sign in sign up
<<
Home , Western honey bee

DOI: 10.2478/v10289-012-0013-y Vol. 56 No. 1 2012 Journal of Apicultural 115

REVIEW OF THE EXPRESSION OF ANTIMICROBIAL PEPTIDE DEFENSIN IN APIS MELLIFERA L.

R u s t e m A. Ilyasov, Louisa R. Gaifullina, E l e n a S. Saltykova, Aleksandr V. Poskryakov, A l e x e i G. N i k o l e n k o

Institute of Biochemistry and Genetics of the Ufa Science Center of the Russian Academy of Sciences, Prospekt Oktyabrya, 71, Ufa, 450054, Bashkortostan Republic, Russia e-mail: [email protected] Received 17 January 2012; accepted 23 April 2012

S u m m a r y Honey bees defensin have a high level of polymorphism and exist as two peptides - defensin 1 and 2. Defensin 1 is synthesized in the salivary glands and is responsible for social . Defensin 2 is synthesized by cells of the fat body and hemolymph is responsible for individual immunity. Defensins are inducible and controlled by the interaction of Toll and Imd signaling pathways and have a broad spectrum of antimicrobial action. The use of chitosan as an immunomodulator has been shown to lead to an increase in the expression levels of defensin and abaecin in the honey organism. Stimulation of the transcriptional activity of the defensin will allow for the control of a ’s immunity level, and reduce the using of antibiotics and other chemicals. Keywords: honey bee, Apis mellifera, defensin, evolution, immunity.

