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Invertebrate iridescent virus 6 (IIV6) is a member of the genus Iridovirus and belongs to the Iridoviridae family. The entirely sequenced dsDNA genome, composed of 212.482 bp, encodes 215 putative open reading frames (ORFs). ORF458R... more
Invertebrate iridescent virus 6 (IIV6) is a member of the genus Iridovirus and belongs to the Iridoviridae family. The entirely sequenced dsDNA genome, composed of 212.482 bp, encodes 215 putative open reading frames (ORFs). ORF458R encodes a putative myristoylated membrane protein. RT-PCR analysis of ORF458R expression in the presence of DNA replication and protein synthesis inhibitors showed that this gene is transcribed in the late phase of the virus infection. Time course analysis showed that transcription of ORF458R initiates between 12 and 24 h p.i. and starts to decrease after this point. Transcription of ORF458R initiated 53 nucleotides upstream of the translation start site and ended 40 nucleotides after the stop codon. Dual luciferase reporter gene assay showed that sequences between -61st and +18th nucleotides are essential for promoter activity. Interestingly, a remarkable decrease in promoter activity, in the presence of sequences between -299th and -143rd nucleotides, suggested a repressor activity between these regions. Our results showed that ORF458R is transcriptionally active, and separately located sequences at its upstream region with promoter and repressor activities regulating its expression. This information on the transcriptional analysis of ORF458R will contribute to our understanding of the molecular mechanisms of IIV6 replication.
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Background: The gypsy moth (Lymantria dispar L., Lepidoptera: Erebidae) is a worldwide pest of trees and forests. Lymantria dispar nucleopolyhedrovirus (LdMNPV) belongs to the Baculoviridae family and is an insect virus specific to gypsy... more
Background: The gypsy moth (Lymantria dispar L., Lepidoptera: Erebidae) is a worldwide pest of trees and forests. Lymantria dispar nucleopolyhedrovirus (LdMNPV) belongs to the Baculoviridae family and is an insect virus specific to gypsy moth larvae. In this study, we describe the complete genome sequences of three geographically diverse isolates, H2 (China), J2 (Japan), and T3 (Turkey), of Lymantria dispar multiple nucleopolyhedrovirus (LdMNPV). Methods: The genomes of isolates H2, J2, and T3 were subjected to shotgun pyrosequencing using Roche 454 FLX and assembled using Roche GS De Novo Assembler. Comparative analysis of all isolates was performed using bioinformatics methods. Results: The genomes of LdMNPV-H2, J2, and T3 were 164,746, 162,249, and 162,614 bp in size, had GC content of 57.25%, 57.30%, and 57.46%, and contained 162, 165, and 164 putative open reading frames (ORFs ≥ 150 nt), respectively. Comparison between the reference genome LdMNPV-5/6 (AF081810) and the genomes...
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This chapter focuses on key virus families that affect wild and cultured insect species and describes current knowledge on the molecular and ecological aspects of these viruses. Insects can be infected by a variety of viruses each with... more
This chapter focuses on key virus families that affect wild and cultured insect species and describes current knowledge on the molecular and ecological aspects of these viruses. Insects can be infected by a variety of viruses each with distinct properties and pathologies. Depending on the taxonomic order to which the insects belong, infections with viruses of certain families are more prominent and strong co-evolutionary relationships between insects and particular virus types are often observed. Viral infections may cause diseases that are often lethal, but viruses can also remain in a persistent or latent stage and only cause a disease outbreak under certain circumstances. We illustrate the impact viruses have on particular groups of insects by providing examples of viral infections in insects in their natural environments, the application of insect viruses in biocontrol programmes, and the effect these viruses have on pollinators. Apart from viruses affecting free-living insects, we also include what is known about virus outbreaks in insects reared on an industrial scale. Furthermore, we include a brief viral disease management section for apiculture and mass rearing of other insects. We conclude this chapter with suggestions on how to obtain the necessary knowledge and tools to better control viral diseases in insects in the future.
