20 localities were randomly selected in Eastern Black Sea Region of Turkey and samples were collected from around the beehives from April to September. Total of 4,640 dead adult worker bees were examined during the st...20 localities were randomly selected in Eastern Black Sea Region of Turkey and samples were collected from around the beehives from April to September. Total of 4,640 dead adult worker bees were examined during the study. Total infection rate in worker bees was 21.23%. Nosema ceranae was identified in all localities with molecular techniques. Temperature and humidity values were measured from around the beehives during field studies. The infection rate ofN. ceranae increased proportionally with increasing temperature and humidity factors. Humidity was more effective than temperature on the infection rate ofN. ceranae. The seasonal activity ofN. ceranae was studied. The highest infection rates were observed in June and July. N. ceranae infection rate was higher in localities that were in low-altitude than in localities that were in high-altitude.展开更多
Propolis collected by stingless bees from various types of plants has been used as an antimicrobial agent in several previous studies. We assessed the effect of propolis produced by a stingless bee, Trigona apicalis, ...Propolis collected by stingless bees from various types of plants has been used as an antimicrobial agent in several previous studies. We assessed the effect of propolis produced by a stingless bee, Trigona apicalis, on Apis florea experimentally infected with Nosema ceranae, a parasite of honeybees. For parasite inoculation each Nosema free-bee was fed 2μL of 50% (w/v) sucrose solution containing N. ceranae spores at 40,000 spores/bee and 0 as a negative control (CO). Treated bees were provided with 0%, 10%, 20% and 50% propolis (w/v) in water, defined as 0P, 10P, 20P and 50P, respectively. We assessed the effects of propolis 14 days post inoculation. All propolis-treated bees had significantly higher survival than untreated bees. However, survival of Nosema-inoculated bees was lower than that of control bees. Bees treated with the highest propolis concentration (50P) had the highest survival ratio. No control bees became infected over the course of the study. However, N. ceranae infection rates of bees treated with 0P, 10P, 20P and 50P were 75 ± 1.4%, 72 ± 5.6%, 69± 4.2% and 47± 1.4%, respectively. In addition, propolis-treated bees had hypopharyngeal gland protein content that was significantly higher than 0P and CO bees. Overall, propolis treatment significantly reduced N. ceranae infection rate and bee mortality and was associated with increased hypopharyngeal gland protein concentration.展开更多
Despite the urgent need for conservation consideration,strategic action plans for the preservation of the Asian honeybee,Apis cerana Fabricius,1793,remain lacking.Both the convergent and divergent adaptations of this ...Despite the urgent need for conservation consideration,strategic action plans for the preservation of the Asian honeybee,Apis cerana Fabricius,1793,remain lacking.Both the convergent and divergent adaptations of this widespread insect have led to confusing phenotypical traits and inconsistent infraspecific taxonomy.Unclear subspecies boundaries pose a significant challenge to honeybee conservation efforts,as it is difficult to effectively prioritize conservation targets without a clear understanding of subspecies identities.Here,we investigated genome variations in 362 worker bees representing almost all populations of mainland A.cerana to understand how evolution has shaped its population structure.Whole-genome single nucleotide polymorphisms(SNPs)based on nuclear sequences revealed eight putative subspecies,with all seven peripheral subspecies exhibiting mutually exclusive monophyly and distinct genetic divergence from the widespread central subspecies.Our results demonstrated that most classic morphological traits,including body size,were related to the climatic variables of the local habitats and did not reflect the true evolutionary history of the organism.Thus,such morphological traits were not suitable for subspecific delineation.Conversely,wing vein characters showed relative independence to the environment and supported the subspecies boundaries inferred from nuclear genomes.Mitochondrial phylogeny further indicated that the present subspecies structure was a result of multiple waves of population divergence from a common ancestor.Based on our findings,we propose that criteria for subspecies delineation should be based on evolutionary independence,trait distinction,and geographic isolation.We formally defined and described eight subspecies of mainland A.cerana.Elucidation of the evolutionary history and subspecies boundaries enables a customized conservation strategy for both widespread and endemic honeybee conservation units,guiding colony introduction and breeding.展开更多