INTRODUCTION 2004), defensin has two isoforms called Antimicrobial peptides (AMP) are royalisin (Fujiwara et al., 1990). important components of the honey bee Kwakman et al. (2010) showed immune system (Hoffmann et al., 1999). antimicrobial properties of honey in This is one of the evolutionarily ancient relation to Bacillus subtilis, methicillin and important protector mechanisms of resistant , . In a short period of time, β-lactamase producing Escherichia coli, AMP can be activated and delivered to ciprofloxacin resistant Pseudomonas the site of infection (Aerts et al., 2008). aeruginosa, and vancomycin resistant At present four types of AMP are found Enterococcus faecium. It was shown in bees: apidaecin presented by twelve that antimicrobial properties of honey isoforms (Casteels et al., 1989), abaecin are provided by the content of hydrogen (Casteels et al., 1990), hymenoptaecin peroxide, methylglyoxal and defensin-1. (Casteels et al., 1993) and defensin On the power of their actions, AMP presented by two isoforms - defensin are comparable with antibiotics and can 1 and 2 (Casteels et al., 1993; Casteels- be used in the development of drugs with Josson et al., 1994; Qu et al., 2008). antifungal and antibacterial properties Several varieties of antimicrobial (Bulet et al., 1999). The use of fungicides molecules are found in honey bee royal and antibiotics in the treatment of honey jelly: major protein (MRJP) bees for diseases leads to the suppression of has five isoforms (MRJP1-5) (Casteels- the bees’ immune system, the emergence of Josson et al., 1994; Klaudiny et al., resistant pathogens, and the contamination 2005; Qu et al., 2008), jelleine has four of bee products (Miyagi et al., 2000). isoforms (Jelleine-I-IV) (Fontana et al., Enhancement of the honey bees immune 116 system by increasing of the expression Fungi. Of the entire AMP of honey bees, level of AMP in bees themselves, is the only defensin has cytotoxic activity against solution to these problems (Bilikova et fungi - fungus of chalkbrood Ascosphaera al., 2001). apis, fungi of aspergillosis Aspergillus Unfortunately, data on the defensins flavus Link and Aspergillus niger Tieghem, having the highest polymorphism and yeast-like fungi Candida albicans and spectrum of activity of all honey bee Aurobasidium pullulans (Chernysh et AMP, are scattered. The data do not give al., 1999; Aronstein et al., 2010). In a complete picture of the role of these particular, Aronstein and Saldivar defensins in the life and anti-infectious (2005) showed an increase in levels of protection of Apis mellifera L. defensin expression in the five-day Activity of defensins against pathogens larvae A. mellifera experimentally infected of honey bees with A. apis. This experiment confirmed Bacteria. Honey bee defensins have the universal mechanism of defensin enough broad-spectrum antibacterial action against fungal infections, since activity. The cytotoxic activity of defensins components of both signaling pathways - against gram-positive bacteria (Casteels- Toll and Imd were involved (Evans and Josson et al., 1994; Bulet and Stocklin, Spivak, 2010). 2005) and several species of gram-negative Protozoa. Microsporidia is bacteria, is known (Mandrioli et al., an obligate parasite in the intestines of 2003). One of the factors the honey bee. Until recently, Nosema is the allocation of royalisin with the royal ceranae was known as a parasite of the jelly for larvae feeding and antibacterial chinese wax bee . Since protection. It has been shown that royalisin 2005, there has been information about is effective primarily against gram- microsporidia N. ceranae parasitizing on negative bacteria and some gram-positive A. mellifera (Higes et al., 2009), which (Klaudiny et al., 2005). In particular, is more pathogenic to honey bees than royalisin is active against American (Klee et al., 2007). This Foulbrood larvae is probably a consequence of long-term (Bilikova et al., 2001; Bachanova et coevolution of A. mellifera with N. apis. al., 2002; Yoshiyama and Kimura, Such a coevolution contributed to the 2010). emergence of certain immune mechanisms The bacteria Lactobacillus is non- in the honey bees to this parasite, including pathogenic to bees. This bacteria also defensin (Antunez et stimulates an increase in the level of gene al., 2009). The microsporidian pathogen expression of abaecin and defensin. The N. ceranae is a new parasite for the honey use of abaecin and defensin as probiotics bee causing a major breach in the gut to enhance honey bee immunity is then and suppressing immunity as a whole, possible (Arbia and Babbay, 2011). expression of defensins in particular (Klee Yoon et al. (2009) showed induction et al., 2007). of defensin gene expression in body Mites and viruses. The weakening of fat of workers from other members of the immunity with ectoparasites is caused : three species of not only by parasitization, but also by the , B. ardens ardens and transmission of viral infections. The most B. hypocrita sapporoensis, in response to common ectoparasites of honey bees are injection of lipopolysaccharides simulating the mites. Among the many species of the action of bacterial infection. Similarly, mites, four species are the most dangerous chitosan also stimulates the defensin gene for honey bees: destructor, Varroa expression in the honeybee, simulating an jaсobsoni, Acarapis woodi, invasion of microorganisms (Saltykova clareae (Grobov and Lihotin, 1989). et al., 2010 a,b). Vol. 56 No. 1 2012 Journal of Apicultural Science 117