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Invertebrate iridescent virus 6 (IIV6) is the type species of the Iridovirus genus in the Betairidovirinae subfamily of the Iridoviridae family. Transcription of the 215 predicted IIV6 genes is temporally regulated, dividing the genes... more
Invertebrate iridescent virus 6 (IIV6) is the type species of the Iridovirus genus in the Betairidovirinae subfamily of the Iridoviridae family. Transcription of the 215 predicted IIV6 genes is temporally regulated, dividing the genes into three kinetic classes: immediate-early (IE), delayed-early (DE), and late (L). So far, the transcriptional class has been determined for a selection of virion protein genes and only for three genes the potential promoter regions have been analyzed in detail. In this study, we investigated the transcriptional class of all IIV6 genes that had not been classified until now. RT-PCR analysis of total RNA isolated from virus-infected insect cells in the presence or absence of protein and DNA synthesis inhibitors, placed 113, 23 and 22 of the newly analyzed viral ORFs into the IE, DE and L gene classes, respectively. Afterwards, in silico analysis was performed to the upstream regions (200 bp) of all viral ORFs using the MEME Suite Software. The AA(A/T)(T/A)TG(A/G)A and (T/A/C)(T/G/C)T(T/A)ATGG motifs were identified in the upstream region of IE and DE genes, respectively. These motifs were validated by luciferase reporter assays as crucial sequences for promoter activity. For the L genes two conserved motifs were identified for all analyzed genes: (T/G)(C/T)(A/C)A(T/G/C)(T/C)T(T/C) and (C/G/T)(G/A/C)(T/A)(T/G) (G/T)(T/C). However, the presence of these two motifs did not influence promoter activity. Conversely, the presence of these two sequences upstream of the reporter decreased its expression. Single nucleotide mutations in the highly conserved nucleotides at the end of the second motif (TTGT) showed that this motif acted as a repressor sequence for late genes in the IIV6 genome. Next, upstream sequences of IIV6 L genes from which we removed this second motif in silico, were re-analyzed for the presence of potential conserved promoter sequences. Two additional motifs were identified in this way for L genes: (T/A)(A/T)(A/T/G)(A/T)(T/C)(A/G)(A/C)(A/C) and (C/G)(T/C)(T/A/C)C(A/T)(A/T)T(T/G) (T/G)(T/G/A). Independent mutations in either motif caused a severe decrease in luciferase expression. Information on temporal classes and upstream regulatory sequences will contribute to our understanding of the transcriptional mechanisms in IIV6.
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In order to detect and identify the most toxic Bacillus thuringiensis strains against pests, we isolated a B. thuringiensis strain (Bn1) from Balaninus nucum (Coleoptera: Curculionidae), the most damaging hazelnut pest. Bn1 was... more
In order to detect and identify the most toxic Bacillus thuringiensis strains against pests, we isolated a B. thuringiensis strain (Bn1) from Balaninus nucum (Coleoptera: Curculionidae), the most damaging hazelnut pest. Bn1 was characterized via morphological, biochemical, and molecular techniques. The isolate was serotyped, and the results showed that Bn1 was the B. thuringiensis serovar, kurstaki (H3abc). The scanning electron microscopy indicated that Bn1 has crystals with cubic and bipyramidal shapes. The Polymerase Chain Reactions (PCRs) revealed the presence of the cry1 and cry2 genes. The presence of Cry1 and Cry2 proteins in the Bn1 isolate was confirmed via SDS-PAGE, at approximately 130 kDa and 65 kDa, respectively. The bioassays conducted to determine the insecticidal activity of the Bn1 isolate were conducted with four distinct insects, using spore-crystal mixtures. We noted that Bn1 has higher toxicity as compared with the standard B. thuringiensis subsp. kurstaki (HD-1). The highest observed mortality was 90% against Malacosoma neustria and Lymantria dispar larvae. Our results show that the B. thuringiensis isolate (Bn1) may prove valuable as a significant microbial control agent against lepidopteran pests.
