Old World and New World Cupressus pollen: morphological and cytological remarks

The aim of this work was to collect new information about pollen morphology and pollen wall structure comparing Cupressus species from the Old World and New World. Cupressus is a nonmonophyletic genus that includes species that appear to be divided into Old World and New World clades. Observations in this study on cypress pollen indicate that grain size and composition of intine β-glucans are different between the considered Old World and New World species. Different from all the other American cypress species, pollen of C. macrocarpa reacted to dying in a similar manner to Old World species. Rehydrated pollen grains collected from 20 Asian, Afro-Mediterranean and American cypress species were measured under a light microscope. The size of the pollen grains and the percentage of intine in relation to the pollen grain diameter were significantly different between Old World and New World species. Pollen wall composition was tested after addition of different dyes to the hydration solution, and subsequent observations were carried out by light and fluorescence microscopy. Lugol and calcofluor staining showed differences in composition of the middle and inner intine layers between New World and Old World species.     

The pollination ecology of Echeandia macrocarpa (Liliaceae)

Field research and observation of the breeding system of the salvadoran extension of theEcheandia macrocarpa complex indicate an entirely mellittophilous syndrome, withBombus ephippiatus Say workers as the primary pollen vectors. Foraging behavior coupled with the floral morphology/ phenology of the host plant suggests a trend towards obligatory out-crossing. The foraging behavior ofB. ephippiatus workers on the host plant and on flowers of co-blooming species is reported to elucidate the dynamic processes that determine the present co-evolutionary status ofE. macrocarpa and its chief pollinator.     

Shared pollinators and pollen transfer dynamics in two hybridizing species, Rhinanthus minor and R. angustifolius

Gene flow between hybridizing plant species depends strongly on pollinator behaviour, which affects pollen transfer among floral types and reproductive isolation. We examined bumblebee behaviour and pollen transfer between two hybridizing Rhinanthus species that are very similar in ecology and floral traits. The two species, Rhinanthus minor and R. angustifolius, shared similar pollinator guilds and assemblages, but pollinator recruitment and flower visitation rates were higher in R. angustifolius sites, probably because of its higher reward levels and better visibility. When presented with Rhinanthus flowers, bumblebees that previously foraged on R. angustifolius were less prone to visit R. minor inflorescences, while R. minor foragers accepted both species in similar proportions. Although Rhinanthus has been cited as a case of mechanical isolation resulting from interactions between bee behaviour and differences in stigma and anther placement, we found no support for efficient mechanical reproductive isolation. Bumblebees that foraged on R. minor flowers carried more pollen, but pollen placement on their bodies was similar to that of bees that visited R. angustifolius, and cross-specific stigmatic pollen deposition was similar in both directions. However, the asymmetry in pollinator handling time between the two species, due to dissimilar pollen rewards, may have lowered relative heterospecific pollen receipt on R. angustifolius, suggesting that net gene flow resulting from pollen transfer dynamics is more likely towards R. minor, although this effect remains weak and will be most likely counterbalanced by context-based labile pollinator preference.     

Pollen production of the genus Cupressus

The study of potential pollen emission in the genus Cupressus was carried out in the city of Córdoba (Spain). Pollen production of 227 trees of the represented species of Cupressus in the area (Cupressus sempervirens, C. macrocarpa and C. arizonica) was studied by counting the number of pollen grains per flower, the number of flowers per branch, and the number of branches per m² of surface area. Total surface area was estimated by considering the shape of the tree as a cone and estimating the average height and radius of each species. Results indicated that there was little variation in the number of pollen grains produced by a flower within each species. Equally, there were few differences in the number of grains per flower among the species, although C. arizonica had slightly fewer than the other species because it possessed fewer stamens per flower. However, there were clear differences in the number of flowers per branch, with C. macrocarpa having the highest floral density, considerably lower in C. arizonica and even lower in C. sempervirens. All of this indicates that the pollen contribution of C. macrocarpa to the atmosphere is far greater than that of C. arizonica and C. sempervirens. The results indicate a different potential emission in each species, and this must be taken into account when determining the quantity of pollen present in the atmosphere. The maximum amount of pollen grains that one individual can spread is provided in this study and would provide useful data in the development of an airborne forecasting system.     

