CLEISTOGAMY IN MICROLAENA POLYNODA (GRAMINEAE): AN EXAMINATION OF SOME MODEL PREDICTIONS

A population of the cleistogamous grass, Microlaena polynoda, was investigated to determine some of the factors responsible for adjusting the balance between reproduction by chasmogamous (CH) and cleistogamous (CL) flowers, the tissue costs associated with the two reproductive modes, the fate of progeny produced by each mode, and the genetic diversity of the progeny. Cleistogamous flower production begins earlier in the season than CH flower production. There is a distinct threshold of low light intensity below which mostly CL flowers are produced. Paternal expenditure per plant is an order of magnitude larger for the CH than the CL component. The opposite relation holds for maternal expenditure. Increased maternal expenditure in the CL component may be due to greater fertilization success and retrieval of paternal costs. Cleistogamous seeds are dispersed later than CH seeds. Following dispersal, the spikelet encloses the CL seed but not the CH seed, and is responsible for inducing dormancy. The ratio of seedlings arising from CH seeds to that from CL seeds in a natural habitat is significantly lower than the ratio of estimated numbers of CH to CL seeds produced. There were no detectable polymorphisms among ten presumptive enzyme loci assayed. Many of the features associated with CH and CL reproduction in M. polynoda are in accord with the theoretical requirements for the evolution of closed flower self-pollination and the maintenance of two distinct methods of reproduction.     

Establishment of chasmogamous and cleistogamous seedlings of an ant-dispersed UNDERSTORY HERB,CALATHEA MICANS (Marantaceae)

This study compares the establishment success of cleistogamous (CL, obligately selfed) and chasmogamous (CH, potentially outcrossed) seeds of the neotropical understory herb, Calathea micans (Marantaceae). In this species, CH and CL seeds are morphologically identical and are both dispersed by ants. I measured dispersal distances and analyzed seedling recruitment and seedling size for ant-planted CH and CL seeds, a protocol made possible by the fact that the seed coat of C. micans can be indelibly marked and remains attached to the base of the seedling for a few months. Seeds were taken by ants from experimental depots placed in natural conditions. In a second experiment, I planted CH and CL seeds in gap and understory sites to determine the effects of seed type and light on germination percentage, date of emergence, and seed and seedling survivorship. Ant-planted CH and CL seedlings did not differ in dispersal distance. CH and CL seeds did not differ significantly in recruitment probability and seedling size in either experiment. However, for the ant-planted seedlings, CH recruitment was higher in the understory than in the gap, while CL recruitment was uniform across light environments. In conclusion, I found a difference in the establishment success of CH and CL seeds in this understory herb, only after dispersal by ants in field conditions.     

RELATIVE PERFORMANCE OF SELFED AND OUTCROSSED PROGENY IN IMPATIENS CAPENSIS

This study compares survival and growth of progeny derived from chasmogamous (CH) and cleistogamous (CL) flowers in Impatiens capensis, a forest annual. When progeny were grown in the field, CH seeds had significantly higher survival rates over winter (64% versus 56%), and the survival advantage of outcrossed progeny was not attributable to seed weight differences. No differences in seedling growth were detected. Greenhouse comparisons revealed no difference in seed survival but a 30% growth advantage to CH seedlings. We found no changes in developmental homeostasis of three leaf shape characters between inbred and outbred progeny, nor was there any difference in variability within CH and CL families. The outcrossing advantage observed in these experiments could not have been caused by avoidance of sib competition. Theory predicts that self-pollinated progenies may be more variable than outcrossed progenies if rare, recessive alleles are important contributors to genetic variances. Electrophoretic markers indicate that progeny derived from CH flowers are predominantly outcrossed (at least 54-97%).     

