Effects of soil nitrogen on diploid advantage in fireweed,Chamerion angustifolium (Onagraceae)

In many ecosystems, plant growth and reproduction are nitrogen limited. Current and predicted increases of global reactive nitrogen could alter the ecological and evolutionary trajectories of plant populations. Nitrogen is a major component of nucleic acids and cell structures, and it has been predicted that organisms with larger genomes should require more nitrogen for growth and reproduction and be more negatively affected by nitrogen scarcities than organisms with smaller genomes. In a greenhouse experiment, we tested this hypothesis by examining whether the amount of soil nitrogen supplied differentially influenced the performance (fitness, growth, and resource allocation strategies) of diploid and autotetraploid fireweed (Chamerion angustifolium). We found that soil nitrogen levels differentially impacted cytotype performance, and in general, diploids were favored under low nitrogen conditions, but this diploid advantage disappeared under nitrogen enrichment. Specifically, when nitrogen was scarce, diploids produced more seeds and allocated more biomass toward seed production relative to investment in plant biomass or total plant nitrogen than did tetraploids. As nitrogen supplied increased, such discrepancies between cytotypes disappeared. We also found that cytotype resource allocation strategies were differentially dependent on soil nitrogen, and that whereas diploids adopted resource allocation strategies that favored current season reproduction when nitrogen was limiting and future reproduction when nitrogen was more plentiful, tetraploids adopted resource allocation strategies that favored current season reproduction under nitrogen enrichment. Together these results suggest nitrogen enrichment could differentially affect cytotype performance, which could have implications for cytotypes’ ecological and evolutionary dynamics under a globally changing climate.     

Cytotype distribution at a diploid-tetraploid contact zone in Chamerion (Epilobium) angustifolium (Onagraceae)

In North America, the geographic distributions of diploid and tetraploid Chamerion (formerly Epilobium) angustifolium overlap in a narrow zone along the southern border of the boreal forest and along the Rocky Mountains. We examined the frequency and distribution of diploid and tetraploid cytotypes in a narrow (5 km) zone of sympatry across an elevational gradient and in putatively uniform diploid and tetraploid reference populations on the Beartooth Pass, in the Rocky Mountains of southern Montana-northern Wyoming. All five reference populations sampled were dominated by a single cytotype, but only one was completely uniform. In the zone of sympatry, 27 transects were sampled every 2 m for a total of 238 plants. Reproductive status (vegetative, flower buds, open flowers) was recorded, and the ploidy of each plant was determined by flow cytometry. Diploid and tetraploid plants predominated (36 and 55%, respectively) but were heterogeneously distributed among the transects. Six of the 27 transects were fixed for a single cytotype (four transects, diploid; two transects, tetraploid), and in seven others either diploids or tetraploids predominated (frequency >75%). Triploids represented 9% of the total sample and occurred most frequently in transects containing both diploids and tetraploids (G = 3.4, df = 2, P = 0.07). Diploids were more often reproductive (in bud, flower, or fruit) than either triploids or tetraploids (G = 12.0, df = 2, P < 0.001) and were the only cytotype to have produced open flowers. These results suggest that the zone of sympatry is best characterized as a mosaic rather than a cline, with diploid and tetraploids in close proximity and that the distribution of polyploidy is regulated by ecological sorting in a heterogeneous physical environment.     

Variation Patterns and Systematics of the Widespread Species Epilobium angustifolium L.(Onagraceae) in China

Cytological, morphological, and geographical studies of Epilobium angustifolium in China reveal two partly distinct, partly overlapping entities that are best treated as subspecies. The diploid (n=18) subsp. angustifolium has smaller, triporate pollen grains, typically glabrous stems and leaves, and smaller leaves and flowers, and occurs in colder, more northern areas, and at higher elevations in overlap zones. The tetraploid (n=36) subsp. circumvagum has larger pollen with a mixture of quadriporate pollen grains, pubescent stems and leaves, and larger leaves and flowers, and occurs in warmer parts of southern China and at lower elevations. We report 11 chromosome counts, six diploid and five tetraploid, includingthe first reported tetraploids in Asia.     

