Are Nectar Sugar Composition and Corolla Tube Length Related to the Diversity of Insects that Visit Asteraceae Flowers?

Abstract: In this work, we analysed interspecific variation in nectar sugar composition, corolla tube length, and the diversity of floral visitors of 35 Asteraceae species. The potential correlations between these variables could arise either as a result of selection to improve pollinator attractiveness or simply as a consequence of phylogenetic constraints. Samples of nectar and flowers, and data on floral visitors, were obtained from living plants in natural populations from Argentina. Asteraceae species showed a large variability in corolla tube length. Nectar of most species presented a larger proportion of hexoses than sucrose. All species were visited by numerous insects belonging to 2 different orders. Results showed that floral traits are not significantly correlated with the diversity of floral visitors. These characters seem to be linked to the phylogeny of the species. Early branching species (species phylogenetically close to the root of the Asteraceae tree) tend to have longer corollas, higher sucrose proportions and lesser diversity of floral visitors than late branching species. Considering that longer corolla tubes and higher nectar sucrose percentages may indicate some specialization in the pollination system, we suggest that there is an evolutionary tendency toward generalist pollination systems within the family.     

Universal and group-specific real-time PCR diagnosis of flavescence dorée (16Sr-V), bois noir (16Sr-XII) and apple proliferation (16Sr-X) phytoplasmas from field-collected plant hosts and insect vectors

Three real‐time PCR–based assays for the specific diagnosis of flavescence dorée (FD), bois noir (BN) and apple proliferation (AP) phytoplasmas and a universal one for the detection of phytoplasmas belonging to groups 16Sr‐V, 16Sr‐X and 16Sr‐XII have been developed. Ribosomal‐based primers CYS2Fw/Rv and TaqMan probe CYS2 were used for universal diagnosis in real‐time PCR. For group‐specific detection of FD phytoplasma, ribosomal‐based primers fAY/rEY, specific for 16Sr‐V phytoplasmas, were chosen. For diagnosis of BN and AP phytoplasmas, specific primers were designed on non‐ribosomal and nitroreductase DNA sequences, respectively. SYBR^® Green I detection coupled with melting curve analysis was used in each group‐specific protocol. Field‐collected grapevines infected with FD and BN phytoplasmas and apple trees infected with AP phytoplasma, together with Scaphoideus titanus, Hyalesthes obsoletus and Cacopsylla melanoneura adults, captured in the same vineyards and orchards, were used as templates in real‐time PCR assays. The diagnostic efficiency of each group‐specific protocol was compared with well‐established detection procedures, based on conventional nested PCR. Universal amplification was obtained in real‐time PCR from DNAs of European aster yellows (16Sr‐I), elm yellows (16Sr‐V), stolbur (16Sr‐XII) and AP phytoplasma reference isolates maintained in periwinkles. The same assay detected phytoplasma DNA in all test plants and test insect vectors infected with FD, BN and AP phytoplasmas. Our group‐specific assays detected FD, BN, and AP phytoplasmas with high efficiencies, similar to those obtained with nested PCR and did not amplify phytoplasma DNA of other taxonomic groups. Melting curve analysis was necessary for the correct identification of the specific amplicons generated in the presence of very low target concentrations. Our work shows that real‐time PCR methods can sensitively and rapidly detect phytoplasmas at the universal or group‐specific level. This should be useful in developing defence strategies and for quantitative studies of phytoplasma–plant–vector interactions.     

Pollen-pistil size correlation and pollen size-number trade-off in species of Argentinian Nyctaginaceae with different pollen reserves

Data on pollen and pistil traits from 14 Argentinean Nyctaginaceae species with starch or lipids as pollen reserves are presented. We expect differences in the traits between these two groups of species, but the same pattern within each group for (a) the relationship between pollen size and pistil length assuming that pollen tube length is predetermined by provisions in the pollen independently of the pollen reserve type, and for (b) a trade-off between size and number because available resources for male function are not unlimited. In particular we expect that (a) species with longer pistils will have larger pollen grains, (b) pollen grain size and the number of pollen grains per flower will be negatively correlated. Significant differences in the mean pollen number per flower and mean pollen size were observed between species with different pollen reserve type but not for pistil length. Then, correlation analyses were performed for species with starchy pollen or with pollen with lipids separately. Pollen size - pistil length correlation was positive and significant for species with starchy pollen but not for species with pollen with lipids. On the other hand, pollen size - number correlation was not significant for both starchy and oil-rich species. Results suggest that pollen reserve type would be a relevant factor that constraint pollen size in species of Nyctaginaceae, and that this pollen trait should be considered when studying pollen-pistil relationships, mainly between species of those families with mixed pollen reserves.     

Flowering phenology of co-occurring Asteraceae: a matter of climate,ecological interactions,plant attributes or of evolutionary relationships among species?