The Effect of Pollen Source vs. Flower Type on Progeny Performance and Seed Predation under Contrasting Light Environments in a Cleistogamous Herb

Dimorphic cleistogamy is a specialized form of mixed mating system where a single plant produces both open, potentially outcrossed chasmogamous (CH) and closed, obligately self-pollinated cleistogamous (CL) flowers. Typically, CH flowers and seeds are bigger and energetically more costly than those of CL. Although the effects of inbreeding and floral dimorphism are critical to understanding the evolution and maintenance of cleistogamy, these effects have been repeatedly confounded. In an attempt to separate these effects, we compared the performance of progeny derived from the two floral morphs while controlling for the source of pollen. That is, flower type and pollen source effects were assessed by comparing the performance of progeny derived from selfed CH vs. CL and outcrossed CH vs. selfed CH flowers, respectively. The experiment was carried out with the herb Ruellia nudiflora under two contrasting light environments. Outcrossed progeny generally performed better than selfed progeny. However, inbreeding depression ranges from low (1%) to moderate (36%), with the greatest value detected under shaded conditions when cumulative fitness was used. Although flower type generally had less of an effect on progeny performance than pollen source did, the progeny derived from selfed CH flowers largely outperformed the progeny from CL flowers, but only under shaded conditions and when cumulative fitness was taken into account. On the other hand, the source of pollen and flower type influenced seed predation, with selfed CH progeny the most heavily attacked by predators. Therefore, the effects of pollen source and flower type are environment-dependant and seed predators may increase the genetic differences between progeny derived from CH and CL flowers. Inbreeding depression alone cannot account for the maintenance of a mixed mating system in R. nudiflora and other unidentified mechanisms must thus be involved.     

FIELD GERMINATION AND SEEDLING GROWTH OF CH AND CL PROGENY OF IMPATIENS CAPENSIS (BALSAMINACEAE)

The germination behavior and early growth of chasmogamous (CH) and cleistogamous (CL) progeny of Impatiens capensis were investigated in two eastern Nebraska stands. Field germination of families of buried seeds was scored in mid-April. In one stand a significantly higher proportion of CL seeds germinated than CH seeds while no significant differences were observed in the second stand. Among-family variation in germination rates was significant in both stands. The time course of field germination was similar for CL and CH seeds. A sample of the germinated CL and CH seedlings, matched in size, was planted individually in pots and grown in the greenhouse for five wk. Early growth of CL and CH seedlings was similar. In addition, CL seedlings were as variable as CH seedlings for four morphological traits. Overall, a large fitness advantage for CH progeny was not detected in the germination and early growth of Impatiens capensis seedlings.     

DISPERSAL OF HETEROMORPHIC SEEDS IN AMPHICARPAEA BRACTEATA (FABACEAE)

The dispersal of the chasmogamous, aerial cleistogamous, and subterranean cleistogamous seeds of Amphicarpaea bracteata was examined. The chasmogamous and aerial cleistogamous seeds are ballistically dispersed. Chasmogamous seeds were dispersed farther than the aerial cleistogamous seeds due to the height advantage of the chasmogamous pods. There was no difference in the firing angle or the initial velocity of the seeds discharged from the two aerial pod types. The subterranean cleistogamous seeds are “dispersed” the shortest distance by the elongation of runners. Differential dispersal of the three types of seeds may be a factor involved in the evolution of cleistogamy in A. bracteata.     

Offspring performance in three cleistogamous Viola species

Seed germination and seedling/juvenile fitness in the cleistogamous perennials Viola hirta, V. mirabilis, and V. riviniana were investigated during three growing seasons, to compare the performance of chasmogamously (CH) and cleistogamously (CL) derived progeny. For V. hirta and V. mirabilis the effects of sibling competition were examined, for V. riviniana the effects of interspecific (grass) competition. Seed abortion and seed weight were also taken into account as fitness measures.In none of the species, seed abortion rate differed between CH and CL capsules. In V. mirabilis and V. riviniana, CL seeds had a lower germination rate than CH seeds. In V. hirta the two seed types did not differ in germinability. Mortality did not differ between the two seedling types in any of the species. In V. hirta and V. riviniana, CL progeny had shorter mean length of largest leaf than CH progeny. In V. mirabilis plant size did not differ between progeny types. Sibling competition had little effect on offspring performance, but grass competition increased mortality and reduced plant size of V. riviniana progeny. The two progeny types did not differ in their response to sibling and grass competition.The differences in performance between progeny types could be attributable to inbreeding depression in the CL phase, but the slightly lower fitness of CL offspring is probably balanced by their lower production costs. It is suggested that a dimorphic reproductive system is maintained in perennial Viola species to maximize total seed output in the face of environmental variation.     

