The Cadherin Protein Is Not Involved in Susceptibility to <i>Bacillus thuringiensis</i> Cry1Ab or Cry1Fa Toxins in <i>Spodoptera frugiperda</i>
It is well known that insect larval midgut cadherin protein serves as a receptor of <i>Bacillus thuringiensis</i> (Bt) crystal Cry1Ac or Cry1Ab toxins, since structural mutations and downregulation of <i>cad</i> gene expression are linked with resistance to Cry1Ac toxin in se...
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2020-06-01
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doaj-25874b64c44a455181392edfa5f99d592020-11-25T02:27:07ZengMDPI AGToxins2072-66512020-06-011237537510.3390/toxins12060375The Cadherin Protein Is Not Involved in Susceptibility to <i>Bacillus thuringiensis</i> Cry1Ab or Cry1Fa Toxins in <i>Spodoptera frugiperda</i>Jianfeng Zhang0Minghui Jin1Yanchao Yang2Leilei Liu3Yongbo Yang4Isabel Gómez5Alejandra Bravo6Mario Soberón7Yutao Xiao8Kaiyu Liu9Institute of Entomology, School of Life Sciences, Central China Normal University, Wuhan 430079, ChinaAgricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518120, ChinaInstitute of Entomology, School of Life Sciences, Central China Normal University, Wuhan 430079, ChinaInstitute of Entomology, School of Life Sciences, Central China Normal University, Wuhan 430079, ChinaInstitute of Entomology, School of Life Sciences, Central China Normal University, Wuhan 430079, ChinaInstituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo. Postal 510-3, Cuernavaca 62250, Morelos, MexicoInstituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo. Postal 510-3, Cuernavaca 62250, Morelos, MexicoInstituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo. Postal 510-3, Cuernavaca 62250, Morelos, MexicoAgricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518120, ChinaInstitute of Entomology, School of Life Sciences, Central China Normal University, Wuhan 430079, ChinaIt is well known that insect larval midgut cadherin protein serves as a receptor of <i>Bacillus thuringiensis</i> (Bt) crystal Cry1Ac or Cry1Ab toxins, since structural mutations and downregulation of <i>cad</i> gene expression are linked with resistance to Cry1Ac toxin in several lepidopteran insects. However, the role of <i>Spodoptera frugiperda</i> cadherin protein (SfCad) in the mode of action of Bt toxins remains elusive. Here, we investigated whether SfCad is involved in susceptibility to Cry1Ab or Cry1Fa toxins. <i>In vivo</i>, knockout of the <i>SfCad</i> gene by CRISPR/Cas 9 did not increase tolerance to either of these toxins in <i>S. frugiperda</i> larvae. <i>In vitro</i> cytotoxicity assays demonstrated that cultured insect TnHi5 cells expressing GFP-tagged SfCad did not increase susceptibility to activated Cry1Ab or Cry1Fa toxins. In contrast, expression of another well recognized Cry1A receptor in this cell line, the ABCC2 transporter, increased the toxicity of both Cry1Ab and Cry1Fa toxins, suggesting that SfABCC2 functions as a receptor of these toxins. Finally, we showed that the toxin-binding region of SfCad did not bind to activated Cry1Ab, Cry1Ac, nor Cry1Fa. All these results support that SfCad is not involved in the mode of action of Cry1Ab or Cry1Fa toxins in <i>S. frugiperda</i>.https://www.mdpi.com/2072-6651/12/6/375<i>Bacillus thuringiensis</i><i>Spodoptera frugiperda</i>cadherinCry1AbCry1Famode of action of Cry toxin |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Jianfeng Zhang Minghui Jin Yanchao Yang Leilei Liu Yongbo Yang Isabel Gómez Alejandra Bravo Mario Soberón Yutao Xiao Kaiyu Liu |
