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      Transcriptional analysis of susceptible and resistant European corn borer strains and their response to Cry1F protoxin

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          Abstract

          Background

          Despite a number of recent reports of insect resistance to transgenic crops expressing insecticidal toxins from Bacillus thuringiensis (Bt), little is known about the mechanism of resistance to these toxins. The purpose of this study is to identify genes associated with the mechanism of Cry1F toxin resistance in European corn borer ( Ostrinia nubilalis Hübner). For this, we compared the global transcriptomic response of laboratory selected resistant and susceptible O. nubilalis strain to Cry1F toxin. We further identified constitutive transcriptional differences between the two strains.

          Results

          An O. nubilalis midgut transcriptome of 36,125 transcripts was assembled de novo from 106 million Illumina HiSeq and Roche 454 reads and used as a reference for estimation of differential gene expression analysis. Evaluation of gene expression profiles of midgut tissues from the Cry1F susceptible and resistant strains after toxin exposure identified a suite of genes that responded to the toxin in the susceptible strain ( n = 1,654), but almost 20-fold fewer in the resistant strain ( n = 84). A total of 5,455 midgut transcripts showed significant constitutive expression differences between Cry1F susceptible and resistant strains. Transcripts coding for previously identified Cry toxin receptors, cadherin and alkaline phosphatase and proteases were also differentially expressed in the midgut of the susceptible and resistant strains.

          Conclusions

          Our current study provides a valuable resource for further molecular characterization of Bt resistance and insect response to Cry1F toxin in O. nubilalis and other pest species.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s12864-015-1751-6) contains supplementary material, which is available to authorized users.

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          Most cited references44

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          Controlling the False Discovery Rate: A Practical and Powerful Approach to Multiple Testing

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            Next-generation transcriptome assembly.

            Transcriptomics studies often rely on partial reference transcriptomes that fail to capture the full catalogue of transcripts and their variations. Recent advances in sequencing technologies and assembly algorithms have facilitated the reconstruction of the entire transcriptome by deep RNA sequencing (RNA-seq), even without a reference genome. However, transcriptome assembly from billions of RNA-seq reads, which are often very short, poses a significant informatics challenge. This Review summarizes the recent developments in transcriptome assembly approaches - reference-based, de novo and combined strategies - along with some perspectives on transcriptome assembly in the near future.
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              Bacillus thuringiensis and its pesticidal crystal proteins.

              During the past decade the pesticidal bacterium Bacillus thuringiensis has been the subject of intensive research. These efforts have yielded considerable data about the complex relationships between the structure, mechanism of action, and genetics of the organism's pesticidal crystal proteins, and a coherent picture of these relationships is beginning to emerge. Other studies have focused on the ecological role of the B. thuringiensis crystal proteins, their performance in agricultural and other natural settings, and the evolution of resistance mechanisms in target pests. Armed with this knowledge base and with the tools of modern biotechnology, researchers are now reporting promising results in engineering more-useful toxins and formulations, in creating transgenic plants that express pesticidal activity, and in constructing integrated management strategies to insure that these products are utilized with maximum efficiency and benefit.
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                Author and article information

                Contributors
                neetha@unl.edu
                hwang4@unl.edu
                seyun2@unl.edu
                emoriyama2@unl.edu
                brad.coates@ars.usda.gov
                nick.miller@unl.edu
                bsiegfried1@unl.edu
                Journal
                BMC Genomics
                BMC Genomics
                BMC Genomics
                BioMed Central (London )
                1471-2164
                29 July 2015
                29 July 2015
                2015
                : 16
                : 1
                : 558
                Affiliations
                [ ]Department of Entomology, University of Nebraska-Lincoln, Lincoln, NE USA
                [ ]Center for Biotechnology, University of Nebraska-Lincoln, Lincoln, NE USA
                [ ]School of Biological Sciences and Center for Plant Science Innovation, University of Nebraska-Lincoln, Lincoln, NE USA
                [ ]USDA-ARS, Corn Insects and Crop Genetics Research Unit, Ames, IA USA
                Article
                1751
                10.1186/s12864-015-1751-6
                4518661
                26220297
                8bce7410-da6b-432e-b5bc-7056590a8358
                © Nanoth Vellichirammal et al. 2015

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 8 May 2015
                : 6 July 2015
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2015

                Genetics
                european corn borer,ostrinia nubilalis,insect resistance,cry1f resistance,bt-toxin,transcriptomics,cry1f response,rna-seq

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