Environmental control of reproductive phenology and the effect of pollen supplementation on resource allocation in the cleistogamous weed, Ruellia nudiflora (Acanthaceae)

• Background and Aims Mixed reproductive strategies may have evolved as a response of plants to cope with environmental variation. One example of a mixed reproductive strategy is dimorphic cleistogamy, where a single plant produces closed, obligately self-pollinated (CL) flowers and open, potentially outcrossed (CH) flowers. Frequently, optimal environmental conditions favour production of more costly CH structures whilst economical and reliable CL structures are produced under less favourable conditions. In this study we explore (1) the effect of light and water on the reproductive phenology and (2) the effect of pollen supplementation on resource allocation to seeds in the cleistogamous weed Ruellia nudiflora.• Methods Split-plot field experiments were carried out to assess the effect of shade (two levels: ambient light vs. a reduction of 50 %) and watering (two levels: non-watered vs. watered) on the onset, end and duration of the production of three reproductive structures: CH flowers, CH fruit and CL fruit. We also looked at the effect of these environmental factors on biomass allocation to seeds (seed weight) from obligately self-pollinated flowers (CL), open-pollinated CH flowers and pollen-supplemented CH flowers.• Key Results CH structures were produced for a briefer period and ended earlier under shaded conditions. These conditions also resulted in an earlier production of CL fruit. Shaded conditions also produced greater biomass allocation to CH seeds receiving extra pollen.• Conclusions Sub-optimal (shaded) conditions resulted in a briefer production period of CH structures whilst these same conditions resulted in an earlier production of CL structures. However, under sub-optimal conditions, plants also allocated more resources to seeds sired from CH flowers receiving large pollen loads. Earlier production of reproductive structures and relatively larger seed might improve subsequent success of CL and pollen-supplemented CH seeds, respectively.     

A TEST OF THE SIB-COMPETITION HYPOTHESIS FOR OUTCROSSING ADVANTAGE IN IMPATIENS CAPENSIS

One of the potential selective mechanisms invoked in discussions of breeding-system evolution is that competition within sibships increases the fitness of outcrossed progeny relative to selfed progeny. We tested this sib-competition hypothesis using cleistogamous (CL) and chasmogamous (CH) seeds of Impatiens capensis in a large greenhouse experiment. The experimental design was a double replacement series which also allowed us to test for inbreeding depression and overall resource partitioning among sibships. We found no evidence for strong inbreeding depression in the study population; although plants from CH seeds had a slight advantage over plants from CL seeds in total flower and pod production, CL plants had slightly higher growth. We also could not detect significant resource partitioning among sibships nor any evidence to support the sib-competition hypothesis for outcrossing advantage. CH sibships were not significantly more variable than CL sibships in any of the phenotypic traits measured. These results suggest that sibling competition may have little importance in the evolution of Impatiens breeding systems.     

ENHANCEMENT OF INBREEDING DEPRESSION BY DOMINANCE AND SUPPRESSION IN IMPATIENS CAPENSIS

We investigated the effect of intraspecific competition on the magnitude of inbreeding depression in Impatiens capensis by planting seeds from chasmogamous (CH) and cleistogamous (CL) flowers in three experimental greenhouse treatments: in individual pots, in flats in dense pure stands according to seed type, and in flats with the two seed types intermixed in a checkerboard array. The size distributions of plants grown in flats were significantly more hierarchical than those of plants grown individually, indicating that larger plants competitively suppressed smaller plants in the high-density treatments. The magnitude of inbreeding depression at high density depended upon the planting arrangement of CL and CH seeds. CH advantage was greatest when CH and CL seedlings were grown in competition with one another, suggesting that fitness differences between outcrossed and inbred individuals were intensified by dominance and suppression. For plants grown individually, the effects of maternal parent, seed weight, and emergence date on seedling size disappeared with plant age, whereas at high density these effects remained at the final harvest. Thus, plant density may influence patterns of natural selection both on mating system and on juvenile traits in natural Impatiens populations.     

