Microsatellites are the most common markers used in population and conservation genetic studies. However, their isolation is laborious and expensive. In some taxa, such as Lepidoptera, it is particularly difficult to isolate microsatellite markers due to the high similarity of the flanking regions of different loci and the presence of null alleles. Here we isolated microsatellites of the endangered butterfly Boloria aquilonaris using 454 GS-FLX Titanium pyro-sequences of biotin enriched DNA libraries and tested the success of cross-amplification on the sister-species B. eunomia. Fifteen polymorphic microsatellite loci were isolated in B. aquilonaris using initially 101 stringently designed primer pairs. Unlike in many other studies of microsatellite isolation in Lepidoptera, few null alleles were detected and only at very low frequencies. Additionally, the raw data set can still be used for the isolation of other microsatellite loci. None of the selected polymorphic loci for B. aquilonaris gave clear banding patterns for B. eunomia, although about 15 other loci gave promising banding patterns for the latter species. Low intra- and inter-specific transferability of developed markers in this study also lends support to the hypothesis that the evolution of the genome of Lepidoptera is dissimilar from that of other organisms., Sofie Vandewoestijne, Camille Turlure, Michel Baguette., and Obsahuje seznam literatury
The use of light traps for controlling insect pests is restricted since they kill both pests and beneficial insects. It may be a possible to reduce the numbers of beneficial insects trapped by adjusting nightly trapping time based on differences recorded in the timing of the nocturnal flight peaks of target pests and beneficials. To test this, insects were collected hourly over night using black light traps at three locations in China from 2003 to 2005. Groups of lepidopteran and coleopteran pests were selected as the target pests that we would control by trapping and groups of beneficial predatory insects the catches of which needed to be reduced. The highest numbers of Coleoptera were caught between 20:00 and 22:00 h and of most Lepidoptera between 02:00 and 04:00 h. The hourly numbers of predatory insects caught by light traps were evenly distributed throughout the night. A model was developed to describe the relationships between the cumulative proportions of insects caught and time of night. The model accurately describes the flight activity of insects that were mainly caught before midnight, after midnight and evenly throughout a night by using different parameters for the three different insect groups. A beneficial-friendly trapping strategy was developed to reduce the numbers of beneficial insects trapped, which was based on differences in the nocturnal flight activity of pests and beneficial insects and validated by a field study in Shandong province. Results show that this trapping strategy reduced the number of beneficial insects caught by 46% and the electricity consumption by 50% compared to the traditional strategy. Thus this strategy is more beneficial-friendly than the traditional trapping strategy for controlling pests., Gang Ma, Chun-Sen Ma., and Obsahuje seznam literatury
Although the effects of host plant quality on the performance of polyphagous herbivores are largely uniform across insect taxa, there are various exceptions to this rule. In particular, there are scattered reports of cases in which the relative quality of different hosts differs among larval instars of a single insect species. Such cases are explained either in terms of differences in the susceptibility of different aged larvae to plant defences or, alternatively, age-specific nutritional demands. Here we report the results of experiments that show that young larvae of the polyphagous common heath moth Ematurga atomaria (Lepidoptera: Geometridae) consistently attain higher weights on common heather Calluna vulgaris than bilberry Vaccinium myrtillus, whereas the rank order of these host plants is reversed in the final larval instar. Phytochemical analyses showed that differences in nutrient content of these plants are not likely to explain the observed pattern. Instead, the results are more consistent with the idea that the greater chemical defence of bilberry has a relatively stronger influence on young than old larvae., Helen Vellau, Siiri-Lii Sandre, Toomas Tammaru., and Seznam literatury
a1_There is little systematic research on the effect of transgenic maize expressing Bacillus thuringiensis (Bt) toxins on insect survival and the response in vivo of their detoxification enzymes. Results of laboratory bioassays indicate that the whorl leaves of transgenic maize lines G03-2396 and G03-2739 significantly affected the survival of neonate and third instar larvae of the Asian corn borer, Ostrinia furnacalis (Güenée). Neonate mortality two days after being fed on G03-2739 was 72.5% and exceeded 92.5% four days later. The percentage mortality of third instar larvae six days after being fed whorl leaves from G03-2396 exceeded 65%. The resistance of whorl leaves to insect attack was greater than that of maize-ears, but less effective against attacks by third instar larvae. Neonate development was more prolonged when they were fed on whorl leaves of both the transgenic maize lines. In contrast, low pupation (<80% of that recorded in the controls), ecolosion and fecundity were recorded following ingestion of maize-ears, with pupal weights 10-14 mg lower than that of controls. The growth rates of third instar larvae recorded three and six days after feeding on whorl leaves were lower, but food utilization, conversion and relative metabolism were not significantly affected. Relative food ingestion, relative growth rate and other nutritional indicators of third instar larvae were significantly lower six days after the ingestion of ears and that of fifth instar larvae after three days. In vivo activity of carboxylesterase was decreased to a greater degree after three days when the larvae were fed on whorl leaves than ears, with no significant effect on fifth instar larvae. Furthermore, in vivo activity of glutathione S-transferase (GST) of third instar larvae was significantly affected following the ingestion of whorl leaves. These results are discussed in the context of the literature on the resistance, a2_of transgenic plants and of improving the resistance of plants to attack by the different larval stages of insects., and Min Juan Shi, Pei Ling Lu, Xiao Li Shi, Yi Zhong Yang.
