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| Characterization and functional analysis of β-1,3-galactosyltransferase involved in Cry1Ac resistance from Helicoverpa armigera (Hübner) |
| ZHANG Li-li, LIANG Ge-mei, GAO Xi-wu, CAO Guang-chun, GUO Yu-yuan |
1、College of Agriculture and Biotechnology, China Agricultural University, Beijing 100193, P.R.China
2、State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R.China |
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摘要 Carbohydrate chains are the principal antigens by which Bacillus thuringiensis (Bt) identify receptor proteins. The interaction between the antigen and Bt causes a pore in the membrane of midgut epithelial cells of insects. Receptor proteins, such as aminopeptidase N and alkaline phosphatase, are glycoproteins. Cadherin is another cell surface receptor protein which has potential glycosylation sites. Glycosyltransferase is very important for the synthesis and modification of receptor proteins. It can indirectly influence the function of Bt. The 1 950 bp full-length cDNA encoding β-1,3-galactosyltransferase was cloned from the the midgut of Helicoverpa armigera by degenerative PCR combined with RACE techniques (GAL-Harm, GenBank accession no.: GQ904195.1) with two potential N-glycosylation sites (157NNTI160 and 272NKTL275). Protein sequence alignments revealed that H. armigera β-1,3-galactosyltransferase shared high identity with β-1,3-galactosyltransferase in other insect species. The expression level of the β-1,3-galactosyltransferase gene in Cry1Ac-resistant H. armigera larvae was 9.2-fold higher than that in susceptible strain. The function of β-1,3-galactosyltransferase was investigated using RNAi technique. The result showed Cry1Ac enhanced the toxicity against the siRNA-treated larvae compared with non-siRNA-treated ones, which indicated β-1,3-galactosyltransferase played an important role for the insecticidal toxicity of Cry1Ac in H. armigera.
Abstract Carbohydrate chains are the principal antigens by which Bacillus thuringiensis (Bt) identify receptor proteins. The interaction between the antigen and Bt causes a pore in the membrane of midgut epithelial cells of insects. Receptor proteins, such as aminopeptidase N and alkaline phosphatase, are glycoproteins. Cadherin is another cell surface receptor protein which has potential glycosylation sites. Glycosyltransferase is very important for the synthesis and modification of receptor proteins. It can indirectly influence the function of Bt. The 1 950 bp full-length cDNA encoding β-1,3-galactosyltransferase was cloned from the the midgut of Helicoverpa armigera by degenerative PCR combined with RACE techniques (GAL-Harm, GenBank accession no.: GQ904195.1) with two potential N-glycosylation sites (157NNTI160 and 272NKTL275). Protein sequence alignments revealed that H. armigera β-1,3-galactosyltransferase shared high identity with β-1,3-galactosyltransferase in other insect species. The expression level of the β-1,3-galactosyltransferase gene in Cry1Ac-resistant H. armigera larvae was 9.2-fold higher than that in susceptible strain. The function of β-1,3-galactosyltransferase was investigated using RNAi technique. The result showed Cry1Ac enhanced the toxicity against the siRNA-treated larvae compared with non-siRNA-treated ones, which indicated β-1,3-galactosyltransferase played an important role for the insecticidal toxicity of Cry1Ac in H. armigera.
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Received: 31 December 2013
Accepted:
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| Fund: This research was supported by the Key Technologies R&D Program of China during the 12th Five-Year Plan period (2012BAD19B05) and the National Natural Science Foundation of China (30971921,31321004). We would like to thank Biotechnology Group in Institute of Plant Protection, Chinese Academy of Agricultural Sciences for provision of Cry1Ac strain. |
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Corresponding Authors:
LIANG Ge-mei, Tel: +86-10-62815929, E-mail: gmliang@ippcaas.cn; GUO Yu-yuan, E-mail: yyguo@ippcaas.cn
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| About author: ZHANG Li-li, E-mail: zhangliliwell@126.com |
Cite this article:
ZHANG Li-li, LIANG Ge-mei, GAO Xi-wu, CAO Guang-chun, GUO Yu-yuan.
2015.
Characterization and functional analysis of β-1,3-galactosyltransferase involved in Cry1Ac resistance from Helicoverpa armigera (Hübner). Journal of Integrative Agriculture, 14(2): 337-346.
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