Why is there variation in mean seed size among plants within single populations? test of the fertilization efficiency hypothesis

We tested the fertilization efficiency hypothesis, which attempts to explain mean seed size variation among plants within single populations, by comparing the patterns of seed size variation between chasmogamous (CH) flowers and cleistogamous (CL) flowers in Impatiens noli-tangere and Viola grypoceras, respectively. The fertilization efficiency hypothesis predicts that larger plants produce larger seeds if the number of pollen grains captured by a plant increases with increased allocation of resources to its attractive structures (e.g., corolla and nectar), but with diminishing gains. Thus, seed size should depend on plant size in seeds from CH flowers because of the diminishing gains of capturing pollen in these flowers, whereas seed size should not depend on plant size in seeds from CL flowers because CL flowers need not capture outcross pollen. We found significant positive correlations between mean seed size per plant and plant size for seeds from CH flowers in both species. However, there was no significant positive correlation between these two factors for seeds from CL flowers of both species. The results of the present investigations were thus consistent with the fertilization efficiency hypothesis.     

CHASMOGAMY/CLEISTOGAMY IN TRIODANIS PERFOLIATA (CAMPANULACEAE): SOME CH/CL COMPARISONS IN FITNESS PARAMETERS

The chasmogamous/cleistogamous (CH/CL) breeding system has evolved independently many times, but its ecological role has been studied in few species. In Triodanis perfoliata, we studied the natural phenology of flowering and seed set, how successful CH flowers were at setting seed, whether seed from CH and CL flowers differed in germination behavior, and whether plants derived from these two seed types differed in vigor. Most notably, we found that the later the CH flowers were produced, the fewer seed they successfully matured. In general the two seed types did not show differences in germination behavior, but for some maternal parents the two seed types differed. For these, the CL seed germinated less readily. Finally, there was some indication that plants from CL seed were not as vigorous as those from CH seed.     

Analysis of mating system and genetic structure in the endangered,amphicarpic plant,Lewton’s polygala (<Emphasis Type="Italic">Polygala lewtonii)</Emphasis>

Polygala lewtonii is a federally endangered, amphicarpic plant with a mixed mating system and three types of flowers: (1) aboveground, chasmogamous flowers (i.e., open-pollinated; CH), (2) aboveground, cleistogamous flowers (i.e., closed, selfing; CL) and (3) CL flowers on belowground stems (amphicarpy). Aboveground seeds are ant-dispersed, whereas belowground seeds are spaced across the length of the rhizome. Here, we collected individuals of P. lewtonii at both range-wide and fine geographic scales and genotyped them at 11 microsatellite loci. We analyzed patterns of genetic diversity and structure to understand: (1) the predominant mating system (selfing or outcrossing), (2) the movement of pollen and seeds across the landscape, and (3) the optimal strategy to conserve the full range of genetic variation. P. lewtonii reproduces predominantly by selfing or bi-parental inbreeding, but reproduction occurred through each of the three flower types. Some individuals produced by selfing/inbreeding were tightly clustered spatially, and were likely produced either by belowground flowers or by aboveground flowers with limited seed dispersal. Other selfed/inbred individuals were spatially separated (maximum of 15 m), and were likely produced by aboveground flowers followed by seed dispersal by ants. Fine-scale patterns of genetic structure indicate that some gene flow is occurring among aboveground CH flowers but both pollen and outcrossed seeds are moving limited distances (maximum of 0.5 km). Because genetic variation is structured at a fine spatial scale, protecting many populations is necessary to fully conserve the genetic variation in P. lewtonii. Conservation seed banking, if accompanied by research on seed germination requirements, may also contribute to the effective protection of genetic variation in P. lewtonii.     

GENETIC CONSEQUENCES OF OUTCROSSING IN THE CLEISTOGAMOUS ANNUAL,IMPATIENS CAPENSIS. II. OUTCROSSING RATES AND GENOTYPIC CORRELATIONS

The genetic consequences of a plant's mating system depend on both the degree of outcrossing and the genetic relationship between mates. We examined the electrophoretic genotypes of seeds derived from cleistogamous (CL) and chasmogamous (CH) flowers in six populations of the facultatively cleistogamous annual, Impatiens capensis. Multilocus estimates of the outcrossing rates for the strongly protandrous CH flowers ranged from 0.29 to 0.71 and were higher than estimates based on single-locus data. Such results suggest that the CH flowers experience variable levels of both geitonogamous self-fertilization and biparental inbreeding. A new and generally applicable technique based on the relative level of inbreeding within progeny groups provided direct estimates of the correlation between the genotypic values of outcrossed mates. These correlations varied widely among populations and contributed up to half of the inbreeding observed among the CH progeny. Such biparental inbreeding biases estimates of the outcrossing rate based on the mixed-mating model downward and influences mating-system evolution by decreasing the “cost of meiosis.”     