Bees infected with V. destructor die when β-lactamase-producing Escherichia coli, exposed to the bacteria Esherichia coli. ciprofloxacin-resistant Pseudomonas These infected bees differ from healthy aeruginosa, and vancomycin-resistant bees in the large number of hemolymph Enterococcus faecium (Kwakman et damaged cells and content of viral al., 2010). These antimicrobial properties particles (Yang et al., 2004). It is shown of honey are provided by the content of that immunosuppressive effect and severe hydrogen peroxide, methyl-glyoxal and the clinical symptoms increase with a rise in antimicrobial peptide defensin-1. the mite infestation level (Williams et al., Defensin 1 gene consists of a 2012 bp 2009). (number in the genebank AY496432) and The humoral immune system of bees contains two introns. The first intron in is affected by V. destructor, reducing the size 571 bp located between 773 and 1345 defensin transcription level (Yang et al., nucleotides, and the second - in the size of 2004; Gregory, et al., 2005). Perhaps, 278 bp located between nucleotides 1525 V. destructor causes immunosuppression and 1804. Excluding introns, the sequence of bees by means of replication of the of the gene is fully aligned with the cDNA deformation of wing virus DWV which of royalisin Ro-K. Sequencing of cDNA is carried by this mites (Genersch and showed a similarity of the gene of royalisin Aubert, 2010). with a defensin 1 gene, except for one site Induction of defensin expression in the in the 377 basis where there was a single honey bee organism nucleotide replacement (C-T) (Casteels- Defensins represent a large family of Josson et al., 1994). cysteine-rich AMP. Honeybee defensins Defensin 2 gene consists of 1950 bp are homologous to formicins of fly (number in the genebank AY588474) and Phormia terranovae and differ from other contains an intron in size 335 bp located peptides by having a peculiar structure as between the 947 and 1283 nucleotides. well as selective activity against gram- At the DNA level there is a high level positive bacteria and some filamentous of similarity of the central module of fungi (Klaudiny et al., 1994; Chernysh defensins. On the other hand, the genes et al., 1999). have a low level of similarity above the At present, four types of AMP were TATA box and TATA box below to the detected in honeybee: apidaecin (Casteels place of processing of the mature peptide et al., 1989), abaecin (Casteels et al., as well as in 3'-noncoding region. 1990), hymenoptaecin (Casteels et al., Genes defensin 1 and defensin 2 genes 1993) and defensins. The defensins are differ significantly in length, intron-exon represented by two peptides, defensin 1 structure and sequence of their pre-pro and defensin 2 with a molecular weight regions. In particular, the second intron of 5.5 and 4.8 kDa, respectively, and of defensin 1, which is not represented at encoded by two genes, defensin 1 and defensin 2 is the first intron found in the defensin 2. Defensin 1 is presented by encoding part of defensin gene. three isoforms - defensin of hemolymph Short aminated C-terminal extensions are and 2 isoforms, found in royal jelly and found only in the defensins named royalisin (Ro-F) with a molecular (Hanzawa et al., 1990; Raj and Dentin, weight of 5525,1 kDa and 5515,5 kDa, 2002). Perhaps these extensions of 11 amino respectively (Klaudiny et al., 2005). acids allow the defensin molecule to take Royalizin differs from the hemolymph an alpha-spiral structure, which stabilizes defensin by two amino acid substitutions the aminated C-terminus (Casteels et al., (Klaudiny et al., 2005). Honey has been 1989). The exact role of these extensions in shown to possess antibacterial properties the function of the peptide is not yet fully against Bacillus subtilis, methicillin- disclosed. Perhaps these extensions are resistant Staphylococcus aureus, the result of exon shuffling (Long, 2001). 118

Because of a recombination of two exons, method, defensin 1 has been shown to be the atypical defensin with an extension expressed in the head and thorax of honey at C-terminus has appeared. Similar bees by the hypopharyngeal, mandibular processes of the emergence of defensin and thoracic salivary glands (Lopez et al., C-terminal extensions also occurred during 2003; Klaudiny et al., 2005) and released the evolution of two species. in the royal jelly (Qu et al., 2008) and Studies concerning the variability honey (Kwakman et al., 2010). There is of the defensin gene fragment in A. m. wide variability in defensin quantity and mellifera populations in the Urals showed level of activity in the royal jelly of different the presence of two alleles of this locus healthy honey bees colonies. One reason (Ilyasov et al., 2008). Allele B of the may be genetic variation in the levels of defensin gene fragment, large in size, met defensin expression. Defensin 1 gene with a frequency of 0.14-0.25, and allele A, polymorphism may play an important role with a smaller size - with a frequency of in the modification of the expression level 0.75-0.86. Such changes in defensin allele of AMP and its antimicrobial and antifungal frequencies may be due to the occurring activity (Bilikova et al., 2001). This may micro evolutionary processes. be important for practical Many variants of defensin isoforms may when selecting honey bees colonies more be due to posttranslational modification resistant to microbial pathogens, especially under the influence of genes in other loci, colonies more resistant to Paenibacillus rather than one-locus mutations (Solbrig larvae larvae (Evans and Spivak, and Solbrig, 1979). Perhaps one of the 2010). The low level of transcription in the causes of the multiplicity of antimicrobial healthy bees and the presence of regulatory activity of bee defensins in the evolution elements in the promotor region of the process, is the variability in processing defensin 2 gene proves that defensin 2 is of defensin precursors which leads to the induced by a pathogenic factor (Casteels appearance of isoforms. et al., 1989; Klaudiny et al., 2005). Thus, The majority of AMP is produced by the we can conclude that the predominant role fat body cells (adipocytes) and hemolymph of defensin 1 is the formation of a social cells (hemocytes) with infections and immune system of the honey bee. This is injuries of the integument, and usually in contrast to the defensins 2 produced by distinguished in the hemolymph adipocytes and hemolymph cells which is a (Hoffmann et al., 1999; Choi et al., component of individual immunity (Fig. 1). 2008). However, using the RT-PCR The wide representation of defensins in

