Deep Sequencing of Organ- and Stage-Specific microRNAs in the Evolutionarily Basal Insect Blattella germanica (L.) (Dictyoptera, Blattellidae)
نویسندگان
چکیده
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.
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Stage-Dependent Tolerance of the German Cockroach, Blattella germanica for Dichlorvos and Propoxur
tage-dependent dichlorvos and propoxur tolerance in a field population of the German cockroach, Blattella germanica Linnaeus (Blatodea: Blattellidae), was investigated in the laboratory using a topical application bioassay. The results showed the 6 week-old nymphs were more tolerant to dichlorvos and propoxur than the other ages tested. LD₅₀ values of dichlorvos and propoxur for the 6 week-old ...
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سابقه و هدف: امروزه تحقیقات برای دست یابی به حشره کش های سازگار با محیط زیست به دلیل توسعه مقاومت به حشره کش های شیمیایی و اهمیت پزشکی و اقتصادی سوسری آلمانی (Blattella germanica) در اماکن شهری کماکان در حال افزایش است. در مطالعه حاضر سمیت تماسی اسانس های سیر (Allium sativum) و نارنگی (Citrus nobilis var. deliciosa) علیه حشرات کامل سوسری آلمانی مورد ارزیابی قرار گرفت. مواد و روش ها: اسانس های گ...
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