HT-SuperSAGE of the gut tissue of a Vip3Aa-resistant Heliothis virescens (Lepidoptera: Noctuidae) strain provides insights into the basis of resistance.

Authors:
Dr. Camilo Ayra-Pardo, PhD
Dr. Camilo Ayra-Pardo, PhD
Henan Provincial Engineering Laboratory of Insect Bio-reactor / Nanyang Normal University
Associate Professor
Plant Biotechnology
Nanyang, Henan | China
Maria E Ochagavia
Maria E Ochagavia
Plant Division Centre for Genetic Engineering and Biotechnology (CIGB)
Ben Raymond
Ben Raymond
School of Biological Sciences
Kansas City | United States
Asim Gulzar
Asim Gulzar
Imperial College London
United Kingdom
Dr. Ryohei Terauchi, PhD
Dr. Ryohei Terauchi, PhD
Iwate Biotechnology Research Center
Head of Division
Kitakami, Iwate | Japan

Insect Sci 2017 Sep 5. Epub 2017 Sep 5.

Department of Life Sciences, Faculty of Natural Sciences, Imperial College London, Berkshire, UK.

Multitoxin Bt-crops expressing insecticidal toxins with different modes of action, for example, Cry and Vip, are expected to improve resistance management in target pests. While Cry1A resistance has been relatively well characterized in some insect species, this is not the case for Vip3A, for which no mechanism of resistance has yet been identified. Here we applied HT-SuperSAGE to analyze the transcriptome of the gut tissue of tobacco budworm Heliothis virescens (F.) laboratory-selected for Vip3Aa resistance. From a total of 1 324 252 sequence reads, 5 895 126-bp tags were obtained representing 17 751 nonsingleton unique transcripts (UniTags) from genetically similar Vip3Aa-resistant (Vip-Sel) and susceptible control (Vip-Unsel) strains. Differential expression was significant (≥2.5 fold or ≤0.4; P < 0.05) for 1989 sequences (11.2% of total UniTags), where 420 represented overexpressed (OE) and 1569 underexpressed (UE) genes in Vip-Sel. BLASTN searches mapped 419 UniTags to H. virescens sequence contigs, of which, 416 (106 OE and 310 UE) were unambiguously annotated to proteins in NCBI nonredundant protein databases. Gene Ontology distributed 345 of annotated UniTags in 14 functional categories with metabolism (including serine-type hydrolases) and translation/ribosome biogenesis being the most prevalent. A UniTag homologous to a particular member of the REsponse to PAThogen (REPAT) family was found among most overexpressed, while UniTags related to the putative Vip3Aa-binding ribosomal protein S2 (RpS2) were underexpressed. qRT-PCR of a subset of UniTags validated the HT-SuperSAGE data. This study is the first providing lepidopteran gut transcriptome associated with Vip3Aa resistance and a foundation for future attempts to elucidate the resistance mechanism.

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http://dx.doi.org/10.1111/1744-7917.12535DOI Listing

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September 2017
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