The mating system and genetic significance of polycarpy in the neem tree (Azadirachta indica)

The objectives of this study were to estimate the outcrossing rate and to explain genetic consequences of the development of seed in the endocarp in a natural population of neem in Bangladesh. Cotyledons of germinated open-pollinated seeds of individual trees were analyzed by starch-gel electrophoresis to examine allozymes. Three loci with clear Mendelian segregation were used to estimate outcrossing rate. A multilocus mixed mating model was used to evaluate the mating system. The population exhibited high outcrossing rates both for multilocus (t_m=0.90±0.024) and mean single-locus (t_s=0.92±0.020) estimates. The difference between these two parameters (t_m–t_s=0±SE 0.038) was insignificant, indicating that there was no ’biparental inbreeding’ in the population. The degree of variance of the estimates of multilocus outcrossing rates decreased when two or more loci were included. In order to elucidate the significance of polycarpy a total of 471 seeds were counted out of 440 endocarps. This mechanism appears to be a possible way of avoiding inbreeding. The results indicated that the studied neem population was predominantly allogamous. Received: 10 January 1999 / Accepted: 10 April 1999     

The mating system in natural and shelterwood stands of Douglas-fir

Summary Mating systems in two pairs of old-growth uncut and adjacent shelterwood stands of Douglas-fir (Pseudotsuga menziesii var. menziesii) were compared by estimating the proportions of viable progenies due to outcrossing (t) with both single-locus and multilocus techniques. Single-locus population estimates (_s), ranging from 0.41 to 1.16, were significantly (P<0.05) heterogeneous among loci in three of four stands; mean single-locus estimates for shelterwoods were not significantly different from those for uncut stands. Multilocus population estimates (_m) ranged from 0.94 to 1.00; again, estimates for shelterwoods were not significantly different from those for uncut stands. Multilocus estimates were slightly higher than mean single-locus estimates for uncut stands but were nearly equivalent for shelterwoods, suggesting that related matings other than selfs may be associated with uncut stands, but not shelterwoods. Individual-tree outcrossing rates (_{m i}), estimated for six trees in each shelterwood, ranged from 0.90 to 1.10 and were significantly heterogeneous among trees at one of the two shelterwoods. Outcrossing was high (> 0.90) in both uncult and shelterwood stands, and no evidence indicated that low parent-tree density had affected stand outcrossing rates.FRL 1918, Forest Research Laboratory. Oregon State University, Corvallis, USA     

用遗传标记监测不同环境和经营措施下的加勒比松森林群体遗传动态

对古巴加勒比松的6个群体（包括天然林、采伐林、母树林和种子园）进行了同功酶分析,根据5个酶系统8个位点的同功酶数据,对各群 交配系统以及群体遗传变异和结构进行了分析,天然林、种子园和母树林的多位点异交率和绝大多数单位点异交率都和完全异交无显著差异,过渡采伐的松树岛群体多位点异交率显著小于完全异交,而只有一半单位点异交率显著小于完全异交,而且该群体单位点平均异交率和多位点异交率均低于其它3个群体的估计值。采伐群体中同功酶变异和基因多样性与天然林群体JAG的相似,但低于其它群体,其近交系数较大,但小于天然林MAN和中国栽培群体的近交系数。中国引种栽培群体无论是同功酶变异还是基因多样性都显著高于古巴群体,与所有古巴群体的遗传距离都显著大于古巴群体之间的遗传距离。结果表明过度采伐导致群体自交程度增加,营建种子园可有效减少近交。自然分布区以外的引种栽培群体遗传变化量大,无论遗传变异和基因多样性都比参试其它群体大。     

Jatropha curcas L. (Euphorbiaceae) exhibits a mixed mating system, high correlated mating and apomixis

