Seed set analysis in three amaranth species (A. cruentus, A. caudatus, and A. lividus) under self- and cross-pollination conditions

Seed set was analyzed in three amaranth species (A. cruentus, A. caudatus, and A. lividus) under conditions of self- and cross-pollination during four vegetative periods in 1995-1998. It was demonstrated, that the populations of these species are polymorphic for the breeding system and that they consist of self-sterile and self-fertile plants. The populations were composed mostly of self-sterile plants and the percentages of self-fertile plants were only 34.7%, 18.7%, and 12.5% for A. cruentus, A. caudatus, and A. lividus respectively. The level of self-fertility was high in the first year of isolation, it sharply decreased during the subsequent years of selfing, due to inbreeding effect. Seed set under self- and cross-pollination is determined by species-specificity of the plants, influence of conditions of their growth, and inbreeding depression. Conditions of growth had the strongest influence on seed set. Index of its influence varied from 0.53 to 0.87 for the cross-pollinated plants and from 0.13 to 0.68 for the self-pollinated ones. Analysis of variance demonstrated a weak effect of the interaction of these factors. Spearman's rank correlation demonstrated a highly significant agreement between the seed set values for the self- and cross-pollinated plants of A. cruentus and A. lividus. The lines derived from the three amaranth species varied widely in the seed set values under both self- and cross-pollination conditions.     

Population differences in self-fertility in the “self-incompatible” milkweed Asclepias exaltata (Asclepiadaceae)

Individual plants of Asclepias exaltata (Asclepiadaceae) typically express an unusual self-incompatibility system under single-gene control. Hand-pollinations performed in six natural populations detected occasional self-fertile plants. The frequency of self-fertile individuals ranged from 0 to 34.0% and differed significantly among populations. Self-fertility appears to be under genetic control, as the ability of most plants (80.0 %) to set fruit following self-pollinations was identical under natural and greenhouse conditions. Seed- and fruit-set, however, were significantly lower from self- vs. cross-pollinations. Allozyme analysis of the population with the highest frequency of self-fertility revealed that adult plants were not significantly inbred. Finally, fruit-set following within-population cross-pollinations did not differ from that following wide, between-population cross-pollinations.     

Pollination biology of Harrisia portoricensis (Cactaceae), an endangered Caribbean species

Traits associated with self-pollination are common features of island plant communities. In this work, we studied the pollination biology and the breeding system of Harrisia portoricensis, an island columnar cactus, to test for the presence of inbreeding and inbreeding depression. For H. portoricensis, which bears flowers with typical outcrossing morphology, the results from 322 h of direct observations and videotaping showed that visits to flowers by animals were uncommon. Controlled pollinations demonstrated that H. portoricensis has a partially self-compatible breeding system that it is not autogamous and thus requires an external mechanism for the movement of pollen to set fruit. We detected differences in seed size, seed mass, germination success, and multiplicative fitness estimates between self- and cross-pollination treatments. We found that progeny resulting from natural and self-pollination treatments showed signs of inbreeding depression compared with progeny resulting from cross-pollination; however, the magnitude of the inbreeding depression was less than 50%. Our combined results suggest that for this species an endogamous breeding system should be favored by natural selection.     

Endogamy costs and reproductive biology of <Emphasis Type="Italic">Laelia autumnalis</Emphasis>, an endemic orchid of Mexico

It has been hypothesized that the fitness of selfed progeny must be lowered by inbreeding depression. Most research into the breeding systems of orchids shows a similar fruit set from outcrossing and self-pollination, but few studies have measured seed production and viability. In five populations of Laelia autumnalis, in central México, we studied the species reproductive system and the early costs of endogamy. We performed spontaneous self-pollination, apomixis, assisted self-pollination, cross-pollination, and exogamous pollination (using pollen from a different population) treatments of bagged flowers and measured fruit set, seed production, and germination. No fruits were produced in the apomixis and spontaneous self-pollination treatments. Reproductive success from self-pollination was lower than that from cross- and exogamous pollination and no difference was found between the latter two treatments. The ratio between self-pollination and cross-pollination success in different traits ranged from 0.80?±?0.18 (fruit set) to 0.22?±?0.13 (seeds with embryo). The latter value suggests a high endogamy costs in the production of viable seeds. The concatenated success of the different traits studied showed that the relative fitness of self-pollination was 63% lower than with cross-pollination. Laelia autumnalis is a self-compatible non-autogamous species in which the cost of endogamy occurs at seed production and cannot be compensated for at other early stages. This also occurs in other orchid species and is likely to be a general pattern.     

