Unknown,Transcriptomics,Genomics,Proteomics

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Host plant genotypes affect miRNA gene expression in Hessian fly larvae

ABSTRACT: MicroRNAs (miRNAs) are small noncoding RNAs that play critical roles in regulating post transcriptional gene expression. Gall midges encompass a large group of insects that are of economic importance and also possess fascinating biological traits. The gall midge Mayetiola destructor, commonly known as the Hessian fly, is a model organism for studying gall midge biology and insect M-bM-^@M-^S host plant interactions. In this study, we systematically analyzed miRNAs from the Hessian fly. Deep-sequencing a Hessian fly larval transcriptome led to the identification of 89 miRNA species that are either identical or very similar to known miRNAs from other insects, and 184 novel miRNAs that have not been reported from other species. Microarray analyses revealed the expression of miRNA genes was strictly regulated during Hessian fly larval development and abundance of many miRNA genes were affected by host genotypes. The identification of a large number of miRNAs for the first time from a gall midge provides a foundation for further studies of miRNA functions in gall midge biology and behavior. Two wheat lines M-bM-^@M-^\MollyM-bM-^@M-^] and M-bM-^@M-^\NewtonM-bM-^@M-^] were used in the experiment. Newton is a susceptible winter wheat that contains no Hessian fly R gene, and Molly is a nearly isogenic line of Newton, but contains the R gene H13. Larvae were collected one and three days after egg hatch from susceptible Newton and resistant Molly plants. Total RNA extracted from the collected larvae was used for microarray analysis. Three biological replications were used for each treatment and at each time point.

ORGANISM(S): Mayetiola destructor

SUBMITTER: Chitvan Khajuria

PROVIDER: E-GEOD-43680 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:In honey bees, Vitellogenin (Vg) is hypothesized to be a major factor affecting hormone signaling, food-related behavior, immunity, stress resistance and lifespan. Likewise microRNAs play important roles in posttranscriptional gene regulation and affect many biological processes thereby showing many parallels to Vg functions. The molecular basis of Vg and microRNA interactions is largely unknown. Here, we exploited the well-established RNA interference (RNAi) protocol for Vg knockdown to investigate its effects on microRNA population in honey bee foragerM-bM-^@M-^Ys brain and fat body tissue. To identify microRNAs that are differentially expressed between tissues in control and knockdown foragers, we used M-BM-5ParafloM-BM-. microfluidic oligonucleotide microRNA microarrays. Our results show 76 and 74 miRNAs were expressed in the brain of control and knockdown foragers whereas 66 and 69 miRNAs were expressed in the fat body of control and knockdown foragers respectively. Target prediction identified potential seed matches for differentially expressed subset of microRNAs affected by Vg knockdown. These candidate genes are involved in a broad range of biological processes including insulin signaling, juvenile hormone (JH) and ecdysteroid signaling previously shown to affect foraging behavior. Thus, here we demonstrate a causal link between Vg expression-variation and variation in the abundance of microRNAs in different tissues with possible consequences for regulation of foraging behavior. We knocked down Vitellogenin (Vg) gene expression (using RNAi) in adult workers to identify potential downstream consequences on the expression of microRNA population in the fat body compared to control group (dsRNA-GFP injected bees). Six biological samples of fat body-derived small RNA fraction were prepared for each treatment group (dsRNA-Vg and dsRNA-GFP). Each biological sample contained pooled RNA from 5 unique individuals. Each fat body pool contained a total of 2 M-BM-5g of small RNA fraction, to which each of the 5 individuals contributed equally (400 ng). Pools were named as M-bM-^@M-^\control forager fat bodyM-bM-^@M-^] (GFFb) and M-bM-^@M-^\knockdown forager fat bodyM-bM-^@M-^] (VFFb), followed by a number from 1 to 6.

Project description:In honey bees, Vitellogenin (Vg) is hypothesized to be a major factor affecting hormone signaling, food-related behavior, immunity, stress resistance and lifespan. Likewise microRNAs play important roles in posttranscriptional gene regulation and affect many biological processes thereby showing many parallels to Vg functions. The molecular basis of Vg and microRNA interactions is largely unknown. Here, we exploited the well-established RNA interference (RNAi) protocol for Vg knockdown to investigate its effects on microRNA population in honey bee foragerM-bM-^@M-^Ys brain and fat body tissue. To identify microRNAs that are differentially expressed between tissues in control and knockdown foragers, we used M-BM-5ParafloM-BM-. microfluidic oligonucleotide microRNA microarrays. Our results show 76 and 74 miRNAs were expressed in the brain of control and knockdown foragers whereas 66 and 69 miRNAs were expressed in the fat body of control and knockdown foragers respectively. Target prediction identified potential seed matches for differentially expressed subset of microRNAs affected by Vg knockdown. These candidate genes are involved in a broad range of biological processes including insulin signaling, juvenile hormone (JH) and ecdysteroid signaling previously shown to affect foraging behavior. Thus, here we demonstrate a causal link between Vg expression-variation and variation in the abundance of microRNAs in different tissues with possible consequences for regulation of foraging behavior. We knocked down Vitellogenin (Vg) gene expression (using RNAi) in adult workers to identify potential downstream consequences on the expression of microRNA population in the brain compared to control group (dsRNA-GFP injected bees). Six biological samples of brain-derived small RNA fraction were prepared for each treatment group (dsRNA-Vg and dsRNA-GFP). Each biological sample contained pooled RNA from 5 unique individuals. Each brain pool contained a total of 1 M-BM-5g of small RNA fraction, to which each of the 5 individuals contributed equally (200 ng). Pools were named as M-bM-^@M-^\control forager brainM-bM-^@M-^] (GFBr) and M-bM-^@M-^\knockdown forager brainM-bM-^@M-^] (VFBr), followed by a number from 1 to 6.

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Host plant genotypes affect miRNA gene expression in Hessian fly larvae

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