RNAi induced knockdown of a cadherin-like protein (EF531715) does not affect toxicity of Cry34/35Ab1 or Cry3Aa to Diabrotica virgifera virgifera larvae (Coleoptera: Chrysomelidae)
Autor: | David McCaskill, Ana María Vélez, Blair D. Siegfried, Murugesan Rangasamy, Jessica Jurzenski, James M. Hasler, Kenneth E. Narva, Haichuan Wang, Tao Xu, Sek Yee Tan, Matthew S. Kelker, Xiaoping Xu, Hong Chen |
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Rok vydání: | 2016 |
Předmět: |
0106 biological sciences
0301 basic medicine Insecticides Bacillus thuringiensis Real-Time Polymerase Chain Reaction Zea mays 01 natural sciences Biochemistry Insecticide Resistance Hemolysin Proteins 03 medical and health sciences Bacterial Proteins Botany Gene expression Animals Molecular Biology Gene knockdown Genetically modified maize Bacillus thuringiensis Toxins biology Cadherin fungi Midgut Cadherins Plants Genetically Modified biology.organism_classification Molecular biology Coleoptera Endotoxins 010602 entomology 030104 developmental biology Western corn rootworm Larva Insect Science Insect Proteins RNA Interference PEST analysis |
Zdroj: | Insect Biochemistry and Molecular Biology. 75:117-124 |
ISSN: | 0965-1748 |
Popis: | The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, is an important maize pest throughout most of the U.S. Corn Belt. Bacillus thuringiensis (Bt) insecticidal proteins including modified Cry3Aa and Cry34/35Ab1 have been expressed in transgenic maize to protect against WCR feeding damage. To date, there is limited information regarding the WCR midgut target sites for these proteins. In this study, we examined whether a cadherin-like gene from Diabrotica virgifera virgifera (DvvCad; GenBank accession # EF531715) associated with WCR larval midgut tissue is necessary for Cry3Aa or Cry34/35Ab1 toxicity. Experiments were designed to examine the sensitivity of WCR to trypsin activated Cry3Aa and Cry34/35Ab1 after oral feeding of the DvvCad dsRNA to knockdown gene expression. Quantitative real-time PCR confirmed that DvvCad mRNA transcript levels were reduced in larvae treated with cadherin dsRNA. Relative cadherin expression by immunoblot analysis and nano-liquid chromatography - mass spectrometry (nanoLC-MS) of WCR neonate brush border membrane vesicle (BBMV) preparations exposed to DvvCad dsRNA confirmed reduced cadherin expression when compared to BBMV from untreated larvae. However, the larval mortality and growth inhibition of WCR neonates exposed to cadherin dsRNA for two days followed by feeding exposure to either Cry3Aa or Cry34/35Ab1 for four days was not significantly different to that observed in insects exposed to either Cry3Aa or Cry34/35Ab1 alone. In combination, these results suggest that cadherin is unlikely to be involved in the toxicity of Cry3Aa or Cry34/35Ab1 to WCR. |
Databáze: | OpenAIRE |
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