Research Interests: Microbiology, Medical Microbiology, Biology, Scanning Electron Microscopy, Scanning Transmission Electron Microscopy, and 15 moreMedicine, Lepidoptera, Bacillus thuringiensis, Pubmed, Animals, Polymerase Chain Reaction, Weevils, Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis, PEST analysis, Larva, Repetitive Strain Injury, Moths, Lymantria Dispar, Cry Gene, and Endotoxins
ABSTRACT A broad survey of the fall webworm (Lepidoptera: Erebidae) populations in agricultural and forested areas in the Central Black Sea Region of Turkey led to the detection of a granulovirus (GV). Hyphantria cunea granulovirus... more
ABSTRACT A broad survey of the fall webworm (Lepidoptera: Erebidae) populations in agricultural and forested areas in the Central Black Sea Region of Turkey led to the detection of a granulovirus (GV). Hyphantria cunea granulovirus (HycuGV)-Hc1 isolate was characterized and tested against third instar larvae of H. cunea. Electron microscopy confirmed typical GV morphology with ovoid granules of approximately 368 ± 16 nm × 201 ± 19 nm. Each granule contained a single rod-shaped virion with a mean size of 43 ± 12 nm × 250 ± 12 nm. The genome was analyzed by restriction endonuclease and estimated to be ∼112 kb. Partial sequencing of the granulin (gran), late expression factor-8 (lef-8) and late expression factor-9 (lef-9) genes also confirmed the identity of the virus as HycuGV. A phylogenetic analysis based on these conserved genes, HycuGV-Hc1 grouped together with the previous HycuGV isolate (A5-1) and Estigmene acrea granulovirus (EsacGV) isolate from the same family. The LC50 of HycuGV-Hc1 was 2.6 × 104 occlusion bodies (OBs/ml). Pot experiments, under field conditions, showed significant differences between virus treated and control groups. This is the first study to describe a novel Turkish HycuGV-Hc1 isolate, and preliminary data suggests that the virus has a significant potential as an effective biopesticide for control of H. cunea.
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A cytoplasmic polyhedrosis virus (CPV) was isolated from the larvae of Thaumetopoea pityocampa and shown to cause an infection of midgut cells. This viral infection revealed several important diagnostic symptoms, including discoloration... more
A cytoplasmic polyhedrosis virus (CPV) was isolated from the larvae of Thaumetopoea pityocampa and shown to cause an infection of midgut cells. This viral infection revealed several important diagnostic symptoms, including discoloration of the posterior midgut, reduced feeding, and extended development time of the larvae. The virus infection is lethal to Thaumetopoea pityocampa, and with the increasing doses kills the larvae within 4-5 days post infection. Electron microscopy studies showed typical cytoplasmic polyhedral inclusion bodies that are icosahedral, and ranged from 2.4 to 5.3 microm in diameter. Electrophoretic analysis of the RNA genome showed that the virus has a genome composed of 10 equimolar RNA segments with the sizes of 3,907, 3,716, 3,628, 3,249, 2,726, 1,914, 1,815, 1,256, 1,058, and 899 bp, respectively. Based on morphology and nucleic acid analysis, this virus was named Thaumetopoea pityocampa cytoplasmic polyhedrosis virus (TpCPV), and belongs to the genus Cypovirus, family Reoviridae.
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Research Interests: Microbiology, Medical Microbiology, Biology, DNA damage, DNA repair, and 15 moreMedicine, Biological Sciences, DNA, Escherichia coli, Animals, Invertebrate Pathology, PEST, Moths, Lymantria Dispar, Amino Acid Sequence, Invertebrate, Photolyase, Dna Synthesis, Molecular Sequence Data, and Medical and Health Sciences
ABSTRACT Chilo iridescent virus (CIV), officially named Insect iridescent virus 6 (IIV6), is the type species of the genus Iridovirus (family Iridoviridae). In this paper we constructed a recombinant CIV, encoding the green fluorescent... more
ABSTRACT Chilo iridescent virus (CIV), officially named Insect iridescent virus 6 (IIV6), is the type species of the genus Iridovirus (family Iridoviridae). In this paper we constructed a recombinant CIV, encoding the green fluorescent protein (GFP). This recombinant can be used to investigate viral replication dynamics. We showed that homologous recombination is a valid method to make CIV gene knockouts and to insert foreign genes. The CIV 157L gene, putatively encoding a non-functional inhibitor of apoptosis (IAP), was chosen as target for foreign gene insertion. The gfp open reading frame preceded by the viral mcp promoter was inserted into the 157L locus by homologous recombination in Anthonomus grandis BRL-AG-3A cells. Recombinant virus (rCIV-Δ157L-gfp) was purified by successive rounds of plaque purification. All plaques produced by the purified recombinant virus emitted green fluorescence due to the presence of GFP. One-step growth curves for recombinant and wild-type CIV were similar and the recombinant was fully infectious in vivo. Hence, CIV157L can be inactivated without altering the replication kinetics of the virus. Consequently, the CIV 157L locus can be used as a site for insertion of foreign DNA, e.g. to modify viral properties for insect biocontrol.