The hierarchical mating system among and within fruits of Jatropha curcas was investigated in a base population using five microsatellite loci, employing mixed mating and correlated mating models. Open-pollinated fruits were collected from 15 randomly selected seed trees, sampling seven fruits per tree for a total of 21 seeds from each tree. We detected multilocus genotypes identical to the mother tree in 13 % of offspring, implying the occurrence of apomixis in J. curcas. The presumed apomictic individuals were excluded from the analysis of the remaining results. Evidence of substantial selfing was provided by the average multilocus outcrossing rate (t _m?=?0.683), showing that the species exhibits a mixed mating system. The outcrossing rate showed a large variation among seed trees, ranging from 0.21 to 1.0, indicating that the species is not self-incompatible. Significant differences were detected between the multilocus and the single locus outcrossing rates (t _m???t _s?=?0.347) that suggested mating among related individuals, possibly because of the presence of individuals from the same progeny (sibs) in the base population. The multilocus paternity correlation was extremely high for the population (r _p(m)?=?0.999), indicating that the progenies were manly composed of full-sibs. As a consequence of selfing and a high paternity correlation, the co-ancestry coefficient within the progeny was higher (Θ?=?0.369) than expected for panmictic populations. Our results indicated that J. curcas produces seeds asexually by apomixis and sexually by a mixed mating system, combining selfing and outcrossing.     

Variation in the mating system of Sitka spruce (Picea sitchensis): evidence for partial assortative mating

Five allozyme polymorphisms were used to analyze the mating system in a Sitka spruce seed orchard in Saanichton, British Columbia. Allelic frequencies differed between the pollen and maternal pools at three of the five loci, with alleles rare in the maternal pool being even rarer in the effective pollen pool. Minor differences in pollen allelic frequencies were observed in the upper vs. lower crown. The multilocus outcrossing rate of the upper crown (t_m = 0.909) exceeded that of the lower crown (t_m = 0.764). Single-locus estimates of the outcrossing rate were significantly heterogeneous, with the lowest estimate of outcrossing, t = 0.773, observed for PGM-2 locus. Analyses of the mating system for the three maternal PGM-2 genotypes revealed heterogeneous pollen allelic frequencies and heterogeneous outcrossing rates, possibly due to assortative mating at this locus.     

Effects of population density on male and female reproductive success in the wind-pollinated, wind-dispersed tree species Betula maximowicziana

The effects of population density on male and female reproductive success of Betula maximowicziana were evaluated in two mixed and two post-fire stands, with various population densities, ranging from 1.9 to 300.0 trees per ha, in central Hokkaido, Japan. First, we investigated ecological determinants of reproductive success (seed set and germination) of both seeds collected from the trees (tree seeds) and dispersed seeds collected from seed traps (dispersed seeds). We then evaluated the effects of population density on seed set and germination of tree seeds and dispersed seeds using a generalized linear mixed model (GLMM). Subsequently, we genotyped 950 seeds collected from mother trees and 940 seeds trapped after dispersal derived from tree seeds and dispersed seeds, respectively, using eleven microsatellite loci. Using the acquired data, we then evaluated the outcrossing rate and effective number of pollen donors (N _ep) of the tree seeds, and the genetic structure of both pollen pools and dispersed seed populations. The seed set and germination rate of dispersed seeds was significantly lower both in the lowest-density stand and in the highest-density stand. The GLMM revealed that seed set and germination rates of dispersed seeds may be maximal at approximately 120 trees per hectare (optimal density). Outcrossing rates were consistently high (t _m = 0.995), regardless of the population density. In contrast, N _ep was lower in the lowest-density stands. Significant genetic structure of the dispersed seed population was found in two low-density stands, probably due to the limitation of overlapping seed shadows.     