[Objective] This study was to clone the GnRH (gonadotropin-releasing hormone), and to investigate its expression in Apis cerana cerana. [Method] The cDNA sequence of GnRHR gene was amplified from Apis cerana cerana ...[Objective] This study was to clone the GnRH (gonadotropin-releasing hormone), and to investigate its expression in Apis cerana cerana. [Method] The cDNA sequence of GnRHR gene was amplified from Apis cerana cerana by using RT-PCR techniques. It was conducted with bioinformatics analysis and the in situ hybridization histochemistry of its expression products was studied. [Result] The sequence analy- sis showed that the full cDNA sequence was 1 050 bp with the open reading frame of 1 050 bp, and it encoded 349 amino acid residues. The deduced amino sequence included 7 transmembrane regions, and the predicted molecular mass and isoelectric point were 40.6 kD and 9.54, respectively. The cluster analysis showed that the GnRHR from ',4. cerana cerana had close relationship to the GnRHR II from other insects. In situ hybridization showed that Bee-GnRHR staining was specifically localized to the brain, intestine, fat body and testis. [Conclusion] The results indicated that the GnRHR provided molecular bond for the reproduction and metabolism for insects, and suggested a functional role for bee-GnRHR signaling in the coupling of reproduction activities and environment conditions.展开更多
Objective: To molecularly identify Nosema species in provinces of Isfahan, Fars,Chaharmahal and Bakhtiari.Methods: One hundred and eighty adult honey bees suspected with nosemosis from provinces of Fars(different coun...Objective: To molecularly identify Nosema species in provinces of Isfahan, Fars,Chaharmahal and Bakhtiari.Methods: One hundred and eighty adult honey bees suspected with nosemosis from provinces of Fars(different counties), Isfahan, and Chaharmahal and Bakhtiari were tested. In order to determine the species of Nosema, previously developed PCR and primers based on 16 S r RNA gene were used. PCR products were puri fi ed and sent to the Korean company of Macrogen for sequencing.Results: Only Nosema ceranae was determined in all samples based on their molecular profile. Sequences of the 16 S r RNA gene were sent to Gen Bank/NCBI(samples accession numbers KP318660–KP318663).Conclusions: This species currently exists in European honeybee apiaries of Apis mellifera in the studied provinces.展开更多
Using culture-independent technique polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and conventional culture techniques, ecological risk of transgenic maize pollen on gut bacteria of the...Using culture-independent technique polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and conventional culture techniques, ecological risk of transgenic maize pollen on gut bacteria of the Chinese honeybee, Apis cerana cerana, was assessed. Honeybees were fed with Bt-transgenic maize pollen, non-transgenic near isoline pollen, linear crylAh gene (800 ng mL^-1) and supercoiled plasmid DNA (800 ng mL^-1) under laboratory conditions. The DGGE profile showed that the number of DGGE bands varied from 10.7 to 14.7 per sample, and the Shannon's index ranged from 0.85 to 1.00. The similarity calculated by PAST was mostly above 92%, indicating no obvious changes among treatments or within replicates. 14 bacterial strains affiliated with Alphaproteobacteria, Gammaproteobacteria, Firmicutes, and Actinobacteria were isolated and characterized on media under aerobic and anaerobic conditions. These results demonstrated that transgenic crylAh maize pollen did not induce significant changes of the honeybee gut bacterial community composition under laboratory conditions.展开更多
By screening the worker (Apis cerana cerana) heads cDNA library using a fragment of the mrjp3 gene of Apis cerana as probe, 120 positive clones were obtained. The clone containing A. cerana cerana MRJP3 (AccMRJP3)...By screening the worker (Apis cerana cerana) heads cDNA library using a fragment of the mrjp3 gene of Apis cerana as probe, 120 positive clones were obtained. The clone containing A. cerana cerana MRJP3 (AccMRJP3) cDNA was selected. Based on the sequencing of the inserts of the positive clone, a sequence of AccMRJP3 cDNA which is 1 887 bp long including a poly (A) tail was obtained. The AccMRJP3 cDNA encompassed an open-reading frame (ORF) with 1 779 bp encoding 593 amino acids. The un-translated regions (UTR) of the 5′ end and 3′end are 46 bp and 160 bp in length, respectively. Similar to AmMRJP3 and AdMRJP3, the putative AccMRJP3 also has a repetitive region. The comparison of the repetitive region of AccMRJP3, AmMRJP3 and AdMRJP3 shows some differences between them.展开更多
