A thioredoxin-like protein (txl) gene was cloned from the bumblebee, Bombus ignitus. The B. ignitus txl (Bitxl) gene spans 1777 bp and consists of three introns and four exons coding for 285 amino acid residues with a conserved active site (CGPC). The deduced amino acid sequence of the Bitxl cDNA was 65% similar to the Drosophila melanogaster txl. Northern blot analysis revealed the presence of Bitxl transcripts in all tissues examined. When H2O2 was injected into the body cavity of B. ignitus workers, Bitxl mRNA expression was up-regulated in the fat body tissue. In addition, the expression levels of Bitxl mRNA in the fat body greatly increased when B. ignitus workers were exposed to low (4°C) or high (37°C) temperatures, or injected with lipopolysaccharide (LPS), which suggests that the Bitxl possibly protects against oxidative stress caused by extreme temperatures and bacterial infection.
A serine protease gene was cloned from the bumblebee, Bombus ignitus. The B. ignitus serine protease (BiSP) gene spans 1702 bp and consists of four introns and five exons coding for 250 amino acid residues. Southern blot analysis of genomic DNA suggested that BiSP gene is a single copy gene. The cDNA encoding BiSP was expressed as a 28-kDa polypeptide in baculovirus-infected insect cells and the recombinant BiSP showed activity in a protease enzyme assay. BiSP was specifically expressed in the midgut of B. ignitus queens, males, and workers, suggesting that the BiSP is a gut enzyme involved in the digestion of dietary proteins.
A global decline in pollinator abundance and diversity has demanded increased research attention to the ecology and genetics of bumblebees. However, as progressively more restrictions are placed on sampling for insects, researchers are increasingly obliged to use archival specimens collected for purposes other than genetic analyses. In this study we assessed the suitability, for population genetic studies, of popular, low-cost methods for preservation and storage of bumblebee specimens. Specimens of Bombus terrestris L. were held under six storage regimes for up to two years. DNA was extracted from the samples using three extraction protocols and the quality of the DNA was examined using PCR amplification of a mitochondrial and a nuclear gene. All extraction and storage methods provided sufficient DNA for successful PCR amplification. However, samples preserved in acetone or at freezing temperatures yielded the highest DNA concentrations. DNA yields from pinned specimens at room temperature declined over time, particularly when using standard extraction techniques. DNA concentrations were significantly lower from specimens preserved in 70% ethanol compared to all other extraction techniques and declined linearly over the two years of storage. These results indicate that two of the most popular insect storage methods (pinning and storage in ethanol) should be avoided for the long-term preservation of genetic material for future studies. We suggest that optimal insect preservation methods should be incorporated into research protocols in order to best capitalise on limited collection opportunities., António S. Moreira ... []., and Obsahuje seznam literatury
Labial gland secretions of 26 males of the bumblebee Bombus lucorum (L.), collected in the Czech Republic, were analysed. The secretions consisted of 60 compounds; ethyl (Z)-9-tetradecenoate was the main component (average 53%). Although the males varied in colour, their labial gland secretions were similar in composition, which indicated they belonged to one species. Chemically the B. lucorum occurring in the Czech Republic correspond to the earlier described "blonde form" of this species.
A fragment encoding melittin cDNA from Apis cerana cerana fused with glutathione S-transferase gene was inserted into the multiple cloning site of the pBacFastHTb to construct a recombinant donor plasmid, pBacHT-GSTAccM, which was transposed to the target bacmid in E. coli (DH10) by Tn7 transposition function. Then the recombinant baculovirus Bacmid-GSTAccM was transfected into Tn-5B1-4 cells of the cabbage looper, Trichoplusia ni, mediated by lipofectin. The expressed protein of about 34 kDa was detected by Western blotting and triple antibody sandwich ELISA, indicating that the recombinant protein is the fusion protein of GSTAccM. Thin layer scanning showed that the expression level of GSTAccM was about 7% of the total cell protein. Purified and recovered recombinant melittin of A. c. cerana showed bioactivity in activating rabbit platelets to aggregate.