去劣疏伐对油松种子园交配系统及遗传多样性影响的研究

 为了解去劣疏伐对种子园群体交配系统和遗传组成的影响程度,采用水平淀粉凝胶电泳方法,对10个等位酶位点上,分别用混合交配模型(MLT)和Biosys-Ⅰ程序,分析了去劣疏伐前后辽宁兴城油松种子园3个年份群体的交配系统和遗传多样性变化。种子园去劣疏伐前(1993年)种子试样多位点异交率为0.962,而疏伐后2个年份(1996年和2000年)试样的多位点异交率明显减小,分别为0.795和0.801。疏伐后种子园的自交率有升高的趋势,从疏伐前的0.038到疏伐后的0.205和0.199。近交系数也有明显变化,疏伐前为0.141,疏伐后分别为0.341和0.239。去劣疏伐后种子园的每个位点的平均等位基因数(A)和多态位点百分率(P)都有不同程度的降低。本文对影响种子园交配系统和遗传多样性的因素作了探讨。     

Fine-scale estimation of outcrossing in western redcedar with microsatellite assay of bulked DNA

Western redcedar (Thuja plicata, Cupressaceae) is a self-fertile conifer with a mixed mating system and significant variation for outcrossing among populations. In this paper, we conducted a fine-scale study of mating system variation to identify correlates of outcrossing in natural populations. We examined variation for outcrossing within and among individual trees, and describe a new method to estimate outcrossing using bulked DNA samples. Bulking (assaying DNA tissues from several individuals simultaneously) increases the experimental power without increasing the experimental effort. We sampled 80 trees from four natural populations in southwestern British Columbia. From each tree, we sampled from up to six crown positions (three heights and inner vs outer branches). From each position, two samples of three seedlings each were bulked before DNA extractions. Using four microsatellite loci, we obtained outcrossing rates for each tree and for each of the six crown positions. We found individual tree selfing rates to increase with tree height in all four populations, but selfing rates did not differ among crown positions. The higher selfing rate of larger trees is probably due to their greater proportional contribution to local pollen clouds. Individual tree outcrossing rates ranged from 22 to 100% and the population outcrossing rates from 66 to 78%. Missed alleles due to bulking and the estimation method used both cause a downward bias in outcrossing rates, so that these estimates are probably lower than the actual outcrossing rates. Nevertheless, the trends we observed are not affected by systematic biases of estimation.     

Temporal patterns of reproduction and outcrossing in weedy populations of Echium plantagineum

Single and multilocus estimates of outcrossing rates were made in three populations of Echium plantagineum. Despite spatial separation, variations in population size (though not density) and reproductive output, no statistically significant difference was detected in outcrossing rates between the populations. Similarly, only slight differences in outcrossing rates were detected within populations when estimates were based on seed collected from flowers open at different times in the flowering season. The earliest flowers tended to have a lower estimated rate of outcrossing. In all cases, multilocus outcrossing rates were high, ranging from 0.81 to 1.05.
In all three populations flower production extended over a period of more than 2 months but the majority of seed was produced by flowers that opened during the first third of the (lowering season. This was largely caused by a high rate of flower production during the early part of the season and not changes in the number of seeds set per flower. The average number of seeds produced per flower varied both between individuals within populations, and between the different populations, but neither of these differences were significant.
Controlled pollination of flowers with self or outcross pollen, applied either singly or together, failed to detect any differences in the likelihood that either type of pollen would give rise to fertile seeds.     

Impacts of forest fragmentation on the mating system and genetic diversity of white spruce (Picea glauca) at the landscape level

We studied the mating system of white spruce (Picea glauca) in a landscape fragmented by agriculture in northern Ontario, Canada. We sampled 23 stands that ranged in size from 1 to >500 trees isolated by 250-3000 m from the nearest other stand. Six polymorphic allozyme loci from four enzyme systems were used to genotype approximately 10 000 embryos from 104 families. We detected no allele frequency heterogeneity in the pollen pool among stands or families (Phi(FT)=-0.025). Overall, estimates of outcrossing were high (t(m)=94% and mean t(s)=91%) but significantly different from unity. Bi-parental inbreeding (t(m)-t(s)=3.2%) was low but significantly different from zero. Allozyme-based outcrossing estimates did not differ significantly among three stand-size classes (SSCs): small (<10 trees), medium (10-100 trees) and large (> or =100 trees). The number of effective pollen donors was high in all SSCs, but was significantly lower in small stands (N(ep)=62.5) than in medium-sized and large stands (both N(ep)=143). The primary selfing rate was significantly higher in medium stands than in large stands. We found no significant difference in genetic diversity measures in the filial (seed) population among SSCs. Overall, these results indicate that white spruce stands in this fragmented landscape are resistant to genetic diversity losses, primarily through high pollen-mediated gene-flow and early selection against inbred embryos. We discuss the importance of using seed data, in conjunction with genetic data, to evaluate the impacts of fragmentation on natural populations.     