The mitogen-activated protein kinase(MAPK)cascade pathway plays an important role in regulating stress responses.The function of the c-Jun NH_(2)-terminal kinase(JNK),a component of the MAPK cascade pathway,in Apis ce...The mitogen-activated protein kinase(MAPK)cascade pathway plays an important role in regulating stress responses.The function of the c-Jun NH_(2)-terminal kinase(JNK),a component of the MAPK cascade pathway,in Apis cerana cerana(Acc)remains unclear.Here,JNK was isolated and identified from Acc.Bioinformatics analyses revealed there is a typical serine/threonine protein kinase catalytic domain in the AccJNK protein.An expression profile analysis showed that AccJNK was significantly induced by pesticide treatments.To further explore the functional mechanisms of AccJNK,a yeast 2-hybrid screen was performed,activator protein-1(AP-1)was screened as the interaction partner of AccJNK,and the interaction relationship was further verified by pull-down assay.Quantitative real-time polymerase chain reaction showed the expression pattern of AccAP-I was similar to that of AccJNK.After a knockdown of AccJNK or AccAP-I by RNA interference,the survival rate of Acc after pesticide treatments increased.Additionally,the expression levels of antioxidant-related genes and the activities of antioxidant enzymes increased,suggesting that the knockdown of AccJNK or AccAP-I increased the antioxidant capacity of bees.Our study revealed that the JNK-mediated MAPK pathway responds to pesticide stress by altering the antioxidant capacity of Acc.展开更多
A clone inserted with 1 104 bp fragment containing a 765bp Open Reading Frame(ORF), encoding a putative 2,3-bisphosphoglycerate(2,3BPG) dependent Phosphoglycerate mutase(dPGAM) that catalyzes the transfer of a phosp...A clone inserted with 1 104 bp fragment containing a 765bp Open Reading Frame(ORF), encoding a putative 2,3-bisphosphoglycerate(2,3BPG) dependent Phosphoglycerate mutase(dPGAM) that catalyzes the transfer of a phosphate group from the C3 carbon atom to the C2 carbon atom of phosphoglycerate, was screened by mass sequencing from the cDNA library of the venom glands of Apis cerana. The deduced amino acid sequence shared high similarities (39%-88%)with the dPGAM of 7 other organisms, but the similarities with the iPGAM of 4 other organisms were low (10%-12%). Moreover, the alignment of Ac-PGAM with the dPGAMs from 7 other organisms showed that all the active site amino acid residues were conserved. This result shows that Ac-PGAM is a typical dPGAM. Thus, this is the second PGAM gene reported in Insecta. Furthermore, phylogenetic analysis showed that the evolutionary tree of PGAMs reflects the systematic relationship of species.展开更多
American foulbrood (AFB) disease is caused by Paenibacillus larvae. Currently, this pathogen is widespread in the European honey bee— Apis mellifera. However, little is known about infectivity and pathogenicity of P....American foulbrood (AFB) disease is caused by Paenibacillus larvae. Currently, this pathogen is widespread in the European honey bee— Apis mellifera. However, little is known about infectivity and pathogenicity of P. lan'ae in the Asiatic cavity-nesting honey bees, Apis cerana. Moreover, comparative knowledge of P. larvae infectivity and pathogenicity between both honey bee species is scarce. In this study, we examined susceptibility, larval mortality, survival rate and expression of genes encoding antimicrobial peptides (AMPs) including defensin, apidaecin, abaecin, and hymenoptaecin in A. mellifera and A. cerana when infected with P. larvae. Our results showed similar effects of P. larvae on the survival rate and patterns of AMP gene expression in both honey bee species when bee larvae are infected with spores at the median lethal concentration (LC5 0 ) for A. mellifera. All AMPs of infected bee larvae showed significant upregulation compared with noninfected bee larvae in both honey bee species. However, larvae of A. cerana were more susceptible than A. mellifera when the same larval ages and spore concentration of P. larvae were used. It also appears that A. cerana showed higher levels of AMP expression than A. mellifera. This research provides the first evidence of survival rate, LC50 and immune response profiles of Asian honey bees, A. cerana, when infected by P. larvae in comparison with the European honey bee, A. mellifera.展开更多
Apis cerana indica foragers were used for the isolation of a full-length α- glucosidase cDNA, and for purification of the active nascent protein by low salt extraction of bee homogenates, ammonium sulphate precipitat...Apis cerana indica foragers were used for the isolation of a full-length α- glucosidase cDNA, and for purification of the active nascent protein by low salt extraction of bee homogenates, ammonium sulphate precipitation and diethylaminoethyl-cellulose and Superdex 200 c hromatographies. The molecular mass of the purified protein was estimated by polyacrylamide gel electrophoresis resolution, and the pH, temperature, incubation, and substrate optima for enzymic activity were determined. Conformation of the purified enzyme as α-glucosidase was performed by BLAST software homology comparisons between matrix assisted laser desorption ionization time of flight mass spectroscopy analysed partial tryptic peptide digests of the purified protein with the predicted amino acid sequences deduced from the α-glucosidase cDNA sequence.展开更多