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Research Interests: Biology and Iridescence
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Research Interests: Biology and Photolyase
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ABSTRACT The wireworm Agriotes lineatus (L.) (Coleoptera: Elateridae) is a serious agricultural pest of various vegetables and fruits throughout the world. To find an effective and safe biological control agent against this pest, we... more
ABSTRACT The wireworm Agriotes lineatus (L.) (Coleoptera: Elateridae) is a serious agricultural pest of various vegetables and fruits throughout the world. To find an effective and safe biological control agent against this pest, we investigated the bacterial flora of A. lineatus. Nineteen different bacterial strains were isolated and identified as Paenibacillus sp. (Ag1), Cellulomonas sp. (Ag2), Bacillus subtilis (Ag3), Staphylococcus sp. (Ag4), Enterococcus mundtii (Ag5), Staphylococcus sp. (Ag6), Sphingobacterium sp. (Ag7), Staphylococcus pasteuri (Ag8), Arthrobacter gandensis (Ag9), Bacillus sp. (Ag10), Chryseobacterium sp. (Ag11), Streptomyces sp. (Ag12), Oerskovia turbata (Ag13), Bacillus thuringiensis (Ag14), Pseudomonas fluorescens (Ag15), Oerskovia jenensis (Ag16), Arthrobacter gandavensis (Ag17), B. thuringiensis (Ag18), and Pseudomonas plecoglossicida (Ag19) based on conventional and molecular tests. A. gandavensis and P. plecoglossicida were isolated for the first time from any insect. The insecticidal effects of these 19 bacterial isolates and the additional 11 isolates belonging to Bacillus genus isolated from different hosts were tested on third instar larvae of A. lineatus. Ag17 (A. gandavensis), Ag18 (B. thuringiensis), and Ag19 (P. plecoglossicida) from the bacterial flora of A. lineatus, and two Bacillus isolates (Bacillus circulans Ar1 from Anoplus roboris and B. thuringiensis subsp. kurstaki BnBt from Balanicus nucum) showed 100% mortality 10 days after treatment. Our results indicate that the bacterial isolates tested in this study may be considered as a possible microbial control agent against A. lineatus.
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Chilo iridescent virus (CIV), officially named invertebrate iridescent virus 6 (IIV6), is a nucleocytoplasmic virus with a ~212-kb linear dsDNA genome that encodes 215 putative open reading frames (ORFs). Proteomic analysis has revealed... more
Chilo iridescent virus (CIV), officially named invertebrate iridescent virus 6 (IIV6), is a nucleocytoplasmic virus with a ~212-kb linear dsDNA genome that encodes 215 putative open reading frames (ORFs). Proteomic analysis has revealed that the CIV virion consists of 54 virally encoded proteins. In this study, we identified the interactions between the structural proteins using the yeast two-hybrid system. We cloned 47 structural genes into both bait and prey vectors, and then analysed the interactions in Saccharomyces cerevisiae strain AH109. A total of 159 protein-protein interactions were detected between the CIV structural proteins. Only ORF 179R showed a self-association. Four structural proteins that have homologues in iridoviruses (118L, 142R, 274L and 295L) showed indirect interactions with each other. Seven proteins (138R, 142R, 361L, 378R, 395R, 415R and 453R) interacted with the major capsid protein 274L. The putative membrane protein 118L, a homologue of the frog virus 3/Ranagrylio virus 53R protein, showed direct interactions with nine other proteins (117L, 229L, 307L, 355R, 366R, 374R, 378R, 415R and 422L). The interaction between 118L and 415R was confirmed by a GST pull-down assay. These data indicate that 415R is a potential matrix protein connecting the envelope protein 118L with the major capsid protein 274L.