Fig. 1. Forms of defensin in individual and social immunity of honey bees. 1 - mandibular gland; 2 - hypopharyngeal gland; 3 -thoracic gland; 4 - dorsal vessel; 5 - fat body; 6 - ventral diaphragm; 7 - midgut; 8 - dorsal diaphragm. Vol. 56 No. 1 2012 Journal of Apicultural Science 119 the honey bee organisms indicates their et al., 1997). Hemocyte mediators, universal and significant functions in such as prostaglandins, can enhance the individual and social immunity. activity of adipocytes and hemocytes The share of AMP synthesized by the (Stanley-Samuelson, 1994). It is hemocytes is negligible. The main part shown that phagocytes with absorbed of AMP is produced by the fat body microorganisms are able to directly attach cells - adipocytes (Hoffmann and to the adipocytes, which also stimulates Richhart, 1997). Initiation of AMP gene the synthesis of AMP (Glupov, 2001). transcriptional activity occurs at the injury In addition, activation of AMP synthesis and gets into the hemolymph of inductors in adipocytes and hemocytes is assumed of a different and origin - bacteria, under the action of mediators of the fungi and fragments peptidoglycanes and endocrine system (Glupov, 2001). lipopolysaccharides of the bacteria cell There are two major NF-kB-mediated walls (Dunn, 1990), some signaling pathways - Imd and Toll - (Zhu and Lu, 1992), and controlling the gene expression of AMP oligomers (Furukawa et al., 1999). in (Osta et al., 2004). The Toll The experimental data obtained under pathway is responsible for the protection the single application of chitosan as an against fungi and gram-positive bacteria, immunomodulator for honey bees showed whereas the Imd pathway primarily a twofold increase of transcriptional provides protection against gram-negative activity of defensin and abaecin genes by bacteria (Hoffmann, 2003; Evans and PCR in real time (Saltykova et al., 2010 Spivak, 2010). At the same time, control a,b) (Fig. 2). of defensin gene expression is carried out Activation of AMP synthesis in by the interaction of Toll and Imd pathways adipocytes and hemocytes is carried (Aronstein et al., 2010) demonstrating out by several mechanisms (Fig. 3). It the importance of this element of the AMP is assumed that the generation of AMP system. Antimicrobial peptides are active at synthesis inducers - lipopolysaccharides, low concentrations. These peptides show a peptidoglycanes and β-1,3-glycans - wide spectrum of activity interacting with involves phagocytic cells releasing into the the cytoplasmic membranes of pathogens hemolymph from the cell wall components and acting in concert with other agents of of absorbed and digested bacteria (Taniani immune response (Osta et al., 2004).