Bee venom phospholipase A2(BvPLA2) is a lipolytic enzyme that catalyzes the hydrolysis of the sn-2 acyl bond of glycerophospholipids to liberate free fatty acids and lysophospholipids.In this work,a new BvPLA2(AccPLA2...Bee venom phospholipase A2(BvPLA2) is a lipolytic enzyme that catalyzes the hydrolysis of the sn-2 acyl bond of glycerophospholipids to liberate free fatty acids and lysophospholipids.In this work,a new BvPLA2(AccPLA2) gene from the Chinese honeybee(Apis cerana cerana) venom glands was inserted into bacmid to construct a recombinant transfer vector.Tn-5B-4(Tn) cells were transfected with the recombinant bacmid DNA for expression.Sodium dodecylsulfate-polyacrylamide gel electrophoresis(SDS-PAGE) analysis revealed a double band with molecular weights of 16 and 18 kDa.Products of hexahistidine AccPLA2 fusion protein accumulated up to 5.32% of the total cellular proteins.The AccPLA2 fusion protein was cross reactive with the anti-AmPLA2(BvPLA2 of the European honeybee,Apis mellifera) polyclonal serum.The reaction resulted in a double glycosylation band,which agrees with the band generated by the native AmPLA2 in Western blot analysis.The PLA2 activity of the total extracted cellular protein in the hydrolyzing egg yolk is about 3.16 μmol/(min·mg).In summary,the recombinant AccPLA2 protein,a native BvPLA2-like structure with corresponding biological activities,can be glycosylated in Tn cells.These findings provided fundamental knowledge for potential genetic engineering to produce AccPLA2 in the pharmaceutical industry.展开更多
Within a honeybee population, due to polyandry, there are super-sister and half-sister relations, thus many sub-families exist. For the Chinese honeybee (Apis cerana cerana F.) a phylogenetic dendrogram has been const...Within a honeybee population, due to polyandry, there are super-sister and half-sister relations, thus many sub-families exist. For the Chinese honeybee (Apis cerana cerana F.) a phylogenetic dendrogram has been constructed in which 4 sub-families are clustered based on DNA fingerprint patterns. And it has been observed that each kind of workers is distributed to several different sub-families.展开更多
文摘20 localities were randomly selected in Eastern Black Sea Region of Turkey and samples were collected from around the beehives from April to September. Total of 4,640 dead adult worker bees were examined during the study. Total infection rate in worker bees was 21.23%. Nosema ceranae was identified in all localities with molecular techniques. Temperature and humidity values were measured from around the beehives during field studies. The infection rate ofN. ceranae increased proportionally with increasing temperature and humidity factors. Humidity was more effective than temperature on the infection rate ofN. ceranae. The seasonal activity ofN. ceranae was studied. The highest infection rates were observed in June and July. N. ceranae infection rate was higher in localities that were in low-altitude than in localities that were in high-altitude.
文摘Propolis collected by stingless bees from various types of plants has been used as an antimicrobial agent in several previous studies. We assessed the effect of propolis produced by a stingless bee, Trigona apicalis, on Apis florea experimentally infected with Nosema ceranae, a parasite of honeybees. For parasite inoculation each Nosema free-bee was fed 2μL of 50% (w/v) sucrose solution containing N. ceranae spores at 40,000 spores/bee and 0 as a negative control (CO). Treated bees were provided with 0%, 10%, 20% and 50% propolis (w/v) in water, defined as 0P, 10P, 20P and 50P, respectively. We assessed the effects of propolis 14 days post inoculation. All propolis-treated bees had significantly higher survival than untreated bees. However, survival of Nosema-inoculated bees was lower than that of control bees. Bees treated with the highest propolis concentration (50P) had the highest survival ratio. No control bees became infected over the course of the study. However, N. ceranae infection rates of bees treated with 0P, 10P, 20P and 50P were 75 ± 1.4%, 72 ± 5.6%, 69± 4.2% and 47± 1.4%, respectively. In addition, propolis-treated bees had hypopharyngeal gland protein content that was significantly higher than 0P and CO bees. Overall, propolis treatment significantly reduced N. ceranae infection rate and bee mortality and was associated with increased hypopharyngeal gland protein concentration.
