Flower color change accelerated by bee pollination in Tibouchina (Melastomataceae)

Floral color changes are common among Melastomataceae and have been interpreted as a warning mechanism for bees to avoid old flowers, albeit increasing long-distance flower display. Here the reproductive systems of Tibouchina pulchra and T. sellowiana were investigated by controlled pollinations. Their pollinators were identified, and experiments on floral color and fragrance changes were conduced to verify if those changes affect the floral visitation. Both Tibouchina species are self compatible. The flowers lasted three days or more, and the floral color changed from white in the 1st day to pink in the following days. Pollen deposition on stigma induced floral color change. The effectiveness of the pollination is dependent on bees’ size; only large bees were regarded as effective pollinators. In experimental tests, the bees in T. pulchra preferred the natural white flowers while the visitors of T. sellowiana were attracted by both natural and mimetic 1st-day flowers (2nd-day flowers with experimentally attached 1st-day flower petals). During the experiments on floral fragrance, the bees visited both natural and mimetic 1st-day flowers (2nd-day flowers with 1st-day flower scents). In both experiments, the bees avoided natural 2nd-day flowers, but seldom visited modified 2nd-day flowers. The attractiveness of T. pulchra and T. sellowiana flowers cannot be attributed exclusively to the color or the fragrance separately, both factors seemingly act together.     

Effects of pollination intensity on Lesquerella fendleri seed set: variation among plants

 The amount of pollen arriving on a flower can be an important determinant of seed production. I investigated the effect of varying pollen loads on seed set of the perennial desert mustard Lesquerella fendleri. To do this, I quantified the dose response relationship between stigmatic pollen load and seed set per fruit using over 400 flowers from 13 greenhouse-grown plants. Seed set per fruit generally increased with pollen up to about 100 pollen grains, then reached a plateau. A negative exponential regression of seed set on pollen load for the pooled data explained less than 10% of the observed variation in seeds per fruit. However, accounting for variation among individual plants in the dose-response relationship increased explained variation to 40%, indicating that plants responded differently to the same amount of available pollen. Plants varied little in the initial slope of the dose-response curve, but differed substantially in the asymptote, which ranged from 3 to 16 seeds. This limit is not imposed by ovule number, and may instead result from variation among plants in vigor, propensity to abort seeds, or in gender specialization. Such variation among plants in dose-response relationships has important consequences for understanding pollination limitation and pollen competition. Received: 29 January 1996 / Accepted: 6 August 1996     

Influence of wind direction on pollen concentration in the atmosphere

The daily pollen concentration in the atmosphere of Badajoz (SW Spain) was analysed over a 6-year period (1993–1998) using a volumetric aerobiological trap. The results for the main pollination period are compared with the number of hours of wind each day in the four quadrants: 1 (NE), 2 (SE), 3 (SW) and 4 (NW). The pollen source distribution allowed 16 pollen types to be analysed as a function of their distribution in the four quadrants with respect to the location of the trap. Four of them correspond to species growing in an irrigated farmland environment (Amaranthaceae-Chenopodiaceae, Plantago, Scirpus, and Typha), five to riparian and woodland species (Salix, Fraxinus, Alnus, Populus, and Eucalyptus), four to urban ornamentals (Ulmus, Arecaceae, Cupressaceae, and Casuarina), and three which include the most frequent pollen grains of widely distributed species (Poaceae, Quercus, and Olea). The results show that the distribution of the sources and the wind direction play a very major role in determining the pollen concentration in the atmosphere when these sources are located in certain quadrants, and that the widely distributed pollen sources show no relationship with wind direction. In some years the values of the correlations were not maintained, which leads one to presume that, in order to draw significant conclusions and establish clear patterns of the influence of wind direction, a continuous and more prolonged study will be required. Received: 6 May 1999 / Revised: 30 March 2000 / Accepted: 31 March 2000     

A test of the effect of floral color change on pollination effectiveness using artificial inflorescences visited by bumblebees

