Relationship between interspecific pollen transfer and pistil length in sympatric congeners,Clerodendrum trichotomum and C. izuinsulare

Interspecific pollen transfer (IPT) often leads to reproductive interference. Although character displacement of reproductive traits in plants is often considered a consequence of reproductive interference, few studies have tested whether intraspecific variation in floral morphology changes the intensity of reproductive interference among pollinator-sharing plants. We investigated whether intraspecific variation in pistil length changes the proportion of IPT (interspecific pollen transfer) in sympatric populations of Clerodendrum trichotomum and C. izuinsulare on the two islands. On Toshima Island, C. izuinsulare flowers with shorter pistils were significantly less likely to receive C. trichotomum pollen, and there was a slight tendency for longer pistiles of C. trichotomum flowers to receive less C. izuinsulare pollen, suggesting that IPT has caused character displacement in pistil length in these congeneric plants. In contrast, we did not detect any relationship between pistil length and the proportion of IPT for the two species on Niijima Island. The discrepancy between the islands may reflect differences in pollinator assemblages.     

Comparison of pollination characteristics between the insular shrub Clerodendrum izuinsulare and its widespread congener C. trichotomum

Impact of pollinator shift on differentiation of floral morphology has attracted the interest of naturalists for many years. A comparative investigation was conducted for determining the pollination characteristics, including pollinator assemblage, floral morphology, flowering phenology, and self‐compatibility, of two closely related Clerodendrum species—insular C. izuinsulare and widespread C. trichotomum. Japanese black swallowtail butterflies were the predominant flower visitors in mainland Japan, whereas diurnal hawk moths were predominantly found on the Izu Islands, a chain of oceanic islands located off the southeastern coast of the main Japanese island of Honshu in the west Pacific Ocean. The corolla tube of C. izuinsulare was longer than that of C. trichotomum, whereas the filaments and petals of C. izuinsulare were shorter than those of C. trichotomum. The flowering season of C. izuinsulare was later than that of C. trichotomum. The self‐compatibility of C. izuinsulare was higher than that of C. trichotomum. These differences might be associated with the low density of Japanese black swallowtail butterflies and dominance of diurnal hawk moths on the Izu Islands.     

Post-pollination reproductive isolation between diurnally and nocturnally flowering daylilies, Hemerocallis fulva and Hemerocallis citrina

To examine whether floral and post-pollination isolation develops independently or not, we conducted a crossing experiment between Hemerocallis fulva and Hemerocallis citrina that shows large floral divergence adapted for diurnal and nocturnal pollinators that have been believed to be fully cross-fertile. Flowers of the two species from sympatric populations were hand-pollinated with conspecific pollen from the same population (control), interspecific pollen from the same area (sympatric cross), and interspecific pollen from the different area (allopatric cross). After capsule dehiscence, the fruit set, seed set per fruit and seed set per flower were determined among three cross categories. The seed sets per flower were 32 and 77% lower in sympatric and allopatric crosses than in the control when H. fulva was the pollen recipient. There was no difference in three reproductive measures among the cross categories when H. citrina was the pollen recipient. This finding indicates that post-pollination isolation does exist between H. fulva and H. citrina, although it is partial, asymmetric, and weakened in sympatry. Our result suggests that floral and post-pollination isolation may develop independently, and reinforcement may not be a general phenomenon in plants.     