基金supported by the National Natural Science Foundation(NSF)of China(32270475)Program of Ministry of Science and Technology of China(2018FY100403)+3 种基金National Special Support Program for High-level Talents(Ten-Thousand Talents Program)2115 Talent Development Program of China Agricultural University through Xin Z.S.L.is supported by Funds for International Cooperation and Exchange of the National Natural Science Foundation of China(3211001043)supported by the NSF of China(31470123)Jilin Science and Technology Program(20030561)through X.L.S.H.P.is supported by the National Mission on Himalayan Studies(NMHS)-Almora,Ministry of Environment,Forest and Climate Change,Government of India,through grant GBPNI/NMHS-2017-18/MG-12。
文摘Despite the urgent need for conservation consideration,strategic action plans for the preservation of the Asian honeybee,Apis cerana Fabricius,1793,remain lacking.Both the convergent and divergent adaptations of this widespread insect have led to confusing phenotypical traits and inconsistent infraspecific taxonomy.Unclear subspecies boundaries pose a significant challenge to honeybee conservation efforts,as it is difficult to effectively prioritize conservation targets without a clear understanding of subspecies identities.Here,we investigated genome variations in 362 worker bees representing almost all populations of mainland A.cerana to understand how evolution has shaped its population structure.Whole-genome single nucleotide polymorphisms(SNPs)based on nuclear sequences revealed eight putative subspecies,with all seven peripheral subspecies exhibiting mutually exclusive monophyly and distinct genetic divergence from the widespread central subspecies.Our results demonstrated that most classic morphological traits,including body size,were related to the climatic variables of the local habitats and did not reflect the true evolutionary history of the organism.Thus,such morphological traits were not suitable for subspecific delineation.Conversely,wing vein characters showed relative independence to the environment and supported the subspecies boundaries inferred from nuclear genomes.Mitochondrial phylogeny further indicated that the present subspecies structure was a result of multiple waves of population divergence from a common ancestor.Based on our findings,we propose that criteria for subspecies delineation should be based on evolutionary independence,trait distinction,and geographic isolation.We formally defined and described eight subspecies of mainland A.cerana.Elucidation of the evolutionary history and subspecies boundaries enables a customized conservation strategy for both widespread and endemic honeybee conservation units,guiding colony introduction and breeding.
基金Supported by the Science and Technology Planning Project of the Education Department of Shaanxi Province(11JK0618)~~
文摘[Objective] This study was to clone the GnRH (gonadotropin-releasing hormone), and to investigate its expression in Apis cerana cerana. [Method] The cDNA sequence of GnRHR gene was amplified from Apis cerana cerana by using RT-PCR techniques. It was conducted with bioinformatics analysis and the in situ hybridization histochemistry of its expression products was studied. [Result] The sequence analy- sis showed that the full cDNA sequence was 1 050 bp with the open reading frame of 1 050 bp, and it encoded 349 amino acid residues. The deduced amino sequence included 7 transmembrane regions, and the predicted molecular mass and isoelectric point were 40.6 kD and 9.54, respectively. The cluster analysis showed that the GnRHR from ',4. cerana cerana had close relationship to the GnRHR II from other insects. In situ hybridization showed that Bee-GnRHR staining was specifically localized to the brain, intestine, fat body and testis. [Conclusion] The results indicated that the GnRHR provided molecular bond for the reproduction and metabolism for insects, and suggested a functional role for bee-GnRHR signaling in the coupling of reproduction activities and environment conditions.
文摘Objective: To molecularly identify Nosema species in provinces of Isfahan, Fars,Chaharmahal and Bakhtiari.Methods: One hundred and eighty adult honey bees suspected with nosemosis from provinces of Fars(different counties), Isfahan, and Chaharmahal and Bakhtiari were tested. In order to determine the species of Nosema, previously developed PCR and primers based on 16 S r RNA gene were used. PCR products were puri fi ed and sent to the Korean company of Macrogen for sequencing.Results: Only Nosema ceranae was determined in all samples based on their molecular profile. Sequences of the 16 S r RNA gene were sent to Gen Bank/NCBI(samples accession numbers KP318660–KP318663).Conclusions: This species currently exists in European honeybee apiaries of Apis mellifera in the studied provinces.