Self-sterility in Epilobium obcordatum (Onagraceae)

Greenhouse pollinations among 23 field-collected plants and their progeny revealed a high and variable degree of self-sterility in Epilobium obcordatum. Field-collected plants averaged 5% seed-set when selfed and 43% seed-set in crosses; greenhouse progeny 11% and 49%, respectively. In two of three pollen chase tests, previously or simultaneously applied self-pollen reduced seed-set to that resulting from pure self-pollination. Based on the relative survivorship of self- and outcross embryos, the mean number of lethal equivalents was determined to be 11.0 (range 2.9 to 17.6) in the field-collected plants, and 7.1 (range 0.62 to 21.8) in the greenhouse progeny. Pollinations among 91 individuals showed a significant negative correlation between seed-set and inbreeding coefficients of the resulting zygotes. Intrapopulation crosses yielded significantly fewer seeds than interpopulation crosses. Early embryonic mutational load, in combination with some level of biparental inbreeding, appears to be responsible for the level of self-sterility in natural populations of this species. A pollen dosage test for an interpopulation cross resulted in a maximum seed set of 47% at 16 tetrads. Additional unmeasured characteristics govern the probability of ovules becoming mature seeds.     

Ovule fates in Epilobium obcordatum (Onagraceae)

We investigated the fates of ovules following self- and cross-pollination of the variably self-sterile Epilobium obcordatum by observing thin sections of ovaries 4, 5, and 10 d postpollination (PP). Three types of unfertilized ovules were observed: apparently normal ovules not entered by pollen tubes, nonfunctional ovules with a nucellus but lacking an embryo sac, and empty ovules with integuments but lacking a nucellus. The average frequencies for all three types were not different for selfs and crosses. Fertilization frequencies did not differ among pollination treatments, averaging 61.4% of the total sample of 4 and 5 d PP ovules. The phenotypes of failing postfertilization ovules at 4 and 5 d PP included those containing preglobular and globular embryos lacking endosperm, ovules with endosperm but no embryo present, and failing embryos in the presence of endosperm. Self- and cross-fertilized 10 d PP ovules also possessed heart-stage and torpedo-stage failing embryos. Early embryo failures (5 d PP) were distributed throughout the length of the ovary, as expected if failures are the consequence of factors intrinsic to each ovule. The sizes of normally developing self- and cross-fertilized embryos were not different at 4 d PP. The 5 d PP cross embryos were larger than selfs. A comparison of 10 d samples was not possible due to attrition of self embryos. Although ovular position did not affect the probability of early embryo failure, it did affect the size of normally developing embryos at 4, 5, and 10 d PP, with embryos at the stylar end larger than those at the basal end.     

The systematics of autopolyploidy in Epilobium latifolium (Onagraceae)

Diploid (2n = 36) plants ofEpilobium latifolium L. have been found in Alaska and the western Cordillera of North America, and tetraploids in Iceland, western Greenland, and southeastern Quebec. Study of the morphology of these chromosome races revealed a character, the number of pores in the pollen grains, by which they could be distinguished with an average probability of 75%. Tetraploids ofE. latifolium tend to have high percentages of 4-pored pollen, while the diploids usually have only 3-pored pollen. On this basis, the ranges of the races were extrapolated from herbarium material. Detailed comparisons of eight other features of the presumptive diploids and tetraploids showed no significant difference between them. The races appear to form worldwide, allopatric, ecogeographic phases. A high level (57.4%) of quadrivalent formation observed during tetraploid meiosis, and other considerations, indicate that the tetraploids arose by autopolyploidy. The recommendation is made that the races not be given formal taxonomic recognition.     

Sources of variation in floral nectar production rate in Epilobium canum (Onagraceae): implications for natural selection

Sources of variation in floral nectar production were investigated in a natural population of Epilobium canum (Onagraceae), a hummingbird-pollinated herbaceous shrub. Field measurements showed significant phenotypic variation among plants in floral nectar production rates. Average variance among flowers within plants was approximately one-third to one-half as great as variance among plants, with coefficients of variation among flowers ranging from 6.5% to 116.7%. A greenhouse experiment using clonally propagated ramets from field plants showed significant genetic variation for nectar production rates; broad sense heritability was estimated to have a maximum value of 0.64. In the greenhouse, plants grown under low water or low light conditions produced approximately 25% less nectar on average than those grown under control conditions. However, significant genotype-environment interactions indicated that genets differed in their responses to the changes in conditions. Rank correlations for genet mean nectar production rates across environmental conditions were low, and in two out of three comparisons were not different from zero. It is concluded that although the opportunity for natural selection on nectar production rates exits in this population, the response to selection will likely be slow, and the opportunity for selection of a narrow-optimum nectar production phenotype may be limited. Received: 9 January 1996 / Accepted: 18 December 1996     

Pollinator-mediated assortative mating in mixed ploidy populations of Chamerion angustifolium (Onagraceae)

