Transcriptome-Based Selection and Validation of Reference Genes for Gene Expression in Goji Fruit Fly (<i>Neoceratitis asiatica</i> Becker) under Developmental Stages and Five Abiotic Stresses
Goji fruit fly, <i>Neoceratitis asiatica</i>, is a major pest on the well-known medicinal plant <i>Lycium barbarum</i>. Dissecting molecular mechanisms of infestation and host selection of <i>N. asiatica</i> will contribute to the determination of best management...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-12-01
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| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/24/1/451 |
| Summary: | Goji fruit fly, <i>Neoceratitis asiatica</i>, is a major pest on the well-known medicinal plant <i>Lycium barbarum</i>. Dissecting molecular mechanisms of infestation and host selection of <i>N. asiatica</i> will contribute to the determination of best management practices for pest fly control. Gene expression normalization by Real-time quantitative PCR (qPCR) requires the selection and validation of appropriate reference genes (RGs). Hence, 15 candidate RGs were selected from transcriptome data of <i>N. asiatica</i>. Their expression stability was evaluated with five algorithms (∆Ct, Normfinder, GeNorm, BestKeeper, and RefFinder) for sample types differing in the developmental stage, sex, tissue type, and in response to five different abiotic stresses. Our results indicated that the RGs <i>β-Actin</i> + <i>GST</i> for sex, <i>RPL32</i> + <i>EF1α</i> for tissue type, <i>RPS13</i>+ <i>EF1α</i> for developmental stages along with odor stimulation, color induction, and starvation-refeeding stresses, <i>EF1α</i> + <i>GAPDH</i> under insecticide stress, <i>RPS13</i> + <i>RPS18</i> under temperature stress, respectively, were selected as the most suitable RGs for qPCR normalization. Overall, <i>RPS18</i> and <i>EF1α</i> were the two most stable RGs in all conditions, while <i>RPS15</i> and <i>EF1β</i> were the least stable RGs. The corresponding suitable RGs and one unstable RG were used to normalize a target odorant-binding protein OBP56a gene in male and female antennae, different tissues, and under odor stimulation. The results of <i>OBP56a</i> expression were consistent with transcriptome data. Our study is the first research on the most stable RGs selection in <i>N. asiatica</i>, which will facilitate further studies on the mechanisms of host selection and insecticide resistance in <i>N. asiatica</i>. |
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| ISSN: | 1661-6596 1422-0067 |