Shifts to earlier selfing in sympatry may reduce costs of pollinator sharing

Coexisting plant congeners often experience strong competition for resources. Competition for pollinators can result in direct fitness costs via reduced seed set or indirect costs via heterospecific pollen transfer (HPT), causing subsequent gamete loss and unfit hybrid offspring production. Autonomous selfing may alleviate these costs, but to preempt HPT, selfing should occur early, before opportunities for HPT occur (i.e., “preemptive selfing hypothesis”). We evaluated conditions for this hypothesis in Collinsia sister species, C. linearis and C. rattanii. In field studies, we found virtually identical flowering times and pollinator sharing between congeners in sympatric populations. Compared to allopatric populations, sympatric C. linearis populations enjoyed higher pollinator visitation rates, whereas visitation to C. rattanii did not differ in sympatry. Importantly, the risk of HPT to each species in sympatry was strongly asymmetrical; interspecies visits comprised 40% of all flower‐to‐flower visits involving C. rattanii compared to just 4% involving C. linearis. Additionally, our greenhouse experiment demonstrated a strong cost of hybridization when C. rattanii was the pollen donor. Together, these results suggest that C. rattanii pays the greatest cost of pollinator sharing. Matching predictions of the preemptive selfing hypothesis, C. rattanii exhibit significantly earlier selfing in sympatric relative to allopatric populations.     

Pollination ofNuphar (Nymphaeaceae) in Europe: Flies and bees rather thanDonacia beetles

Nuphar (Nymphaeaceae) comprises a small holarctic group of aquatic perennials whose flowers are pollinated by flies, bees, and beetles. We studied pollination in different populations of the two European speciesN. lutea andN. pumila in Norway and in Germany. Flowers are self-compatible and protogynous, preventing automatic selfing, and insect pollination is required for seed production. Sympatric populations were studied in Vest-Agder county in Norway to determine whetherN. lutea andN. pumila have the same or different pollinators. Allopatric populations ofN. lutea in Germany and Norway were then compared to determine whether their pollinator spectra differ as would be expected in an open flower with seemingly easily accessible pollen and nectar. Results of the present as well as previous studies of the pollination ofN. lutea andN. pumila show that both species are mainly pollinated by flies, including apparentNuphar specialists, such as the scatophagidHydromyza livens and the ephydridsHydrellia andNotiphila, the last also a long-known pollinator ofN. advena in Florida. Pollinator overlap between sympatric heterospecific populations was small, while allopatric conspecific populations had similar visitor and pollinator spectra. We found no evidence of pollination byDonacia beetles as reported from some North American populations ofNuphar. Dedicated to Univ.-Prof. DrF. Ehrendorfer on the occasion of his 70th birthday     

Lack of evidence on hybrid swarm in the sympatric population of Pinus mugo and P. sylvestris

Natural hybridisation between the taxa from Pinus mugo complex and P. sylvestris was postulated in several sympatric populations of the species in Europe. However, due to the absence of precise methods for identification of hybrid seeds and hybrid trees, the frequency of hybridisation and its influence on the genetic structure of relict P. mugo populations has not been clarified so far. In the present study, the species-specific chloroplast DNA (cpDNA) marker and isozymes were applied to test the hybridisation hypothesis in a postulated hybrid swarm population of the species from “Bór na Czerwonem” reserve at the northern foothills of Tatra Mts., Poland. The cpDNA marker was used to identify hybrids within two groups of polycormic (multi-stemmed) and monocormic (single-stemmed) pines from this population. Allelic frequencies at isozyme loci from both groups were compared to frequencies found in allopatric populations of the species. Additionally, cpDNA haplotypes of seedlings derived from open pollinated seeds were studied to detect the possibility of successful cross-pollination. The mixture of seedlings with P. sylvestris and P. mugo cpDNA haplotypes was derived from one parental tree that indicates hybridisation. However, all the mature polycormic pines had cpDNA haplotypes species specific to P. mugo and the isozyme frequencies were similar to frequencies found in three allopatric populations of P. mugo from Tatra Mts. (mean genetic distance, Dn=.027). The differences were much larger in comparisons with monocormic pines from this area (Dn=.085) and two P. sylvestris samples from distant allopatric populations (Dn=.077). Nearly all monocormic pines had cpDNA species specific to P. sylvestris and isozyme frequencies similar to other populations of this species (mean Dn=.004). Only one P. sylvestris-like monocormic tree had cpDNA of P. mugo and can be considered as a hybrid. The results do not provide evidence supporting the hybrid swarm hypothesis. Rather, the results suggest that mature hybrids have a low frequency within this population and (rare) hybridisation is not reciprocal but unidirectional with P. mugo as pollen donor.     