Establishment of polyploid individuals within diploid populations is theoretically unlikely unless polyploids are reproductively isolated, pre-zygotically, through assortative pollination. Here, we quantify the contribution of pollinator diversity and foraging behaviour to assortative pollen deposition in three mixed-ploidy populations of Chamerion angustifolium (Onagraceae). Diploids and tetraploids were not differentiated with respect to composition of insect visitors. However, foraging patterns of the three most common insect visitors (all bees) reinforced assortative pollination. Bees visited tetraploids disproportionately often and exhibited higher constancy on tetraploids in all three populations. In total, 73% of all bee flights were between flowers of the same ploidy (2x–2x, 4x–4x); 58% of all flights to diploids and 83% to tetraploids originated from diploid and tetraploid plants, respectively. Patterns of pollen deposition on stigmas mirrored pollinator foraging behaviour; 73% of all pollen on stigmas (70 and 75% of pollen on diploid and tetraploid stigmas, respectively) came from within-ploidy pollinations. These results indicate that pollinators contribute to high rates of pre-zygotic reproductive isolation. If patterns of fertilization track pollen deposition, pollinator–plant interactions may help explain the persistence and spread of tetraploids in mixed-ploidy populations.     

Unfertilized ovules of Epilobium obcordatum (Onagraceae) continue to grow in developing fruits

To determine whether unfertilized ovules continue to grow when in an ovary containing fertilized ovules, we measured ovule lengths in developing fruits of Epilobium obcordatum that were harvested 4, 5, 8, and 10 d post pollination. We found that unfertilized ovules that were in the presence of fertilized ovules continued to grow and that there was a broad range of overlap in their sizes at all sampling times. This effect was found for two types of unfertilized ovules that occur throughout the length of the ovary: normal, unfertilized ovules, apparently bypassed by pollen tubes; and sterile ovules lacking an embryo sac. In addition, there is a position effect within developing fruits. Both fertilized and unfertilized ovules are larger at the stylar end. In six samples resulting from pollination with a single pollen tetrad, a total of 18 embryos were found, and the effect on unfertilized ovules, greatest at the stylar end, diminished with distance from the ovules with embryos. Our results are consistent with the interpretation that diffusible hormones produced by developing seeds cause nearby unfertilized ovules to grow. We conclude that caution is necessary when attempting to infer ovule fertilization histories from the appearances of ovules in developing and mature fruits. What are often inferred to be aborted seeds, in many cases, may not be seeds at all. They may be enlarged, unfertilized ovules.     

Hybridization in Epilobium (Onagraceae): the effect of clearance and re-establishment of fen carr

A population of hybridizing Epilobium in a clearing in fen carr was observed for two years. Traditional and statistical techniques were used to identify hybrids, of which there appeared to be three, involving three parents. The identified hybrids showed correlation in abundance with that of their parents and with the successional status of the vegetation. Even what purported to be a pure E. obscurum population showed characteristics of E. palustre , suggesting a hybrid swarm, but there was evidence that selection was gradually restoring the population to its presumed pre-clearance state.     

Contrasting movement patterns of nectar-collecting and pollen-collecting bumble bees (Bombus terricola) on fireweed (Chamaenerion angustifolium) inflorescences

Abstract. 1. Movements of nectar and pollen-foraging bumble bees on inflorescences of Chamaenerion angustifolium (L.) J. Holub (fireweed or rosebay willow herb) were compared with predictions based on reward distributions and optimality principles. Observations suggest that nectar and pollen-gathering bumble bees behave according to the same set of reward maximization criteria when foraging from flowers of this species.
2. Both kinds of foragers matched their arrival points with the vertical positions on inflorescences in which the densities of their respective food resources were greatest. For nectar-foragers, this point was located at the lowest tier of flowers, whereas for pollen-foragers it was found in the middle of the inflorescences. Nectar and pollen-foraging bees both moved upward on inflorescences following gradients from high to low reward availability.
3. Nectar-foragers responded to decreases in inflorescence size over the season by reducing the number of flower visits made on each raceme. Number of flowers visited by pollen-foragers was low throughout and reflected the scarcity of male-phase flowers on racemes. Flower revisitation rates were low for both kinds of workers, but were slightly higher for those collecting pollen.     

Genetic variability in Chamaenerion angustifolium (L.) Scop. (Onagraceae) occurring on contrasting soils

Isoenzyme analysis of 5 populations of Chamaenerion angustifolium (L.) Scop. from different habitats revealed that only two of the nine enzymes scored were polymorphic. The estimated number of polymorphic loci was 2 out of 16. The only significant difference found was with regard to the gene and genotype frequencies of glucose-6-phosphate dehydrogenase (G6PDH). This difference could be demonstrated for established populations, occurring on contrasting soils but not for their seed populations.The genetic similarity between seed populations even in a sample obtained from only four established individuals was striking and its consequences for the life strategy and colonizing ability of the species are discussed. Genotypic differentiation due to age structure was not present. Genotype-dependent selection occurs in the seedling phase only.     