Mating system analysis in a natural population of Acacia nilotica subspecies leiocarpa

The mating-system was investigated in a natural population of the tetraploid taxon Acacia nilotica ssp. leiocarpa using open-pollinated seeds from 15 families. Six-day-old germinated seeds were used for starch-gel electrophoresis. Three enzyme systems (ADH, EST-1, and 6PGD) were scored. Isozyme banding patterns and segregation of isozyme variants within families suggest that the species is an autotetraploid displaying tetrasomic inheritance. Estimates of single-locus (t_s=0.358) and multilocus (t_m=0.384) population outcrossing rates were homogeneous, and indicate substantial selfing in the population. Heterogeneity of outcrossing estimates among loci and families were marked, suggesting departure from the assumptions of the mixed-mating model. Implications of the result for the utilization of germplasm in tree-improvement programmes are noted.     

Genetic diversity in a seed production population vs. natural populations of Sitka Spruce

Isozyme analysis was applied to estimate the level of variation and the genetic structure of a seed-production population (i.e., seed orchard) and 10 range-wide natural populations of Sitka spruce (Picea sitchensis (Bong.) Carr.). Gene diversity and heterozygosity estimates were comparatively high in both the seed orchard and the natural populations studied. The seed orchard population showed a significantly higher number of alleles per locus and percentage of polymorphic loci. Though not significant, mean heterozygosity of the seed orchard was higher than that observed for all natural populations. Genetic distance analysis indicated that the seed-orchard population was genetically similar to three natural populations from which the parent trees were selected. Parent trees sampling breadth has been identified as the major cause for the observed increased level. The impact of recurrent selection and seed orchard biology and management on maintaining the genetic diversity is discussed.     

Outcrossing rates and relatedness estimates in pecan (Carya illinoinensis) populations

Estimates of single and multilocus outcrossing rates as well as relatedness among progeny of individual seed trees were obtained for 14 populations of pecan [Carya illinoinensis (Wangenh.) K. Koch]. Mean outcrossing estimates were not significantly different from 1.0 and relatedness values indicate that most progeny within families are half sibs. Biparental inbreeding was insignificant in all study sites, and inbreeding coefficients indicated that populations were close to inbreeding equilibrium.     

EFFECTS OF POPULATION STRUCTURE AND CONE PRODUCTION ON OUTCROSSING RATES IN ENGELMANN SPRUCE AND SUBALPINE FIR

Variation in the mating systems of Engelmann spruce and subalpine fir was examined in two adjacent subpopulations in the Colorado Front Range. Multilocus outcrossing rates were estimated from analysis of allozyme variation in embryo and megagametophyte tissue. The overall multilocus outcrossing rate of 0.93 for spruce was significantly higher than the outcrossing rate of 0.89 for fir. Outcrossing rates varied from 0.73 to 0.97 for spruce and from 0.65 to 0.94 for fir when trees of each species were grouped according to age, size, spatial distribution, maternal heterozygosity, cone production, and year of sampling. In both spruce and fir, trees with higher levels of male-cone production and/or more clumped spatial distribution had lower outcrossing rates. Temporal variation in outcrossing rates within subpopulations, examined only in spruce, was less than variation between subpopulations. Male-cone production was significantly correlated with tree size, and regression analyses revealed that the highest outcrossing rates are expected on trees that are medium to large in size, rather than the largest trees in the forest. The results show that, while there is some temporal variation, patterns of male cone production and spatial distribution of individuals are the most important factors influencing outcrossing levels in these wind-pollinated forest trees.     