spellingShingle |
Jianfeng Zhang Minghui Jin Yanchao Yang Leilei Liu Yongbo Yang Isabel Gómez Alejandra Bravo Mario Soberón Yutao Xiao Kaiyu Liu The Cadherin Protein Is Not Involved in Susceptibility to <i>Bacillus thuringiensis</i> Cry1Ab or Cry1Fa Toxins in <i>Spodoptera frugiperda</i> Toxins <i>Bacillus thuringiensis</i> <i>Spodoptera frugiperda</i> cadherin Cry1Ab Cry1Fa mode of action of Cry toxin |
author_facet |
Jianfeng Zhang Minghui Jin Yanchao Yang Leilei Liu Yongbo Yang Isabel Gómez Alejandra Bravo Mario Soberón Yutao Xiao Kaiyu Liu |
author_sort |
Jianfeng Zhang |
title |
The Cadherin Protein Is Not Involved in Susceptibility to <i>Bacillus thuringiensis</i> Cry1Ab or Cry1Fa Toxins in <i>Spodoptera frugiperda</i> |
title_short |
The Cadherin Protein Is Not Involved in Susceptibility to <i>Bacillus thuringiensis</i> Cry1Ab or Cry1Fa Toxins in <i>Spodoptera frugiperda</i> |
title_full |
The Cadherin Protein Is Not Involved in Susceptibility to <i>Bacillus thuringiensis</i> Cry1Ab or Cry1Fa Toxins in <i>Spodoptera frugiperda</i> |
title_fullStr |
The Cadherin Protein Is Not Involved in Susceptibility to <i>Bacillus thuringiensis</i> Cry1Ab or Cry1Fa Toxins in <i>Spodoptera frugiperda</i> |
title_full_unstemmed |
The Cadherin Protein Is Not Involved in Susceptibility to <i>Bacillus thuringiensis</i> Cry1Ab or Cry1Fa Toxins in <i>Spodoptera frugiperda</i> |
title_sort |
cadherin protein is not involved in susceptibility to <i>bacillus thuringiensis</i> cry1ab or cry1fa toxins in <i>spodoptera frugiperda</i> |
publisher |
MDPI AG |
series |
Toxins |
issn |
2072-6651 |
publishDate |
2020-06-01 |
description |
It is well known that insect larval midgut cadherin protein serves as a receptor of <i>Bacillus thuringiensis</i> (Bt) crystal Cry1Ac or Cry1Ab toxins, since structural mutations and downregulation of <i>cad</i> gene expression are linked with resistance to Cry1Ac toxin in several lepidopteran insects. However, the role of <i>Spodoptera frugiperda</i> cadherin protein (SfCad) in the mode of action of Bt toxins remains elusive. Here, we investigated whether SfCad is involved in susceptibility to Cry1Ab or Cry1Fa toxins. <i>In vivo</i>, knockout of the <i>SfCad</i> gene by CRISPR/Cas 9 did not increase tolerance to either of these toxins in <i>S. frugiperda</i> larvae. <i>In vitro</i> cytotoxicity assays demonstrated that cultured insect TnHi5 cells expressing GFP-tagged SfCad did not increase susceptibility to activated Cry1Ab or Cry1Fa toxins. In contrast, expression of another well recognized Cry1A receptor in this cell line, the ABCC2 transporter, increased the toxicity of both Cry1Ab and Cry1Fa toxins, suggesting that SfABCC2 functions as a receptor of these toxins. Finally, we showed that the toxin-binding region of SfCad did not bind to activated Cry1Ab, Cry1Ac, nor Cry1Fa. All these results support that SfCad is not involved in the mode of action of Cry1Ab or Cry1Fa toxins in <i>S. frugiperda</i>. |
topic |
<i>Bacillus thuringiensis</i> <i>Spodoptera frugiperda</i> cadherin Cry1Ab Cry1Fa mode of action of Cry toxin |
url |
https://www.mdpi.com/2072-6651/12/6/375 |
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