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Strain GKT was isolated from the Kumbet plateu of Giresun in Turkey. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain GKT belonged to genus Janthinobacterium and 16S rRNA gene sequence similarities with all... more
Strain GKT was isolated from the Kumbet plateu of Giresun in Turkey. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain GKT belonged to genus Janthinobacterium and 16S rRNA gene sequence similarities with all type strains of the genus Janthinobacterium were 98.89%–99.78%. The calculated pairwise average nucleotide identity (ANI) values between strain GKT and all type strains of Janthinobacterium species were in the range of 79.8%–93.2%. In addition, digital DNA–DNA hybridization (dDDH) values were in the range of 23.0%–51.7%. Major fatty acids are C10:03OH, C12:0, C16:1ω7c, C16:0, and C18:1ω7c, and polar lipids included phosphatidylethanolamine, phosphatidylglycerol, also one unidentified phospholipid and one unidentified aminophospholipid. The respiratory quinone of strain GKT was determinated to be Q-8. The genome sizes of strain GKT was 6 197 538 bp with 63.16% G + C ratio. Strain GKT is Gram-stain-negative, aerobic, rod-shaped, and motile. A violet pigm...
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: In this study, the bacterial flora of Hyphantria cunea Drury. (Lep., Arctiidae) were investigated during three hazelnut seasons from 1998 to 2000. Four different bacteria were found in dead and living larvae. They were isolated and... more
: In this study, the bacterial flora of Hyphantria cunea Drury. (Lep., Arctiidae) were investigated during three hazelnut seasons from 1998 to 2000. Four different bacteria were found in dead and living larvae. They were isolated and identified as Bacillus thuringiensis, Escherichia freundii, Micrococcus sp. and Streptococcus sp. Laboratory experiments carried out to determine the insecticidal activities of these isolates showed that E. freundii and Micrococcus sp. did not have any insecticidal effect on second – third instar larvae of H. cunea. However, B. thuringiensis and Streptococcus sp. had 56 and 38% effects, respectively. Crystals and spores from B. thuringiensis were also purified and the crystals, spores and crystals–spore mixture were tested separately against the larvae of H. cunea. It was found that the insecticidal activities of the crystals, spores and crystal–spore mixture were 37.5, 25 and 62.5%, respectively, on second – third instar larvae of H. cunea. These results indicate that the crystal–spore mixture has 6.5% more insecticidal effect than that of the vegetative cells of the B. thuringiensis isolate.
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Baculoviruses have become a noteworthy research and study material for various applications in recent years. They are used as insecticides in agriculture because of their high host specifity. Recombinant baculoviruses are being used in... more
Baculoviruses have become a noteworthy research and study material for various applications in recent years. They are used as insecticides in agriculture because of their high host specifity. Recombinant baculoviruses are being used in biotechnology for the expression of related genes under the control of strong polh and p10 genes promoters. They have attracted attention as gene therapy vectors in medicine. Baculoviruses can enter the mammalian cells; however, they neither replicate nor show any pathogenic effect. Also baculoviruses are model organisms for molecular biology studies of gene structure and organization. Although baculoviruses are used commonly, their genome replication is not understood. Most of the research data about this subject have been obtained during recent years. Human beings have just begun to benefit from the use of baculoviruses. Baculoviruses will serve science in the future as mentioned above. We believe that this review will contribute to the appreciation of the field in our country in regard to baculoviruses which are popular all over the world. For this reason, in this review after giving information about the biology of baculoviruses, replication of baculoviruses has been explained in detail.
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Research Interests: Molecular Biology, Biology, Apoptosis, Medicine, Gene Silencing, and 15 moreBiological Sciences, Cell line, Animals, Programmed cell death, Gene, Amino Acid Profile, Amino Acid Sequence, Open Reading Frame, Gene expression profiling, Iridovirus, Cydia pomonella, Molecular Sequence Data, Medical and Health Sciences, Nucleopolyhedrovirus, and Defense mechanism
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Insect viruses are biological control agents that cause their illness or dead by infecting the insects. Recently, these viruses have great interest at modern biotechnological applications. Insect viruses that have high host specificity,... more
Insect viruses are biological control agents that cause their illness or dead by infecting the insects. Recently, these viruses have great interest at modern biotechnological applications. Insect viruses that have high host specificity, have been used against various agricultural and forest pest as an alternative to chemical pesticides. Studies done with these viruses have been used as model for high organizational organisms. Many genes that has industrial, agricultural, medical and economical importance have been produced at great amounts at expression systems developed from these viruses. Also, recently these viruses are being used as gene therapy vector. At this review paper, we will pay attention on subjects especially at insect viruses, and potential of the usage of baculoviruses at various biotechnological studies.