Determination of double bond position in higher hydrocarbons

Hydrocarbons including alkenes are in addition to wax esteres, alcohols, ketones and acids one of the main compounds of insect cuticle. Hydrocarbons protect insect body against dry, water and UV radiation. In some species they play an important role in mutual communicative and recognition. Hydrocarbons profile can in some cases serve as fingerprint defining species specificity. Therefore, the position of double bonds in cuticular hydrocarbons is important. This work describes utilization of catalytic hydroxylation with polymer-supported osmium tetroxide as a derivatization technique to determine double bond position. Reaction conditions of hydroxylation and mass spectrometric fragmentation of derivatized alkenes were optimized for standard containing one double bond. These conditions were applied to real samples from the american cockroach Periplaneta americana and blowfly Neobellieria bullata.Uhlovodíky, včetně alkenů jsou vedle voskových esterů, alkoholů, ketonů a kyselin jednou z hlavních sloučenin hmyzí kutikuly. Jejich praktický význam spočívá především v ochraně hmyzího těla před vysoušením, vodou a UV zářením. U vybraných druhů hmyzu plní funkci chemické komunikace a druhového rozpoznávání. Profil uhlovodíků může rovněž v některých případech sloužit jako tzv. otisk prstu definující specifitu druhu. Proto hraje poloha dvojné vazby alkenů obsažených ve svrchní části hmyzího těla důležitou roli. Tato práce popisuje využití katalytické hydroxylace oxidem osmičelým vázaným na polymerním nosiči, jako derivatizační metodu k určení polohy dvojné vazby. Byly optimalizovány reakční podmínky derivatizace na standardu obsahujícím jednu dvojnou vazbu a vzniklý derivát byl podroben fragmentačním experimentům. Optimální podmínky derivatizace a fragmentací nenasycených uhlovodíků byly aplikovány na reálných vzorcích ze švába amerického Periplaneta americana a masařky Neobellieria bullata.Department of Analytical ChemistryKatedra analytické chemieFaculty of SciencePřírodovědecká fakult

Experimental loss of generalist plants reveals alterations in plant-pollinator interactions and a constrained flexibility of foraging

Species extinctions undermine ecosystem functioning, with the loss of a small subset of functionally important species having a disproportionate impact. However, little is known about the effects of species loss on plant-pollinator interactions. We addressed this issue in a field experiment by removing the plant species with the highest visitation frequency, then measuring the impact of plant removal on flower visitation, pollinator effectiveness and insect foraging in several sites. Our results show that total visitation decreased exponentially after removing 1–4 most visited plants, suggesting that these plants could benefit co-occurring ones by maintaining high flower visitor abundances. Although we found large variation among plant species, the redistribution of the pollinator guild affected mostly the other plants with high visitor richness. Also, the plant traits mediated the effect of removal on flower visitation; while visitation of plants which had smaller inflorescences and more sugar per flower increased after removal, flower visitors did not switch between flower shapes and visitation decreased mostly in plants visited by many morpho-species of flower visitors. Together, these results suggest that the potential adaptive foraging was constrained by flower traits. Moreover, pollinator effectiveness fluctuated but was not directly linked to changes of flower visitation. In conclusion, it seems that the loss of generalist plants alters plant-pollinator interactions by decreasing pollinator abundance with implications for pollination and insect foraging. Therefore, generalist plants have high conservation value because they sustain the complex pattern of plant-pollinator interactions

Minority cytotypes in European populations of the Gymnadenia conopsea complex (Orchidaceae) greatly increase intraspecific and intrapopulation diversity

Background and Aims Patterns of ploidy variation among and within populations can provide valuable insights into the evolutionary mechanisms shaping the dynamics of plant systems showing ploidy diversity. Whereas data on majority ploidies are, by definition, often sufficiently extensive, much less is known about the incidence and evolutionary role of minority cytotypes. Methods Ploidy and proportions of endoreplicated genome were determined using DAPI (4',6-diamidino-2-phenylindole) flow cytometry in 6150 Gymnadenia plants (fragrant orchids) collected from 141 populations in 17 European countries. All widely recognized European species, and several taxa of less certain taxonomic status were sampled within Gymnadenia conopsea sensu lato. Key Results Most Gymnadenia populations were taxonomically and/or ploidy heterogeneous. Two majority (2x and 4x) and three minority (3x, 5x and 6x) cytotypes were identified. Evolution largely proceeded at the diploid level, whereas tetraploids were much more geographically and taxonomically restricted. Although minority ploidies constituted <2 % of the individuals sampled, they were found in 35 % of populations across the entire area investigated. The amount of nuclear DNA, together with the level of progressively partial endoreplication, separated all Gymnadenia species currently widely recognized in Europe. Conclusions Despite their low frequency, minority cytotypes substantially increase intraspecific and intrapopulation ploidy diversity estimates for fragrant orchids. The cytogenetic structure of Gymnadenia populations is remarkably dynamic and shaped by multiple evolutionary mechanisms, including both the ongoing production of unreduced gametes and heteroploid hybridization. Overall, it is likely that the level of ploidy heterogeneity experienced by most plant species/populations is currently underestimated; intensive sampling is necessary to obtain a holistic pictur