Mating system and multilocus associations in a natural population of Pseudotsuga menziesii (Mirb.) Franco

Summary Arrays of open-pollinated seeds were assayed for allozyme polymorphisms at ten loci (Aat2, Est1, G6pd, Idh, Mdh2, Mdh3, Pgm, Sod, 6Pgd1, 6Pgd2) to obtain estimates of the outcrossing rate and assess multilocus association in a natural population of coastal Douglas-fir, Pseudotsuga menziesii (Mirb.) Franco. The allele frequencies in the samples of adult trees and pollen-gamete pool were similar. Maximum-likelihood estimators of the outcrossing rate for individual loci and two multilocus models were derived using counting methods. The single-locus maximum likelihood estimates (MLEs) of the outcrossing rate were significantly heterogeneous; they varied over a more than two-fold range from 0.404 to 0.935, with an average MLE of 0.741. Both multilocus MLEs of the outcrossing rate were 0.887. The sample of trees was in random mating equilibrium when assessed on a pairwise-locus basis using Burrows' composite measure of gametic disequilibrium, with one exception (Mdh2 Sod) that was attributable to a rare gametic class. In the sample of pollen gametes, 5 of the 45 pairwise-locus associations were nominally significant at the 0.05 level: Idh Est1, Mdh2 Sod, Aat2 Est1, Aat2 Mdh3, and Est1 Mdh3. These apparent associations were attributable in most cases to the relative excess of uncommon or rare paternal gametes of discernibly outcrossed embryos. An additional two-locus association was identified for Mdh2 Pgm which was marginally significant for the major partition of the contingency table that excluded paternal gametes with the rare allele Mdh2 ² .     

Mating system and pollen gene flow in Mediterranean maritime pine

Mating systems define the mode of gene transmission across generations, helping to determine the amount and distribution of genetic variation within and among populations of plant species. A hierarchical analysis of Mediterranean maritime pine mating system (61 mother trees from 24 plots, clustered in three populations) was used to identify factors affecting mating patterns and to fit pollen dispersal kernels. Levels of ovule and seed abortion, multi- and single-locus outcrossing rates and correlated paternity were estimated from progeny arrays and correlated with ecological stand variables and biometric tree measures. Pollen dispersal kernels were fitted using TwoGener and KinDist indirect methods and simulations were carried out to identify relevant factors affecting correlated paternity. Maritime pine showed high outcrossing rates (t(m) and t(s) approximately 0.96) and relatively low levels of correlated paternity [an r(p) of 0.018 (Ritland's estimate) or 0.048 (Hardy's estimate)], although higher than in other anemophilous tree species. Mating system parameters had high variation at the single-tree level (99-100%) but no stand or population effect was detected. At the single-tree level, outcrossing rates were correlated with tree (diameter and height) and crown size. In addition, correlated paternity showed a significant negative correlation with tree height, height to crown base and height to the largest crown width, probably reflecting the importance of the trees' 'ecological neighborhoods'. Indirectly estimated pollen dispersal kernels were very leptokurtic (exponential-power distributions with beta<0.5), with mean dispersal distances from 78.4 to 174.4 m. Fitted dispersal kernels will be useful in building explicit simulation models that include dispersal functions, and which will contribute to current conservation and management programs for maritime pine. Nevertheless, the numerical simulations showed that restricted dispersal, male fertility and phenological overlap could only partially explain the observed levels of correlated paternity; so other factors may also be relevant for the management of this valuable forest tree species.     

Evidence for selective fruit abortion in Banksia spinulosa (Proteaceae)

