Floral divergence and temporal pollinator partitioning in two synchronopatric species of <Emphasis Type="Italic">Vigna</Emphasis> (Leguminosae-Papilionoideae)

Exclusivity of pollinators, temporal partitioning of shared pollinators and divergence in pollen placement on the shared pollinators’ bodies are mechanisms that prevent interspecific pollen flow and minimize competitive interactions in synchronopatric plant species. We investigated the floral biology, flower visitors, pollinator effectiveness and seasonal flower availability of two syntopic legume species of the genus Vigna, V. longifolia and V. luteola, in ‘restinga’ vegetation of an island in southern Brazil. Our goal was to identify the strategies that might mitigate negative consequences of their synchronous flowering. Vigna longifolia and V. luteola were self-compatible, but depended on pollinators to set seeds. Only medium to large bees were able to trigger the ‘brush type’ pollination mechanism. Vigna longifolia, with its asymmetrical corolla and hugging mechanism, showed a more restrictive pollination system, with precise sites of pollen deposition/removal on the bee’s body, compared to V. luteola, with its zygomorphic corolla and cymbiform keel. There was a daily temporal substitution in flower visitation by the main pollinators. Vigna longifolia and V. luteola had overlapping flowering phenology but the densities of their flowers fluctuated, resulting in a seasonal partitioning of flower visitation. The differences in corolla symmetry and mainly the temporal partitioning among pollinators throughout the day and the flowering season proved to be important factors in maintaining the synchronopatry of V. longifolia and V. luteola.     

Convergence of anti-bee pollination mechanisms in the Neotropical plant genus <Emphasis Type="Italic">Drymonia</Emphasis> (Gesneriaceae)

The neotropical plant genus Drymonia displays a remarkable variety of floral shapes and colors. One feature that is particularly important to coevolution with pollinators involves the variable shapes and widths of corolla tubes. To evaluate the evolutionary context for changes in corolla shape, we constructed a phylogeny of 50 of the 75 species of Drymonia using molecular markers from plastid (trnK-matK) and nuclear regions (ITS and ETS). Mapping tube shapes on the phylogeny supports open, bell-shaped (campanulate) corolla shape as the ancestral character state for Drymonia, with multiple independent origins of constriction in the corolla tube. Corollas with constrictions take one of three tube shapes: a constricted flower opening and throat with a large, expanded pouch on the lower surface (hypocyrtoid); a constricted flower opening and throat lacking an expanded pouch on the lower surface (urceolate); or a constricted opening and throat where the sides of the corolla appear laterally compressed. Fieldwork demonstrates euglossine bees (mostly Euglossa spp. and Epicharis spp.) visit campanulate corollas while hummingbirds visit corollas that are constricted. Results support eight independent origins of constricted corolla tubes from ancestors with campanulate corolla tubes: 3 hypocyrtoid clades, 3 laterally compressed clades, and 3 urceolate clades (one of which represents a shift from a hypocyrtoid ancestor). Constricted corollas are associated with shifts from the ancestral condition of poricidal anther dehiscence, which presents pollen to pollinators in multiple small doses, to the derived condition of longitudinal anther dehiscence, which presents all pollen to pollinators simultaneously. The association of hummingbird pollination with constricted corolla tubes suggests that narrowing evolved as a barrier mechanism that prohibits the visitation of flowers by bees.     

Bee pollination and evidence of substitutive nectary in <Emphasis Type="Italic">Anadenanthera colubrina</Emphasis> (Leguminosae-Mimosoideae)

