Böcek virüsleri, böcekleri enfekte ederek, onların hastalanmaları veya ölmelerine neden olan biyolojik etmenlerdir. Son yıllarda, bu virüsler modern biyoteknoloji uygulamalarında önemli ölçüde ilgi ...uyandırmaktadır. Yüksek konukçu seçiciliklerine sahip böcek virüsleri, çeşitli tarım ve orman zararlılarına karşı kimyasal pestisitlerin alternatifi olarak kullanılmaktadır. Bu virüslerle yapılan çalışmalar yüksek organizasyonlu canlılar için model olarak kullanılmaktadır. Endüstriyel, zirai, tıbbı ve ekonomik öneme sahip ilgi duyulan birçok gen, bu virüslerden geliştirilmiş ekspresyon vektörlerinde bol miktarda üretilmektedir. Ayrıca, son yıllarda bu virüsler gen terapi vektörü olarak da kullanılmaktadır. Bu derleme makalede, başta genel böcek virüsleri olmak üzere bakülovirüslerin çeşitli biyoteknolojik çalışmalarda kullanılma potansiyelleri üzerinde durulacaktır
The fall webworm (Hyphantria cunea) impacts a wide variety of crops and cultivated broadleaf plant species. The pest is native to North America, was introduced to Europe and has since spread further ...as far as central Asia. Despite several attempts to control its distribution, the pest continues to spread causing damage all over the world. A naturally occurring baculovirus, Hyphantria cunea granulovirus (HycuGV-Hc1), isolated from the larvae of H. cunea in Turkey appears to have a potential as microbial control agent against this pest. In this report we describe the complete genome sequence and organization of the granulovirus isolate (HycuGV-Hc1) that infects the larval stages and compare it to other baculovirus genomes. The HycuGV-Hc1 genome is a circular double-stranded DNA of 114,825 bp in size with a nucleotide distribution of 39.3% G + C. Bioinformatics analysis predicted 132 putative open reading frames of (ORFs) ≥ 150 nucleotides. There are 24 ORFs with unknown function. Seven homologous repeated regions (hrs) and two bro genes (bro-1 and bro-2) were identified in the genome. Comparison to other baculovirus genomes, HycuGV-Hc1 revealed some differences in gene content and organization. Gene parity plots and phylogenetics confirmed that HycuGV-Hc1 is a Betabaculovirus and is closely related to Plutella xylostella granulovirus. This study expands our knowledge on the genetic variation of HycuGV isolates and provides further novel knowledge on the nature of granuloviruses.
•The HycuGV-Hc1 DNA genome has 114,825 bp, GC content of 39.3% and 132 putative ORFs.•Seven homologous regions (hrs) were identified in the genome.•The phylogenetic tree showed that HycuGV-Hc1 to be closely related to the PlxyGV.•HycuGV-Hc1 is infective for H. cunea larvae which impacts a wide variety of crops.
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 LC
50
of HycuGV-Hc1 was 2.6 × 10
4
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.
The lackey moth,
Malacosoma neustria
(Linnaeus, 1758), a worldwide pest, causes extensive economic losses particularly on hazelnut, plum, oak, poplar, and willow trees. A baculovirus, Malacosoma ...neustria nucleopolyhedrovirus (ManeNPV-T2), has been isolated from the larvae collected in Turkey and appears to have a potential as a microbial control agent. In this study, we describe the complete genome sequence of ManeNPV-T2 and compare it to other sequenced baculovirus genomes. The ManeNPV-T2 genome is a circular double-stranded DNA molecule of 130,202 bp, has 38.2% G + C, and is predicted to contain 131 putative open reading frames (ORFs) each with a coding capacity of more then 50 amino acids. There are 27 ORFs with unknown function of which 6 are unique to ManeNPV-T2. Eleven homologous regions (hrs) and two
bro
genes (
bro-a
and
bro-b
) were identified in the genome. There are two homologues of
chaB
and nicotinamide riboside kinase-1 genes, separated from themselves with a few nucleotides. Additionally,
ac145
, thought to be per os infectivity factor (pif) gene, is also found as two homologues. All 38 core genes are found in the ManeNPV-T2 genome. The phylogenetic tree of ManeNPV-T2 in relation to 50 other baculoviruses whose genomes have been completely sequenced showed ManeNPV-T2 to be closely related to the group II NPVs. This study expands our knowledge on baculoviruses, describes the characterization ManeNPV, and ultimately contributes to the registration of this virus as a microbial pesticide.
The gypsy moth (
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).
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.