Transgenic lines of silver birch (Betula pendula) carrying the sugar beet chitinase IV gene were used to study the effects of the heterologous expression of a transgenic chitinase on the performance of lepidopteran herbivores. The effect of wounding the leaves of birch on the performance of lepidopteran larvae and the growth of trees was also studied. Larvae of Orgyia antiqua L., Lymantriidae, and Phalera bucephala L., Notodontidae, were separately fed on the leaves of transgenic and wild-type birch, and their performance measured using nutritional indices. The relative growth rate (RGR) of O. antiqua larvae fed transgenic leaves was significantly lower than that of larvae fed wild-type leaves. Furthermore, there is little evidence that transgenic chitinase affects survival but it was lowest for the group of larvae fed leaves with the highest expression of chitinase IV. Wounding did not have a significant effect on the performance of the larvae or on the growth of the branches of the trees. The growth of branches of particular transgenic lines, however, was significantly associated with tree line. The performance of P. bucephala larvae fed leaves of transgenic and wild-type birches did not differ. The leaves used in both experiments from transgenic trees were shorter than those from wild-type trees. Using transgenic birch expressing sugar beet chitinase IV to improve the resistance of birch to fungal diseases can have negative effects on O. antiqua larvae feeding on the leaves of these birches. P. bucephala, however, was not similarly affected, which indicates that these two ecologically similar lepidopteran species may differ in their response to transgenic chitinase., Liisa Vihervuori ... [et al.]., and Obsahuje seznam literatury
In order to determine when during the development of H. armigera CYP6B6 is expressed, we measured the width of the head capsule, the length of the body and weight of the different life stages of the cotton bollworm reared under laboratory condition. The larvae took about 20 d to complete their development under these conditions, the pre-pupae about 4 d, pupae about 9 d, and the adults lived for about 9 d and the eggs took about 4 d to hatch. Knowing this it was possible to determine the pattern of cytochrome P450 CYP6B6 expression at different stages in the life cycle of H. armigera using real-time quantitative polymerase chain reaction (RT-qPCR). The CYP6B6 gene was expressed in all the developmental stages of the cotton bollworm. The results indicate that the level of expression was lower in the egg, pre-pupal, pupal and adult stages than in larvae. The level of expression of CYP6B6 decreased from the 1st to the 3rd instar larva, with the lowest level recorded in the 3rd instar larvae and then increased reaching its highest value in 6th instar larvae. A similar trend in the expression of CYP6B6 was detected using immunohistochemistry. As the results show, more efficient control of the cotton bollworm can be achieved if insecticides are applied when the larvae of H. armiger are in the 3rd instar, because at that stage the eat little food and there is a low level of detoxifying enzymes in the larvae., Xuetao Zhang ... [et al.]., and Obsahuje seznam literatury
The aim of this study was to investigate whether haemocytes of Galleria mellonella (Lepidoptera: Pyralidae) larvae produce reactive oxygen species (ROS) like human blood phagocytes. The production of ROS was measured first using luminol-enhanced chemiluminescence of un-stimulated and stimulated (four activators with different modes of action) haemolymph or isolated haemocytes. However, spontaneous and activated production of ROS remained at the background level. In subsequent experiments an ultrasensitive fluorescence method using Amplex Red reagent to detect hydrogen peroxide (H2O2) was used. After optimization, Amplex Red was successfully used for determining H2O2 production by both un-stimulated and stimulated haemocytes. To determine the affect of pH and ions on the measurement, several diluent solutions were tested. This revealed that Ca2+ and Mg2+ ions are less important for the reaction in insect than mammalian cells. Among the activators tested, phorbol myristate acetate (PMA) and calcium ionophore (Ca-I) had the best stimulatory effect on insect samples, while opsonised zymosan particles (OZP) was the best activator for human phagocytes. In conclusion, the haemocytes of G. mellonella produce H2O2 as an important innate immunity factor, but under different conditions and in different amounts, which probably results in them being less effective in killing microbes than human phagocytes. and Ondřej Vašíček, Ivana Papežíková, Pavel Hyršl.