Fig. 2. Expression level changes of AMP defensin and abaecin in the honey bee organism in two experiments (biological replicates). Transcript level for a gene with low transcriptional variation mgst (microsomal glutathione-S-transferase) (Evans and Wheeler 2000) was used to normalize against variable mRNA levels (Saltykova et al., 2010 a,b). 120

Fig. 3. Activation of AMP synthesis in insect adipocytes and hemocytes. I - generation of inductors of AMP synthesis; II - secretion of hemocyte mediators; III - attachment of hemocyte to the fat body. 1 - the surface of the body; 2 - phagocyte, 3 - adipocyte; 4 - microorganism; 5 - cell wall components of the microorganism; 6 - hemocyte mediators; 7 - mediators of the endocrine system. The general principle of the AMP is Nosema apis). The uniqueness of this reduced not only to inconsistency of peptide of Hymenoptera among other membranes of pathogenic cells, but it is as well as the multiplicity mediated by other mechanisms. Defensins of antimicrobial activity of honey bee kill pathogenic cells, penetrating through defensins is determined by the variability their cytoplasmic membrane (Cociancich in the processing of defensin precursors et al., 1993). Defensins cause the formation leading to the appearance of multiple of channels in the plasma membrane of molecular forms. The action of the target cells through which cytoplasmic defensin mechanism is reduced to a breach K+ outflows, and consequently, there of the integrity and permeability of the is a partial depolarization of the plasma cytoplasmic membrane of pathogenic membrane, reduction of ATP content organisms and provides a wide range of in the cytoplasm and inhibition of action. respiratory processes. Experiments in vitro Defensin 1, contained in royal jelly and demonstrated that defensins of Phormia honey and synthesized in the salivary and Aeschna also cause a disturbance of glands, is involved in the formation of the membrane permeability of Plasmodium social immunity of colonies. Defensin 2, gallinaceum sporozoites followed by a synthesized by cells of the fat body and change in morphology and loss of the hemolymph, is responsible for individual parasitoid mobility (Shahabuddin et al., immunity of the honey bee. The wide 1998). multifunctional activity of defensins is characterized by principles of social life CONCLUSIONS that emerged during the evolution of some Honey bee defensins have a high level of Hymenoptera species. polymorphism, and exists in two forms - The use of immunomodulators leads to a defensin 1 and 2. Defensin gene expression marked increase in the level of expression is inducible and regulated by the Toll and of antimicrobial peptides in the organism Imd signaling pathways, which lead to of the honey bee. This allows for the its versatility action against pathogens control of the level of honey bee colony of the honey bees (Paenibacillus larvae, immunity and allows for a reduction in the Melissococcus pluton, Ascosphaera apis, use of antibiotics and other chemicals. Vol. 56 No. 1 2012 Journal of Apicultural Science 121