Pollen-pistil interactions in Populus: β-galactosidase activity associated with pollen-tube growth in the crosses P. nigra x P. nigra and P. nigra x P. alba

Summary In order to understand the nature of interspecific barriers in Populus, we have explored pollen/pistil interactions in intra- and interspecific crosses Populus nigra x P. nigra and P. nigra x P. alba. The kinetics of pollen-tube growth demonstrated that P. nigra and P. alba pollen tubes have distinct behavioral patterns inside P. nigra pistils. P. alba pollen tubes exhibit an unique S-shaped growth curve and an arrested growth site near the sylodium. P. nigra pollen tubes exhibit two growth phases, in the stigmatic tissues and in the ovarian cavity respectively. P. nigra and P. alba curves diverge 5 h after controlled pollination and could be related to a change in the physiology of the P. nigra pollen tube, which shifts from an autotrophic to a heterotrophic type of nutrition. Protein analysis of pollinated stigmatic extracts (0,6 and 20 h after pollination) revealed qualitative and quantitative differences that are related to the presence of either P. nigra or P. alba pollen tubes inside the stigmatic tissues. Increasing numbers of protein bands were detectable from 0 to 20 h after pollination only in intraspecific cross. Glycoproteins were detected, and the differences observed were dependent of the cross. -Galactosidase activity was found in pollinated stigmas, but an increase in its activity (one isozyme of pHi 4.2) between 6 h and 20 h after pollination was detected only in the intraspecific cross. This enzyme could play a role in heterotrophic pollen-tube nitrition, and its activity could be the final result of a series of interactions started by the initial pollen-stigma dialog.     

The pollination biology of two species of Helicteres (Malvaceae) with different mechanisms of pollen deposition

Species of Helicteres are pollinated mainly by hummingbirds and bats. Most species pollinated by hummingbirds have a mechanism of depositing pollen on the top of the pollinator's head such as is shown in this work for Helicteres sacarolha. H. brevispira has an unusual mechanism of depositing pollen grains under the tail or on the abdomen of the hummingbirds. The top of the birds’ head may be considered an efficient place to transport pollen grains, because it is plain, easily accessible for deposition and donation of pollen and not easily accessible for grooming, while the tail is movable and inclined. Thus, H. brevispira pollinators may carry or transfer fewer amounts of pollen grains than H. sacarolha pollinators from one flower to another. If a large amount of pollen grains is lost, a higher quantity of flower or pollen has to be produced to guarantee reproductive success. Plants of H. brevispira set higher number of flowers and pollen grains per flower than H. sacarolha and have also higher rates of fruit and flower abortion. Thus, pollination efficiency of H. brevispira may be reached by high pollen and flower production. Flower change mechanism presented in this species may be also involved with the optimization of pollinator feeding and pollination efficiency.     

Calling song and selective phonotaxis in the field crickets,Gryllus firmus andG. pennsylvanicus (Orthoptera: Gryllidae)

The field cricket species, Gryllus firmusand G. pennsylvanicus,occur in a mosaic hybrid zone that roughly parallels the eastern slope of the Appalachian mountains in the northeastern United States. It is important to know what role, if any, the calling song plays in mate choice in sympatric and allopatric populations. In this report, we present results on the variability of calling song properties along transects across this hybrid zone. We also present the results of experiments on phonotactic selectivity of females from an allopatric population of G. firmus.The male calling song of allopatric G. firmuswas significantly slower in temporal rhythm (i. e., chirp and pulse repetition rates) and lower in pitch (i.e., dominant frequency) than that of allopatric G. pennsylvanicus.Calling song properties of males recorded in the hybrid zone varied considerably in temporal and spectral properties. In two-stimulus (choice) phonotaxis experiments, allopatric females of G. firmuspreferred synthetic calling songs with conspecific pulse repetition rates over songs that had lower and higher pulse rates. This preference persisted even when the sound pressure levels of alternative stimuli were unequal. Therefore, allopatric females of G. firmuscan discriminate between conspecific and heterospecific calling songs. Whether or not this same selectivity is present in sympatric populations remains unclear. Investigations of phonotactic selectivity in other allopatric and sympatric populations of both species are currently under way.     