Silk production quantity is the most economically important characteristic of the domesticated silkworm moth, Bombyx mori. It is controlled by multiple loci. The cocoon and silk production quantity of silkworm strains Jingsong and Lan10 have significantly diverged. A backcross population (BC1) was bred using Jingsong and Lan10 as parents to identify quantitative trait loci (QTLs) for silk quality. In this research, a genetic linkage map of the silkworm was constructed using the BC1 mapping population. The map contained 85 sequence-tagged site markers, 80 simple sequence repeat markers, and 16 single nucleotide polymorphisms. A linkage map was constructed from the data, which consisted of 181 markers distributed over 28 expected linkage groups and spans 2147.1 cM in total length. Fourteen QTLs were detected for cocoon filament length, whole cocoon weight, pupae weight, filament weight, and cocoon shell weight. The 14 QTLs were distributed in 5 linkage groups (linkage groups 1, 14, 18, 23 and 25) based on the constructed linkage map. In addition, five QTLs, which had the highest logarithm (base 10) of odds (LOD) values, were located on the first chromosome, three of which located at the same region in linkage group 1. These results represent an important foundation for the map-based cloning of QTLs and marker-assisted selection for improving the silk quality of economically important silkworm strains., Bing Li ... [et al.]., and Obsahuje seznam literatury
a1_Cities contain only a low representation of natural and semi-natural habitats, existing in fragments surrounded by built-up areas. In 2003-2004, we surveyed butterflies and Zygaenidae moths in 21 reserves and 4 parks within the city of Prague, Czech Republic, situated from the periphery to city centre. A total of 85 species (47% of the Czech fauna of the study groups) was detected, 22 of them being of conservation concern. Ordination analyses of the local assemblages revealed that the richest sites were large, situated far from the city centre, on alkaline bedrocks, south- to southwest oriented, and hosting high numbers of vegetation types and vascular plant species. We then used generalised linear models to fit responses of individual species to the main ordination gradient, corresponding to increasing urbanisation. Out of 60 species that met criteria for the modelling, none responded positively to urbanisation. Twenty displayed negative linear response; these urban avoiders contained a surplus of mesophilous species presumably preferring rural landscapes. Further 29 species (suburban adaptable) responded in domed manners, peaking at the city periphery. Prevailing among them were xerophilous specialists inhabiting large grassland reserves at the Prague outskirts. Finally, eleven urban tolerant species did not respond to urbanisation at all, containing three highly mobile species, three xerothermophilous specialists, and five species utilising shrubs or trees and finding suitable conditions even in urban parks. Suburban adaptable butterflies apparently benefit from such suburban environments as gardens, road or railway verges, collectively increasing the connectivity of remnants of seminatural habitats. The fact that arboreal species persist even in urban parks, whereas common grassland species are absent there, implies that the quality of urban habitats might be increased by a more sensitive management of, a2_urban green spaces, such as leaving aside small temporary fallows or adapting lawns mowing schemes., and Martin Konvicka, Tomas Kadlec.
Which factors influence the occurrence, population size and density of species in fragmented habitat patches are key questions in population and conservation ecology. Metapopulation theory predicts that larger and less isolated habitat patches should positively influence species occurrence and population size. However, recent studies have shown that habitat quality, human activity and permeability of the landscape surrounding habitat patches may be also important. In this paper we test the relative effects of habitat patch characteristics, human settlement and landscape permeability on the occurrence, local population size and density of the Chalk-hill Blue Polyommatus coridon a charismatic butterfly inhabiting calcareous grasslands in a fragmented landscape in southern Poland. Patch occupancy rate (corrected for the butterfly detection probability) was 0.45. Habitat patch area, proximity of human settlement and cover of larval food plants positively affected occurrence of the Chalk-hill Blue. Local population size of the Chalk-hill Blue was positively affected by patch area and proximity of human settlement, and negatively by patch isolation. Local density was higher in patches located close to human settlement. Our study is one of the few showing a positive effect of human settlement on a grassland specialist butterfly although the mechanism hidden behind this phenomenon is unknown and requires further examination. In order to maintain local populations of the Chalk-hill Blue in southern Poland, conservation action should be focused on large, closely connected calcareous grasslands. Moreover, extensive management of this habitat should be maintained by local inhabitants who are better placed to undertake such work. and Zuzanna M. ROSIN, Piotr SKÓRKA, Magdalena LENDA, Dawid MORON, Tim H. SPARKS, Piotr TRYJANOWSKI.