ACKNOWLEDGMENTS Casteels P., Ampe C., Jacobs F., This work is supported by RFBR 08- Tempst P. (1993) - Functional and chemical 04-97039-r_povolgie_a and 11-04-97078- characterization of hymenoptaecin, an r_povolgie_a. antibacterial polypeptide that is infection- inducible in the honey bees (Apis mellifera). J. REFERENCES Biol. Chem., 268: 7044-7054. Casteels P., Ampe C., Jacobs F., Aerts A. M., Francois I. E., Vaek M., Tempst P. (1989) - Apidaecins: Cammue B. P., Thevissen K. (2008) - antibacterial peptides from honey bees. The The mode of antifungal action of plant, insect EMBO Journal, 8: 2387-2391. and defensins. Cell. Mol. Life Sci., 65: 2069-2079. Casteels P., Ampe C., Riviere L., Damme J. V., Elicone C., Fleming M., Antunez K., Martin-Hernandez R., Jacobs F., Tempst P. (1990) - Isolation Prieto L., Meana A., Zunino P., Higes and characterization of abaecin, a major M. (2009) - Immune suppression in the honey antibacterial peptide in the honey bees (Apis bees (Apis mellifera) following infection by mellifera). Eur. J. Biochem., 187: 381-386. (Microsporidia). Environ. Microbiol., 11(9): 2284-2290. Casteels-Josson K., Zhang W., Capaci T., Casteels P., Tempst P. Arbia K. A., Babbay B. (2011) - Management (1994) - Acute transcriptional response of the strategies of honey bees diseases. J. Entomol., honey bees peptide-antibiotics gene repertoire, 8 (1): 1-15. required posttranslational conversion of the Aronstein K. A., Murray K. D., precursor structures. J. Biol. Chem., 269: Saldivar E. (2010) - Transcriptional 28569-28575. responses in honey bees larvae infected with Chernysh S. I., Gordya N. A., chalkbrood fungus. BMC Genomics, 11: 1-12. Filatova N. A. (1999) - Sacrificial Aronstein K. A., Saldivar E. (2005) - mechanisms of insects: the rate of molecular Characterization of a honey bees Toll related and phenotypic evolution. Genetics Research, receptor gene Am18w and its potential 12: 52-59. involvement in antimicrobial immune defense. Choi Y. S., Choo Y. M., Lee K. S., Yoon Apidologie, 36: 3-14. H. J., Kim I., Je Y. H., Sohn H. D., Bachanova K., Klaudiny J., Jin B. R. (2008) - Cloning and expression Kopernicky J., Simuth J. (2002) - profiling of four antibacterial peptide genes Identifcation of honey bees peptide active from the bumblebee Bombus ignites. Comp. against Paenibacillus larvae larvae through Biochem. Physiol., 150: 141-146. bacterial growth-inhibition assay on Cociancich S., Ghazi A., Hetru C., polyacrylamide gel. Apidologie, 33: 259-269. Hoffmann J. A., Letellier L. (1993) - Bilikova K., Gusui W., Simuth J. (2001) Insect defensin, an inducible antibacterial - Isolation of a peptide fraction from honey peptide, forms voltage-dependent channels bees royal jelly as a potential antifoulbrood in Micrococcus luteus. J. Biol. Chem., 268: factor. Apidologie, 32: 275-283. 19239-19245. Bulet P., Hetru C., Dimarcq J. L., Dunn P. E. (1990) - Humoral immunity in Hoffmann D. (1999) - Antimicrobial insects. Immune strategy appeart to correspond peptides in insects; structure and function. to life-history characteristics. Bioscience, DeComposition Immunology, 23: 329-344. 40(10): 738-744. Bulet P., Stocklin R. (2005) - Insect Evans J. D., Spivak M. (2010) - Socialized antimicrobial peptides: structure, properties medicine individual and communal disease and gene regulation. Protein & Peptide barriers in honey bees. J. Invertebr. Pathol., Letters, 12: 3-11. 103: 562-572. 122