We analyzed the flowering phenodynamics of 43 Asteraceae species co-occurring in natural populations of Chaco Serrano forests in central Argentina. We explored the potential influence of factors such as photoperiod and climate (variations in temperature, rainfall, and frost), animal-plant interactions (richness of floral visitors, frequency of visits), some plant attributes (plant growth form, seed dispersal mechanism), and evolutionary relationships among species on flowering phenodynamics. Cluster Analysis (CA) and Principal Component Analysis (PCA) were the multivariate statistical methods used to analyze emerging patterns associated with these co-occurring species. Null-model analyses were used to evaluate whether flowering times are aggregated, segregated, or random. Results showed that flowering phenology was significantly correlated with the seasonal variation in temperature, photoperiod, rainfall, and frost. The multivariate statistical methods separated all the species in three groups: 1) species with short flowering time, large plant floral display, high frequency of visits by a large number of species of floral visitors, anemochorous fruits, and shrubby growth form, with a tendency to a segregated flowering pattern; 2) species with long flowering time, small plant floral display, low frequency of visits by few insect species, anemochorous fruits, and herbaceous growth form; and 3) species with long flowering time, small plant floral display, intermediate values for frequency of visits and number of species of floral visitors, seed dispersal mechanisms other than anemochory, and herbaceous growth form. In addition, all but one species belonging to early-branching tribes (tribes phylogenetically close to the root of the Asteraceae tree) were grouped together and clustered in the same region of the two-dimensional PCA ordination. All species belonging to the late-branching tribes (Asteroideae subfamily tribes) included in group 1 were separated from the other Asteroideae species in the PCA. In conclusion, it seems that climatic factors restrict the phenological period of most species, and that plant attributes and taxonomic membership are strongly related to flowering phenodynamics in this group of Asteraceae studied.     

Nectar regulation in Euphorbia tithymaloides L., a hummingbird‐pollinated Euphorbiaceae

Floral sexual phases can differ in nectar production and might be under selective pressure by pollinators. We studied Euphorbia tithymaloides, which has inflorescences that are initially female and then hermaphroditic. Volume and concentration of nectar were measured in both stages. Nectar production and the effect of extractions were determined using sets of bagged inflorescences; inflorescences in the hermaphroditic phase had higher values of nectar concentration, volume and sugar mass than inflorescences in the female phase. Nectar resorption was detected in senescent inflorescences. To test for homeostatic nectar regulation, artificial nectar was added and the response assessed after 24 h. The experiments showed that concentration and sugar mass are regulated within a narrow range, and the homeostatic points differ between the two sexual phases. These differences in nectar can be detected by hummingbirds, which prefer the female stage. Resorption and secretion seem to be part of a homeostatic mechanism by which nectar attributes are maintained to optimise sugar recovery.     

A cell surface integral membrane glycoprotein of 85,000 mol wt (gp85) associated with triton X-100-insoluble cell skeleton

The Triton X-100-insoluble skeleton of baby hamster kidney BHK cells consists of the nucleus, intermediate-size filaments, and actin fibers. By transmission electron microscopy, membrane fragments were found to be associated with these insoluble structures. When radioiodinated or [3H]glucosamine-labeled cells were extracted with 0.5% Triton, most plasma membrane glycoproteins were solubilized except for a glycoprotein with a molecular weight of 85,000 (gp85) that remained associated with the insoluble skeletons. Immunoprecipitation with a specific antiserum indicated that the gp85 is not a proteolytic degradation product of fibronectin, an extracellular matrix glycoprotein insoluble in detergent. A monoclonal antibody of BHK cells specific for gp85 was produced. Immunofluorescence analysis with this monoclonal antibody indicated that gp85 is not associated with the extracellular matrix, but is confined to the cell membrane. Both in fixed and unfixed intact cells, fluorescence was concentrated in dots preferentially aligned in streaks on the cell surface. Gp85 was found to behave as an integral membrane protein interacting with the hydrophobic core of the lipid bilayer since it was extracted from membrane preparations by ionic detergents such as SDS, but not by 0.1 N NaOH (pH 12) in the absence of detergents, a condition known to release peripheral molecules. Association of gp85 with the cell skeleton was unaffected by increasing the Triton concentration up to 5%, but it was affected when actin filaments were dissociated or when a protein-denaturing agent (6 M urea) was used in the presence of Triton, suggesting that protein-protein interactions are involved in the association of gp85 with the cell skeleton. We conclude that gp85 is an integral plasma membrane glycoprotein that might have a role in cell surface-cytoskeleton interaction.     

Influence of secondary dispersal by ants on invasive processes of exotic species with fleshy fruits

Biological Invasions - Invasive alien plant species (IAPS) are severely changing ecosystems on earth. Studying the interactions that allow IAPS to establish and spread in the new regions is...     

Increased expression of HSP70 by colon cancer cells is not always associated with access to the dendritic cell cross-presentation pathway

Dendritic cells (DCs) are highly specialized antigen-presenting cells endowed with the unique ability to not only present exogenous antigens upon exposure to MHC II, but also to cross-present these upon exposure to MHC I. This property was exploited to generate the tumor-specific CD8 cytotoxic lymphocyte (CTL) response in DCs-based cancer vaccine protocols. In this context, the source of tumor antigens remains a critical challenge. A crude tumor in the context of danger signals is believed to represent an efficient source of tumor antigens (TAs) for DCs loading. In our previous work, increased DCs cross-presentation of antigens from necrotic gastric carcinoma cells paralleled up-regulation of the heat shock protein hsp70. We studied the expression of hsp70 on primary colon carcinoma cells and its relevance in the cross-priming of anti-tumor CTL by tumor-loaded DCs. Hsp70 was expressed on all three of the tumors studied, but was never detected in the peritumoral normal mucosa (NM). The uptake of the tumor induced a trend towards down-modulation of the monocyte-specific marker CD14, but had no effect on the chemokine receptors CCR4 and CCR7. The IFN-γ enzyme-linked immunospot assay (ELIspot) showed cross-priming of CTL by tumor-loaded but not NM-loaded DCs in four of the six cases studied. The CTL response generated in DC+tumor cultures was directed towards the tumor, but not towards NM, and it was characterized by refractoriness to polyclonal (Ca ionophores, PKC activators) stimuli. Of the three CTL-generating tumors, only one expressed hsp70. This data indicates a tumor-specific expression of hsp70, but does not support its relevance in the DC cross-presentation of TAs.