Early-inbreeding depression inVaccinium myrtillus andV. vitis-idaea

Summary Fertility parameters were estimated inVaccinium myrtillus andV. vitis-idaea after self- and cross-pollinations performed in growth chamber. We showed a drastic decrease in fertility after self-pollination as compared to cross-pollination. Number of plump seeds per berry was compared with previous field data. In both species, growth room conditions improved plump seed number after cross-pollination but not after self-pollination. In addition, in order to enhance resources supply to young developing fruits, cytokinin application was tested inV. myrtillus but no effect was detected. We hypothesize that the partial self-sterility is due to inbreeding depression based on the expression of recessive lethals.     

The role of a mixed mating system in the reproduction of a Mediterranean subshrub (Fumana hispidula, Cistaceae)

Knowledge about mixed mating systems can improve our understanding of the evolutionary dynamics of reproductive systems. Here we report a study of the pollination system (floral and reproductive biology, pollen limitation and stigmatic pollen load, floral visitors and inbreeding depression) of Fumana hispidula, a Mediterranean subshrub belonging to a species group with a strong selfing tendency. Autonomous self-pollination, hand self- and hand cross-pollination, open pollination and controls yielded fruits (0.28, 0.65, 0.68, 0.38, and 0.67, respectively); however, autonomous self-pollination resulted in the lowest fruit set. All individuals were fully self-compatible but we found great inter-individual variation in autonomous self-pollination ability, which was probably related to a variable expression of herkogamy degree. Inbreeding depression was low (0.040 for fruit set, 0.015 for seed set and ?0.026 for seed mass). The pollen supplementation experiment did not reveal pollen limitation and pollinators were seen visiting the flowers during the observation periods. These results support the idea that F. hispidula has a mixed mating strategy, which represents a successful reproduction mode in their patchy habitats.     

Pollination and self-interference in Nothofagus

Interference between male and female functions within a monoecious plant may hinder crossing and decrease seed set. We assessed the probability of self-pollination and the effect of self-pollination on cross-pollination for two self-incompatible species: Nothofagus obliqua and N. nervosa. The probability of self-pollination was studied by tracking the phenologies of staminate and pistillate flowers, including an analysis of stigmatic receptivity. Pure and mixed pollinations were performed in order to evaluate the effect of self-pollination upon cross-pollination. Phenological observations suggest that self-pollination is highly likely in both species. Compared to pure cross-pollination, the application of self-pollination prior to cross-pollination resulted in lower numbers of germinated pollen grains for both species, and also in a lower production of viable seeds in N. nervosa. The low proportion of viable seeds often observed in natural populations of N. obliqua and N. nervosa may be related to self-pollination.     

Fertility differences among floral morphs following selfing in tristylous Eichhornia paniculata (Pontederiaceae): inbreeding depression or partial incompatibility?

Reduction in seed set following self- vs. cross-pollination in flowering plants can result from abortion of selfed offspring owing to inbreeding depression and/or partial self-incompatibility. Previous studies on tristylous Eichhornia paniculata (Pontederiaceae) indicate that reduced seed set following self-pollination generally occurs in the short- (S), but not the long-(L) or mid-styled (M) morphs. To determine whether this pattern results from morph-specific differences in inbreeding depression owing to the sheltering of deleterious alleles at the S locus and/or partial self-incompatibility, we conducted controlled hand-pollinations of the floral morphs and measured seed set and levels of seed abortion. There were no significant differences in fertilization success and seed set following self-, illegitimate, and legitimate pollinations in the L and M morphs. In contrast, in the S morph self-, intramorph and intermorph illegitimate pollinations resulted in significant reduction in seed set in comparison with legitimate pollination. This indicates that the reduced seed set observed in self-pollination is the result of partial incompatibility rather than inbreeding depression. Significantly reduced fertilization success and low levels of ovule abortion in illegitimate pollinations of S plants also supported this conclusion. Reduced fertility in the S morph may have implications for the observed loss of this morph from natural populations and the evolutionary breakdown of tristyly.     