Floral color change has been recognized as a pollination strategy, but its relative effectiveness has been evaluated insufficiently with respect to other floral traits. In this study, effects of floral color change on the visitation pattern of bumblebees were empirically assessed using artificial flowers. Four inflorescence types were postulated as strategies of flowering behavior: type 1 has no retention of old flowers, resulting in a small display size; type 2 retains old flowers without nectar production; type 3 retains old flowers with nectar; and type 4 retains color-changed old flowers without nectar. Effects of these treatments varied depending on both the total display size (single versus multiple inflorescences) and the pattern of flower-opening. In the single inflorescence experiment, a large floral display due to the retention of old flowers (types 2–4) enhanced pollinator attraction, and the number of flower visits per stay decreased with color change (type 4), suggesting a decrease in geitonogamous pollination. Type-4 plants also reduced the foraging time of bees in comparison with type-2 plants. In the multiple inflorescence experiment, the retention of old flowers did not contribute to pollinator attraction. When flowering occurred sequentially within inflorescences, type-4 plants successfully decreased the number of visits and the foraging time in comparison with type-2 plants. In contrast, floral color change did not influence the number of visits, and it extended the foraging time when flowering occurred simultaneously within inflorescences but the opening of inflorescences progressed sequentially within a plant. Therefore, the effectiveness of floral color change is highly susceptible to the display size and flowering pattern within plants, and this may limit the versatility of the color change strategy in nature.     

Leaf damage by herbivores affects attractiveness to pollinators in wild radish, Raphanus raphanistrum

 We carried out two experiments to determine the effect of leaf damage on plant attractiveness to pollinators using wild radish, Raphanus raphanistrum (Brassicaceae), a self-incompatible annual herb. Pairs of plants from 36 full-sib families were grown in pots in the greenhouse. One member of each pair was damaged by Pieris rapae larvae that were allowed to remove half of the leaf area of each of the first four rosette leaves. The plants were subsequently taken out for pollinator observations once a week from the beginning of flowering in late June until the end of August. We conducted two experiments to examine how foliar damage affected visitation by pollinators. In the first experiment, numbers of pollinators visiting plants were compared between damaged and control sibling plants. In the second experiment, the number of open flowers during observations was controlled to be the same for both damaged and undamaged sibs. Damage significantly decreased the number and size of flowers during the first observations in late June. Damaged plants received fewer visits by native bees during the first week of observations. Since damage did not affect native bee visits when the number of open flowers was equalized between treatments, flower number was probably the main cue attracting native bees to plants. In the experiment without flower number control, syrphid flies, the other abundant pollinator taxon, spent more time per flower on the undamaged than on the damaged plants. When flower number was controlled, flies probed significantly more flowers during each visit on the undamaged than on the damaged plants and had higher visitation rates to undamaged plants early in the season. Since syrphid flies preferred undamaged plants both with and without flower number control, they apparently used cues apart from flower number for visitation. The difference between undamaged and damaged plants in floral characteristics and pollinator visitation vanished within a few weeks after the start of flowering. This result suggests that early damage may not have a strong fitness effect through reduction in mating success. However, poor weather conditions can cause early mortality of plants in the field, and nutrient depletion and competition decrease fruit set of later flowers. Therefore, conditions exist under which visitation to early flowers may affect plant fitness. Received: 30 July 1996 / Accepted: 10 February 1997     

Expression of a polygalacturonase enzyme in germinating pollen of <Emphasis Type="Italic">Brassica napus</Emphasis>

Penetration of pollen tubes through stigmatic tissues in Brassica napus L. may involve the release of cell wall modifying enzymes from the pollen tube tip. We examined the expression of a pectin-degrading polygalacturonase (PG) enzyme in unpollinated and early and late pollinated stigmas via immunoblotting and immuno-light microscopy using a PG polyclonal antibody. Immunoblotting analysis indicated that PG enzyme was present at low levels in unpollinated stigmas and at high levels in pollinated stigmas. The level of PG did not detectably increase between early and late pollinated stigmas. Immuno-light microscopy demonstrated that PG enzyme was present in ungerminated pollen grains, stigmatic papillae and in the tip of pollen tubes growing into the papillar wall. This latter evidence suggests that PG enzyme may play an important role in papillar cell wall penetration during pollination although other interpretations of the role of pollen PG should not be discounted. Received: 9 November 2000 / Accepted: 7 December 2000     

