Transcriptional analysis of Tranosema rostrale ichnovirus (TrIV) genes, with emphasis on the rep genes family

Authors: Rasoolizadeh, Asieh
Advisor: Cloutier, ConradCusson, Michel
Abstract: The endoparasitic wasp Tranosema rostrale transmits an ichnovirus (“TrIV”) to its lepidopteran host, Choristoneura fumiferana, during parasitization. This virus, which has a segmented dsDNA genome and can replicate only in the wasp’s ovaries, is essential to the survival of the immature wasp within its host. In a prior study, 86 putative open reading frames (ORFs) were identified in the TrIV genome, including 35 that could be assigned to previously recognized ichnoviral gene families. The balance displayed no similarity to known genes. In an effort to assess (i) the accuracy of the TrIV genome annotation and (ii) the relative importance of each gene family in the success of parasitism by T. rostrale, a temporal and tissue-specific qPCR transcriptional analysis was conducted in infected C. fumiferana hosts and T. rostrale wasp ovaries. The majority (91%) of putative ORFs assigned to known gene families were observed to be expressed in infected larvae, albeit at widely varying levels, but this proportion was lower (67%) for a sample of 12 unassigned ORFs. Among the seven known gene families present in the TrIV genome, the rep family is the numerically most important one, with 17 members; all of these were shown to be expressed in infected larvae and/or wasp ovaries. In infected caterpillars, however, two of them, F1-1 and F1-2, had much more abundant transcripts than the others. The rep transcriptional profile was markedly different in wasp ovaries, where the C166-1 gene generated the most abundant rep transcripts, suggesting that different members of this family may have host-specific functions. Relative abundance of genome segments was highest for the two segments bearing the three most highly expressed rep genes, but the correlation between these two variables was poor for the other rep genes, suggesting that some other factors are involved in the regulation of rep gene expression in infected larvae. Inter-gene differences were also observed in the relative abundance of TrIV rep transcripts in different C. fumiferana tissues, pointing to tissue-specific roles or specialized functions for individual members of this gene family. In comparing rep transcript levels to those of genes belonging to other known TrIV gene families, a TrV (TrV1) and a rep (F1-1) gene clearly outnumbered all other genes examined in infected caterpillars, pointing to the likely importance of these two gene families in host subjugation by T. rostrale. In wasp ovaries, the transcriptional profile was dominated by a rep gene and a member of a newly described family identified among previously unassigned ORFs; these genes encode secreted proteins displaying a novel cysteine motif.
Document Type: Mémoire de maîtrise
Issue Date: 2009
Open Access Date: 16 April 2018
Grantor: Université Laval
Collection:Thèses et mémoires

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