Differences in the location of subcotyledonary buds among Epilobium angustifolium L., E. dodonaei Vill. and E. fleischeri Hochst. (Onagraceae) and effects on architecture and population structure

STÖCKLIN, J., 1992. Differences in the location of subcotyledonary buds among Epilobium angustifolium L., E. dodonaei Vill. and E. fleischeri Hochst. (Onagraceae) and effects on architecture and population structure . Morphological and architectural features in the two closely related pioneer plant species Epilobium dodonaei and E. fleischeri are examined in cultivation and in the field and compared with E. angustifolium. In E. angustifolium , the aerial shoot system is renewed every year from buds on horizontal roots and results in a horizontal spread and a clonal growth form. In E. dodonaei , bud formation is restricted to the hypocotyl and in larger plants to the transitional region between root and shoot. Consequently this species shows no vegetative mobility and develops a shrub-like habit. The alpine E. fleischeri combines the habit of E. angustifolium and E. dodonaei and may either develop successive generations of shoots from the transitional region between root and shoot and/or exploit new areas by horizontal roots and the formation of shoots from root buds. The simple difference in the location of renewal buds is accentuated by cumulative growth. The study shows that E. dodonaei and E. fleischeri , which often are considered as subspecies, are separated by fundamental differences in their architectural models. The ecological and demographic implications of these differences are discussed.     

The effect of protandry on siring success in Chamerion angustifolium (Onagraceae) with different inflorescence sizes

Abstract Protandry, a form of temporal separation of gender within hermaphroditic flowers, may reduce the magnitude of pollen lost to selfing (pollen discounting) and also serve to enhance pollen export and outcross siring success. Because pollen discounting is strongest when selfing occurs between flowers on the same plant, the advantage of protandry may be greatest in plants with large floral displays. We tested this hypothesis with enclosed, artificial populations of Chamerion angustifolium (Onagraceae) by experimentally manipulating protandry (producing uniformly adichogamous or mixed protandrous and adichogamous populations) and inflorescence size (two-, six-, or 10-flowered inflorescences) and measuring pollinator visitation, seed set, female outcrossing rate, and outcross siring success. Bees spent more time foraging on and visited more flowers of larger inflorescences than small. Female outcrossing rates did not vary among inflorescence size treatments. However, seed set per fruit decreased with increasing inflorescence size, likely as a result of increased abortion of selfed embryos, perhaps obscuring the magnitude of geitonogamous selfing. Protandrous plants had a marginally higher female outcrossing rate than adichogamous plants, but similar seed set. More importantly, protandrous plants had, on average, a twofold siring advantage relative to adichogamous plants. However, this siring advantage did not increase linearly with inflorescence size, suggesting that protandry acts to enhance siring success, but not exclusively by reducing between-flower interference.     

Competition and the compensatory regulation of fruit and seed set in the perennial herb Epilobium dodonaei (Onagraceae)

I studied the effects of competition and soil fertilization on variation of seed yield components of the outcrossing perennial Epilobium dodonaei Vill. using a randomized complete block design. Fertilization as a main effect was not significant. Competition, however, reduced the number of flower buds per shoot (-29%), the number of ovules per fruit (-12%), and the number of ripe fruits per shoot (-51%). In addition, competition caused an increase in abortion of flower buds. Consequently the fruit/flower bud ratio decreased from 0.71 to 0.47. Average seed mass was not reduced significantly by competition. Some of the negative effects of competition on fruit production were mitigated by fertilization. However, competition considerably reduced the proportion of late-aborted seeds, which resulted in an increase of the seed/ovule ratio from 0.31 to 0.49 (+58%). As a consequence of this compensation, the reduction in the number of seeds per shoot due to competition was not significant. At the level of the genet, competition had a strong effect on seed yield due to decreases in the number of shoots produced. I discuss patterns of regulation at consecutive levels of reproduction. Shoots of plants suffering from competition initially invested less in reproductive structures than the control and showed a higher abortion rate of less costly structures early in reproductive development, but also had a lower abortion rate later in their development. The observed compensatory pattern in yield components illustrates the fine-tuning regulation capacity at different levels of reproductive development in plants.     