Fruit set is consistently low in the genus Banksia (Proteaceae). One explanation for low fruit:flower ratios is that excess flowers allow plants to selectively abort inferior progeny thereby increasing the average quality of the seed crop. We examined whether self-pollinated fruits were aborted selectively in a partially self-compatible population of Banksia spinulosa. Inflorescences were divided in half and pollinated with cross pollen on both sides, self pollen on both sides or cross pollen on one side and self pollen on the other. Seed production was reduced significantly by 38% on fully self-pollinated compared to fully cross-pollinated inflorescences, indicating partial self-incompatibility or inbreeding depression. On inflorescences pollinated with both cross and self pollen, selfed seed set was reduced by 63% compared to crossed seed set. On the selfed half of these inflorescences, fewer fruits and seeds were produced, and more fruits had aborted seeds than on randomly selected halves of fully self-pollinated inflorescences. These results suggest that self-fertilized embryos were developed in the absence of outcrossed progeny but were selectively aborted when outcrossed progeny were present on inflorescences. To determine the amount of outcrossing occurring in the study population, outcrossing rates (t) of naturally pollinated plants were examined by electrophoretic assessment of progeny arrays. Values of I did not differ significantly from one, indicating complete outcrossing. High outcrossing, despite the high probability of self pollen receipt under natural conditions, is consistent with the conclusion that selective abortion occurs in B. spinulosa.     

Gene flow estimation with microsatellites in a Malagasy seed orchard of Eucalyptus grandis

Eucalyptus grandis has a mixed-mating reproductive system. Malagasy Eucalyptus seed orchards were established 15 years ago with two aims both based on panmixia: open-pollinated seed production and genetic improvement. The panmixia hypothesis has never been confirmed in the seed orchard. From a seedling seed-orchard stand comprising 349 trees and using data obtained with six selected microsatellite markers, paternity analysis was performed for 724 offspring collected on 30 adult trees. Paternity assignment, based on exclusion procedures and likelihood-ratio method, was achieved with high accuracy; the exclusion probability value was 0.997. The outcrossing rate was very high (96.7%). More than 50% of potential male trees (199 out of 349) in the seed orchard contributed to pollination for 440 offspring of 30 progenies (8.6% of the basic population). The pollination rate from outside the seed orchard was high (39.2%), but might be due to the small size of this seed orchard. This study showed that "panmixia-like pollination" can be assumed.     

Genetic variability of maternal plants and embryos of their seeds in the populations of <Emphasis Type="Italic">Pinus kochiana</Emphasis> Klotzsch ex Koch in Crimea

Samples of trees and seed embryos of five Pinus kochiana Klotzsch ex Koch populations in Crimea were comparatively studied at 12 allozymic loci for its genetic variability and mating system. It has been shown that the embryos of seeds reproduce the allelic diversity of maternal plants, but the level of their observed heterozygosity (H_o) is significantly lower, i.e., 0.286 vs. 0.189, respectively. Unlike maternal trees, embryos are characterized by a significant divergence between the observed distribution of genotypes and the theoretically expected one according to the Hardy–Weinberg law for the majority of analyzed loci. The frequency of cross pollination in populations varied from 0.384 to 0.673 according to single-locus (t_s) estimates and from 0.639 to 0.841 according to multilocus (t_m) estimates.     

Fine-scale genetic structure enhances biparental inbreeding by promoting mating events between more related individuals in wild soybean (Glycine soja; Fabaceae) populations

Outcrossing between genetically distant individuals in a plant population enhances allelic heterozygosity-an important source for genetic diversity and adaptive evolution. Fine-scale spatial genetic structure (FSGS) can interfere with outcrossing by promoting mating between more related individuals. To test the influence of FSGS on outcrossing, FSGS and outcrossing rates were analyzed with four wild soybean (Glycine soja) populations from different habitats, using simple sequence repeat (SSR) fingerprints. Spatial autocorrelation analysis indicated variable FSGS (15.44-25.87 m) in all four populations. Multilocus mixed-mating analysis of 1605 progeny indicated substantial variation in single-locus outcrossing (T(s) = 6.3-12.6%) although the total outcrossing rates as estimated by multilocus outcrossing (T(m) = 12.8-17%) did not vary significantly among populations. The comparison between FSGS and outcrossing rates demonstrated that strong FSGS with large genetic patch size can enhance biparental inbreeding by promoting mating between more related individuals in a population. The results suggest that patch size management can aid in situ conservation by avoiding formation of strong FSGS and encouraging true outcrossing among individuals.