Pollen precedence and stigma closure: a mechanism of competition for pollination between Delphinium nelsonii and Ipomopsis aggregata

Summary Previous experiments showed that the sympatric herbs Delphinium nelsonii and Ipomopsis aggregata compete for hummingbird pollination and that deleterious effects of the former species on seed set of the latter involve interspecific pollen transfer. However, seed set was not reduced when pollen of both species was applied simultaneously to I. aggregata stigmas. Hence a competitive effect may require arrival of foreign pollen before conspecific pollen. To explore this possibility we subjected I. aggregata flowers to a competition treatment in which they received D. nelsonii pollen 6 h before I. aggregata pollen, or to a control in which they received only the conspecific pollen. Foreign pollen precedence decreased mean seed set by almost 50%, which is consistent with effects observed in previous experiments. Reduced seed set can be explained by the fact that foreign pollen often caused stigma lobes to close together within 1.5–6 h, reducing subsequent receptivity. Stigma closure was also elicited by conspecific pollen, but not by mechanical stimulation, and was influenced by size of the pollen load and identity of the plant being pollinated.     

Nectar flow and pollinators' efficiency in two co-occurring species ofCapparis (Capparaceae) in Israel

Two closely related species ofCapparis, C. ovata andC. spinosa, which are exposed to the same pollination environment were studied. The nectar volume and concentration ofC. ovata are higher than inC. spinosa. Both species have similar pollination efficiency, but the relative contribution of each pollinator is different.C. ovata is pollinated mainly hy hawkmoths while the principal pollinators inC. spinosa are bees.     

Temperature Effects on the Temporal Properties of Calling Songs in the Crickets <Emphasis Type="Italic">Gryllus fultoni</Emphasis> and <Emphasis Type="Italic">G. vernalis</Emphasis>: Implications for Reproductive Isolation in Sympatric Populations

Two closely related wood-cricket species, Gryllus fultoni (Orthoptera: Gryllidae) and G. vernalis, produce similar calling songs, consisting of 3-pulse chirps. Analysis of field and laboratory recordings of calling songs showed that, after correction to a common temperature, there was a divergence in chirp and pulse rates between far allopatric populations of G. fultoni and populations sympatric with G. vernalis. To determine whether the divergence in calling songs potentially provides reproductive isolation between G. fultoni and G. vernalis throughout the temperature range over which these insects sing, we recorded calling songs of lab-reared G. fultoni and G. vernalis populations between 18 and 28°C. Mean chirp rate significantly differed between sympatric and far allopatric G. fultoni populations as well as between sympatric G. fultoni and G. vernalis populations. Although there was a significant difference in mean pulse rate between sympatric G. fultoni and G. vernalis populations, pulse rate did not differ between sympatric and far allopatric G. fultoni populations in the laboratory study. Considering the magnitudes of differences in calling song characters discriminated by females of G. fultoni and the mean differences and the variability in calling song characters between the two species, the joint difference in chirp and pulse rates may be adequate for species discrimination over most of the range at which these crickets breed.     

Possible tobacco progenitors share long-tongued hawkmoths as pollen vectors

The putative ancestors of the allopolyploid hybrid Nicotiana tabacum have distinct flower features, apparently suited either for hawkmoth or bat pollination. This suggests that progenitors were reproductively isolated by mechanical and ethological barriers. However, the present data show that in natural populations pollen vectors could be shared by two of the possible progenitors. Pollen vectors of one of the possible male progenitors (N. otophora) were short- and long-tongued hawkmoths and a nectar-feeding bat, while those of the female ancestor (N. sylvestris) were only long-tongued hawkmoths. The latter are then the most likely vectors responsible for the presumed spontaneous hybridization. These data also suggest that interspecific pollen transfer occurred more likely in one direction.     

