Structural and ultrastructural alterations of Malpighian tubules of Anticarsia gemmatalis (Hübner) (Lepidoptera: Noctuidae) larvae infected with different Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) recombinant viruses

Cordeiro, Bruno; Tibúrcio, Victor Hugo S.; Hallwass, Mariana; Paes, Hugo Costa; Ribeiro, Bergmann Morais; Báo, Sônia Nair

doi:10.1016/j.jip.2008.01.001

Cited by 10 publications

(8 citation statements)

References 46 publications

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“…Rohrmann (2013) attributes the resistance of specific cell types to the expression of beta-N-acetylglucosaminidase-2 and aminoacylase proteins, which influence the activity of the fusion protein of the GP64 viral envelope, changing the infective capacity of the BV. Cordeiro et al (2008) also reported the absence of infection in the MT cells of Anticarsia gemmatalis (Lepidoptera: Noctuidae) larvae that were infected with recombinant AgMNPV baculovirus. However, they also described the premature degeneration of the MT with morphological changes in the epithelium, which was possibly due to apoptosis resulting from impaired oxygenation and nutrient transport to the MT.…”

Section: Resultsmentioning

confidence: 98%

Ampullae infection of the malpighian tubules of Bombyx mori by Alphabaculovirus

Santorum¹,

Beu²,

Ribeiro³

et al. 2017

Biosci. J.

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ABSTRACT:Alphabaculovirus is a genus of the entomopathogenic virus, whose species Bombyx mori nucleopolyhedrovirus (BmNPV) infects the silkworm, Bombyx mori, which is an important insect in the sericulture industry. A geographic isolate of BmNPV was identified in the state of Paraná, Brazil. It was infecting B. mori larvae and various organs and target tissues were identified, however, there was no information about the infection of Malpighian tubules (MT). The MT comprises the excretory system of B. mori and acts in the elimination of toxic substances and in hydroelectrolytic homeostasis. Thus, the present study examined the susceptibility and cytopathology of B. mori MT to BmNPV. To this end, hybrid fifth instar larvae were inoculated with a virus suspension at different days post-inoculation (dpi). MT segments were collected and divided into the ampullae, proximal, medial and distal regions. These were processed for light microscopy and transmission electron analysis. The MT regions revealed differences in susceptibility to BmNPV and the ampullae in its transition area was infected from the sixth dpi; the other regions did not reveal any evidence of infection. The transition area of the ampullae has not been previously described in Lepidoptera and its cytopathology revealed a hypertrophic nucleus with viroplasm, followed by the formation and development of viral polyhedra, which are common characteristics of infections by Alphabaculovirus. Thus, infection of the ampullae of the MT of B. mori by BmNPV, together with other known targets, compromises the metabolic balance of the insect, which results in consequences for silk production and damage to the sericulture sector.

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Section: Resultsmentioning

confidence: 98%

Ampullae infection of the malpighian tubules of Bombyx mori by Alphabaculovirus

Santorum¹,

Beu²,

Ribeiro³

et al. 2017

Biosci. J.

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“…The constructs containing the promoters fused with the fluc gene (pIE1FLUC, pLEF1FLUC and pGP64FLUC) were subcloned by restriction enzyme digestion with the enzymes Apa I and Sac I, followed by T4 DNA polymerase blunting and DNA ligation into the homologous recombination vector p2100, which contains a unique Eco RV site 158 bp upstream of the polyhedrin ( polh ) ORF start codon [43]. The resulting plasmids, p2100prGP64FLUC, p2100prLEF1FLUC and p2100prIE1FLUC, contain the promoter- fluc constructs and a complete polh gene within a 2100 bp fragment of the polh gene locus of AgMNPV.…”

Section: Methodsmentioning

confidence: 99%

Real-Time Expression Analysis of Selected Anticarsia gemmatalis multiple nucleopolyhedrovirus Gene Promoters during Infection of Permissive, Semipermissive and Nonpermissive Cell Lines