Variation in style length and anther–stigma distance in Ixiolirion songaricum (Amaryllidaceae)

Variation in style length and anther–stigma distance (ASD) was investigated in 18 populations of Ixiolirion songaricum, an early-spring ephemeral perennial herb in northern Xinjiang. The effect of ASD on autonomous self-pollen deposition and seed set was assessed using bagging experiments. Seed production under autonomous self-pollination, cross-pollination and natural pollination was determined by manual pollination experiments. Stigmas of multiple flowers within an individual were sandwiched between two separate anther levels, and no individuals possessed approach or reverse herkogamy. Style length varied continuously among flowers, resulting in a wide range of variation in ASD. There were fewer flowers with a small ASD than with a large ASD in populations. ASD was negatively correlated with autonomous self-pollen deposition and seed set, but it was positively correlated with natural pollination seed set. Seed production under natural pollination was higher than that under autonomous self-pollination, but it was smaller than that under cross-pollination. Variation in style length and ASD among flowers was not caused by style or stamen elongation in I. songaricum, which differ from other species reported in literature that have continuous variation in style length and ASD.     

Is Eucalyptus Cryptically Self-incompatible?

BACKGROUND AND AIMS: The probability that seeds will be fertilized from self- versus cross-pollen depends strongly on whether plants have self-incompatibility systems, and how these systems influence the fate of pollen tubes. METHODS: In this study of breeding systems in Eucalyptus urophylla and Eucalyptus grandis, epifluorescence microscopy was used to study pollen tube growth in styles following self- and cross-pollinations. KEY RESULTS: Pollen tubes from self-pollen took significantly longer than those from cross-pollen to grow to the base of the style in both E. urophylla (120 h vs. 96 h) and E. grandis (96 h vs. 72 h). In addition, both species exhibited reduced seed yields following self-pollination compared with cross-pollination. CONCLUSIONS: The present observations suggest that, in addition to a late-acting self-incompatibility barrier, cryptic self-incompatibility could be a mechanism responsible for the preferential out-crossing system in these two eucalypt species.     

THE CONSEQUENCES OF SELF-POLLINATION IN ASCLEPIAS EXALTATA,A SELF-INCOMPATIBLE MILKWEED

We investigated the effect of self-pollination on fruit- and seed-set in Asclepias exaltata, a self-incompatible milkweed. A total of 1,380 hand-pollinations were performed on 138 flowers in each of five treatments. In each treatment, two pollinia were placed in adjacent stigmatic chambers, which transmitted pollen tubes to the same ovary. Treatments 1 and 2 involved simultaneous placement of two self-pollinia (treatment 1) or two cross-pollinia (treatment 2) into adjacent stigmatic chambers. No fruits were produced after self-pollinations, but 23% of the cross-pollinated flowers matured fruits. The three remaining experimental pollination treatments were self-pollination preceding cross-pollination by 24 hr (treatment 3), simultaneous self- and cross-pollination (treatment 4), and cross-pollination preceding self-pollination by 24 hr (treatment 5). Compared with that in flowers receiving only cross-pollen, fruit-set in treatments 3–5 was reduced 81%, 49%, and 29%, respectively. Seed-set was also significantly reduced in flowers receiving self-pollen 24 hr in advance of the cross-pollen. Using genetic markers, we observed that only 0.6% of seeds resulted from self-pollination. Our data strongly suggest that self-pollination in milkweeds not only wastes pollen but also greatly reduces the number of ovules and ovaries that might otherwise mature fruits and seeds after cross-pollination.     