Comparison of pollen grain morphological features of selected species of the genus Crataegus (Rosaceae) and their spontaneous hybrids

The aim of our study was to verify, on the basis of statistical analyses of nine quantitative morphological features of pollen grains, the hypothesis that pollen grains of three parental species of Crataegus (C. laevigata, C. monogyna, C. rhipidophylla) differed from the pollen of three spontaneous hybrids of these species (C. × macrocarpa, C. × media, C. × subsphaericea). Contrast analysis revealed that a majority of the pollen features of hybrid species were characterized by significantly higher mean values than those of parental species. Analysis of pollen shape classes indicated that the parental species clearly differed from each other in contrast with hybrids, which were characterized by a similar proportion of pollen in individual pollen shape classes. Statistical analyses showed that the pollen grains of two parental species, C. laevigata and C. monogyna, were most similar to one another. Pollen grains of typical C. rhipidophylla were similar to the pollen of hybrids and the mean values of almost all studied pollen features [P, E, Exp, Exp/P, Le, d, d/E (PAI)] of C. rhipidophylla var. rhipidophylla were intermediate between those of C. monogyna and C. rhipidophylla var. lindmanii. This corroborates Zieliński's conjecture that C. rhipidophylla is probably an old, conserved hybrid between C. monogyna and C. calycina (= C. lindmanii = C. rhipidophylla var. lindmanii). According to the analysis of canonical variables, C. × macrocarpa and C. × media pollen grains were most similar. C. × subsphaericea and C. rhipidophylla var. rhipidophylla and C. rhipidophylla var. lindmanii formed another pair (group), and C. monogyna and C. laevigata constituted separate ‘single species groups’. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 172 , 555–571.     

Geographic differentiation of cortical monoterpenoids of Pseudotsuga macrocarpa

Cortical essential oil from Pseudotsuga macrocarpa was analyzed for monoterpenoid compounds. Only minor differences were apparent between samples collected in different locations. Pseudotsuga macrocarpa and P. menziesii appeared to be quite distinct chemically. No evidence was obtained of any gene exchange between the two species in the southern California coastal region, where they grow a short distance from each other.     

Pollen morphology and taxonomy of the subtribe Diocleinae (Leguminosae: Papilionoideae: Phaseoleae)

The pollen morphology of the thirteen genera currently included in the subtribe Diocleinae is described. Nine pollen types with a number of subtypes are recognized. These are arranged into two main groups. The first comprises the macromorphologically anomalous genera Calopogonium, Herpyza and Pachyrhizus which, although united by some pollen characters, are distinct both from each other and from the rest of the subtribe. The second group, consisting of the remaining genera which can to some extent be distinguished from each other by their pollen morphology, nevertheless appear to make up both macromorphologically and palynologically a more natural Diocleinae. The six pollen types in the second group are further divided into two slightly overlapping subgroups. Luzonia and Macropsychanthus are palynologically somewhat distinct but, together with Dioclea and Canavalia, have generally rather large, oblate pollen grains with a complete or perforate tectum, in contrast with the pollen of the genera Camptosema, Collaea, Cratylia and much of Galactia which is smaller, spheroidal with a reticulate tectum.Dioclea has some species with isopolar pollen and others with heteropolar pollen grains and this more or less correlates with subgeneric divisions within the genus. Galactia from the Caribbean area has similar but much smaller pollen. The taxonomic significance of this parallel between the pollen morphology of Dioclea and Caribbean Galactia is unclear. The pollen of Canavalia appears to show no significant variation.A possible correlation between the pollen subgroups with ecological habitat and pollinators is discussed.     

Specialist bees collect Asteraceae pollen by distinctive abdominal drumming (<Emphasis Type="Italic">Osmia</Emphasis>) or tapping (<Emphasis Type="Italic">Melissodes</Emphasis>, <Emphasis Type="Italic">Svastra</Emphasis>)

Four species of western US Osmia (3 Cephalosmia) that are Asteraceae specialists (mesoleges) were observed using a stereotypical means of collecting pollen—abdominal drumming—to gather pollen from 21 flowering species representing nine tribes of Asteraceae. Abdominal drumming is a rapid dorso-ventral motion of the female’s abdomen (467 pats/min) used to directly collect and place pollen in the bee’s ventral scopa. A co-occurring generalist, O. lignaria, never drummed Asteraceae flowers for pollen, but instead used its legs to harvest pollen. Observed drumming by several other osmiines is noted. A different pollen-harvesting behavior, abdominal tapping, is described for two eucerine bees (Melissodes agilis and Svastra obliqua), both oligolectic for the Asteraceae. The behavior also involves a dorso-ventral motion, but they tap their distal abdominal venter against disk flowers at a slower tempo (304 taps/min). These females’ distal sternites have distinctly dense and long hair brushes for acquiring pollen by this behavior. Brief accounts of similar abdominal pollen gathering behaviors by other megachilids are summarized.     

