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.     

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.     

Comparisons of breeding systems between two sympatric species, Nastanthus spathulatus (Calyceraceae) and Rhodophiala rhodolirion (Amaryllidaceae), in the high Andes of central Chile

Alpine vegetation occurs over a wide range of ecological conditions. Thus, the breeding systems of alpine plants are likely to be diverse and vary from one geographical area to another. The reproductive characteristics of Nastanthus spathulatus (Calyceraceae) and Rhodophiala rhodolirion (Amaryllidaceae), species with contrasting floral morphology, were studied in the high Andes of Chile, which has a Mediterranean-type climate. Natural and supplemental open pollination, and cross pollination and self-pollination trials were carried out in the field. Flower visitors were quantified by field and video observations. Both species had high outcrossing properties, and Nastanthus was strongly self-incompatible. Rhodophiala could form some seed by self-pollination, but fruit and seed sets were much lower after self-pollination compared with outcrossing. The phenology and flower/inflorescence forms of these species supported the view that alpine flowers are comparatively long lived and that the floral display contributes to a large proportion of the plant biomass. Rhodophiala was well attended by a native bee species ( Megachile sauleyi ) that was appropriately sized for efficient pollination. Although no flower visitors were observed on Nastanthus and wind pollination was discarded experimentally, a high proportion of the flowers produced seeds under natural pollination. Therefore, the seed set was not severely pollen limited in these species. Including previously published information, breeding systems are now known for 12 species on this Mediterranean alpine site and current knowledge suggests an emphasis on outcrossing breeding systems.     

Effects of floral display size and biparental inbreeding on outcrossing rates in Delphinium barbeyi (Ranunculaceae)

Floral display size represents a tradeoff between the benefits of increased pollinator visitation and the quantity of pollen received vs. the costs of increased self-pollination and reduced pollination quality. Plants with large floral displays often are more attractive to pollinators, but pollinators visit more flowers per plant. Intraplant foraging movements should increase self-pollination through geitonogamy, lowering outcrossing rates in large plants. Local genetic structure should also increase inbreeding and decrease outcrossing estimates, if pollinators move between neighboring, related plants. These predictions were tested in a population of larkspurs (Delphinium barbeyi) in Colorado. Allozymes were used to estimate outcrossing rates of plants varying in display size. Floral displays varied widely (2-1400 flowers; 1-26 inflorescences per plant), and outcrossing rate decreased significantly with increasing display size. Large, multistalked plants self over twice as frequently as single-stalked plants (46 vs. 21%). Local population structure is significant, and biparental inbreeding depresses outcrossing in plants surrounded by genetically similar neighbors. Protandry, coupled with stereotypical bottom-up pollinator foraging, reduces self-fertilization by autogamy or geitonogamy within inflorescences. Selfing is predominantly (>60%) by geitonogamy between inflorescences in large plants. Geitonogamy may be a significant cost to plants with large floral displays if inbreeding depression and/or pollen and ovule discounting results. If so, floral display size, particularly inflorescence number, may be under contrasting selection for pollination quantity vs. quality.     

Conservation implications of the reproductive ecology of Agalinis acuta (Scrophulariaceae)

Reproductive ecology of Agalinis acuta was investigated by examining potential for self-fertilization before and at anthesis, reproductive output from outcrossed vs. selfed matings, and effects of browsing, plant size, and conspecific plant density on seed and fruit production. These features of a plant species can provide indirect information pertinent to conservation such as patterns and maintenance of genetic diversity, risk associated with inbreeding depression, and changes in pollinator abundance or effectiveness. The species is self-compatible, with 97% of selfed flowers setting fruit; pollinators were not required for reproduction. However, seed set in self-pollinated fruits averaged 17-20% less than that in open-pollinated fruits. Geitonogamous and facilitated selfing are possible throughout anthesis and autonomous selfing is possible late in anthesis as corollas abscise. Delaying self-pollination until after outcrossing opportunities likely limits selfing rates and thus reduces risks associated with inbreeding but allows reproduction in absence of pollinators. Supplementing pollen on open-pollinated flowers yielded no additional seed set over controls. Neither early-season browsing of primary stems nor conspecific plant density had significant effects on number of fruits per plant, on fruit size, or on number of seeds from open-pollinated flowers. Currently, reproduction appears to be high (about 2400 seeds/plant), and future risks due to lack of genetic diversity are likely low.     