The breeding system of Lycium cestroides: a Solanaceae with ovarian self-incompatibility

To determine the breeding system of Lycium cestroides,several treatments were performed: self-, cross-, and geitonogamous pollinations, autonomous self-pollination, and a control (flowers exposed to natural pollinators). Production and quality of fruits and seeds as well as pollen tube growth were evaluated for each treatment. Experimental pollinations indicate that L. cestroides is self-incompatible since fruits were obtained only under cross-, and open-pollination treatments. However, in self- and geitonogamous hand pollinations, as well as in autonomous self-pollination, pollen tubes developed successfully and reached the ovules. The speed of pollen tube growth did not differ significantly among the different hand-pollinated flowers (cross-, self-, and geitonogamous). These facts indicate the presence of an ovarian self-incompatibility system. Significant differences were observed in fruit set, fruit size, and seed number per fruit between cross-pollinated flowers and open pollination (control). These results could be explained in terms of quantity and quality of pollination in each case. Received: 2 November 2000 / Revision accepted: 26 January 2001     

Female reproductive success decreases with display size in monkshood, Aconitum kusnezoffii (Ranunculaceae)

Background and Aims

Reduction in female fitness in large clones can occur as a result of increased geitonogamous self-fertilization and its influence through inbreeding depression. This possibility was investigated in the self-compatible, bee-pollinated perennial herb Aconitum kusnezoffii which varies in clone size.

Methods

Field investigations were conducted on pollinator behaviour, flowering phenology and variation in seed set. The effects of self-pollination following controlled self- and cross-pollination were also examined. Selfing rates of differently sized clones were assessed using allozyme markers.

Key Results

High rates of geitonogamous pollination were associated with large display size. Female fitness at the ramet level decreased with clone size. Fruit and seed set under cross-pollination were significantly higher than those under self-pollination. The pre-dispersal inbreeding depression was estimated as 0·502 based on the difference in seed set per flower between self- and cross-pollinated flowers. Selfing rates of differently sized clones did not differ.

Conclusions

It is concluded that in A. kusnezoffii the negative effects of self-pollination causing reduced female fertility with clone size arise primarily from a strong early-acting inbreeding depression leading to the abortion of selfed embryos prior to seed maturation.Key words: Early-acting inbreeding depression, Aconitum kusnezoffii, clone size, female reproductive success, geitonogamy     

Differential ovule development following self- and cross-pollination: the basis of self-sterility in Narcissus triandrus (Amaryllidaceae)

Self-pollination results in significantly lower seed set than cross-pollination in tristylous Narcissus triandrus. We investigated structural and functional aspects of pollen–pistil interactions and ovule–seed development following cross- and self-pollination to assess the timing and mechanism of self-sterility. Ovule development within an ovary was asynchronous at anthesis. There were no significant differences in pollen tube behavior following cross- vs. self-pollination during the first 6 d of growth, regardless of style morph type. Double fertilization was significantly higher following cross- vs. self-pollination. Aborted embryo development was not detected following either pollination type up to seed maturity. Prior to pollen tube entry, a significantly greater number of ovules ceased to develop following self- vs. cross-pollination. These results indicate that self-sterility in N. triandrus operates prezygotically but does not involve differential pollen tube growth typical of many self-incompatibility (SI) systems. Instead, low seed set following self-pollination is caused by a reduction in ovule availability resulting from embryo sac degeneration. We hypothesize that this is due to the absence of a required stimulus for normal ovule development. If this is correct, current concepts of SI may need to be broadened to include a wider range of pollen–pistil interactions.     

Is self-compatibility the main breeding system in borage (Borago officinalis L.)?