Anadenanthera colubrina (Vell.) Brenan (Leguminosae-Mimosoideae) is a widely-distributed tree in seasonally dry tropical forests of South America that was classified previously as lacking nectaries. However, some studies have stated that its flowers produce nectar, while others analyzed the composition of unifloral honey produced from A. colubrina flowers, raising the question about nectar production in the species. We studied the pollination and reproductive biology of A. colubrina var. cebil (Griseb.) Altschul in a natural population in the Caatinga, northeastern Brazil. Reproductive phenology, sexual system, floral biology, resource, and pollinators were investigated. We analyzed the breeding system through controlled pollinations for addressing its dependence on pollen vectors for reproduction. Anadenanthera colubrina flowered in the dry season, flower heads are heteromorphic, with staminate flowers at the base and perfect flowers at the apex of the inflorescence, characterizing andromonoecy. Anthesis is diurnal. We observed small drops of nectar at the apex of the petals of some flowers per inflorescence. Together with observations on flower visitor behavior and histochemical tests, we propose that A. colubrina produces floral nectar at the apex of the corolla, characterizing a substitutive nectary (sensu Vogel). This is the first record of substitutive nectary in the Mimosoideae and the first record of andromonoecy in the genus. Bees were the main pollinators (higher frequency), although other insects such as wasps, butterflies, and small beetles were also observed collecting nectar and/or pollen. The species is self-incompatible, thus depending on insect pollen vectors, mainly bees, for reproduction.     

Morphological variation in pollen grains of <Emphasis Type="Italic">Mucuna</Emphasis> (Leguminosae): new biogeographic and evolutionary patterns

Mucuna comprises 105 species with an overall pantropical distribution and is divided into three subgenera: M. subg. Mucuna, M. subg. Stizolobium and M. subg. Macrocarpa. Although phylogenetic studies have supported the occurrence of three main clades, evolutionary relationships among them are not fully resolved. The objective of this study was to examine pollen grain morphology from representatives of all three subgenera and map these onto the phylogenetic trees generated by analysis of other characters. Pollen grain surface, form, size, and aperture number were compared. A Bayesian inference tree using matK sequences was constructed. The results indicate that the representatives of M. subg. Macrocarpa have the smallest pollen grains in the genus (a synapomorphic character here identified for this subgenus) and that species of subgenus Mucuna (those with umbelliform inflorescences) have the largest pollen grains. Additional morphological diversity of the pollen grain surface was noted: reticulate and/or micro-reticulate (in all three subgenera), perforate, gemmate or verrucose (only in M. subg. Mucuna). For all studied taxa, the pollen grains are triaperturate, except for two species of M. subg. Mucuna, which have tetraperturate pollen. The phylogenetic tree obtained using the matK marker resolved M. subg. Stizolobium as the earliest diverging lineage in Mucuna. Based on this phylogeny, a reticulate ornamentation pattern of the pollen surface may represent the ancestral state for the genus, while the larger pollen size and the foraminate, gemmate, and verrucose ornamentations are derived characteristics within the genus. These putative derived ornamentations have been observed only in neotropical species.     

Flowering phenology and potential pollen emission of three <Emphasis Type="Italic">Artemisia</Emphasis> species in relation to airborne pollen data in Poznań (Western Poland)

Artemisia pollen is an important allergen in Europe. In Poznań (Western Poland), three Artemisia species, A. vulgaris, A. campestris and A. absinthium, are widely distributed. However, the contributions of these species to the total airborne pollen are unknown. The aim of the study was to determine the flowering phenology and pollen production of the three abovementioned species and to construct a model of potential Artemisia pollen emission in the study area. Phenological observations were conducted in 2012 at six sites in Poznań using a BBCH phenological scale. Pollen production was estimated by counting the pollen grains per flower and recalculating the totals per inflorescence, plant and population in the study area. Airborne pollen concentrations were obtained using a Hirst-type volumetric trap located in the study area. Artemisia vulgaris began to flower the earliest, followed by A. absinthium and then A. campestris. The flowering of A. vulgaris corresponded to the first peak in the airborne pollen level, and the flowering of A. campestris coincided with the second pollen peak. The highest amounts of pollen per single plant were produced by A. vulgaris and A. absinthium. A. campestris produced considerably less pollen, however, due to its common occurrence, it contributed markedly (30 %) to the summation of total of recorded pollen. A. vulgaris is the most important pollen source in Poznań, but the roles of two other Artemisia species cannot be ignored. In particular, A. campestris should be considered as an important pollen contributor and likely might be one of the main causes of allergic reactions during late summer.     