An exception to Darwin's syndrome: floral position,protogyny, and insect visitation in Besseya bullii (Scrophulariaceae)

Darwin pointed out that plants with vertical inflorescences are likely to be outcrossed if the inflorescence is acropetalous (flowers from the bottom up), the flowers are protandrous (pollen is dispersed before stigmas are receptive), and pollinators move upward on the inflorescence. This syndrome is common in species pollinated by bees and flies, and very few exceptions are known. We investigated flowering phenology and pollinator behavior in Besseya bullii (Scrophulariaceae) and found that it did not fit Darwin's syndrome. The vertical inflorescence was acropetalous but the flowers were distinctly protogynous, so flowers with newly receptive stigmas appeared on the inflorescence above those with dehiscing anthers. A number of small insects visited B. bullii; bees in the family Halictidae (Augochlorella striata and Dialictus spp.) were most common. When insects moved between gender phases within inflorescences, they moved up more often than down (61% versus 39% of observations, respectively) but this difference was only marginally significant. Most visits were to male-phase flowers only, and this preference was more pronounced for pollen-foraging insects than for nectar-foraging insects. B. bullii was self-compatible, so its flowering characteristics potentially could result in considerable self-pollination. However, an average of 38% of the lowermost flowers opened before any pollen was available on the same inflorescence; these solo females had a high probability of outcrossing (though fruit set was relatively low in the bottom portion of the inflorescence). Upper flowers may also be outcrossed because downward insect movement was not uncommon. Therefore protogyny in B. bullii may not necessarily lead to more selfing than would protandry.     

Cytogenetic analysis of interspecific sunflower hybrids and molecular evaluation of their progeny

Meiotic cells of transgenic asymmetric somatic hybrid (ASH) plants obtained by fusion of microprotoplasts of the donor species Helianthus giganteus or Helianthus maximiliani and recipient protoplasts of Helianthus annuus were investigated. Over 85% of the ASH meiocytes showed regular bivalent chromosome pairing; however, several anomalies like anaphase bridges, laggard chromosomes, univalent and multivalent pairing were also observed. Pollen viability of the ASH plants ranged from 79.2 to 95% with a strong negative correlation to chromosome number which varied between 34 and 42. Molecular investigation of ASH progeny using RAPD markers revealed the presence of donor genotype markers in 68% of the offspring. These results suggest that asymmetric somatic hybridization offers an efficient alternative method to overcome sexual barriers for gene flow and the genetic improvement of H. annuus by introgression of economical important traits from wild Helianthus species. Received: 16 July 2000 / Accepted: 25 September 2000     

Geographic and population variation in pollinating seed-consuming interactions between senita cacti (Lophocereus schottii) and senita moths (Upiga virescens)

Interspecific interactions can vary within and among populations and geographic locations. This variation can subsequently influence the evolution and coevolution of species interactions. We investigated population and geographic variation in traits important to pollinating seed-consuming interactions between the senita cactus (Lophocereus schottii) and its obligate pollinating moth (Upiga virescens), both of which are geographically restricted to the Sonoran Desert. Female moths actively pollinate senita flowers and oviposit onto flowers. Their larvae consume developing seeds and fruit of flowers pollinated by females. Traits important to this interaction include fruit set from moth pollination, fruit survivorship, and costs of fruit consumption by larvae. We studied these traits for five populations at two widely separated geographic locations. On average, 37% of flowers set fruit, 22% of flowers produced mature fruit, and larvae consumed 25% of immature fruit pollinated by female senita moths. Senita cactus and senita moth interactions were strongly mutualistic in all populations that we studied. Although one population had statistically lower fruit set and fruit production than the other four, all five populations were qualitatively similar in fruit production, costs, and patterns of fruit survivorship. Hand-pollination experiments suggested that fruit set was resource-limited in all but this one population. Apparent pollen limitation in the one population explains the quantitative differences in fruit set and fruit survivorship among the populations. As predicted by theory and exemplified by the senita mutualism, specialized and/or obligate interactions vary little among populations and geographic locations. Received: 5 March 1999 / Accepted: 25 June 1999     

