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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).
Insect Biochem Mol Biol. 2016 08; 75:117-24.IB

Abstract

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.

Authors+Show Affiliations

Dow AgroSciences, 9330 Zionsville Rd, Indianapolis, IN 46268, United States.Dow AgroSciences, 9330 Zionsville Rd, Indianapolis, IN 46268, United States.University of Nebraska-Lincoln, 103 Entomology Hall, Lincoln, NE 68583, United States.University of Nebraska-Lincoln, 103 Entomology Hall, Lincoln, NE 68583, United States.Dow AgroSciences, 9330 Zionsville Rd, Indianapolis, IN 46268, United States.Dow AgroSciences, 9330 Zionsville Rd, Indianapolis, IN 46268, United States.Dow AgroSciences, 9330 Zionsville Rd, Indianapolis, IN 46268, United States.University of Nebraska-Lincoln, 103 Entomology Hall, Lincoln, NE 68583, United States.University of Nebraska-Lincoln, 103 Entomology Hall, Lincoln, NE 68583, United States.Dow AgroSciences, 9330 Zionsville Rd, Indianapolis, IN 46268, United States.Dow AgroSciences, 9330 Zionsville Rd, Indianapolis, IN 46268, United States.Dow AgroSciences, 9330 Zionsville Rd, Indianapolis, IN 46268, United States.University of Florida, Entomology and Nematology Department, Charles Steinmetz Hall, PO Box 110620, Gainesville, FL 32611-0620, United States. Electronic address: bsiegfried1@ufl.edu.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

27334721

Citation

Tan, Sek Yee, et al. "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)." Insect Biochemistry and Molecular Biology, vol. 75, 2016, pp. 117-24.
Tan SY, Rangasamy M, Wang H, et al. 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). Insect Biochem Mol Biol. 2016;75:117-24.
Tan, S. Y., Rangasamy, M., Wang, H., Vélez, A. M., Hasler, J., McCaskill, D., Xu, T., Chen, H., Jurzenski, J., Kelker, M., Xu, X., Narva, K., & Siegfried, B. D. (2016). 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). Insect Biochemistry and Molecular Biology, 75, 117-24. https://doi.org/10.1016/j.ibmb.2016.06.006
Tan SY, et al. 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). Insect Biochem Mol Biol. 2016;75:117-24. PubMed PMID: 27334721.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - 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). AU - Tan,Sek Yee, AU - Rangasamy,Murugesan, AU - Wang,Haichuan, AU - Vélez,Ana María, AU - Hasler,James, AU - McCaskill,David, AU - Xu,Tao, AU - Chen,Hong, AU - Jurzenski,Jessica, AU - Kelker,Matthew, AU - Xu,Xiaoping, AU - Narva,Kenneth, AU - Siegfried,Blair D, Y1 - 2016/06/19/ PY - 2016/02/27/received PY - 2016/06/14/revised PY - 2016/06/15/accepted PY - 2016/6/24/entrez PY - 2016/6/24/pubmed PY - 2017/7/15/medline KW - Cadherin KW - Cry34/35Ab1 KW - Cry34Ab1 KW - Cry35Ab1 KW - Cry3Aa KW - Diabrotica virgifera virgifera KW - RNAi SP - 117 EP - 24 JF - Insect biochemistry and molecular biology JO - Insect Biochem Mol Biol VL - 75 N2 - 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. SN - 1879-0240 UR - https://www.unboundmedicine.com/medline/citation/27334721/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__ DB - PRIME DP - Unbound Medicine ER -