Evans J. D., Wheeler D. E. (2000) - Hoffmann J. A. (2003) - The immune response Expression profiles during honeybee caste of . Nature, 426: 33-38. determination. Biology, 2(1): research Hoffmann J. A., Kafatos F. C., 0001.1- 0001.6. Janawey C. A., Ezekovitz R. A. B. Fontana R., Mendes M. A., (1999) - Phylogenetic perspectives in innate de Souza B. M., Konno K., immunity. Science, 284: 1313-1318. Cesar L. M. M., Malaspina O., Hoffmann J. A., Richhart J. M. (1997) - Palma M. S. (2004) - Jelleines: a family of Drosophila immunity. Trends in Cell Biology, antimicrobial peptides from the Royal Jelly 7: 309-316. of honey bees (Apis mellifera). Peptides, 25: Ilyasov R. A., Poskryakov A. V., 919-928. Nikolenko A. G. (2008) - Polymorphism of Fujiwara S., Imai J., Fujiwara M., antimicrobial peptides in honey bee population Yaeshima T., Kawashima T., in the Urals. Biodiversity: Problems and Kobayashi K. (1990) - A potent antibacterial perspectives of preservation. Proceedings of protein in royal jelly. J. Biol. Chem., 265: the International Scientific Conference, Penza, 11333-11337. 13-16 may, 2008, Volume 2. pp: 247-248. Furukawa S., Taniai K., Yang J., Klaudiny J., Hanes J., Kulifajova J., Shono T., Yamakawa M. (1999) - Albert S., Simuth J. (1994) - Molecular Induction of gene expression of antibacterial cloning of two cDNAs from the head of the by chitin oligomers in the silkworm, nurse honey bee (Apis mellifera L.) for coding . Insect Mol. Biol., 8(1): 145-148. related proteins of royal jelly. J. Apic. Res., 33: Genersch E., Aubert M. (2010) - Emerging 105-111. and re-emerging viruses of the honey bees Klaudiny J., Albert S., Bachanova K., (Apis mellifera L.). Vet. Res., 41(6): 54-74. Kopernicky J., Simuth J. (2005) - Two Glupov V. V. (2001) - Mechanisms of structurally different defensin genes, one of resistance of insects, in: Glupov V. V. (ed.) them encoding a novel defensin isoform, are Insect Pathogens: structural and functional expressed in honey bees Apis mellifera. Insect aspects, Moscow, Kruglyi god, 2001, pp 475- Biochemi. Mol. Biol., 35: 11-22. 557. Klee J., Besana A. M., Genersch E., Gregory G., Evans J. D., Rinderer T., Gisder S., Nanetti A., Tam D. Q. (2007) de Guzman L. (2005) - Conditional immune- - Widespread dispersal of the microsporidian gene suppression of honey bees parasitized by Nosema ceranae, an emergent pathogen of the Varroa mites. J. Insect Sci., 5: 1-5. western honey bee, Apis mellifera. J. Invertbr. Grobov O. F., Lihotin A. K. (1989) - Diseases Pathol., 96: 1-10. and pests of bees. Moscow: Agropromisdat. p: Kwakman H. S., te Velde A. A., 239. de Boer L., Speijer D., Vandenbroucke- Hanzawa H., Shimada I., Kuzuhara T., Grauls C. M. J. E., Zaat S. A. J. (2010) - Komano H., Kohda D., Inagaki F., How honey kills bacteria. The FASEB Journal, Natori S., Arata Y. (1990) - 1H nuclear 24(7): 2576-2582. magnetic resonance study of the solution Long M. (2001) - Evolution of novel genes. conformation of an antibacterial protein, Curr. Opinion Genet. Dev., 11: 673-680 sapecin. FEBS Letters, 269: 413-420. Lopez L., Morales G., Ursic R., Higes M., Martin-Hernandez R., Wolff M., Lowneberger C. (2003) - Gonzalez-Porto A. V., Garcia- Isolation and characterization of a novel insect Palencia P., Meana A., del Nozal M. J. defensin from Rhodnius prolixus, a vector of (2009) - Honey bees colony collapse due Chagas disease. Insect Biochem. Mol. Biol., 33: to Nosema cernae in professional . 439-447. Environmental Microbiology Reports, 1: 110- 113. Vol. 56 No. 1 2012 Journal of Apicultural Science 123