Better safe than sorry? A Fabaceae species exhibits unfavourable pollen properties for developing bee larvae despite its hidden anthers

Empirical evidence suggests that pollen chemistry plays an important role in shaping the pollen host spectra of many bee species. Although the underlying mechanisms are poorly understood, pollen diets of several plant taxa have experimentally been found to impede larval development of unspecialized bees. The pollen of all plant taxa, for which such a detrimental effect on bee larval development has been observed so far, is freely accessible in the flowers and thus easily harvestable for flower visitors, suggesting that this pollen might be chemically protected in order to reduce its loss to pollen-feeding animals. In the present study, we compared larval performance of five solitary bee species on pollen diets of the two Fabaceae species Onobrychis viciifolia and Lotus corniculatus, which have their anthers concealed inside the flowers, with that on control diets composed of host pollen provisions. As the complex flower morphology of the two Fabaceae species already considerably narrows the spectrum of pollen harvesting bee taxa, which might supersede costly chemical protection of the pollen, we expected bees that usually do not exploit Fabaceae to develop well on Onobrychis and Lotus pollen diets. Larval survival on the Onobrychis pollen diet was successful for all five bee species tested. In contrast, larval survival on the Lotus pollen diet was reduced in three species despite the fact that Lotus flowers are more difficult to exploit for pollen than Onobrychis flowers. We conclude that there is no trade-off between pollen concealment and pollen defence in Lotus and that pollen of morphologically complex flowers with a restricted visitor spectrum is not necessarily an easy-to-use nutritional source.     

Pollen evolution and its taxonomic significance in Cuscuta (dodders, Convolvulaceae)

The pollen morphology of 148 taxa (135 species and 13 varieties) of the parasitic plant genus Cuscuta (dodders, Convolvulaceae) was examined using scanning electron microscopy. Six quantitative characters were coded using the gap-weighting method and optimized onto a consensus tree constructed from three large-scale molecular phylogenies of the genus based on nuclear internal transcribed spacer (ITS) and plastid trn-LF sequences. The results indicate that 3-zonocolpate pollen is ancestral, while grains with more colpi (up to eight) have evolved only in two major lineages of Cuscuta (subg. Monogynella and clade O of subg. Grammica). Complex morphological intergradations occur between species when their tectum is described using the traditional qualitative types—imperforate, perforate, and microreticulate. This continuous variation is better expressed quantitatively as “percent perforation,” namely the proportion of perforated area (puncta or lumina) from the total tectum surface. Tectum imperforatum is likely the ancestral condition, while pollen grains with increasingly larger perforation areas have evolved multiple times. The reticulated tectum, unknown in other Convolvulaceae, has evolved in Cuscuta only in two lineages (subg. Monogynella, and clade O of subg. Grammica). Overall, the morphology of pollen supports Cuscuta as a sister to either the “bifid-style” Convolvulaceae clade (Dicranostyloideae) or to one of the members of this clade. Pollen characters alone are insufficient to reconstruct phylogenetic relationships; however, palynological information is useful for the species-level taxonomy of Cuscuta.     

Pollen presentation, pollen-ovule ratios, and other reproductive traits in Mediterranean Legumes (Fam. Fabaceae - Subfam. Faboideae)

We studied the allocation of reproductive resources in 32 Mediterranean legume species. Correlations between main parameters were analysed at interspecific level. For each species we described the type of pollen presentation and examined its systematic significance. The high P/O values observed reflect low pollen transfer efficiency, typical of xenogamous species. Lowest P/Os were found in species with brush or explosive tripping mechanism, while highest ones were associated to primary pollen presentation. No trade-off was observed between P/O and the volume of a single pollen grain, nor between pollen number and pollen grain volume. Perennials showed higher P/Os compared to annuals, in accordance with the assumption that annual species are often self-compatible while most perennials are outcrossers. Fruit and seed set, evaluated in eight species under different experimental conditions, showed a close dependence on insects, and the existence of self-compatibility in three species (Colutea arborescens, Coronilla emerus, Hedysarum coronarium). In Colutea arborescens and Coronilla emerus pollen-pistil interaction and pre-zygotic barriers to selfing were investigated through stigma receptivity tests and in-vivo pollen germination trials. Results demonstrated the importance of the stigmatic cuticle in reducing levels of autogamy. Productivity tests confirmed the utility of P/O ratio as indirect measure of species' breeding system.     