Effects of habitat isolation on pollinator communities and seed set

Destruction and fragmentation of natural habitats is the major reason for the decreasing biodiversity in the agricultural landscape. Loss of populations may negatively affect biotic interactions and ecosystem stability. Here we tested the hypothesis that habitat fragmentation affects bee populations and thereby disrupts plant-pollinator interactions. We experimentally established small ”habitat islands” of two self-incompatible, annual crucifers on eight calcareous grasslands and in the intensively managed agricultural landscape at increasing distances (up to 1000 m) from these species-rich grasslands to measure effects of isolation on both pollinator guilds and seed set, independently from patch size and density, resource availability and genetic erosion of plant populations. Each habitat island consisted of four pots each with one plant of mustard (Sinapis arvensis) and radish (Raphanus sativus). Increasing isolation of the small habitat islands resulted in both decreased abundance and species richness of flower-visiting bees (Hymenoptera: Apoidea). Mean body size of flower-visiting wild bees was larger on isolated than on nonisolated habitat islands emphasizing the positive correlation of body size and foraging distance. Abundance of flower-visiting honeybees depended on the distance from the nearest apiary. Abundance of other flower visitors such as hover flies did not change with increasing isolation. Number of seeds per fruit and per plant decreased significantly with increasing distance from the nearest grassland for both mustard and radish. Mean seed set per plant was halved at a distance of approximately 1000 m for mustard and at 250 m for radish. In accordance with expectations, seed set per plant was positively correlated with the number of flower-visiting bees. We found no evidence for resource limitation in the case of mustard and only marginal effects for radish. We conclude that habitat connectivity is essential to maintain not only abundant and diverse bee communities, but also plant-pollinator interactions in economically important crops and endangered wild plants. Received: 7 May 1999 / Accepted: 19 July 1999     

Floral biology of the dioecious species Laurus nobilis L. (Lauraceae)

The present study examines the cytological, physiological, chemical and ecological characteristics of pollen and nectar offered by male and female flowers of the dioecious plant Laurus nobilis. The various phases of floral phenology and the insect pollinators were observed. We used cytological methods to determine anther, pollen and nectary structure. Nectar sugar composition was evaluated by HPLC. Pollen viability in time was compared with cytoplasmic and intine water content. Pollen presentation was found to be reversible by opening and closing of anther valves, determined by hydration of the mechanical layer of the anther. Pollen, covered by pollenkitt, was presented for dispersal for 3 consecutive days and during this time the intine and cytoplasm lost water and pollen viability diminished. At germination exine ruptured together with the outermost layer of the intine. Nectaries of male flowers were observed on the anther filament and on staminodes of female flowers. The nectar consisted almost entirely of sucrose and was more concentrated in male flowers. Secreted through stomata, nectar was presented in a thin layer. In the study area, the main pollinators (about half the total number of all visits) were hymenopterans. Pollen is of the recalcitrant type due to its high water content (>30%) but its viability is long-lasting because the intine is thick and stores water, keeping the cytoplasm of the vegetative cell hydrated and viable, and because anther valves may close under adverse conditions, protecting the pollen. Insects are attracted by male and female flowers similarly, males offer nectar and pollen, whilst females only nectar.     

Floral rewards and pollination in Cytiseae (Fabaceae)

 Cytiseae have been reported to be mostly nectar-lacking, yet some taxa secrete nectar from extrastaminal nectaries. We studied the pollination biology of four shrubby species of Cytiseae (Cytisophyllum sessilifolium (L.) Lang, Spartium junceum L., Genista radiata (L.) Scop., Genista cilentina Valsecchi) which differ for ecology, distribution and population size. All species resulted obliged xenogamous, insect visits being necessary for successful pollination. Bumblebees and solitary bees are the principal pollinators, but also many beetles, some hover-flies, and few bugs visit flowers. Pollinator specificity is low, and this may be the reason of the scarce seed set compared to the number of ovules. Pollen is the main reward, but traces of glucose were detected in all species, at the base of vexillum or on the reproductive column. Nectar production is irregular in time, and apparently unpredictable. We suppose that nectar may play a role in attracting pollinators determining their right position for a successful pollination. Received August 12, 2002; accepted November 25, 2002 Published online: June 2, 2003     