基金supported by the National Basic Research Program of China (2007CB109203 and 2009CB118902)
文摘Using culture-independent technique polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and conventional culture techniques, ecological risk of transgenic maize pollen on gut bacteria of the Chinese honeybee, Apis cerana cerana, was assessed. Honeybees were fed with Bt-transgenic maize pollen, non-transgenic near isoline pollen, linear crylAh gene (800 ng mL^-1) and supercoiled plasmid DNA (800 ng mL^-1) under laboratory conditions. The DGGE profile showed that the number of DGGE bands varied from 10.7 to 14.7 per sample, and the Shannon's index ranged from 0.85 to 1.00. The similarity calculated by PAST was mostly above 92%, indicating no obvious changes among treatments or within replicates. 14 bacterial strains affiliated with Alphaproteobacteria, Gammaproteobacteria, Firmicutes, and Actinobacteria were isolated and characterized on media under aerobic and anaerobic conditions. These results demonstrated that transgenic crylAh maize pollen did not induce significant changes of the honeybee gut bacterial community composition under laboratory conditions.
基金the National Natural Science Foundation of China(30200206) Zhejiang Provincial Natural Science Foundation of China(302113).
文摘By screening the worker (Apis cerana cerana) heads cDNA library using a fragment of the mrjp3 gene of Apis cerana as probe, 120 positive clones were obtained. The clone containing A. cerana cerana MRJP3 (AccMRJP3) cDNA was selected. Based on the sequencing of the inserts of the positive clone, a sequence of AccMRJP3 cDNA which is 1 887 bp long including a poly (A) tail was obtained. The AccMRJP3 cDNA encompassed an open-reading frame (ORF) with 1 779 bp encoding 593 amino acids. The un-translated regions (UTR) of the 5′ end and 3′end are 46 bp and 160 bp in length, respectively. Similar to AmMRJP3 and AdMRJP3, the putative AccMRJP3 also has a repetitive region. The comparison of the repetitive region of AccMRJP3, AmMRJP3 and AdMRJP3 shows some differences between them.
基金supported by the Shandong Provincial Natural Science Foundation(No.ZR2019MC050)special funds for the Efficient Ecological Agriculture Innovation Project of the Taishan Industry Leading Talent Program(No.LJNY202003)the earmarked fund for the China Agriculture Research System of Ministry of Finance and Ministry of Agriculture and Rural Affairs(No.CARS-44).
文摘The mitogen-activated protein kinase(MAPK)cascade pathway plays an important role in regulating stress responses.The function of the c-Jun NH_(2)-terminal kinase(JNK),a component of the MAPK cascade pathway,in Apis cerana cerana(Acc)remains unclear.Here,JNK was isolated and identified from Acc.Bioinformatics analyses revealed there is a typical serine/threonine protein kinase catalytic domain in the AccJNK protein.An expression profile analysis showed that AccJNK was significantly induced by pesticide treatments.To further explore the functional mechanisms of AccJNK,a yeast 2-hybrid screen was performed,activator protein-1(AP-1)was screened as the interaction partner of AccJNK,and the interaction relationship was further verified by pull-down assay.Quantitative real-time polymerase chain reaction showed the expression pattern of AccAP-I was similar to that of AccJNK.After a knockdown of AccJNK or AccAP-I by RNA interference,the survival rate of Acc after pesticide treatments increased.Additionally,the expression levels of antioxidant-related genes and the activities of antioxidant enzymes increased,suggesting that the knockdown of AccJNK or AccAP-I increased the antioxidant capacity of bees.Our study revealed that the JNK-mediated MAPK pathway responds to pesticide stress by altering the antioxidant capacity of Acc.