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 of LdMNPV-H2, J2, and T3 revealed differences in gene content. Compared with LdMNPV-5/6, ORF5, 6, 8, 10, 31, and 67 were absent in LdMNPV-H2, ORF5, 13, and 66 were absent in LdMNPV-J2, and ORF10, 13, 31, and 67 were absent in LdMNPV-T3. In addition, the gene encoding the mucin-like protein (ORF4) was split into two parts in isolates H2 and T3 and designated ORF4a and ORF4b. Phylogenetic analysis grouped isolates H2 and J2 in a different cluster than isolate T3, which is more closely related to the Turkish and Polish isolates. In addition, H2 was found to be closely related to a South Korean LdMNPV isolate.
This study provided a more detailed overview of the relationships between different geographic LdMNPV isolates. The results showed remarkable differences between groups at the genome level.
Türkiye'de araziden toplanan Malacosoma neustria larvalarından izole edilen Malacosoma neustria
nukleopolihedrovirus (ManeNPV) plak yöntemiyle saflaştırıldı. Elde edilen genetik varyantın in vitro ...replikasyonal
özellikleri Malacosoma disstria'dan geliştirilmiş Md203 hücre suşunda karakterize edildi. ManeNPV'nin Md203
hücrelerindeki replikasyon döngüsü sitopatik etkilerin (CPEs) tespiti, polihedral inklüzyon yapıların (PIB) oluşumu,
tomurcuk virusların (BV) üretimi, viral DNA'nın replikasyonu ve polihedrin proteinin ifadesine göre çalışıldı.
Enfeksiyondan sonra 24 ve 36 saat içinde ManeNPV ile enfekte Md203 hücrelerinde sitopatik etkilerin oluştuğu ve
enfeksiyonun 4. gününde hücrelerin çoğunun kültür kabı tabanından ayrıldığı tespit edildi. Enfekte hücrelerde
granülleşme, sitoplazmik uzantılarını kaybederek yuvarlaklaşma, nukleus irileşmesi ve bölünen hücre sayısının azalması
gibi tipik NPV sitopatik etkileri gözlendi. Ektrasellular virus üretimi enfeksiyondan 24 saat sonra başladı. Ancak, BV
miktarındaki önemli artışın enfeksiyondan 48 saat sonra meydana geldiği belirlendi. İlk PIB oluşumu ise enfeksiyondan
36 saat sonra tespit edildi. Md203 hücreleri BV'ler ile yeniden enfekte edildiklerinde hücrelerde yavru virus oluştuğu
belirlendi. Ayrıca, enfeksiyondan 24 saat sonra viral DNA sentezinin önemli seviyede arttığı gözlendi. Sonuçlar,
ManeNPV'nin ilerki çalışmalar için in vitro sistemde çoğaltılabileceğini ve böylece, virusun etkili bir biyoteknolojik
materyal ve mikrobiyal insektisid haline dönüştürülebilme potansiyeline sahip olduğunu göstermektedir.
A plaque-purified genotypic variant, derived from a field isolate of the Malacosoma neustria
nucleopolyhedrovirus (ManeNPV) from Turkey was characterized based on in vitro replication properties in a cell line,
Md203 derived from Malacosoma disstria. The life cycle of ManeNPV was studied based on the cytopathic effects (CPEs),
polyhedral inclusion body (PIB) formation, budded virus (BV) production, viral DNA replication and polyhedrin protein
expression in ManeNPV-infected Md203 cells. Infection of Md203 cells with ManeNPV resulted in cytopathic effects
within 24 and 36 h post infection and most cells detachted from the bottom of the culture plate on the fourth day. Typical
NPV cytopathic effects (CPE) like granulated and rounded cells, nuclear hypertrophy and impairment in cell proliferation,
and lost cell shape with the extendings were observed. Budded viruses were detected at 24 h p.i. However, significant
increase in BV production was observed after 48 h p.i. PIB was firstly detected at 36 h p.i. When BV's were used for
reinfection of the Md203 cells, the result showed that these cells support production of viable virus progeny. We also
observed a significant level of viral DNA synthesis by 24 h p.i. The results indicate that ManeNPV can be propagated at
in vitro system for further studies and thus has a potential for development into an effective biotechnological and
microbial insecticidal agent.
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.
► Nineteen bacterial strains from Agriotes lineatus were isolated and identified. ► Virulence of A. lineatus and different Bacillus isolates was tested on A. lineatus. ► Arthrobacter gandavensis, Bacillus thuringiensis and Pseudomonas plecoglossicida caused 100% mortality. ► Among Bacillus isolates, Bacillus circulans and B. thuringiensis caused 100% mortality. ► Some strains showed potential for biological control.
PDF