Endothermy by flowers of Rhizanthes lowii (Rafflesiaceae)

Rhizanthes lowii (Beccari) Harms (Rafflesia- ceae) is a parasitic plant that grows in the understory of the rainforest in South-East Asia. This plant does not have leaves, stems, or photosynthetic tissue and is characterised by the emission of a strong odour that attracts the natural pollinators, carrion flies. Flowers that volatilise odorous compounds and attract carrion flies, beetles and other insects are often thermogenic. Here we present evidence of both thermogenesis and thermoregulation in R. lowii from microclimate and tissue temperatures measured during different stages of flower development in R. lowii, in natural conditions in Brunei, Borneo. Endothermy was detected in young and mature buds as well as in blooming flowers and even in decaying tissues 3 or more days after blooming. Tissue temperatures were maintained at 7–9 K above air temperature, in both female and male flowers, at all stages of floral development. Received: 29 August 1999 / Accepted 9 January 2000     

Movement coordination and signalling in ravens (Corvus corax): an experimental field study

Vagrant non-breeding ravens frequently attract conspecifics to rich ephemeral food sources. There, grouping may allow them to overcome the defence of territorial breeders. Here, we focus on ravens making use of regular food supplies in a game park, where they divert food from the provision of park animals. We investigated if ravens foraging in the Cumberland game park (Grünau, Austria) are attentive towards one another when they experience some unpredictability in food provisioning. We confronted a group of 30–50 ravens with two different treatments. Ten minutes ahead of the feeding of either wolves or wild boars we showed buckets containing pieces of meat to the ravens flying overhead. In the reliable cue treatment (RCT), the meat was placed next to one of the two enclosures, whereas in the unreliable cue treatment (UCT), the buckets were placed simultaneously in front of both enclosures though only in one of the enclosures were the animals fed 10 min later. Thus, during RCT but not during UCT, ravens could predict where food would become available. Only during UCT, ravens moved in large groups between the two feeding sites. Many ravens moving at the same time in the same direction may indicate some co-ordination in space and time, which is most likely achieved by social attraction among individuals. Furthermore, the number of ravens approaching and leaving, respectively, a feeding site cross-correlated with a temporary increase in the rate of a food-associated call, the yell. This suggests that in addition to watching each other, calling may have contributed to group formation. Possible benefits of group formation during food inspection are discussed. Received: 9 February 2000 / Received in revised form: 30 May 2000 / Accepted: 1 June 2000     

Main role of self-pollination rate on reproductive allocations in pseudogamous apomicts

 Most apomicts are hermaphroditic and pseudogamous (pollination is necessary to trigger parthenogenesis). In these plants, fitness depends on the number of progeny obtained by maternal reproduction. We determined the evolutionary stable strategy for male and female sex allocation. We show that the efficiency of pollination determines male and female resource allocations. Predictions are made of these allocations, of pollen/ovule ratio and of seed-set. We show that self-compatibility in apomicts is necessary for the maintenance of an apomictic population, and thus can account for the association between the loss of self-incompatibility and pseudogamous apomixis. In contrast to sexuals, male investment in pseudogamous apomicts increases with the rate of self-pollination. Received: 15 June 1996 / Accepted: 20 September 1996     

Effects of pollination intensity on the vigor of the sporophytic and gametophytic generation of Cucurbita texana

 We examined the effects of pollen competition (pollen load size) on sporophytic vigor and gametophytic performance in Cucurbita texana, a wild gourd, while controlling for alternative interpretations of the data. Under field conditions we compared the vigor of progeny produced from large and small pollen loads and examined the in vitro performance of the pollen produced by the progeny. We found that the progeny from large pollen loads germinated faster and had a greater reproductive output (male flowers and fruits) than progeny produced from small pollen loads. In addition, we found that the pollen produced on plants derived from large pollen loads grew faster in vitro than the pollen produced on plants derived from small pollen loads. These findings indicate that pollen competition affects the performance of the resulting sporophytic generation and the microgametophytes they produce. Received: 26 January 1997 / Revision accepted: 25 June 1997     