Inbreeding, outbreeding, and heterosis in the yellow pitcher plant, Sarracenia flava (Sarraceniaceae), in Virginia

The yellow pitcher plant, Sarracenia flava, is an insectivorous plant restricted to fire-maintained wetland ecosystems in southeastern Virginia. Only four natural sites remain in the state totaling fewer than 100 clumps. Plants from sites located in Dinwiddie, Greensville, Prince George, Sussex counties, and the city of Suffolk were tested for the effects of self-pollination, intrasite outcrossing, and intersite outcrossing on offspring quantity (total seed number and total seed mass) and offspring quality (avarage seed mass, germination, and growth).Self-pollination resulted in significantly lower offspring quantity and quality. Total seed number and total seed mass for self-pollinated capsules were approximately one-fourth that of outcrossed capsules. Germination, survivorship, and growth over 5 yr were also significantly lower for offspring from self-pollinated capsules. Together, these results suggest strong inbreeding depression in this species.Relative to offspring from intrasite crosses, offspring from intersite crosses were significantly larger after 5 yr of growth. This suggests that restoration efforts for Virginia S. flava will be most successful when plants from multiple sites are used.     

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.     

Effects of pollen quantity and quality on reproduction and offspring vigor in the rare plant Scorzonera humilis (Asteraceae)

We studied the effects of pollinator exclusion, interparental distance, and supplementary hand pollination on reproduction and progeny vigor in Scorzonera humilis (Asteraceae), a rare plant of fragmented, nutrient-poor grasslands. Caged flowers produced no seeds and selfed flowers only very rarely, indicating that S. humilis is mainly self-incompatible. Seed production, seed mass, and seed germination following between-population crosses were consistently, but not significantly, higher than after within-population crosses. Seed set increased with local density of conspecifics, indicating that the reduced plant density in fragmented populations may reduce plant reproductive success. Seed set was pollen limited in all four populations studied. Supplementary hand-pollination strongly increased the survival of offspring, indicating that either pollinators transferred pollen from related individuals resulting in inbreeding depression in spite of the incompatibility system or that higher pollen loads increased pollen competition and the selectivity among gametes. In one of the populations, adding pollen from a different population strongly increased progeny fitness compared with both natural pollination and pollen supplementation from the same population. The results indicate that S. humilis is sensitive to inbreeding and that pollen limitation can reduce both the number and quality of offspring.     

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.     

Autogamy in Boronia falcifolia: consequences for inbreeding and persistence in fragmented coastal heaths

Coastal heath is becoming increasingly fragmented, resulting in reduced pollinator abundance affecting species viability. The reproductive ecology and inbreeding of a common coastal heath species, Boronia falcifolia (Rutaceae), was investigated. Pollination observations and experiments were carried out at one site, and inbreeding was assessed using genetic markers at eight populations. Four pollination treatments were assessed: cross-pollination, self-pollination, autogamous pollination and open (insect) pollination. Boronia falcifolia was found to be highly self-compatible, frequently setting seeds from autogamous pollination. There were no significant differences between the autogamous, self and cross pollen treatments for any measures of reproductive success (pollen tube growth, fruit set, swollen locules and seed production). Insect activity resulted in fewer seeds per fruit than hand cross pollination, although pollinator visits were few. All populations studied were highly inbred. Autogamous self-pollination is widespread in B. falcifolia and this combined with its ability to vegetatively regenerate enhances its potential for survival when pollinators are few.     

Reproductive biology of the invasive exotic shrub, Rhododendron ponticum L. (Ericaceae)

The reproductive biology of an exotic species will affect its ability to become naturalized and invasive in non-native habitats. Rhododendron ponticum is an ecologically damaging exotic weed in the British Isles, which spreads predominantly by seed. I investigated how inbreeding and outcrossing affect seed production and germination in a wild population of this species in Ireland. Experimental manipulations revealed low fruit and seed set when insects were excluded from flowers, suggesting that this species has limited capability for spontaneous autogamy. Hand-pollination treatments showed that, although flowers are self-compatible (with self and same plant pollen), higher levels of seed set occur following outcrossing (xenogamy). There was no significant difference in rate of germination of seeds from inbred or outcrossed treatments. The addition of xenogamous pollen to open flowers did not increase fruit or seed set, suggesting that flowers in this population are not pollen limited: native generalist pollinators, mainly bumblebees ( Bombus spp.), are providing an adequate pollinator service. This work demonstrates that outcrossing increases seed set and pollinators are required to facilitate this. Hence, generalist native pollinators can promote invasion by exotic plants. Native pollinators can clearly play an important part in alien species invasion.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 155 , 373–381.     