Palynological survey of the subtribe Elephantopinae (Asteraceae, Vernonieae)

Despite the taxonomic value of pollen morphology within the tribe Vernonieae, a detailed study of the pollen of the subtribe Elephantopinae is still lacking. The pollen morphology of ten species, representing three of the four genera of the subtribe, Elephantopus, Pseudoelephantopus and Orthopappus, was studied with LM, SEM, and TEM. The pollen of all the species studied was found to be echinolophate, although the differences in aperture features and both sculpture and exine structure allowed recognizing two pollen types. The species of Elephantopus (except for E. elongatus) and Pseudoelephantopus share the regular or more or less regular ridges pattern, the colporate condition but with the ectoapertures little developed, and the acaveate exine structure. From the sculpture and the apertures, the pollen of Caatinganthus harleyi was found to be similar to that of Elephantopus and Pseudoelephantopus. The pollen morphology of the monotypic Orthopappus, which is shared with that of E. elongatus, was further investigated for the first time. We found that it differs from that of the other species in having an irregular pattern of ridges, colporate condition but with a well-developed ectoapertures, and caveate exine. Additional studies of the exine structure and apertures features, coupled with molecular phylogeny, are needed to understand the evolution of pollen characters and re-evaluate the intergeneric relationships within the tribe.     

Floral resource distribution and fitness consequences for two solitary bee species in agricultural landscapes

Floral resources are crucial for wild pollinators. Identifying the spatio-temporal floral resource use of wild pollinators and effects of resource distribution on their development might help to promote them and their pollination services to crops in agricultural landscapes.We established populations of Osmia cornuta and Osmia bicornis, two solitary wild bees, in 24 agricultural landscapes with varying floral resource availability. Based on their pollen use, we mapped the landscape-scale distribution of the visited plants, estimated pooled specific floral resource availabilities and measured its effects on reproductive output.Woody semi-natural habitats such as hedgerows provided the majority of pollen sources for both Osmia species. Pollen use differed strongly between the two species. The offspring of both Osmia increased with availability of pooled specific pollen resources. In accordance with their preferred pollen types, offspring of O. cornuta increased with increasing cover of trees and shrubs of the Rosaceae family, and that of O. bicornis with increasing cover of Papaver rhoeas, Ranunculus acris and Quercus spp. as well as with the proximity to oilseed rape. In spite of their specific responses to pollen resources, the offspring of both species decreased with the distance to forest. The floral resource availability did not significantly affect the proportion of adult females and the weight of the offspring. As forest does not appear to be a main foraging habitat for both species, the benefit of forest proximity indicates an additional role of forest in addition to food availability.Specific flowering plants and forests should thus be conserved and enhanced to maintain and support O. cornuta, O. bicornis and likely other wild bee populations in agricultural landscapes. The combined information of land cover and detailed floral resource availability gives a deeper understanding into population processes in agricultural landscapes.     

Strength and variability of postmating reproductive isolating barriers between four European white oak species

The identification and quantification of the relative importance of reproductive isolating barriers is of fundamental importance to understand species maintenance in the face of interspecific gene flow between hybridising species. Yet, such assessments require extensive experimental fertilisations that are particularly difficult when dealing with more than two hybridising and long-generation-time species such as oaks. Here, we quantify the relative contribution of four postmating reproductive isolating barriers consisting of two prezygotic barriers (gametic incompatibility, conspecific pollen precedence) and two postzygotic barriers (germination rate, early survival) from extensively controlled pollinations between four oak species (Quercus robur, Quercus petraea, Quercus pubescens and Quercus pyrenaica) that have been shown to frequently hybridise in natural populations. We found high variation in the strength of total reproductive isolation between species, ranging from total reproductive isolation to advantage toward hybrid formation. As previously found, Q. robur pollen was unable to fertilise Q. petraea due to a strong reproductive isolating mechanism. On the contrary, Q. pubescens pollen was more efficient at fertilising Q. petraea than conspecific pollen. Overall, prezygotic barriers contribute far more than postzygotic barriers to isolate species reproductively, suggesting a role for reinforcement in the development of prezygotic barriers. Conspecific pollen precedence reduced hybrid formation when pollen competition was allowed; however, presence of conspecific pollen did not totally prevent hybridization. Our results suggest that pollen competition depends on multiple ecological and environmental parameters, including species local abundance, and that it may be of uppermost importance to understand interspecific gene flow among natural multispecies populations.     