Effects of RNases on rejection of pollen from Nicotiana tabacum and N. plumbaginifolia

S-RNases are implicated in both inter- and intra-specific pollen rejection in Nicotiana. At least two mechanisms contribute to S-RNase dependent rejection of pollen from self compatilble species such as Nicotiana plumbaginifolia and N. tabacum. Cloned S-RNases from self incompatible N. alata expressed in styles of self compatible N. tabacum cause rejection of N. tabacum pollen through a factor-independent mechanism. However, rejection of N. plumbaginifolia pollen occurs only when S-RNases are expressed in conjunction with non-S-RNase factors from N. alata (factor-dependent pollen rejection). Here, we compared the pollen rejection activity of four RNases in these two systems. Recombinant RNase expression levels in the factor-dependent N. plumbaginifolia system were insufficient to cause pollen rejection. However, three S-RNases were active in the factor-independent N. tabacum pollen rejection system. This system shows the broadest specificity of any system so far examined. However, RNaseI from E. coli could not cause N. tabacum pollen rejection. Thus, RNase activity alone is not sufficient for pollen rejection. Our results suggest that S-RNases are specially adapted to function in pollen rejection. Received: 15 December 2000 / Accepted: 1 May 2001     

Floral organogenesis of Chloranthus sessilifolius, with special emphasis on the morphological nature of the androecium of Chloranthus (Chloranthaceae)

 Floral organogenesis of Chloranthus sessilifolius K. F. Wu is described. The inflorescence primordium is dome-like in the beginning and then elongates, and bract primordia initiate almost decussately. Each floral primordium, arising from the axil of a bract, soon becomes a scale-like structure, with three primordia of androecial lobes originating from its abaxial part, and the gynoecial primordium in adaxial position. As the androecial lobes become more distinct, four thecae are already in differentiation, and the gynoecial primordium appears as a shallow disc. The androecial lobes do not extend their length until the thecae approach maturity and the stigma is differentiated. The androecial lobes are united at all the stages of development, and the entire androecium falls off as a unit at the end of anthesis. Based on these results, combined with published evidence from neobotany, palaeobotany and phylogenetic studies, the morphological nature of the androecium of Chloranthus is further discussed. Our studies support the viewpoint that the androecial structure of Chloranthus may have arisen by splitting of a single stamen with 2 marginal thecae. Received May 2, 2001 Accepted December 18, 2001     

Selective fruit filling in relation to pollen load size in Alstroemeria aurea (Alstroemeriaceae)

 We conducted an experiment in a natural population of Alstroemeria aurea, a clonal perennial, to determine (1) if reproduction was resource limited, and (2) if fruits would be selectively filled based on differences in pollination intensity when pollen loads were adequate for full seed set. Under these conditions, differences in pollination intensity are unlikely to affect seed number, but could affect seed quality, providing an interesting test of the gametophytic competition hypothesis. To test for resource limitation, percent fruit maturation, number of seeds per fruit and average seed weight were compared to paired controls for ramets in which all but one fruit was removed. To test the effect of pollination intensity on selective resource allocation, three types of pollination treatments were performed: (1) all flowers of the single inflorescence received a low pollen load, (2) all flowers received a high pollen load, (3) alternate flowers of the inflorescence received either a high or a low pollen load. We determined the percentage of fruit that reached maturity, counted the number of seeds and ovules and calculated the average seed weight for all capsules in each treatment. Resources appeared to limit reproduction in this population since seed number and weight were significantly higher than in controls when competing capsules were removed. At the whole ramet level, a four fold difference in pollen loads had no significant effect on any of the parameters measured. However, when pollination intensity varied within an inflorescence, the number of seeds per fruit increased by about 10% in flowers that received the higher pollen load. We observed the same trend in each of 2 years, but the increase was significant in only 1 year. The differences, although not great, were only slightly smaller than when all competing fruits were removed, and were consistent with selective resource allocation based on pollination intensity independent of seed set. Received: 28 September 1997 / Accepted: 30 April 1998