Variability of metric and qualitative traits of <Emphasis Type="Italic">Nitraria</Emphasis> species in relation to ecological and climatic conditions of Siberian habitats

The metric traits of shrubs, leaves, seeds, and flower parts have been studied in 19 Siberian populations of Nitraria sibirica Pall. and N. schoberi L. According to the obtained data, N. schoberi is characterized by larger shrubs (150–179 cm in height), leaves (15–30 mm in length, 5.7–8.8 mm in width), and seeds (up to 10 mm in length) and has rather wide (~2.5 mm) corolla petals. The size of anthers exceeds that of N. sibirica 1.2–1.5 times. The height of N. sibirica shrubs does not exceed 80 cm; the length of lamina, seed, and petals of corolla is 8–10, 3.7–7.4, and ~3.5 mm, respectively; and the size of anthers is 0.5–1 mm. Irrespective of habitat conditions, the stability of metric traits typical for both species has been shown to be rather high. The only dependence on climatic parameters is observed for the height of N. sibirica, which forms creeping forms in continental regions, especially in the mountains of the Altai Republic. Vegetative organs (laminas and young shoots) of N. sibirica are always more pubescent than those of N. shoberi.     

The genus ixocactus (Loranthaceae,s.s.): Description of its first species

The first species ofIxocactus Rizzini,I. hutchisonii Kuijt, is described and illustrated. The genus is unique in the family in that it is squamate and lacks inflorescences. Other remarkable features are the dense covering of spines, tetracolpate pollen, the small number of pollen grains per anther, the small size of the flower, and the occurrence of this parasite on stems and roots of other mistletoes.Ixocactus is believed to be related toOryctanthus.     

Differentiation of the pollen size in five representatives of <Emphasis Type="Italic">Taraxacum</Emphasis> sect. <Emphasis Type="Italic">Palustria</Emphasis>

Measurements of the pollen size in 5 species of Taraxacum sect. Palustria at three levels of ploidy: 2n = 3x = 24 (T. paucilobum), 2n = 4x = 32 (T. vindobonense, T. trilobifolium), 2n = 5x = 40 (T. mendax) and one taxon of unknown number of chromosomes 2n = ? (T. portentosum) are presented in this paper. Obtained results indicate a lack of distinct positive correlation between the pollen size and ploidy in the studied group of plants. Distinct relationship was, however, found between ploidy and the range of pollen size and shape variability. Most variable were the pollen grains of triploid T. paucilobum and the least — those in pentaploid T. mendax. Ranges of pollen variability in tetraploid T. trilobifolium and T. vindobonense and in T. portentosum of unknown number of chromosomes showed intermediate values.     

Shakers and head bangers: differences in sonication behavior between Australian <Emphasis Type="Italic">Amegilla murrayensis</Emphasis> (blue-banded bees) and North American <Emphasis Type="Italic">Bombus impatiens</Emphasis> (bumblebees)

Many bees collect pollen by grasping the anthers of a flower and vibrating their flight muscles at high frequencies—a behavior termed sonication, or buzz-pollination. Here we compare buzz-pollination on Solanum lycopersicum (cherry tomatoes) by two bees that fill similar niches on different continents—in Australia, Amegilla murrayensis (blue-banded bee), and in North America, Bombus impatiens (bumblebee). We collected audio recordings of buzz-pollination and quantified the frequency and length of buzzes, as well as the total time spent per flower. We found that A. murrayensis buzzes at significantly higher frequencies (~350 Hz) than B. impatiens (~240 Hz) and flaps its wings at higher frequencies during flight. There was no difference in the length of a single buzz, but A. murrayensis spent less time on each flower, as B. impatiens buzzed the flower several times before departing, whereas A. murrayensis typically buzzed the flower only once. High-speed videos of A. murrayensis during buzz-pollination revealed that its physical interaction with the flower differs markedly from the mechanism described for Bombus and other bees previously examined. Rather than grasping the anther cone with its mandibles and shaking, A. murrayensis taps the anther cone with its head at the high buzzing frequencies generated by its flight muscles. This unique behavior, combined with its higher buzzing frequency and reduced flower visit duration, suggests that A. murrayensis may be able to extract pollen more quickly than B. impatiens, and points to the need for further studies directly comparing the pollination effectiveness of these species.     

