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Pestic Biochem Physiol ; 157: 186-195, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31153467


The sugarcane shoot borer Chilo infuscatellus (Snellen) is known for causing severe damage to sugarcane yield in China. Methods have been developed to control this pest, including Cry toxin pesticide and transgenic Bt plants. In order to investigate the molecular mechanism of the Cry toxin binding process and provide a basis for understanding the insect's resistance mechanism, we used a high throughput sequencing platform to perform a de novo transcriptome assembly across different larval developmental stages and analyzed Cry toxin receptors based on our assembled transcripts. We cloned twelve Cry toxin receptor genes including 1 cadherin (Cad), 7 aminopeptidase-Ns (APNs), 3 alkaline phosphatases (ALPs), and 1 ATP-binding cassette transporter subfamily C2 (ABCC2), and three of them with full length. The sublethal dosage of Cry1Ac toxin was applied to sugarcane shoot borer and identified some Cry toxin receptor genes that were significantly induced after 48 h of exposure. Furthermore, quantitative RT-PCR was conducted to detect the expression profiles of these genes. Our transcriptome sequence data provided a valuable molecular resource for further study and the identified Cry toxin receptor data gave insights for improved research into the mechanism of Bt resistance.

Proteínas de Bactérias/metabolismo , Proteínas Hemolisinas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Saccharum/metabolismo , Animais , Proteínas de Bactérias/genética , Endotoxinas/genética , Endotoxinas/metabolismo , Proteínas Hemolisinas/genética , Resistência a Inseticidas/genética , Mariposas , Plantas Geneticamente Modificadas/genética , Saccharum/genética
Pestic Biochem Physiol ; 156: 56-62, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31027581


Glutamate-gated chloride channels (GluCls) mediate inhibitory synaptic transmission in invertebrate nervous systems, and only one GluCl gene has been found in insects. Therefore, insect GluCls are one of the major targets of insecticides including avermectins. In the present study, a 1347 bp full-length cDNA encoding a 449-amino acid protein (named MsGluCl, GenBank ID: MK336885) was cloned from the oriental armyworm, Mythimna separata, and characterized two alternative splicing variants of MsGluCl. The protein shares 76.9-98.6% identity with other insect GluCl isoforms. Spatial and temporal expression analysis revealed that MsGluCl was highly expressed in the 3rd instar and adult head. Dietary ingestion of dsMsGluCl significantly reduced the mRNA level of MsGluCl and decreased abamectin mortality. Thus, our results reveal that MsGluCl could be the molecular target of abamectin and provide the basis for further understanding the resistance mechanism to abamectin in arthropods.

Processamento Alternativo/genética , Canais de Cloreto/metabolismo , Clonagem Molecular/métodos , Mariposas/genética , Animais , Canais de Cloreto/genética , DNA Complementar/genética , DNA Complementar/metabolismo , Inseticidas/farmacologia , Ivermectina/análogos & derivados , Ivermectina/farmacologia , Mariposas/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
Vet Res Commun ; 34(1): 25-32, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20024621


Challenge tests with Artemia four different development stages (nauplii, metanauplii, pseudoadults and adults) to white spot syndrome virus was carried out by immersion challenge and virus-phytoplankton adhesion route in order to asses the possibility of Artemia acting as a vector of WSSV to penaeid shrimp Litopenaeus vannamei postlarvae. The WSSV succeeded in infecting four stages Artemia, and nested-PCR detection for WSSV revealed positive results to virus-phytoplankton adhesion route. No mass mortalities were observed in penaeid shrimp postlarvae fed with WSSV-positive Artemia which exposed to WSSV by virus-phytoplankton adhesion route, whereas WSSV DNA detected in penaeid shrimp postlarvae by nested-PCR. By contrary, no WSSV-positive was detected in any animal fed with WSSV-negative Artemia. These results indicated that Artemia could serve as a vector in WSSV transmission.

Artemia/virologia , Transmissão Vertical de Doença Infecciosa/veterinária , Penaeidae/virologia , Fitoplâncton/virologia , Vírus da Síndrome da Mancha Branca 1/isolamento & purificação , Animais , Vetores Artrópodes/virologia , DNA Viral , Estágios do Ciclo de Vida , Vírus da Síndrome da Mancha Branca 1/genética
J Invertebr Pathol ; 97(1): 33-9, 2008 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-17698077


White Spot Syndrome Virus (WSSV) is now one of the most devastating and virulent viral agents threatening the penaeid shrimp culture industry and has been responsible for serious economic losses for shrimp farms worldwide. One remarkable characteristic of WSSV is its wide reservoir range, which contributes to its wide geographical distribution. Among epizootiological surveys, there is substantial evidence for WSSV-positive copepods found in shrimp farming ponds. Therefore, copepods are suspected to be the vector of WSSV. In the present study, nested-PCR analysis showed positive results in the harpacticoid copepod Nitocra sp. exposed to WSSV by virus-phytoplankton adhesion route. Oral route and intramuscular injection were used to test the pathogenicity of WSSV isolated from the WSSV-positive Nitocra sp. For the oral route of infection, Marsupenaeus japonicus postlarvae were fed with WSSV-positive copepods. The shrimp postlarvae in the infected treatment became WSSV-positive and occurred 52.50+/-5.00% mortality which was significant higher (P <0.05) than that in the control treatment (20.00+/-0.00%) when postlarvae were fed with WSSV free copepods. In the intramuscular injection challenge, M. japonicus juveniles were injected with the copepods inoculum extracted from the WSSV-positive Nitocra sp., and showed 72.50+/-9.57% mortality which was also significant higher (P <0.05) than that in the control treatment (22.50+/-5.00%) when juveniles were received mock injection of a tissue homogenate prepared from WSSV-negative Nitocra sp. Based on these laboratory challenge studies, it was confirmed that the copepods can serve as a vector in WSSV transmission.

Copépodes/virologia , Vetores de Doenças , Penaeidae/virologia , Viroses/transmissão , Vírus da Síndrome da Mancha Branca 1/fisiologia , Animais , Bioensaio , Reação em Cadeia da Polimerase
J Invertebr Pathol ; 94(2): 144-8, 2007 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-17069846


To test the possibility that shrimp pond rotifer resting eggs and hatched rotifers could transmit white spot syndrome virus (WSSV) to crayfish (Procambarus clarkii), we injected crayfish with rotifer and resting egg inocula that were WSSV-positive only by dot-blot analysis of PCR products. No crayfish became WSSV-positive after challenge with the resting egg inoculum. However, 1/15 crayfish became WSSV-positive after challenge with the rotifer inoculum. The results demonstrated that rotifers constitute a potential risk for WSSV transmission to crayfish and other cultivated crustaceans. However, the actual quantitative risk of transmission in an aquaculture setting depends on many variables that remain untested.

Aquicultura , Astacoidea/virologia , Vetores de Doenças , Rotíferos/virologia , Vírus da Síndrome da Mancha Branca 1/patogenicidade , Animais , DNA Viral/análise , DNA Viral/genética , Transmissão de Doença Infecciosa , Immunoblotting , Óvulo/virologia , Reação em Cadeia da Polimerase , Replicação Viral , Vírus da Síndrome da Mancha Branca 1/genética , Vírus da Síndrome da Mancha Branca 1/isolamento & purificação