Reward partitioning in Capparis spp. along ecological gradient

Summary The genus Capparis is represented in Israel by three taxons of the section spinosa. The pollinators of the three taxa are hawkmoths, crepuscular bees of the genus Proxylocopa and various daily bees. However, the efficiency of the different pollinators is very varied with respect to the individual taxons. Hawkmoths were frequently observed as pollinators of C. spinosa var. aravensis and C. ovata, but rarely on C. spinosa var. aegyptia. All the other bees, on the other hand, were found abundantly on all three taxons, all over the country. The three taxa all provide different rewards for their particular pollinators. Bees visiting C. spinosa var. aegyptia are provided with a lot of pollen but a relatively small amount of nectar, with a low sugar content. C. spinosa var. aravensis and C. ovata (the flowers visited more by hawkmoths), have less pollen but a higher amount of nectar which has a higher concentration of sugar. In drier habitats the emphasis of the reward provided by the flowers is nectar, whilst in the Mediterranean habitats it is the pollen which is the reward, without any connection to the systematic origin of the plant.     

Breeding systems and hybridization inPetrorhagia sect.Kohlrauschia (Caryophyllaceae)

In the diploid speciesP. prolifera, the evolution of autogamy has resulted in the presence of distinct selfing and outcrossing races. The change in breeding system toward autogamy is associated with a reduction in the size of floral features and a lower pollen production. The outbreeding populations have only been found in Greece, while the selfing populations have colonized much of Europe. The two races appear to be reproductively isolated from one another. —Petrorhagia sect.Kohlrauschia contains four species.P. glumacea is outbreeding and shows unilateral incompatibility with the small-flowered race ofP. prolifera and interfertility with the large-flowered race; althoughP. glumacea is sympatric with the large-flowered race, they do not appear to hybridize in the wild. Amongst the allopatric species, internal breeding barriers are also found. These may take the form of hybrid sterility or seed incompatibility.P. velutina is autogamous and reproductively isolated from all the other species of the section regardless of whether they are sympatric or allopatric. It is suggested that the evolution of autogamy has been an important factor in the spread ofP. velutina, the tetraploidP. nanteuilii, and the small-flowered race ofP. prolifera in Europe.     

Hybridisation processes in sympatric populations of pines Pinus sylvestris L., P. mugo Turra and P. uliginosa Neumann

Natural hybridisation was postulated between the closely related pine species Pinus sylvestris and the P. mugo complex, however no clear evidence on propagation of mature hybrids in nature has been documented so far. To test the hybridisation hypothesis we applied chloroplast DNA (cpDNA) markers and isozymes in the analyses of 300 individuals representing the variety of morphological forms in the sympatric populations of P. sylvestris, P. mugo and P. uliginosa at the peat bog complex in the Sudety Mts., Poland. Additionally, the haplotypes of paternally inherited cpDNA of 149 open pollinated progeny derived from seeds were compared to the haplotypes of parental trees to access the intensity and direction of contemporary hybridisation. The morphologically highly variable polycormic (multi-stemmed) hybrids between P. mugo and P. uliginosa were identified. The second group of hybrids was found among the monocormic (single-stemmed) P. sylvestris-like individuals carrying the cpDNA from P. mugo complex. Hybrids of P. sylvestris as a pollen donor and P. mugo or P. uliginosa as a mother were not found, either in the group of examined trees, or among the open pollinated progeny. The results indicate that numerous hybrids can exist in the sympatric population of the species studied and that gene flow can successfully proceed from P. mugo complex to P. sylvestris. Hybridisation and ecological selection seems to play a significant role in diversification and evolution of the investigated species.     

Comparative responses of tetraploid wheats pollinated with Zea mays L. and Hordeum bulbosum L.

