eastern distributions of these two groups of populations suggest that immigration into the region may have involved both eastern and southern geographic sources. However, the geographic boundaries between the two groups of populations are diffuse, and the relatively low between-population component of genetic diversity (G_ST = 16.4%) suggests a history of extensive gene dispersal by pollen.  © 2002 The Linnean Society of London, Biological Journal of the Linnean Society , 2002, 77 , 23–34.     

Pollen dispersal and gene flow within and into a population of the alpine monocarpic plant Campanula thyrsoides

Background and Aims

Gene flow by seed and pollen largely shapes the genetic structure within and among plant populations. Seed dispersal is often strongly spatially restricted, making gene flow primarily dependent on pollen dispersal within and into populations. To understand distance-dependent pollination success, pollen dispersal and gene flow were studied within and into a population of the alpine monocarpic perennial Campanula thyrsoides.

Methods

A paternity analysis was performed on sampled seed families using microsatellites, genotyping 22 flowering adults and 331 germinated offspring to estimate gene flow, and pollen analogues were used to estimate pollen dispersal. The focal population was situated among 23 genetically differentiated populations on a subalpine mountain plateau (<10 km²) in central Switzerland.

Key Results

Paternity analysis assigned 110 offspring (33·2 %) to a specific pollen donor (i.e. ‘father’) in the focal population. Mean pollination distance was 17·4 m for these offspring, and the pollen dispersal curve based on positive LOD scores of all 331 offspring was strongly decreasing with distance. The paternal contribution from 20–35 offspring (6·0–10·5 %) originated outside the population, probably from nearby populations on the plateau. Multiple potential fathers were assigned to each of 186 offspring (56·2 %). The pollination distance to ‘mother’ plants was negatively affected by the mothers'' degree of spatial isolation in the population. Variability in male mating success was not related to the degree of isolation of father plants.

Conclusions

Pollen dispersal patterns within the C. thyrsoides population are affected by spatial positioning of flowering individuals and pollen dispersal may therefore contribute to the course of evolution of populations of this species. Pollen dispersal into the population was high but apparently not strong enough to prevent the previously described substantial among-population differentiation on the plateau, which may be due to the monocarpic perenniality of this species.     

Fat-tailed gene flow in the dioecious canopy tree species Fraxinus mandshurica var. japonica revealed by microsatellites

Pollen flow, seed dispersal and individual reproductive success can be simultaneously estimated from the genotypes of adults and offspring using stochastic models. Using four polymorphic microsatellite loci, gene flow of the wind-pollinated and wind-seed-dispersed dioecious tree species, Fraxinus mandshurica var. japonica, was quantified in a riparian forest, in northern Japan. In a 10.5-ha plot, 74 female adults, 76 male adults and 292 current-year seedlings were mapped and genotyped, together with 200 seeds. To estimate dispersal kernels of pollen and seeds, we applied normal, exponential power, Weibull, bivariate t-distribution kernels, and two-component models consisting of two normal distribution functions, one with a small and one with a large variance. A two-component pollen flow model with a small contribution (26.1%) from short-distance dispersal (sigma = 7.2 m), and the rest from long-distance flow (sigma = 209.9 m), was chosen for the best-fitting model. The average distance that integrated pollen flows inside and outside the study plot was estimated to be 196.8 m. Tree size and flowering intensity affected reproduction, and there appeared to be critical values that distinguished reproductively successful and unsuccessful adults. In contrast, the gene flow model that estimated both pollen and seed dispersal from established seedlings resulted in extensive seed dispersal, and the expected spatial genetic structures did not satisfactorily fit with the observations, even for the selected model. Our results advanced small-scale individual-based parentage analysis for quantifying fat-tailed gene flow in wind-mediated species, but also clarified its limitations and suggested future possibilities for gene flow studies.     

Primary pollinator exclusion has divergent consequences for pollen dispersal and mating in different populations of a bird‐pollinated tree

Pollination by nectarivorous birds is predicted to result in different patterns of pollen dispersal and plant mating compared to pollination by insects. We tested the prediction that paternal genetic diversity, outcrossing rate and realized pollen dispersal will be reduced when the primary pollinator group is excluded from bird‐pollinated plants. Pollinator exclusion experiments in conjunction with paternity analysis of progeny were applied to Eucalyptus caesia Benth. (Myrtaceae), a predominantly honeyeater‐pollinated tree that is visited by native insects and the introduced Apis mellifera (Apidae). Microsatellite genotyping at 14 loci of all adult E. caesia at two populations (n = 580 and 315), followed by paternity analysis of 705 progeny, revealed contrasting results between populations. Honeyeater exclusion did not significantly impact pollen dispersal or plant mating at Mount Caroline. In contrast, at the Chiddarcooping site, the exclusion of honeyeaters led to lower outcrossing rates, a threefold reduction in the average number of sires per fruit, a decrease in intermediate‐distance mating and an increase in near‐neighbour mating. The results from Chiddarcooping suggest that bird pollination may increase paternal genetic diversity, potentially leading to higher fitness of progeny and favouring the evolution of this strategy. However, further experimentation involving additional trees and study sites is required to test this hypothesis. Alternatively, insects may be effective pollinators in some populations of bird‐adapted plants, but ineffective in others.