DNA barcoding surveys of small insects usually extract DNA from either a complete insect or a leg. Little is known about how to optimize DNA quantity and quality from different insect parts while preserving a morphological voucher. Here, we quantify DNA yield from different body parts (antenna, hind leg, forewing, hind wing and abdomen) of the micro-moth Cameraria ohridella (Lepidoptera: Gracillariidae) using fluorescent nucleic acid stain (PicoGreen). Samples were preserved in 100% ethanol or dried for three weeks. Our experiment was designed to encompass practical sampling options during fieldwork. DNA quality was assessed by PCR amplification of the mitochondrial COI barcode fragment. In addition, we compared PCR amplification using Platinum® Taq and Qiagen DNA Polymerase and quantified sequence success of amplified DNA. We show that overall, dry parts showed higher eluted DNA yields. PCR and sequencing success rate were slightly higher for dry tissue than ethanol-preserved parts. We also show that Platinum® Taq yielded the highest PCR success rate and that all dry tissues are sequenceable. The optimal strategy for DNA barcoding surveys is therefore to mount micro-Lepidoptera specimens in the field for morphological analysis and sample tissues (hind legs are favoured) from dried samples at a later time (several weeks) in the lab for DNA barcoding using preferentially Platinum® Taq. If larger amounts of DNA are required (i.e. for nuclear gene sequencing), several legs from one side of the specimen or the abdomen should be preserved in pure ethanol., Carlos Lopez-Vaamonde ... [et al.]., and Obsahuje seznam literatury
This study of the proteins in the silk of the summer and winter cocoons of the horse chestnut leaf miner Cameraria ohridella revealed they can inhibit protease activity. The inhibitory activity of the summer silk was higher against both the fungal proteinase K and bacterial subtilisin than that of the winter silk. Also, the winter silk was more effective in inhibiting proteinase K than subtilisin. Further, it was demonstrated that some of the silk proteins are glycosylated probably by mannose carbohydrates. An electron microscopy study of the cocoons revealed the presence of silk fibres with different diameters in some pupal chambers. and Veronika Hněvsová, Dalibor Kodrík, František Weyda.
Poor synchronisation is considered to be one important reason for the ineffective control of the invasive horse chestnut leafminer by native parasitic Hymenoptera. Parasitoids hibernating in dry horse chestnut leaves break diapause early in spring and presumably leave the vicinity, since no hosts are available when they emerge. As a consequence, the percentage parasitism of the first generation of the leafminer in early summer is low. The experiments presented below were designed to test this hypothesis. Horse chestnut saplings were brought on in a greenhouse and infested artificially with C. ohridella prior to parasitoid emergence in the field. These saplings were then exposed to parasitoid attack under natural conditions to eliminate the synchronisation problem. In addition, the parasitoid complexes of other leafmining hosts, which appear early in the season, were analysed. The results confirm that the most important parasitoids of the horse chestnut leafminer are active early in the season, long before the larvae of the first generation of the host are present. Nevertheless, poor synchronisation with the invasive host did not significantly influence the abundance of the most important parasitoid in the complex, Minotetrastichus frontalis, and consequently had no impact on the parasitism of C. ohridella. Nevertheless, a detailed analysis of the parasitoid community shows that certain species are affected by poor synchronisation, while others are probably limited by ecological parameters, such as a specialisation to foraging in particular strata of vegetation. Several leafmining flies were found early in spring infesting other host plants commonly planted in urban green areas, including species with a parasitoid complex similar to that of C. ohridella. The role of these alternative hosts in the food web associated with the horse chestnut leafminer should be subject to further study.
A set of six polymorphic microsatellite markers was developed for the highly invasive horse chestnut leaf-mining moth, Cameraria ohridella. For 96 positive clones, six polymorphic microsatellites with conserved flanking regions were identified. The average number of alleles per locus was eight (range from three to 12). Two of these loci showed significant heterozygosity deficits due to null alleles. The remaining four loci did not depart from Hardy-Weinberg equilibrium. In addition, all six loci were amplified for 20 other gracillariid species belonging to eight different genera, including another invasive species Phyllonorycter platani. These are the first microsatellite markers described for a species within the lepidopteran family Gracillariidae.