First report of fern (Culcita macrocarpa) spore consumption by a small mammal (Apodemus sylvaticus)

Few vertebrates are known to consume ferns regularly. Several species of mammals consume leaves to some extent but the consumption of fern spores is much rare. In Galicia (Northwest Spain) we studied the seasonal variation in the consumption of Culcita macrocarpa fertile leaves (i.e. with spores) in two populations (Capelada and Eume), assessed whether consumption rate increased with fern population size, and evaluated whether the consumer was a spore predator or a spore disperser. Consumption began in December and finished by mid February, and occurred before spore release, which happened later in Capelada than in Eume, probably influenced by differences in altitude. The consumer was identified as Apodemus sylvaticus by DNA analysis of its droppings and by capture of live animals. Throughout Galicia there was a significant increase in fern consumption rate as the population size of C. macrocarpa increased. Germination tests from droppings were carried out in 14 dishes but only in two dishes 1% and 0.3% of the spores germinated. Our results suggest that woodmouse can disperse spores of C. macrocarpa, although most of the spores were digested.     

Pollination partners of Mucuna macrocarpa (Fabaceae) at the northern limit of its range

Mucuna macrocarpa is a plant found in tropical and subtropical regions that requires an “explosive opening.” Explosive opening is the process that exposes the stamen and pistil from the opening of the carina. This process is needed for cross pollination; however, the plant cannot open itself and opening by an animal is needed. The most common opener of Mucuna flowers is several nectar‐eating bats (e.g., Syconycteris), but the flying fox, Pteropus dasymallus, is the only opener of M. macrocarpa on the subtropical island of Okinawajima. Here, we present the explosive openers and possible pollinators in the northernmost and temperate Kamae region, Kyushu, Japan, where nectar‐eating bats are absent. The Japanese macaque, Macaca fuscata, and the Japanese marten, Martes melampus, were the explosive openers observed during our survey in Kamae. Martens opened flowers using their snout in a manner similar to that of the flying fox, whereas macaques opened flowers using their hands. This is the first time that an animal has been observed opening these flowers with its hands rather than snout. In total, 97% (n = 283) of explosively opened flowers were opened by macaques, and the macaque largely contributed to the overall flower opening. Because many pollen grains become attached to the explosive openers, they are considered to be primary pollinators. Furthermore, two bee species, Apis cerana japonica and Bombus ardens ardens, also visited opened flowers and collected pollen, and they were possibly secondary pollinators.     

Comparative pollen morphology of Brachylena, Tarchananthus and two species of Tubulifloridites (Asteraceae) from the Eocene, Knysna Lignite of South Africa

Two fossil taxa Tubulifloridites antipodica and T. viteauensis recovered from the Eocene Knysna Lignite of South Africa were examined with scanning (SEM) and transmission electron microscopy (TEM). The details of their sculpturing and wall structure are similar to the same species of fossil dispersed pollen taxa recovered from southwestern Africa and South America. Fifteen species of the woody South African taxa, Brachylaena (9 species) and Tarchonanthus (6 species) were investigated with SEM and TEM. All of the taxa are tricolporate, spherical to slightly prolate, microechinate to echinate and have a bilayered columellate infrastructure, except B.ilicifolia, which has a single columellate infrastructural level with the “granularization” of the outer portion of the infrastructural layer or the inner layer of the tectum. There is a similar distribution of plesiomorphic and derived pollen characters in a number of aster subfamilies and tribes suggesting a similar evolutionary progression of pollen, and pollen wall character evolution was occurring synchronously in a variety of aster subfamilies during the middle Tertiary and that these unique pollen features may be important to the evolution and diversification of the Asteraceae.     

A new species of Amanita section Lepidella from South China

Amanita macrocarpa from Guangdong Province of China is described. The new species is a large, brownish orange to light brown mushroom, characterized by numerous pyramidal warts on the pileus, yellowish lamellae, a large annulus at the middle of the stipe, and amyloid ellipsoidal basidiospores. Phylogenetic analyses of the new species and related taxa based on the large subunit of nuclear ribosomal DNA (LSU) and the internal transcribed spacer (ITS) sequences are provided. Morphological and molecular data show that A. macrocarpa is a member of Amanita, section Lepidella, and clearly different from any known taxon of the section.