Micromorphological and histochemical attributes of flowers and floral reward in <Emphasis Type="Italic">Linaria vulgaris</Emphasis> (Plantaginaceae)

The self-incompatible flowers of Linaria vulgaris have developed a range of mechanisms for attraction of insect visitors/pollinators and deterrence of ineffective pollinators and herbivores. These adaptive traits include the flower size and symmetry, the presence of a spur as a “secondary nectar presenter,” olfactory (secondary metabolites) and sensual (scent, flower color, nectar guide—contrasting palate) signals, and floral rewards, i.e. pollen and nectar. Histochemical tests revealed that the floral glandular trichomes produced essential oils and flavonoids, and pollen grains contained flavonoids, terpenoids, and steroids, which play a role of olfactory attractants/repellents. The nectary gland is disc-shaped and located at the base of the ovary. Nectar is secreted through numerous modified stomata. Nectar secretion began in the bud stage and lasted to the end of anthesis. The amount of produced nectar depended on the flower age and ranged from 0.21 to 3.95 mg/flower (mean?=?1.51 mg). The concentration of sugars in the nectar reached up to 57.0%. Both the nectar amount and sugar concentration demonstrated a significant year and population effect. Pollen production was variable between the years of the study. On average, a single flower of L. vulgaris produced 0.31 mg of pollen. The spectrum of insect visitors in the flowers of L. vulgaris differed significantly between populations. In the urban site, Bombus terrestris and Apis mellifera were the most common visitors, while a considerable number of visits of wasps and syrphid flies were noted in the rural site.     

Growth and photosynthetic responses in Jatropha curcas L. seedlings of different provenances to watering regimes

Seedlings from four provenances of Jatropha curcas were subjected to 80, 50, and 30% of soil field capacity in potted experiments in order to study their responses to water availability. Our results showed that with the decline of soil water availability, plant growth, biomass accumulation, net photosynthetic rate, stomatal conductance (g_s), and transpiration rate (E) decreased, whereas leaf carbon isotope composition (δ¹³C), leaf pigment contents, and stomatal limitation value increased, while maximal quantum yield of PSII photochemistry was not affected. Our findings proved that stomatal limitation to photosynthesis dominated in J. curcas under low water availability. The increase of δ¹³C should be attributed to the decrease in g_s and E under the lowest water supply. J. curcas could adapt to low water availability by adjusting its plant size, stomata closure, reduction of E, increasing δ¹³C, and leaf pigment contents. Moreover, effects of provenance and the interaction with the watering regime were detected in growth and many physiological parameters. The provenance from xeric habitats showed stronger plasticity in the plant size than that from other provenances under drought. The variations may be used as criteria for variety/provenance selection and improvement of J. curcas performance.     

Morphological and genetic characterization of a new cytoplasmic male sterility system (<Emphasis Type="Italic">oxa</Emphasis> CMS) in stem mustard (<Emphasis Type="Italic">Brassica juncea</Emphasis>)

Key message

oxa CMS is a new cytoplasmic male sterility type in Brassica juncea.

Abstract

oxa CMS is a cytoplasmic male sterility (CMS) line that has been widely used in the production and cultivation of stem mustard in the southwestern China. In this study, different CMS-type specific mitochondrial markers were used to confirm that oxa CMS is distinct from the pol CMS, ogu CMS, nap CMS, hau CMS, tour CMS, Moricandia arvensis CMS, orf220-type CMS, etc., that have been previously reported in Brassica crops. Pollen grains of the oxa CMS line are sterile with a self-fertility rate of almost 0% and the sterility strain rate and sterility degree of oxa CMS is 100% due to a specific flower structure and flowering habit. Scanning electron microscopy revealed that most pollen grains in mature anthers of the oxa CMS line are empty, flat and deflated. Semi-thin section further showed that the abortive stage of anther development in oxa CMS is initiated at the late uninucleate stage. Abnormally vacuolated microspores caused male sterility in the oxa CMS line. This cytological study combined with marker-assisted selection showed that oxa CMS is a novel CMS type in stem mustard (Brassica juncea). Interestingly, the abortive stage of oxa CMS is later than those in other CMS types reported in Brassica crops, and there is no negative effect on the oxa CMS line growth period. This study demonstrated that this novel oxa CMS has a unique flower structure with sterile pollen grains at the late uninucleate stage. Our results may help to uncover the mechanism of oxa CMS in Brassica juncea.