Nectary and gender‐biased nectar production in dichogamous Chamaenerion angustifolium (L.) Scop. (Onagraceae)

In dichogamous plants, nectar characteristics (i.e. nectar amount and its composition) can differ between sexual phases. In the present study, we investigated the structural organization of the floral nectary, nectar production and carbohydrate composition in the protandrous Chamaenerion angustifolium (L.) Scop. (Onagraceae). The receptacular nectary consisted of an epidermis with numerous nectarostomata, several layers of photosynthetic secretory parenchyma, and subsecretory parenchyma. Nectariferous tissue was not directly vascularized and starch grains were rarely observed in the secretory cells, occurring exclusively in the guard cells of modified stomata. The nectar was released via nectarostomata. The floral nectar was hexose rich (32.8/39.1/28.1% glucose/fructose/sucrose) and the total concentration was constant throughout the anthesis (47% on average). However, contrasting patterns in nectar amount and carbohydrate composition between the floral sexual phases were observed. On average, female‐phased flowers produced 1.4‐fold more nectar than male‐phased flowers, and although the nectar was sucrose rich during the male phase, it was hexose rich during the female phase, suggesting sucrose hydrolysis.     

Vapour Phase Activation,Intracellular Motility and Germination in the Tripororate Pollen of Epilobium angustifolium

The germinability of newly released pollen of Epilobium angustifolium in liquid medium is low, but it is enhanced by a short period in an atmosphere of 85–95% RH without access to liquid water or solutes. During this period the thickened pectic layers of the intine at the germination apertures of the grains are eroded and prospective tube tips emerge. Concomitantly polarised movement of organelles begins in the vegetative cells, and wall precursor vesicles (P-particles) accumulate at the apertures. In pollen held continuously in the humid environment transition to cylindrical growth follows, with the establishment of the inverse-fountain pattern of circulation in the apical part of the tubes.     

Reproductive allocation and the fitness consequences of selfing in two sympatric species of Epilobium (Onagraceae) with contrasting mating systems

Selfing has evolved repeatedly in outcrossing taxa, and theory predicts that an increase in the level of self-fertilization should occur in concert with changes in reproductive allocation and the magnitude of inbreeding depression. Here we characterize the mating system of two sympatric congeners, Epilobium ciliatum and E. angustifolium, and compare the taxa for 1) reproductive allocation patterns and 2) the fitness consequences of self-fertilization. For E. ciliatum, autogamy rates were high, pollinator visitation was low, and electrophoresis revealed no genetic variation at 11 putative isozyme loci. For E. angustifolium, autogamy rates were low, pollinator visitation was relatively high, and electrophoresis generated an outcrossing rate estimate of t = 0.64 (SE = 0.08). The pollen/ovule ratio was ten times higher for E. angustifolium than for E. ciliatum, due to a decline in pollen production in the selfing species. The proportion of total flower biomass allocated to female function was significantly greater in E. ciliatum, while that allocated to male function and attractive structures was greater in E. angustifolium. We quantified the fitness consequences of selfing at three life stages: seed number, percent germination, and mature biomass. Relative performance (RP) measures indicated less inbreeding depression for E. ciliatum than for E. angustifolium at all stages; differences in RP between the species were significant for seed number and cumulative total, but not for germination or biomass. RP was correlated among life history stages for only one comparison, suggesting that the genetic basis of inbreeding depression differs among life history stages. Variation among maternal parents for RP was significant at almost all stages in both species, with the exception of seed number in E. ciliatum. The striking variation among maternal parents in E. angustifolium, ranging from strong inbreeding depression to strong outbreeding depression, may reflect both variation in the history of inbreeding and the long-distance migration of individuals from different populations.     

The mechanisms of the restriction of pollinator range in the fireweed (Chamaenerion angustifolium) and 2 species of geranium (Geranium palustre) and G. pratense)

Flowers of Chamaenerion angustifolium, Geranium palustre and G. pratense are visited by more than 100 insect species. For all plants the spectrum of visitors is similar. However the role of insects species in pollination is different and depends on the stamen and stigma length, flexibility of pedicle and feeding behaviour of insect inside flower. The possibility to take and to transport pollen grains usually increase with the size of insect. The nature of this correlation is determined by stamen and stigma length. Pollinators of G. palustre with short stamens and stigmas are smaller than those of Ch. angustifolium and G. pratense. On the other hand, more flexible pedicle of G. palustre prevent the flowers from visits of big insects. Three plants studied by the authors are not equally attractive for different insect groups. Dense in fluorescence of Ch. angustifolium and G. pratense that usually are lifter under the grass are very attractive for foraging social insects (honey bee, bumblebee). Flies avoid long distance travelling and prefer single flowers located not far from each other. For instance G. palustre is more attractive for flies not for social bees. It is pollinated mainly by flies and solitary bees with average weight of 10-70 mg. The main pollinators of Ch. angustifolium and G. pratense are honey bees, bumblebees and wasps with average mass exceeding 70 mg.