The polygenic postzygotic self-incompatible system reported in borage (Borago officinalis L.) is refuted. The behaviour of pollen tubes after self- and cross-pollination, analysis of the crossability coefficient, seed set after continuous generations of self-pollination and the effects of inbreeding depression in borage were studied. Evidence of self-compatibility was established. The influence of protandry in a self-compatible system is also discussed. Received: 15 July 1999 / Accepted: 3 November 1999     

Polyploidy and polyembryony in Anemopaegma (Bignonieae, Bignoniaceae)

Polyploidy is a key process in plant evolution, with the asexual formation of embryos representing a way through which polyploids can escape sterility. The association between polyploidy and polyembryony is known to occur in Bignoniaceae. In this study, we investigate polyembryony in four polyploid species of Anemopaegma: A. acutifolium, A. arvense, A. glaucum and A. scabriusculum as well as in one diploid species, A. album. Polyembryony was observed only in polyploid species. We used seed dissection and germination tests to compare the number of polyembryonic seeds. We tested how the pollen source influences the number of polyembryonic seeds and the number of embryos per seed and tested the correlation between the number of viable seeds per fruit and mean number of embryos per seed. The number of polyembryonic seeds observed by seed dissection was higher than the number of polyembryonic seeds determined by the germination test, with the number of embryos produced per seed being higher than the number of seedlings. The dissection of seeds of A. glaucum indicated that a higher number of polyembryonic seeds and a higher number of embryos were present in seeds from cross-pollination than in seeds from self-pollination. On the other hand, germination tests indicated that a higher number of polyembryonic seeds were present in fruits from self-pollination than from cross-pollination. The mean number of embryos per seed was not influenced by the number of viable seeds per fruit in fruits from open pollination. These results indicate a positive relationship between polyembryony and polyploidy in Anemopaegma.     

INBREEDING EFFECTS IN CLARKIA TEMBLORIENSIS (ONAGRACEAE) POPULATIONS WITH DIFFERENT NATURAL OUTCROSSING RATES

Inbreeding depression is commonly observed in natural populations. The deleterious effects of forced inbreeding are often thought to be less pronounced in populations with self-pollinating mating systems than in primarily outcrossing populations. We tested this hypothesis by comparing the performance of plants produced by artificial self- and cross-pollination from three populations whose outcrossing rate estimates were 0.03, 0.26, and 0.58. Outcrossing rates and inbreeding coefficients were estimated using isozyme polymorphisms as genetic markers. Analysis of F statistics suggests that biparental inbreeding as well as self-fertilization contribute to the level of homozygosity in the seed crop. Biparental inbreeding will reduce the heterozygosity of progeny produced by outcrossing, relative to random outcrossing expectations, and hence will reduce the effects of outcrossing versus self-fertilization. Heterotic selection may increase the average heterozygosity during the life history. Selfed and outcrossed seeds from all three populations were equally likely to germinate and survive to reproduce. However, inbreeding depression was observed in fecundity traits of plants surviving to reproduction in all three populations. Even in the population whose natural self-fertilization rate was 97%, plants grown from seed produced by self-pollination produced fewer fruits and less total seed weight than plants grown from outcrossed seed. There was no detectable inbreeding depression in estimated lifetime fitness. Inbreeding effects for all reproductive yield characters were most severe in the accession from the most outcrossing population and least severe in the accession from the most self-fertilizing population.     

Effects of pollen shortage and self-pollination on seed production of an endangered tree, Magnolia stellata