Mandrioli M., Bugli S., Saltini S., Shahabuddin M., Fields I., Bulet P., Genedani S., Ottaviani E. (2003) - Hoffmann J. A., Miller L. (1998) - Molecular characterization of a defensin in Plasmodium gallinaceum: differential killing the IZD-MB-0503 cell line derived from of some stages of the parasite by immunocytes of the insect Mamestra brassicae insect defensin. Experimental Parasitology, 89 (Lepidoptera). Biology of the Cell, 95: 53-57. (1): 103-112. Miyagi T., Peng Ch. Y. S., Chuang R. Y., Solbrig O. T., Solbrig D. J. (1979) - An Mussen E. C., Spivak M. S., Doi R. H. Introduction to Population Biology and (2000) - Verification of oxytetracycline- Evolution. Addison Wesley Publishing resistant pathogen Company, Reading, Massachusets. p: 468. Paenibacillus larvae in the . J. Stanley-Samuelson D. W. (1994) - Invertebr. Pathol., 75: 95-96. Prostaglandins, related eicosanoids in insects. Osta M. A., Christophides G. K., Advances in Insect Physiology, 24: 115-212. Vlachou D., Kafatos F. C. (2004) - Innate Taniani K., Wago H., Yamakawa M. immunity in the malaria vector Anopheles (1997) - In vitro phagocytosis of Escherichia gambiae: comparative and functional coli and release of lipopolysaccharide by genomics. J. Exp. Biol., 207(15): 2551-2563. adhering hemocytes of the silkworm, Bombyx Qu N., Jiang J., Sun L., Lai C., Sun L., mori. Biochem. Biophys. Res. Commun., 231: Wu X. (2008) - Proteomic Characterization of 623-627. Royal Jelly Proteins in Chinese (Apis cerana Williams G. R., Rogers R. E. L., cerana), European (Apis mellifera) Honey Kalkstein A. L., Taylor B. A., bees, Biochemistry, 1: 1-12. Shutler D., Ostiguy N. (2009) - Deformed Raj P. A., Dentino A. R. (2002) - Current wing virus in western honey bees (Apis status of defensins and their role in innate mellifera) from Atlantic Canada, the first and adaptive immunity. FEMS Microbiology description of an overtlyinfected emerging Letters, 206: 9-18. queen. J. Invertebr. Pathol., 101: 77-79. Saltykova E. S., Gaifullina L. R., Yang D., Biragyn A., Hoover D. M., Ilyasov R. A., Nikolenko A. G. (2010a) Lubkowski J., Oppenheim J. J. (2004) - Chitosan action on the main antibacterial - Multiple roles of antimicrobial defensins, peptides induction of the honey bee. Modern cathelicidins, and eosinophil-derived perspectives in chitin and chitosan studies. neurotoxin in host defense. Annual Review of Proceedings of the Xth International Immunology, 22: 181-215. Conference, Nizhny Novgorod, 29 June-2 July, Yoon H. J., Sohn M. R., Young M. C., 2010. pp. 308-310. Jianhong L., Hung D. S., Byung R. J. Saltykova E. S., Ilyasov R. A., (2009) - Defensin gene sequences of three Gaifullina L. R., Poskryakov A. V., different bumblebees, Bombus spp. Journal of Yamidanov R. S., Nikolenko A. G. Asia-Pacific Entomology, 12: 27-31. (2010b) - Changes in the expression level of Yoshiyama M., Kimura K. (2010) - antimicrobial peptides in the honeybee (Apis Characterization of antimicrobial peptide mellifera mellifera L.) organism. Modern genes from Japanese honey bees Apis cerana beekeeping. Concerns, experiences, new japonica (Hymenoptera: Apidae). Applied technologies: Proceedings of the International Entomology and Zoology, 45(4): 609-614. Scientific-practical Conference. Yaroslavl, 12- Zhu P., Lu Z. (1992) - Studies on the 13 August, 2010. pp. 159-160. antibacterial substances of Pieris rapae induced by deltamethrin and trichlorfon. 19 Int. Congr. Entomol, Beijing, p: 594. 124

Ekspresja peptydU przeciwbakteryjneGO - defensyny u pszczoły miodnej Apis mellifera L. - praca przeglądowa

I l y a s o v R . A., Gaifullina L. R., Saltykova E. S., Poskryakov A. V., Nikolenko A. G.

Streszczenie Pszczoła miodna posiada w wysokim stopniu zróżnicowaną defensynę występującą w postaci dwóch peptydów - defensyny 1 oraz 2. Defensyna 1 jest syntetyzowana w gruczołach ślinowych i odpowiada za odporność społeczną pszczół, natomiast defensyna 2 syntetyzowana jest przez komórki ciała tłuszczowego oraz w hemolimfie i odpowiada za odporność indywidualną. Defensyny są indukowalne. Regulowane są poprzez współoddziaływanie szlaków sygnalizacyjnych Toll i Imd. Defensyny posiadają szerokie spektrum działania przeciwbakteryjnego. Wykazano, że zastosowanie chitozanu jako immunomodulatora prowadzi do podniesienia poziomu ekspresji defensyny i abaecyny w organizmie pszczoły miodnej. Stymulowanie aktywności transkrypcyjnej genów defensynowych pozwoli kontrolować poziom odporności rodzin pszczelich oraz zmniejszyć zastosowanie antybiotyków i innych leczniczych środków chemicznych. Słowa kluczowe: pszczoła miodna, Apis mellifera, defensyna, ewolucja, odporność.