Consequences of a mixed reproductive system in the hog peanut,Amphicarpaea bracteata, (Fabaceae)

Summary The forest annual, Amphicarpaea bracteata L. can reproduce via aerial chasmogamous, aerial cleistogamous, and subterranean cleistogamous flowers. Both plant size and light intensity influenced the utilization of the three modes of reproduction. chasmogamous and aerial cleistogamous flower number and the ratio of chasmogamous flowers to the total number of aerial flowers increased with plant size. The latter demonstrated a shift to xenogamy and outbreeding in larger plants. Light intensity indirectly influenced reproductive modes through its infuence on plant size. Seed set by both types of aerial flowers was low and unrelated to plant size. Subterranean seed number and the total dry weight of subterranean seeds per plant increased with size. The subterranean seeds of Amphicarpaea bracteata are thirty-four times larger than the aerial seeds (fresh weight). Under field conditions, subterranean seeds had greater germination after one year than acrial seeds. The plants arising from subterranean seeds were significantly larger and more fecund than those from aerial seeds. Seeds produced by aerial cleistogamous, hand selfpollinated chasmogamous, and naturally pollinated chasmogamous flowers had equivalent germination rates and produced plants of equal size and fecundity. This suggests that the outbred progeny from chasmogamous flowers have no advantage over the inbred progeny from aerial cleistogamous flowers.     

Cleistogamy in Scutellaria indica (Labiatae): effective mating system and population genetic structure

Scutellaria indica is a perennial herb with both chasmogamous (CH) and cleistogamous (CL) flowers on the same plant in some populations, and only CL flowers in other populations. Actual seed production by CH and CL flowers was investigated in populations of S. indica. The average seed set of CL flowers was 19 times higher than CH flowers, indicating much greater fertilization success. The CL seeds were also significantly heavier than the CH seeds. However, the resource cost of producing a CH flower was much higher than that of producing a CL flower. The CH flower was approximately seven times larger, and its pollen/ovule ratio was approximately five times higher than flowers. The level and pattern of genetic diversity at both allozyme and random amplified polymorphic DNA (RAPD) levels were consistent with a predominantly selfing system in the species. The average amount of within-population genetic variation was extremely low (A = 1.025, P = 2.36%, HO = 0.001 and HE = 0.008 based on allozyme data, and P = 8.94% and HE = 0.03 based on RAPD data). At the species level, the estimates of total gene diversity (HT) were 0. 101 based on allozyme data and 0.139 based on RAPD data. A very high level of genetic differentiation occurred between populations (allozyme GST = 0.92 and RAPD GST = 0.81). Genetic drift coupled with predominant cleistogamous selfing apparently played the major role in determining the population genetic structure in S. indica. Although the features associated with CH and CL flower and seed production seem to be sufficient for the evolution of complete cleistogamy in S. indica, random fixation of alternative alleles for dimorphic or complete cleistogamy in small populations could maintain the multiple strategy of chasmogamous and cleistogamous reproduction in the species.     

Reproductive investment in a cleistogamous morph of <Emphasis Type="Italic">Polygonum jucundum</Emphasis> (Polygonaceae)

Cleistogamy, a breeding system with permanently closed and self-pollinated flowers, is expected to assure reproductive success at a lower cost. Previous studies have inferred the occurrence of cleistogamous flowers in Polygonum, but there are no detailed studies on their reproductive investment compared with that of the chasmogamous flowers in this genus. Here, we studied a cleistogamous morph of P. jucundum to investigate the investment in pollen number, tepal and nectary size. The number of pollen grains per flower was counted with a light microscope. Nectaries and perianths were observed via scanning electron microscope and light microscopy, photographed and measured via ImageJ. The perianths of the cleistogamous flowers, as well as the pollen numbers and nectary sizes, were significantly smaller than those of the chasmogamous flowers. The pollen numbers of the CL flowers were seven times lower than those in the CH flowers. The tepal areas of the CL flowers were, on average, approximately 38% those of the CH flowers. The nectary areas of the CH flowers were almost twice those of the CL flowers. In addition, the nectaries of the cleistogamous flowers were degenerated and inconspicuous, in distinct contrast with the well-developed and conspicuous nectaries of the chasmogamous flowers. Self-fertilization was completed and produced seeds. The cleistogamous P. jucundum, compared with chasmogamous individuals, exhibited lower costs in male function, pollinator attraction and reward structure investment. This cleistogamy appears to be favourable for the plant reproduction under suboptimal conditions.     

