Deep sequencing of organ- and stage-specific micrornas in the evolutionarily basal insect Blattella germanica (L.) (Dictyoptera, Blattellidae)

Cristino, Alexandre S., Tanaka, Erica D., Rubio, Mercedes, Piulachs, Maria-Dolors and Belles, Xavier (2011) Deep sequencing of organ- and stage-specific micrornas in the evolutionarily basal insect Blattella germanica (L.) (Dictyoptera, Blattellidae). PLoS One, 6 4: e19350.1-e19350.11. doi:10.1371/journal.pone.0019350


Author Cristino, Alexandre S.
Tanaka, Erica D.
Rubio, Mercedes
Piulachs, Maria-Dolors
Belles, Xavier
Title Deep sequencing of organ- and stage-specific micrornas in the evolutionarily basal insect Blattella germanica (L.) (Dictyoptera, Blattellidae)
Formatted title
Deep sequencing of organ- and stage-specific micrornas in the evolutionarily basal insect Blattella germanica (L.) (Dictyoptera, Blattellidae)
Journal name PLoS One   Check publisher's open access policy
ISSN 1932-6203
Publication date 2011-04-01
Year available 2011
Sub-type Article (original research)
DOI 10.1371/journal.pone.0019350
Open Access Status DOI
Volume 6
Issue 4
Start page e19350.1
End page e19350.11
Total pages 11
Place of publication San Francisco, CA, United States
Publisher Public Library of Science
Language eng
Subject 1300 Biochemistry, Genetics and Molecular Biology
1100 Agricultural and Biological Sciences
Abstract Background: microRNAs (miRNAs) have been reported as key regulators at post-transcriptional level in eukaryotic cells. In insects, most of the studies have focused in holometabolans while only recently two hemimetabolans (Locusta migratoria and Acyrthosiphon pisum) have had their miRNAs identified. Therefore, the study of the miRNAs of the evolutionarily basal hemimetabolan Blattella germanica may provide valuable insights on the structural and functional evolution of miRNAs.
Formatted abstract
Background: microRNAs (miRNAs) have been reported as key regulators at post-transcriptional level in eukaryotic cells. In insects, most of the studies have focused in holometabolans while only recently two hemimetabolans (Locusta migratoria and Acyrthosiphon pisum) have had their miRNAs identified. Therefore, the study of the miRNAs of the evolutionarily basal hemimetabolan Blattella germanica may provide valuable insights on the structural and functional evolution of miRNAs.
Methodology/Principal Findings: Small RNA libraries of the cockroach B. germanica were built from the whole body of the last instar nymph, and the adult ovaries. The high throughput Solexa sequencing resulted in approximately 11 and 8 million reads for the whole-body and ovaries, respectively. Bioinformatic analyses identified 38 known miRNAs as well as 11 known miRNA*s. We also found 70 miRNA candidates conserved in other insects and 170 candidates specific to B. germanica. The positive correlation between Solexa data and real-time quantitative PCR showed that number of reads can be used as a quantitative approach. Five novel miRNA precursors were identified and validated by PCR and sequencing. Known miRNAs and novel candidates were also validated by decreasing levels of their expression in dicer-1 RNAi knockdown individuals. The comparison of the two libraries indicates that whole-body nymph contain more known miRNAs than ovaries, whereas the adult ovaries are enriched with novel miRNA candidates.
Conclusions/Significance: Our study has identified many known miRNAs and novel miRNA candidates in the basal hemimetabolan insect B. germanica, and most of the specific sequences were found in ovaries. Deep sequencing data reflect miRNA abundance and dicer-1 RNAi assay is shown to be a reliable method for validation of novel miRNAs.
Keyword Multidisciplinary Sciences
Science & Technology - Other Topics
MULTIDISCIPLINARY SCIENCES
Q-Index Code C1
Q-Index Status Confirmed Code
Grant ID CGL2008-03517/BOS
2005 SGR 00053
Institutional Status UQ
Additional Notes Article # e19350

Document type: Journal Article
Sub-type: Article (original research)
Collections: Queensland Brain Institute Publications
Official 2012 Collection
 
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Citation counts: TR Web of Science Citation Count  Cited 55 times in Thomson Reuters Web of Science Article | Citations
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Created: Sat, 10 Dec 2011, 02:08:50 EST by Debra McMurtrie on behalf of Queensland Brain Institute