Horse chestnut leaf miner (Cameraria ohridella) has achieved ecological success by colonizing the entire European range of its primary host, horse chestnut (Aesculus hippocastanum). This insect has attracted the attention of scientists, but its ecology is poorly understood. Here, we investigated the effects of varying degrees of light availability on the leaf morphology of horse chestnut saplings and the performance of C. ohridella. A pot experiment under greenhouse conditions was performed in which the photosynthetic photon flux density (PPFD) was reduced from full light by 50% (high light - HL) or 80% (low light - LL). Insect performance parameters were quantified (i.e., pupal mass, efficiency of conversion of utilised leaf tissue (ECU) and potential fecundity). Compared with HL leaflets those from LL were characterised by higher contents of nitrogen and water but lower total phenolics. The oxidative capacity of phenolics (at pH ≈ 10, common in the lepidopteran gut) was low and did not differ in the two treatments. Compared with those collected from HL leaves, the mines of those collected from leaves of plants grown under LL conditions were larger in area but the leaf mass utilized by larvae was similar. Pupae were heavier in LL than in HL conditions, and ECU was higher in LL. The potential fecundity of females was not sensitive for experimental treatment. We conclude that (1) reduced light had a strong beneficial effect on the performance of C. ohridella and (2) phenolics in A. hippocastanum leaf tissues are a poor defence against this herbivore.
The horse chestnut leaf miner, Cameraria ohridella Deschka & Dimić, is a species of unknown origin that recently invaded most of Europe, causing serious damage to horse chestnut trees, Aesculus hippocastanum. Parasitism was studied over a period of three years in the region of Plzeň in the Czech Republic. Additional collections were made in Slovakia and Slovenia. The parasitoid complex, dominated by polyphagous idiobiont parasitoids of the family Eulophidae, is similar to that found in other studies in Europe. Minotetrastichus frontalis (Nees) was the most abundant parasitoid found, except in Slovakia where Pediobius saulius (Walker) dominated. One parasitoid species, the eulophid Cirrospilus diallus (Walker) was recorded for the first time from C. ohridella. A new method is proposed to calculate stage-specific and total parasitism rates. Parasitism rates of spinning larvae and pupae were higher than of feeding larvae; however, total parasitism was low. We estimated that between 1% and 17% of moths died from parasitism during the larval and pupal stages whereas the rate of mortality caused by other factors varied from 7% to 62%, depending on the locality, year and generation. The proposed method for calculating total parasitism, based on the integration of stage-specific parasitism rates, which takes into account the mortality not directly attributed to parasitism, is discussed and compared with other methods commonly used in studies on C. ohridella.
Horse chestnut trees in many regions of Europe have suffered from epidemic infestations of C. ohridella for more than ten years. There has been no obvious decrease in the infestation level anywhere on the continent. One reason is, that the native natural enemies have not been able to control mass outbreaks of the leafminer. Parasitoid Hymenoptera have very little impact on the first generation of the moth in early summer, regardless of the number of parasitoids that overwintered in horse chestnut leaves. This study revealed that there is a considerable time lag between the emergence of the parasitoids from the leaflitter in spring and the appearance of suitable host instars in early summer. The poor control of C. ohridella by natural enemies may partly be due to the poor synchronisation between the life cycles of the introduced host and native parasitoid wasps.
Performance of the invasive horse-chestnut leaf miner, Cameraria ohridella Deschka & Dimic, 1986 (Lepidoptera: Gracillariidae), was studied on two host plants: the white-flowering horse-chestnut Aesculus hippocastanum L. and the Ohio buckeye Aesculus glabra Willd. C. ohridella developed successfully on both host plants; however, mine density and survival were much higher on A. hippocastanum than on A. glabra. The pupal mass and potential fecundity were strongly affected by the host plant on which the larvae fed. On A. hippocastanum pupae were significantly heavier and females more fecund than those on A. glabra. Furthermore, on both host plants there was a significant positive correlation between the number of oocytes in ovaries and pupal body mass, and as a consequence, heavier females produced more eggs. Our study demonstrates that the mine density, survival, pupal mass and potential fecundity were significantly lower on A. glabra than on A. hippocastanum. The observed lower performance of C. ohridella on the exotic host plant was assumed to be due to its poor food quality (nutritional and chemical composition)., Urszula Walczak, Edward Baraniak, Piotr Zduniak., and Obsahuje bibliografii