Ten different tetraploid wheat (Triticum turgidum) genotypes were pollinated with maize (Zea mays). Fertilization was achieved in all ten genotypes and no significant difference in fertilization frequency between the tetraploid wheat genotypes was detected. A mean of 41.1% of pollinated ovaries contained an embryo. All these crosses were characterized by the elimination of the maize chromosomes, and the resulting embryos were haploids. Six of the tetraploid wheat genotypes were also pollinated with Hordeum bulbosum. Fertilization frequencies with H. bulbosum were much lower (mean=13.4%), and significant differences between the tetraploid wheat genotypes were detected. Observation of pollen tube growth revealed that part of the incompatibility reaction between tetraploid wheats and H. bulbosum was due to an effect similar to that of the Kr genes, namely pollen tube growth inhibition. These results indicate that pollinations with maize may have potential as a broad spectrum haploid production system for tetraploid wheats. Present address: Agriculture Canada, Research Branch, Central Experimental Farm, Bldg 50, Ohawa, Ontario, Canada K1A OC6     

Gene flow and spatio-temporal genetic variation among sympatric populations of Tetranychus kanzawai (Acari: Tetranychidae) occurring on different host plants, as estimated by microsatellite gene diversity

We investigated spatio-temporal genetic variation in allele frequency and estimated gene flow among sympatric populations of Tetranychus kanzawai on different host plants by the use of microsatellite markers. In the analysis of spatial genetic variation, no isolation by distance was detected among the populations. Gene flow between populations on Hydrangea macrophylla and those on other host plants was relatively restricted, whereas the populations on Akebia quinata and Clerodendrum trichotomum were almost panmictic. Our study on temporal genetic variation showed (1) that population differentiation was slightly reduced during the period from April to May owing to frequent gene flow among populations; and (2) that population differentiation was greatly enhanced from May to October because of bottleneck effects. Genetic differentiation among T. kanzawai populations was caused by the effect of host plants rather than by the effect of geographic distance among populations, suggesting possibility of sympatric host race formation in this species.This revised version was published online in May 2005 with a corrected cover date.     

Character displacement in sailfin mollies, <Emphasis Type="Italic">Poecilia latipinna</Emphasis>: allozymes and behavior

Synopsis We analyzed variation in allozymes and mating preferences in 12 populations across much of the range of the sailfin molly, Poecilia latipinna. Sailfin mollies can be sympatric with its sexual parasite Amazon mollies, P. formosa. Amazon mollies must co-exist and mate with bisexual males of closely related species (including sailfin mollies) to induce embryogenesis but inheritance is strictly maternal. Where sailfin and Amazon mollies are sympatric there is evidence of reproductive character displacement as males show a significantly stronger mating preference for sailfin molly females over Amazon mollies compared to preferences of males from allopatric populations. From the allozyme data we found a moderate amount of genetic variation across all populations but this variation did not reveal significant partitioning between sympatric and allopatric populations. Additionally, we found no evidence for isolation by distance as genetic distance was not significantly correlated with geographic distance. While allozyme variation also did not significantly correlate with male mating preferences, there was a significant correlation between male mating preferences and geographic distance. This correlation between mating preferences and geographic distance may have arisen from coevolution with Amazon mollies resulting in reproductive character displacement. Taken together, the distribution of genetic and behavioral variation among sympatric and allopatric populations suggests that behavioral evolution has outpaced evolution at the allozyme loci we examined in P. latipinna.     

Electrophoretic analysis of the phylogeny of the self-pollinating populations ofClarkia xantiana

Electrophoretic variation in isozymes coded by 40 loci was examined in two self-pollinating populations, one with white and the other with pink flowering plants, and four outcrossing populations ofClarkia xantiana (Onagraceae) native to California. The study was carried out to test theMoore & Lewis (1965) hypothesis that the pink selfer originated from the sympatric outcrossing population and then gave rise to the white selfer. The hypothesis could be rejected if one or the other selfer was more similar genetically to an allopatric population than to the sympatric one. Both selfers were monomorphic at all loci whereas the outcrossing populations were polymorphic at nearly half of them. The two selfers had the same genes at 32 loci but had different ones at eight loci. The pink selfer was not more similar to the allopatric populations than to the sympatric one, consistent with theMoore & Lewis model. The evidence also supported their proposal that the white selfer originated from the pink one and not independently. The electrophoretic evidence was valuable because it permitted qualitative comparisons (presence versus absence of particular alleles) between the selfer and the several outcrossing populations; such analysis was not previously possible because the latter populations are morphologically and cytologically similar.