Morgado

Ardisson-Araújo

Ribeiro

2017

Viruses

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Baculovirus infection follows a transcriptionally controlled sequence of gene expression that occurs by activation of different viral gene promoter sequences during infection. This sequence of promoter activation may be disrupted by cellular defenses against viral infection, which might interfere with viral progeny formation. In this work, the activity of the ie1, gp64, lef-1, vp39, p6.9 and polh promoters of the Anticarsia gemmatalis multiple nucleopolyhedrovirus was assessed during infection of permissive, semipermissive and nonpermissive cell lines by a novel methodology that detects reporter protein luminescence in real-time. This technique allowed us to characterize in rich detail the AgMNPV promoters in permissive cell lines and revealed differential profiles of expression in cells with limited permissivity that correlate well with limitations in viral DNA replication. Semipermissive and nonpermissive cell lines presented delays and restrictions in late and very late promoter expression. Cells undergoing apoptosis did not inhibit late gene expression; however, viral progeny formation is severely affected. This work demonstrates the application of the real-time luminescence detection methodology and how the promoter expression profile may be used to diagnose cellular permissivity to baculovirus infection.

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“…The isolated recombinant E. coli was amplified in L-Broth medium, followed by plasmid DNA extraction on a large scale according to the alkaline lysis protocol [23]. The p2100 vector was constructed as described in Cordeiro et al (2008) [24]. In brief, this vector was constructed from PCR amplifications using the AgMNPV HindIII-G DNA fragment cloned into plasmid pGEM3Z (Maruniak et al 1999), generating two fragments of 682 and 1439 bp, using the oligonucleotides pairs polAgR/SphIF and polAgF/PstIR, respectively (Table S1).…”

Section: Methodsmentioning

confidence: 99%

A Recombinant Anticarsia gemmatalis MNPV Harboring chiA and v-cath Genes from Choristoneura fumiferana Defective NPV Induce Host Liquefaction and Increased Insecticidal Activity

et al. 2013

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One of the interesting features of Anticarsia gemmatalis multiple nucleopolyhedrovirus isolate 2D (AgMNPV-2D) genome is the absence of chitinase (chiA) and cathepsin (v-cath) genes. This characteristic may be responsible for the lack of liquefaction and melanization in A. gemmatalis larvae killed by AgMNPV-2D infection. This study aimed to test the hypothesis that CHIA and V-CATH proteins from Choristonera fumiferana DEF multiple nucleopolyhedrovirus (CfDEFNPV) are able to liquefy and melanize the cuticle of A. gemmatalis larvae infected by a recombinant AgMNPV containing chiA and v-cath genes inserted in its genome. A fragment from the CfDefNPV genome containing chiA and v-cath genes was inserted into the genome of AgMNPV-2D. The recombinant virus (vAgp2100Cf.chiA/v-cath) was purified and used to infect insect cells and larvae. Transcripts of v-cath and chiA genes were detected along the infection of insect cells by qRT-PCR, from early to late phases of infection. The analysis of A. gemmatalis larvae killed by vAgp2100Cf.chiA/v-cath infection confirmed the hypothesis proposed. The vAgp2100Cf.chiA/v-cath showed higher insecticidal activity against third instar A. gemmatalis larvae when compared to AgMNPV-2D. The mean time to death was also lower for the vAgp2100Cf.chiA/v-cath when compared to AgMNPV-2D at 10 days post infection. Occlusion body production was higher in A. gemmatalis larvae infected with vAgp2100Cf.chiA/v-cath when compared to AgMNPV-2D. Enzyme assays showed higher chitinase and cysteine protease activities in insect cells and insects infected with vAgp2100Cf.chiA/v-cath when compared to AgMNPV-2D. The introduction of chiA and v-cath genes into the genome of AgMNPV improves its insecticidal activity against A. gemmatalis larvae and this recombinant virus could be used as an alternative to the wild type virus to control this important insect pest.

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Structural and ultrastructural alterations of Malpighian tubules of Anticarsia gemmatalis (Hübner) (Lepidoptera: Noctuidae) larvae infected with different Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) recombinant viruses

Cited by 10 publications

References 46 publications

Ampullae infection of the malpighian tubules of Bombyx mori by Alphabaculovirus

Ampullae infection of the malpighian tubules of Bombyx mori by Alphabaculovirus

Real-Time Expression Analysis of Selected Anticarsia gemmatalis multiple nucleopolyhedrovirus Gene Promoters during Infection of Permissive, Semipermissive and Nonpermissive Cell Lines

A Recombinant Anticarsia gemmatalis MNPV Harboring chiA and v-cath Genes from Choristoneura fumiferana Defective NPV Induce Host Liquefaction and Increased Insecticidal Activity

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