文摘A clone inserted with 1 104 bp fragment containing a 765bp Open Reading Frame(ORF), encoding a putative 2,3-bisphosphoglycerate(2,3BPG) dependent Phosphoglycerate mutase(dPGAM) that catalyzes the transfer of a phosphate group from the C3 carbon atom to the C2 carbon atom of phosphoglycerate, was screened by mass sequencing from the cDNA library of the venom glands of Apis cerana. The deduced amino acid sequence shared high similarities (39%-88%)with the dPGAM of 7 other organisms, but the similarities with the iPGAM of 4 other organisms were low (10%-12%). Moreover, the alignment of Ac-PGAM with the dPGAMs from 7 other organisms showed that all the active site amino acid residues were conserved. This result shows that Ac-PGAM is a typical dPGAM. Thus, this is the second PGAM gene reported in Insecta. Furthermore, phylogenetic analysis showed that the evolutionary tree of PGAMs reflects the systematic relationship of species.
文摘American foulbrood (AFB) disease is caused by Paenibacillus larvae. Currently, this pathogen is widespread in the European honey bee— Apis mellifera. However, little is known about infectivity and pathogenicity of P. lan'ae in the Asiatic cavity-nesting honey bees, Apis cerana. Moreover, comparative knowledge of P. larvae infectivity and pathogenicity between both honey bee species is scarce. In this study, we examined susceptibility, larval mortality, survival rate and expression of genes encoding antimicrobial peptides (AMPs) including defensin, apidaecin, abaecin, and hymenoptaecin in A. mellifera and A. cerana when infected with P. larvae. Our results showed similar effects of P. larvae on the survival rate and patterns of AMP gene expression in both honey bee species when bee larvae are infected with spores at the median lethal concentration (LC5 0 ) for A. mellifera. All AMPs of infected bee larvae showed significant upregulation compared with noninfected bee larvae in both honey bee species. However, larvae of A. cerana were more susceptible than A. mellifera when the same larval ages and spore concentration of P. larvae were used. It also appears that A. cerana showed higher levels of AMP expression than A. mellifera. This research provides the first evidence of survival rate, LC50 and immune response profiles of Asian honey bees, A. cerana, when infected by P. larvae in comparison with the European honey bee, A. mellifera.
文摘Apis cerana indica foragers were used for the isolation of a full-length α- glucosidase cDNA, and for purification of the active nascent protein by low salt extraction of bee homogenates, ammonium sulphate precipitation and diethylaminoethyl-cellulose and Superdex 200 c hromatographies. The molecular mass of the purified protein was estimated by polyacrylamide gel electrophoresis resolution, and the pH, temperature, incubation, and substrate optima for enzymic activity were determined. Conformation of the purified enzyme as α-glucosidase was performed by BLAST software homology comparisons between matrix assisted laser desorption ionization time of flight mass spectroscopy analysed partial tryptic peptide digests of the purified protein with the predicted amino acid sequences deduced from the α-glucosidase cDNA sequence.
基金Project (No 2007AA10Z324) supported by the National High-Tech Research and Development Program (863) of China
文摘Bee venom phospholipase A2(BvPLA2) is a lipolytic enzyme that catalyzes the hydrolysis of the sn-2 acyl bond of glycerophospholipids to liberate free fatty acids and lysophospholipids.In this work,a new BvPLA2(AccPLA2) gene from the Chinese honeybee(Apis cerana cerana) venom glands was inserted into bacmid to construct a recombinant transfer vector.Tn-5B-4(Tn) cells were transfected with the recombinant bacmid DNA for expression.Sodium dodecylsulfate-polyacrylamide gel electrophoresis(SDS-PAGE) analysis revealed a double band with molecular weights of 16 and 18 kDa.Products of hexahistidine AccPLA2 fusion protein accumulated up to 5.32% of the total cellular proteins.The AccPLA2 fusion protein was cross reactive with the anti-AmPLA2(BvPLA2 of the European honeybee,Apis mellifera) polyclonal serum.The reaction resulted in a double glycosylation band,which agrees with the band generated by the native AmPLA2 in Western blot analysis.The PLA2 activity of the total extracted cellular protein in the hydrolyzing egg yolk is about 3.16 μmol/(min·mg).In summary,the recombinant AccPLA2 protein,a native BvPLA2-like structure with corresponding biological activities,can be glycosylated in Tn cells.These findings provided fundamental knowledge for potential genetic engineering to produce AccPLA2 in the pharmaceutical industry.
文摘Within a honeybee population, due to polyandry, there are super-sister and half-sister relations, thus many sub-families exist. For the Chinese honeybee (Apis cerana cerana F.) a phylogenetic dendrogram has been constructed in which 4 sub-families are clustered based on DNA fingerprint patterns. And it has been observed that each kind of workers is distributed to several different sub-families.