scholarly journals RNA Interference as a Method for Target-Site Screening in the Western Corn Rootworm,Diabrotica Virgifera Virgifera

2010 ◽  
Vol 10 (162) ◽  
pp. 1-16 ◽  
Author(s):  
Analiza P. Alves ◽  
Marcé D. Lorenzen ◽  
Richard W. Beeman ◽  
John E. Foster ◽  
Blair D. Siegfried
Keyword(s):  
Rna Interference ◽  
Diabrotica Virgifera Virgifera ◽  
Western Corn Rootworm ◽  
Target Site ◽  
Corn Rootworm ◽  
Diabrotica Virgifera

scholarly journals Detection of alternative splicing in Diabrotica virgifera virgifera LeConte, in association with Bt resistance using RNA-seq and PacBio Iso-Seq

2020 ◽  
Author(s):  
Zixiao Zhao ◽  
Christine G. Elsik ◽  
Bruce E. Hibbard ◽  
Kent S. Shelby
Keyword(s):  
Alternative Splicing ◽  
Matrix Protein ◽  
Diabrotica Virgifera Virgifera ◽  
Western Corn Rootworm ◽  
Corn Rootworm ◽  
Diabrotica Virgifera ◽  
Rna Seq ◽  
Bt Resistance ◽  
Bt Toxins ◽  
Alternatively Spliced

AbstractBackgroundAlternative splicing is one of the major mechanisms that increases transcriptome diversity in eukaryotes, including insect species that have gained resistance to pesticides and Bt toxins. In western corn rootworm (Diabrotica virgifera virgifera LeConte), neither alternative splicing nor its role in resistance to Bt toxins has been studied.ResultsTo investigate the mechanisms of Bt resistance we carried out single-molecule real-time (SMRT) transcript sequencing and Iso-seq analysis on resistant, eCry3.1Ab-selected and susceptible, unselected, western corn rootworm neonate midguts which fed on seedling maize with and without eCry3.1Ab for 12 and 24 hours. We present transcriptome-wide alternative splicing patterns of western corn rootworm midgut in response to feeding on eCry3.1Ab-expressing corn using a comprehensive approach that combines both RNA-seq and SMRT transcript sequencing techniques. We found that 67.73% of multi-exon genes are alternatively spliced, which is consistent with the high transposable element content of the genome. One of the alternative splicing events we identified was a novel peritrophic matrix protein with two alternative splicing isoforms. Analysis of differential exon usage between resistant and susceptible colonies showed that in eCry3.1Ab-resistant western corn rootworm, expression of one isoform was significantly higher than in the susceptible colony, while no significant differences between colonies were observed with the other isoform.ConclusionOur results provide the first survey of alternative splicing in western corn rootworm and suggest that the observed alternatively spliced isoforms of peritrophic matrix protein may be associated with eCry3.1Ab resistance in western corn rootworm.

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