     

More than protein? Bee–flower interactions and effects of disturbance regimes revealed by rare pollen in bee nests

Bees and their host flower populations were studied by identifying pollen to species or genus, from trap nests where bees were reared. Rare plant species in bee diets, and disturbance regimes, have not previously been researched and are emphasized here. Two focal bee groups with one species each (Megachilidae and Apidae) were studied in a 500,000-ha tropical reserve in the Yucatán Peninsula nine complete years. The number of rare or major pollen species in nests had no statistical correlation; thus, rare pollen analysis complements study of major brood provisions. We found most nests (87 % Megachile zaptlana, 93 % Centris analis) contained rare pollen; only 12 % of the 438 nests contained major pollen alone. Rare pollen sometimes indicated an energy source rather than a scarce protein resource. Trichome nectar of Cydista, along with Ipomoea and Caesalpinia, were nectar sources. Malpighiaceae, despite lacking nectar, often provided the complete Centris diet. Considering rare pollen, only Centris responded to drought, or competition from immigrant honeybees. Neither bee responded to hurricanes. Drought years coincided with low bee populations; Centris nests contained more rare species then. After feral Africanized honeybees colonized, Centris had more major species and fewer rare. Some herbarium vouchers from the study area contained exotic pollen, demonstrating in situ floral contamination and ecological generalization by bees, but this rarely occurred in plants found among the bee diets. Megachile and Centris responded differently to competition and resource scarcity, and plausibly evolved under different disturbance regimes, yet appeared well adapted to hurricane disturbance.     

Taxonomic significance of pollen morphology for species delimitation in <Emphasis Type="Italic">Psidium</Emphasis> (Myrtaceae)

Previous studies reported Psidium as one of the most difficult genera to delimit within the American Myrtaceae. Even though palynology has improved the taxonomy of Angiosperms, information about the usefulness of pollen morphology for taxonomic purposes in Myrtaceae remains contradictory. Here, we investigate the significance of pollen morphology for Psidium taxonomy with specific focus on its usefulness for determining species groups of taxonomic significance. Pollen traits observed by light and scanning electron microscopy were quantified and examined using cluster and ordination analyses. Average size of pollen grains was visualized by boxplots. Pollen grains of Psidium are isopolar, oblate, peroblate or oblate-spheroidal, 3-syncolporate or 4-syncolporate. The sexine ornamentation is rugulate, granulate or spinulose-granulate and differs between the mesocolpium and apocolpium. Cluster analysis revealed four distinct groups: Psidium cauliflorum (G1) and Psidium oligospermum (G3) as single-species groups; Psidium brownianum, P. oblongatum, P. ovale, P. sartorianum, P. guajava, Psidium sp. 1, Psidium sp. 2 (G2), and Psidium cattleianum, P. longipetiolatum, P. guineense, P. myrtoides (G4). Supported by ordination analysis, three traits better explained these groups: type of exine ornamentation, size of P-EV and pollen shape. The used approach efficiently distinguished related species, as well as explained species groups of taxonomic significance suggesting pollen morphology to be a significant source of information for taxonomic studies in Psidium.     

Pollen morphology of selected crop plants from southern China and testing pollen morphological data in an archaeobotanical study