Interactions with mycorrhizal fungi in two closely related hybridizing orchid species

The nuclear ribosomal DNA was used to identify the orchid mycorrhizal fungi found in roots of Orchis xbivonae and its parental species Orchis anthropophora and Orchis italica. Polymerase chain reaction products were sequenced and identified using the expanded database.We determined that closely related Tulasnellaceae are mycorrhizal in the three orchid taxa, suggesting that the mycorrhizal partner does not impair hybrid survival. This study demonstrates that O. xbivonae displays few differences in comparison with its two parental species in identity of its associated mycorrhizal fungi, it is a short- -term by-product of the hybridizing behavior of common pollinators, and thus it will not easily origin descendents with potential new genetic combinations and/or ecological preferences

Genetic differentiation and admixture between sibling allopolyploids in the Dactylorhiza majalis complex

Allopolyploidization often happens recurrently, but the evolutionary significance of its iterative nature is not yet fully understood. Of particular interest are the gene flow dynamics and the mechanisms that allow young sibling polyploids to remain distinct while sharing the same ploidy, heritage and overlapping distribution areas. By using eight highly variable nuclear microsatellites, newly reported here, we investigate the patterns of divergence and gene flow between 386 polyploid and 42 diploid individuals, representing the sibling allopolyploids Dactylorhiza majalis s.s. and D. traunsteineri s.l. and their parents at localities across Europe. We make use in our inference of the distinct distribution ranges of the polyploids, including areas in which they are sympatric (that is, the Alps) or allopatric (for example, Pyrenees with D. majalis only and Britain with D. traunsteineri only). Our results show a phylogeographic signal, but no clear genetic differentiation between the allopolyploids, despite the visible phenotypic divergence between them. The results indicate that gene flow between sibling Dactylorhiza allopolyploids is frequent in sympatry, with potential implications for the genetic patterns across their entire distribution range. Limited interploidal introgression is also evidenced, in particular between D. incarnata and D. traunsteineri. Altogether the allopolyploid genomes appear to be porous for introgression from related diploids and polyploids. We conclude that the observed phenotypic divergence between D. majalis and D. traunsteineri is maintained by strong divergent selection on specific genomic areas with strong penetrance, but which are short enough to remain undetected by genotyping dispersed neutral markers.UE FWF; P22260UE: Y66

The mirror crack'd: both pigment and structure contribute to the glossy blue appearance of the mirror orchid, Ophrys speculum.

The Mediterranean orchid genus Ophrys is remarkable for its pseudocopulatory pollination mechanism; naïve male pollinators are attracted to the flowers by olfactory, visual and tactile cues. The most striking visual cue is a highly reflective, blue speculum region at the centre of the labellum, which mimics the corresponding female insect and reaches its strongest development in the mirror orchid, O. speculum. We explored the structure and properties of the much-discussed speculum by scanning and transmission electron microscopic examination of its ultrastructure, visible and ultraviolet (UV) angle-resolved spectrophotometry of the intact tissue, and mass spectrometry of extracted pigments. The speculum contrasts with the surrounding labellar epidermis in being flat-celled with a thick, smooth cuticle. The speculum is extremely glossy, reflecting intense white light in a specular direction, but at more oblique angles it predominantly reflects blue and UV light. Pigments in the speculum, dominantly the cyanidin 3-(3''-malonylglucoside), are less diverse than in the surrounding regions of the labellar epidermis and lack quercetin copigments. Several physical and biochemical processes interact to produce the striking and much-discussed optical effects in these flowers, but the blue colour is not produced by structural means and is not iridescent

Are food-deceptive orchid species really functionally specialized for pollinators?

Food-deceptive orchid species have traditionally been considered pollination specialized to bees or butterflies. However, it is unclear to which concept of specialization this assumption is related; if to that of phenotypic specialization or of functional specialization. The main aim of this work was to verify if pollinators of five widespread food-deceptive orchid species (Anacamptis morio (L.) R.M. Bateman, Pridgeon & M.W. Chase, Anacamptis pyramidalis (L.) Rich., Himantoglossum adriaticum H. Baumann, Orchis purpurea Huds. and Orchis simia Lam.) predicted from the phenotypic point of view matched with the observed ones. We addressed the question by defining target orchids phenotypic specialization on the basis of their floral traits, and we compared the expected guilds of pollinators with the observed ones. Target orchid pollinators were collected by conducting a meta-analysis of the available literature and adding unpublished field observations, carried out in temperate dry grasslands in NE Italy. Pollinator species were subsequently grouped into guilds and differences in the guild spectra among orchid species grouped according to their phenotype were tested. In contradiction to expectations derived from the phenotypic point of view, food-deceptive orchid species were found to be highly functionally generalized for pollinators, and no differences in the pollinator guild spectra could be revealed among orchid groups. Our results may lead to reconsider food-deceptive orchid pollination ecology by revaluating the traditional equation orchid-pollination specialization