BACKGROUND AND AIMS: Pollen limitation is a significant determinant of seed production, and can result from both insufficient pollen quantity (pollen shortage) and quality (mainly relating to self-pollination). For animal-pollinated tree species with large floral displays, pollen limitation may be determined by a balance between increased pollen quantity due to increased attractiveness for pollinators, countered by increased self-pollination due to increased geitonogamy. The contributions of pollen shortage and self-pollination on seed production were quantitatively examined in the natural pollination of an insect-pollinated, dichogamous, endangered tree, Magnolia stellata, which has a large, showy floral display. METHODS: Manual self- and cross-pollinations were conducted to determine the effects of selfing on seed production. The outcrossing rate was measured using microsatellite analyses of open-pollinated seeds, and the embryo mortality rate caused by self-pollination was indirectly estimated. The frequency of ovule mortality due to pollen shortage was also inferred using the embryo mortality and ovule survival rates from natural pollination. KEY RESULTS: The average fruit set, seed set per fruit, and ovule survival rate per tree from hand cross-pollination were 1.37, 3.15, and 3.34 times higher than those from hand self-pollination, respectively, indicating that self-pollination causes inbreeding depression for fruit and seed set. The multilocus-outcrossing rate (t(m)) was intermediate, 0.632, and the primary selfing rate was 0.657. This indicates that frequent geitonogamous selfing occurs. The ovule mortality rate due to pollen shortage and the embryo mortality rate due to self-pollination were estimated to be 80.8 % and 45.9 %, respectively. CONCLUSIONS: It is concluded that seed production of M. stellata is strongly limited by both pollen shortage and self-pollination. Inefficient beetle-pollination and the automimicry system via asynchronous flowering might be responsible for the high level of pollen shortage and frequent geitonogamy. This is despite a large, showy floral display and the dichogamous system of the species.     

Embryonic inbreeding depression varies among populations and by mating system in Witheringia solanacea (Solanaceae)

? Premise of the study: Embryonic inbreeding depression is a key influence on mating system evolution and can be difficult to estimate in self-incompatible species. A pollen chase experiment was used to estimate the magnitude of embryonic inbreeding depression in Costa Rican Witheringia solanacea, a species polymorphic for self-incompatibility (SI). In a pollen chase experiment, bud self-pollinations are followed after anthesis by outcross pollinations, with a comparable pair of outcross pollinations used as a control. Lowered seed set for the self-precedence treatment indicates embryonic inbreeding depression. ? Methods: Embryonic inbreeding depression was assayed for self-compatible (SC) individuals and for SI plants from two populations that differ quantitatively in the onset and enzymatic activity of their SI response. Microsatellite markers were used to assay the selfing rate of a sample of surviving progeny from the prior self-pollination treatment. ? Key results: SC individuals showed no evidence of embryonic inbreeding depression. In SI plants, prior self-pollination reduced seed number by 28-70%, depending on population. Microsatellite genotyping revealed that embryonic inbreeding depression was even more severe than estimated by the phenotypic data: for mature fruits resulting from self-pollination precedence, the majority of the progeny were the result of outcross fertilization. ? Conclusions: Lineage-specific purging of recessive lethals has accompanied the evolution of SC in this species. SI populations show contrasting levels of embryonic inbreeding depression, with nearly complete embryonic lethality upon selfing in the Monteverde population. In the face of high embryonic inbreeding depression, an increase in selfing rate can evidently occur only under severe pollen limitation.     

Restricted seed contribution and clonal dominance in a free-floating aquatic plant <Emphasis Type="Italic">Utricularia australis</Emphasis> R. Br. in southwestern Japan

Utricularia australis R. Br. is an aquatic angiosperm species common in natural and irrigation ponds in temperate regions. This species reproduces both sexually and vegetatively, but in southwestern Japan the occurrence of male-sterile populations, in which plants produce no pollen and propagate only vegetatively, has been recorded. We studied the reproductive contribution of seeds in normal pollen-producing populations using isozyme analyses, a pollination experiment under culture and field observations. Although seedlings obtained from controlled mating indicated segregation of isozyme, polymorphism of the isozyme genotype was detected mainly among populations, but rarely within each pond population. This suggested clonal dominance and rarity of seed or seedling survival in natural populations. In the pollination experiment, the mean seed set ratio in cross-pollination between plants of the same isozyme genotype (7.6%) did not differ significantly from self-pollination (7.6%), but was lower than cross-pollination between plants of different genotypes (45.7%). The low ratio in crossing between the same genotype plants was ascribed to the clonality of the parents. In general, these results corresponded with the low ratios in seed setting observed in natural populations (7.9–13.7%). All the male-sterile populations we surveyed showed the same genotype, thus male sterility in the study area was considered to have the same origin.