Brazilian native meliponines are currently threatened by increased human impacts. The assessment of their genetic variation by microsatellite DNA markers can assist in the conservation of populations and help in the planning and establishment of efficient management strategies. The purpose of this study was to develop the first set of microsatellite markers for Melipona fasciculata, selected from partial genome assembly of Illumina paired-end reads. Primer pairs were designed for each detected locus at their flanking regions. Bee samples were genotyped from two different populations of Northeastern Brazil for marker characterization and validation. A total of 17 microsatellite loci displayed polymorphism. Mean HE and HO heterozygosities were 0.453 and 0.536, respectively. PIC across all loci ranged from 0.108 to 0.714. A genetic diversity analysis revealed high values for population differentiation estimates (FST = 0.194, RST = 0.230, and Dest = 0.162) within the investigated region. PCoA and Bayesian clustering showed a separation of the species into two distinct clusters. These microsatellite markers have demonstrated strong potential for population-level genetic studies. Moreover, the preliminary analysis of the genetic diversity in M. fasciculata provides provisional evidence of significant population differentiation between the two studied populations., Geice Ribeiro Da Silva, Isis Gomes De Brito Souza, Fabia De Mello Pereira, Bruno De Almeida Souza, Maria Teresa Do Rego Lopes, Paul Bentzen, Fabio Mendonça Diniz., and Obsahuje bibliografii
Novel microsatellite markers for the solitary bee, Anthophora plumipes, were identified and characterised using 454 GS-FLX Titanium pyrosequencing technology. Thirty seven loci were tested using fluorescently labelled primers on a sample of 20 females from Prague. The number of alleles ranged from 1 to 10 (with a mean of 4 alleles per locus), resulting in an observed heterozygosity ranging from 0.05 to 0.9 and an expected heterozygosity from 0.097 to 0.887. None of the loci showed a significant deviation from the Hardy-Weinberg equilibrium and only two loci showed the significant presence of null alleles. No linkage between loci was detected. We further provide information on a single multiplex PCR consisting of 11 of the most polymorphic loci. This multiplex approach provides an effective analytical tool for analysing genetic structure and carrying out parental analyses on Anthophora populations. Most of the 37 loci tested also showed robust amplification in five other Anthophora species (A. aestivalis, A. crinipes, A. plagiata, A. pubescens and A. quadrimaculata). The result of this study demonstrates that next generation sequencing technology is a valuable method for isolating quality microsatellites in non-model species of solitary bees. and Obsahuje seznam literatury
The endemic large carpenter bee, Xylocopa (Koptortosoma) ogasawarensis Matsumura (Hymenoptera: Apidae), on the oceanic Ogasawara (Bonin) Islands, located in the northwestern Pacific Ocean, approximately 1000 km south of the Japanese mainland, is a generalist flower visitor. Although the flower-visiting behaviour of X. ogasawarensis females has been frequently recorded, the behaviour of the males in this species has rarely been studied. I observed the territorial behaviour of males on flowers of the native plant species Scaevola sericea Vahl (Campanulales: Goodeniaceae) in a coastal area of Hahajima in early July 2007. Each male chose a particular inflorescence, hovered near it (mean distance, 239 mm from the inflorescence) and patrolled around it for several minutes (mean time, 331 s). Therefore, X. ogasawarensis males exhibit resource-based (i.e., flowers) territoriality. Males frequently attacked other males when defending their territories. However, males did not defend their territories against flower visitors of other species (i.e., introduced honeybees). Therefore, male territorial behaviour in X. ogasawarensis may be related to intrasexual competition for sites visited by females. This is the first report describing the male territorial behaviour of X. ogasawarensis.
The ribosomal protein S6 kinase (S6K) plays a pivotal role in developmental processes and cell survival by participating in protein synthesis relevant signaling pathways. In the present study, an S6K gene (AccS6K-p70) was isolated and characterized from the Chinese honeybee, Apis cerana cerana (Fabricius) (Hymenoptera: Apidae), an important economic insect in the agricultural industry. The cDNA of AccS6K-p70 was 1683 bp in length and predicted to encode a protein of 467 amino acid residues. Sequence and structure analysis showed that there was a conserved catalytic domain in AccS6K-p70, whilst a phosphorylation site was found in the conserved part of the catalytic domain. Development relevant transcription factor binding sites found in the 5’-flanking region of AccS6K-p70 suggest that AccS6K-p70 might be involved in A. c. cerana development. Furthermore, quantitative PCR revealed that the expression levels of AccS6K-p70 were higher in head and thorax than in other tissues. The AccS6K-p70 was highly expressed in both larvae and adults compared with that in pupae, whilst expression of the gene was significantly down-regulated by hydrogen peroxide (H2O2) (although initially and slightly increased by it) and pyriproxyfen (a juvenile hormone analogue insecticide) stresses. These results suggest that AccS6K-p70 may play critical roles in developmental processes and cell survival in A. c. cerana, whilst both oxidative stress and pyriproxyfen may impair S6K-p70 mediated developmental processes by down-regulation of AccS6K-p70 expression., Yingqi Cai ... [et al.]., and Obsahuje seznam literatury
The effects of three factors operating during pre-emergence development period on some characteristics of drones were
studied. Weight of newly emerged drones, length of forewing, length of tibia, length of femur, length and width of basitarsus and
number of spermatozoa in drones from colonies in which the workers had access to drone brood (A), the size of brood cells differed
(B) or the colony had a queen or was queenless (C), were determined. For this purpose, 9 colonies were chosen at random from the test apiary and prepared so that each contained one empty Langstroth frame with six small sub-frames containing drone combs, three of which were used to test the effect of one level and the other three the other level of each factor. The results showed that, colony
status had a greater effect on the parameters measured than either the size of the brood cells or whether the workers had access to
sealed brood cells. Most of the parameters were significantly different in C, whereas only a few in B and non in A were significantly
different. The distance between discriminant scores in each experiment enhanced the previous results, as it was highest in C, followed by B and then A. Optimal drone characteristics may be recorded for colonies in which the queens were induced to lay unfertilized eggs in newly built drone combs, and then removed and the drone brood reared in a queen less colony.