Effect of pollen load size and source (self, outcross) on seed and fruit production in highbush blueberry cv. 'Bluecrop' (VACCINIUM CORYMBOSUM; Ericaceae)

Reproductive fitness of a plant is ultimately determined by both number and quality of seed offspring. This is determined by sexual selection of pollen microspores and ovules during pollination and fertilization. These processes may include pollen competition and seed abortion, which reduce the number of microspores and ovules available for final seed production. Thus, even an excess of pollen microspores to ovules does not result in fertile seeds equal to ovule number. We investigated pollen requirements of highbush blueberry (Vaccinium corymbosum cultivar 'Bluecrop') for maximal seed production and how fertile seed number translates into fruit quality, since fruit quality would ultimately determine the dispersal of its offspring. We demonstrate that individual blueberry flowers with a mean of 106 ovules reach their maximum fruit set and mass and minimum time to ripen when 125 outcross pollen tetrads pollinate a flower, compared to 10 or 25. Three hundred tetrads resulted in the increase of fertile seeds, but did not result in a further increase of fruit mass or fruit set, or decrease in time to ripen. We also examined the effect of pure and mixed loads of self and outcross pollen (25 and 125 tetrads), and found no differences in fertile seed number, fruit mass, or percentage fruit set when pollen loads were either 25 self or outcross pollen tetrads, although number of days to ripen was significantly shorter by 8 d with 25 outcross tetrads. When the pollen load of 125 tetrads consisted of self or a 50:50 mixture of self and outcross pollen, fruit mass, days to ripen, and percentage fruit set were not different from loads of 125 outcross pollen. In addition, a pollen load of 25 outcross tetrads resulted in fertile seed number and fruit quality in between that of 25 self, and 125 self, 125 mixed, or 125 outcross tetrads. Large, small, and flat seed types were identified, and only large seeds (length = 1.7 mm) were fertile. These results improve our understanding of pollen load size and source requirements of a crop plant and the limits to pollen transfer when translated to fruit growth.     

A clarification of pollen discounting and its joint effects with inbreeding depression on mating system evolution

Given the predominance of outcrossing by angiosperms, large costs must often overwhelm the genetic benefit of selfing derived from contributing two haploid genomes to each off-spring rather than one. In addition to the well-studied genetic cost of inbreeding depression, selfing imposes a mating cost whenever self-pollination reduces opportunities for pollen export. Because self-pollination is a heterogeneous process, pollen discounting and its evolutionary consequences vary with pollination conditions. In this article we model self-pollination as comprising discounting and nondiscounting components, and we consider the consequences of this heterogeneity for outcross siring success. Aided by this depiction of pollination, we then compare previous theoretical representations of pollen discounting and consider their relative virtues. Finally, we consider conditions that would allow a population to be invaded by a variant with different pollination characteristics. This analysis exposes the pollination conditions implicit in standard results of mating system theory. It also identifies associations between four possible changes in pollination expected in different reproductive environments, including the incidence of positive or negative correlations between self-pollination and pollen export. These results emphasize the benefits of expanding the theory of plant reproduction to recognize explicitly when and how pollination mechanisms affect mating outcomes.     

Do consecutive flower visits within a crown diminish fruit set in mass‐flowering Hancornia speciosa (Apocynaceae)?

Hancornia speciosa is a self-incompatible, mass-flowering, sphingophilous fruit crop (mangaba) of northeast and central Brazil. The flowers have a precise pollination apparatus, which optimizes pollen transfer between flower and pollinator. While the pollination mechanism avoids self-pollination, mass-flowering promotes geitonogamy. During a flower visit, almost half of the exogenous pollen grains adhering to the proboscis are deposited on the stigma surface. A pollination experiment with a nylon thread simulating six consecutive flower visits within a crown revealed that only the first two flowers visited (positions 1 and 2) are highly likely to set fruit. Super-production of flowers, and consequently obligate low fruit set, seem to be part of the reproductive strategy of the obligate outcrossing plant, Hancornia speciosa.     

华北驼绒藜自然种群结实的花粉和资源限制

运用人工授粉、补充无机营养、去叶处理以及疏花处理研究了华北驼绒藜自然种群花粉和资源有效性对结实的影响,并进一步探讨了该植物的选择性败育现象和繁殖对策。结果表明:花粉来源而不是花粉数量对华北驼绒藜结实存在显著影响;同枝授粉结籽率和结实率均低于自然授粉,异株授粉则显著高于同枝授粉和自然授粉;异株授粉种子千粒重显著高于自然授粉和同株异枝授粉,极显著高于同枝授粉。华北驼绒藜结实存在资源限制,补充施肥提高了单枝开花数和结籽率。随摘除叶片数量的增加,结籽率明显降低,摘除叶片处理与对照之间均形成显著差异,摘除1/2叶、3/4叶以及去全叶处理使种子千粒重显著低于对照。人工疏花实验表明华北驼绒藜存在选择性败育现象。华北驼绒藜可根据花粉和资源的可利用性来调整性分配、授粉方式和结实以获得最大适合度,具有复杂的繁殖对策,表现出较好的环境适应性。     

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     

Mating system,outcrossing distance effects and pollen availability in the wind-pollinated treeline species Polylepis australis BITT. (Rosaceae)