Identification of pollen grains of cultivated plants is essential in archaeobotanical studies. In this study, we investigated the pollen morphology of 30 species which are representatives of most of the crop plants in southern China, using a light microscope. Our results show that the pollen grains of these species or genera can generally be identified by their size, aperture(s) and exine sculpture. We found that: (1) some cultivated cereals can be distinguished from wild species of Poaceae according to their size frequency combined with their morphological features; (2) the lengths of the equatorial diameter (E), polar axis (P) and the greatest dimension of the lumina (the size of the network sculpturing) of the exine reticulum may be diagnostic features to distinguish some brassicaceous vegetables. There are significant differences between the E and P values among Brassica campestris (B. rapa, oilseed rape, Chinese cabbage), B. alboglabra (B. oleracea var. alboglabra, gai lan, Chinese kale), B. parachinensis (B. rapa var. parachinensis, choy sum, Chinese flowering cabbage) and B. chinensis (B. rapa ssp. chinensis, pak choi), but moderate differences in the longer axis length of the reticulum lumina, which provide potential for identifying species on the basis of pollen grains. We compared the P values and the longer axis length of the lumina of modern specimens of Brassicaceae pollen grains with those of fossil pollen extracted from the Ming-Qing cultural layer in the Fuqikou site at Chongqing, China, and found that the fossil pollen grains of Brassicaceae probably represent vegetable plants related to B. parachinensis. Moreover, we measured the diameters of rice pollen grains from modern paddy fields to assess the pollen size frequency and found that the size range from ~?34 to 38 µm is closely associated with rice pollen in southern China, which can be used to detect pollen signals of human activities in archaeobotanical investigations.     

Characterization and Expression Pattern of a Putative Pectin Methylesterase Gene, <Emphasis Type="Italic">BcMF27</Emphasis>, in <Emphasis Type="Italic">Brassica campestris</Emphasis> ssp. <Emphasis Type="Italic">chinensis</Emphasis>

Pectin methylesterases (PMEs) play an important role in modifying cell wall. PMEs catalyze the de-esterification of pectin, an important compound of cell wall, to affect fertility in plant reproduction. However, little especially molecular mechanism about pectin methylesterase is studied in recent years despite its importance to reproductive development in flower plant. Here the bioinformatics analysis of BcMF27 (Brassica campestris Male Fertility 27) (BRAD: Bra000541 GenBank: KT600012) sequence isolated from Brassica campestris L. ssp. chinensis showed its highly and characteristically conserved structure as a pectin methylesterase. Transient expression analysis in the onion epidermal cells revealed the product of BcMF27 was a transmembrane protein. Real-time RT-PCR and in situ hybridization suggested that BcMF27 was expressed in pollen grain and pollen tube. This study demonstrates that BcMF27 encodes a transmembrane pollen- and pollen tube-specific PME gene, and is also considered to help further understand the biological function of pectin methylesterases and the molecular mechanism of pollen development, pollen tube growth as a genic tool.     

<Emphasis Type="Italic">Cannabis</Emphasis> is indigenous to Europe and cultivation began during the Copper or Bronze age: a probabilistic synthesis of fossil pollen studies

Conventional wisdom states Cannabis sativa originated in Asia and its dispersal to Europe depended upon human transport. Various Neolithic or Bronze age groups have been named as pioneer cultivators. These theses were tested by examining fossil pollen studies (FPSs), obtained from the European Pollen Database. Many FPSs report Cannabis or Humulus (C/H) with collective names (e.g. Cannabis/Humulus or Cannabaceae). To dissect these aggregate data, we used ecological proxies to differentiate C/H pollen, as follows: unknown C/H pollen that appeared in a pollen assemblage suggestive of steppe (Poaceae, Artemisia, Chenopodiaceae) we interpreted as wild-type Cannabis. C/H pollen in a mesophytic forest assemblage (Alnus, Salix, Populus) we interpreted as Humulus. C/H pollen curves that upsurged and appeared de novo alongside crop pollen grains we interpreted as cultivated hemp. FPSs were mapped and compared to the territories of archaeological cultures. We analysed 479 FPSs from the Holocene/Late Glacial, plus 36 FPSs from older strata. The results showed C/H pollen consistent with wild-type C. sativa in steppe and dry tundra landscapes throughout Europe during the early Holocene, Late Glacial, and previous glaciations. During the warm and wet Holocene Climactic Optimum, forests replaced steppe, and Humulus dominated. Cannabis retreated to steppe refugia. C/H pollen consistent with cultivated hemp first appeared in the Pontic-Caspian steppe refugium. GIS mapping linked cultivation with the Copper age Varna/Gumelni?a culture, and the Bronze age Yamnaya and Terramara cultures. An Iron age steppe culture, the Scythians, likely introduced hemp cultivation to Celtic and Proto-Slavic cultures.