FITNESS CONSEQUENCES OF OUTCROSSING IN IMPATIENS CAPENSIS: TESTS OF THE FREQUENCY-DEPENDENT AND SIB-COMPETITION MODELS

Using field and greenhouse experiments, we tested two hypotheses that could account for the maintenance of outcrossing in Impatiens capensis. Seedlings derived from cleistogamous (CL) and chasmogamous (CH) flowers were grown under competitive conditions while flanked by neighbors that were either related or unrelated. In both experiments, CH progeny sometimes expressed more phenotypic variability than CL progeny. In the greenhouse experiment, CH progeny attained the same weight as CL progeny, and the relatedness of neighboring plants did not affect the growth of either type. In the field experiment, CL and CH progeny performed similarly when grown with related competitors. However, CH progeny were somewhat larger when planted with nonsibs, while CL progeny were somewhat smaller under those conditions. Thus, there is no evidence that either frequency-dependent selection or the avoidance of competition among siblings favors the maintenance of outcrossing in this species. We also modeled the relative variability of selfed and outcrossed progeny under several reproductive systems. When females mate with one male (progeny are full sibs), selfed progeny are often more variable than outcrossed progeny. When females engage in both selfing and outcrossing, variation among progeny is frequently maximized at an intermediate selfing rate. The sib-competition mechanism, under a range of genetic models, is not apt to promote outcrossing, since selfed progeny are commonly more variable than outcrossed progeny.     

A herbivory-induced increase in the proportion of floating seeds in an invasive plant

It is important to determine the factors prompting seed dispersal because for plant species seed dispersal is the only opportunity to disperse into a new habitat. Previous studies showed that the maternal stress, such as high density and low nutrient levels, induces the adaptive plastic increase of the dispersal ability in seed heteromorphic plants. In this study, we examined whether herbivory can change the relative proportion of dispersal-related seed heteromorphism (floating or non floating seeds) in an invasive weed Ambrosia artemisiifolia. Because A. artemisiifolia often distributes in the riparian habitat, floating seeds might contribute to the long distance dispersal by hydrochory. Floating ability and seed weight were compared between plants damaged by a specialist herbivore Ophraella communa and undamaged plants. The damaged plants produced lighter and more likely floating seeds than the undamaged plants. However, multi-regression analysis revealed that the probability of floating was affected by seed weight but was not affected by herbivore treatment (damaged vs. undamaged plants). These results suggest that the increased proportion of floating seeds was not a direct response to the herbivore signal but an indirect response through the herbivore's effect on the reduction of seed weight. Plants damaged by herbivores might not only decrease seed production and quality but also increase the dispersal ability. These responses in dispersal ability against the herbivores might contribute to the spread of invasive plants.     

QUANTITATIVE VARIATION OF PROGENY FROM CHASMOGAMOUS AND CLEISTOGAMOUS FLOWERS IN THE GRASS DANTHONIA SPICATA

Progeny from chasmogamous (CH) and cleistogamous (CL) flowers of the grass Danthonia spicata were raised in their native habitat and in the greenhouse in order to determine how genetic variation was distributed among families and between CH and CL progeny within families. Twelve quantitative characters were measured on clones from individuals known to have arisen from either CH or CL flowers on a particular plant. Significant genetic variation existed for all characters measured. Most genetic variation was between families and two morphologically similar groups of families were identified. Relatively little genetic variation was found within families (approximately 5% of the total phenotypic variance). In field-raised plants, variance component analysis suggested that CL progeny were genetically more similar to each other than were CH progeny from the same plant. Levene's test of the average deviation of CH and CL progenies from their group means was nonsignificant but suggested there was a trend (0.05 < P < 0.10) for CH progeny to be more variable than CL progeny in the field but less variable in the greenhouse. The amount and distribution of genetic variation in the study population indicates that selective differentials would be larger among families than within families.