Fragmentation may negatively affect plant fitness through pollen limitation and increased levels of inbreeding. Effects of fragmentation may vary with regard to life form and breeding system, and few studies exist for wind-pollinated trees. We examined the effects of hand-selfing, varying outcrossing distances and pollen addition on seed mass and germination rate of Polylepis australis BITT. (Rosaceae), a wind-pollinated treeline species endemic to Argentina. We also investigated pollen germination on the stigma and pollen tube growth to determine compatibility resulting from selfing and outcrossing. Selfing reduced seed germination rates with significant differences between open pollination and outcrosses at 30 km. In addition, we found a tendency for pollen germination and pollen tube growth to decrease following selfing. Between-fragment crosses resulted in a trend of higher reproductive output than within-fragment crosses, whereas values were similar between open pollination and between-fragment crosses. Pollen addition did not increase reproductive success neither in small nor in larger fragments. Our results suggest that highly isolated P. australis forests have a potential for inbreeding depression through selfing and within-fragment crosses. However, the results also indicate that pollen flow between P. australis forest fragments is still effective at the current fragmentation level, counteracting negative effects on seed quality resulting from reproductive isolation.     

Dose-response relationships between pollination and fruiting refine pollinator comparisons for cranberry (Vaccinium macrocarpon [Ericaceae])

Comparisons of pollinator efficacy using pollen received on stigmas can be refined by incorporating experimental dose-response relationships for pollen deposition and fruiting responses. A range of discrete pollen doses applied to cranberry stigmas resulted in decelerating curvilinear responses for fruiting, berry size, and seed set. Minimum thresholds and maximum asymptotes bounded reproductive responses to incremental stigmatic pollen loads. Four bee species were compared for their pollination efficacies on commercial cranberries, using counts of pollen received by stigmas during single bee visits to previously virgin flowers. Differences between these bee species were found to be exaggerated when raw pollen counts were used for comparison because foragers of some species often delivered pollen in excess of that needed to maximize fruit and seed production. Sixfold differences between species in mean pollen deposition translated into 1.5-2-fold differences in predicted cranberry fruit set and size. Implications for pollen tube competition and agricultural production are discussed.     

交配距离与父本数量对野慈姑有性繁殖的影响

植物的有性繁殖是生活史和进化的核心, 母本及环境对繁殖的影响固然重要, 父本的贡献亦不容忽视。父本来源与多样性对坐果结籽和后代质量的影响明显, 但由于不同物种或种群的繁殖特点和进化历程, 往往也会呈现其独特性。该研究旨在探究挺水植物野慈姑(Sagittaria trifolia)是否存在自交或远交衰退, 以及父本数量对其繁殖和后代表现的影响。通过人工控制授粉, 设置自交、近距离异交(<50 km)、远距离异交(＞200 km) 3种交配距离以及单、双两类父本数量, 共计5种授粉处理, 观测野慈姑坐果概率、单果种子数量、种子面积、萌发率(2018和2019年)、幼苗芽长(2018和2019年)共7项指标。结果显示: 不同距离的交配对野慈姑的坐果概率、单果种子数量、种子面积、萌发率、幼苗芽长没有显著影响, 野慈姑未出现明显的自交衰退或远交衰退现象。父本数量的增加对野慈姑的结实数量(坐果概率、单果种子数量、种子面积)无影响, 但结实质量方面, 双父本处理的种子萌发率略高于单父本处理。综上所述, 交配距离与父本数量对野慈姑的繁殖表现影响较小, 这可能与其频繁的自交历史、远距离的基因扩散以及高度相似的水生环境有关; 同时, 该研究基于20余个自然种群的基因型, 研究结果也表明了野慈姑物种水平有性繁殖的优异和稳定性。     

Pollen dispersal in Yucca filamentosa (Agavaceae): the paradox of self-pollination behavior by Tegeticula yuccasella (Prodoxidae)

We investigated pollen dispersal in an obligate pollination mutualism between Yucca filamentosa and Tegeticula yuccasella. Yucca moths are the only documented pollinator of yuccas, and moth larvae feed solely on developing yucca seeds. The quality of pollination by a female moth affects larval survival because flowers receiving small amounts of pollen or self-pollen have a high abscission probability, and larvae die in abscised flowers. We tested the prediction that yucca moths primarily perform outcross pollinations by using fluorescent dye to track pollen dispersal in five populations of Y. filamentosa. Dye transfers within plants were common in all populations (mean ± 1 SE, 55 ± 3.0%), indicating that moths frequently deposit self-pollen. Distance of dye transfers ranged from 0 to 50 m, and the mean number of flowering plants between the pollen donor and recipient was 5 (median = 0), suggesting that most pollen was transferred among near neighbors. A multilocus genetic estimate of outcrossing based on seedlings matured from open-pollinated fruits at one site was 94 ± 6% (mean ± 1 SD). We discuss why moths frequently deposit self-pollen to the detriment of their offspring and compare the yucca-yucca moth interaction with other obligate pollinator mutualisms in which neither pollinator nor plant benefit from self-pollination.