Selection and Validation of Reference Genes for Quantitative Real-Time PCR Analysis in Cockroach Parasitoid <i>Tetrastichus hagenowii</i> (Ratzeburg)
Parasitoid wasps play a crucial role in the efficient control of pests, a substantial menace to human health and well-being. <i>Tetrastichus hagenowii</i> (Ratzeburg) stands out as the most effective egg parasitoid wasp for controlling American cockroaches, but accurate and stable refere...
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MDPI AG
2024-09-01
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| Serija: | Insects |
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| Online pristup: | https://www.mdpi.com/2075-4450/15/9/668 |
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| author | Renke Dong Fengming Cao Jincong Yu Yuan Yuan Jiahui Wang Zining Li Chunxue Zhu Sheng Li Na Li |
| author_facet | Renke Dong Fengming Cao Jincong Yu Yuan Yuan Jiahui Wang Zining Li Chunxue Zhu Sheng Li Na Li |
| author_sort | Renke Dong |
| collection | DOAJ |
| description | Parasitoid wasps play a crucial role in the efficient control of pests, a substantial menace to human health and well-being. <i>Tetrastichus hagenowii</i> (Ratzeburg) stands out as the most effective egg parasitoid wasp for controlling American cockroaches, but accurate and stable reference genes for quantitative real-time polymerase chain reaction of <i>T. hagenowii</i> genes are still lacking. In this study, we assessed seven candidate nuclear genes, including <i>α-tubulin</i> (<i>α-TUB</i>), <i>elongation factor-1-alpha</i> (<i>EF-1α</i>), <i>β-actin</i> (<i>Actin</i>), <i>ribosomal protein 49</i> (<i>RP49</i>), <i>glyceraldehyde-3-phosphate dehydrogenase</i> (<i>GAPDH</i>), <i>nicotinamide adenine dinucleotide</i> (<i>NADH</i>), and <i>elongation factor 2</i> (<i>EF2</i>) of <i>T. hagenowii</i>. By analyzing expression stability with four algorithms (Delta Ct, geNorm, NormFinder, and BestKeeper), as well as comprehensive ranking with RefFinder, we identified <i>α-TUB</i> as the most stable reference gene for the larval, pupal, female adult, and male adult stages. Subsequently, we estimated the transcript levels of vitellogenin (<i>Vg</i>) and cuticle protein (<i>CP</i>) after normalization with <i>α-TUB</i> across various developmental stages. Significantly higher expression levels of <i>CP</i> and <i>Vg</i> were observed in pupae and female adults, respectively, consistent with previous findings in other insects. This study offers a reliable reference gene for normalizing transcription levels of <i>T. hagenowii</i> genes. |
| first_indexed | 2025-03-20T08:32:05Z |
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| institution | Directory Open Access Journal |
| issn | 2075-4450 |
| language | English |
| last_indexed | 2025-03-20T08:32:05Z |
| publishDate | 2024-09-01 |
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| series | Insects |
| spelling | doaj.art-27be0b1ca8bb4e44bcc8fc3a6a714a2c2024-09-27T13:47:10ZengMDPI AGInsects2075-44502024-09-0115966810.3390/insects15090668Selection and Validation of Reference Genes for Quantitative Real-Time PCR Analysis in Cockroach Parasitoid <i>Tetrastichus hagenowii</i> (Ratzeburg)Renke Dong0Fengming Cao1Jincong Yu2Yuan Yuan3Jiahui Wang4Zining Li5Chunxue Zhu6Sheng Li7Na Li8Guangdong Provincial Key Laboratory of Insect Developmental Biology and Applied Technology, Institute of Insect Science and Technology, School of Life Sciences, South China Normal University, Guangzhou 510631, ChinaGuangdong Provincial Key Laboratory of Insect Developmental Biology and Applied Technology, Institute of Insect Science and Technology, School of Life Sciences, South China Normal University, Guangzhou 510631, ChinaGuangdong Provincial Key Laboratory of Insect Developmental Biology and Applied Technology, Institute of Insect Science and Technology, School of Life Sciences, South China Normal University, Guangzhou 510631, ChinaGuangdong Provincial Key Laboratory of Insect Developmental Biology and Applied Technology, Institute of Insect Science and Technology, School of Life Sciences, South China Normal University, Guangzhou 510631, ChinaGuangdong Provincial Key Laboratory of Insect Developmental Biology and Applied Technology, Institute of Insect Science and Technology, School of Life Sciences, South China Normal University, Guangzhou 510631, ChinaGuangdong Provincial Key Laboratory of Insect Developmental Biology and Applied Technology, Institute of Insect Science and Technology, School of Life Sciences, South China Normal University, Guangzhou 510631, ChinaGuangdong Provincial Key Laboratory of Insect Developmental Biology and Applied Technology, Institute of Insect Science and Technology, School of Life Sciences, South China Normal University, Guangzhou 510631, ChinaGuangdong Provincial Key Laboratory of Insect Developmental Biology and Applied Technology, Institute of Insect Science and Technology, School of Life Sciences, South China Normal University, Guangzhou 510631, ChinaGuangdong Provincial Key Laboratory of Insect Developmental Biology and Applied Technology, Institute of Insect Science and Technology, School of Life Sciences, South China Normal University, Guangzhou 510631, ChinaParasitoid wasps play a crucial role in the efficient control of pests, a substantial menace to human health and well-being. <i>Tetrastichus hagenowii</i> (Ratzeburg) stands out as the most effective egg parasitoid wasp for controlling American cockroaches, but accurate and stable reference genes for quantitative real-time polymerase chain reaction of <i>T. hagenowii</i> genes are still lacking. In this study, we assessed seven candidate nuclear genes, including <i>α-tubulin</i> (<i>α-TUB</i>), <i>elongation factor-1-alpha</i> (<i>EF-1α</i>), <i>β-actin</i> (<i>Actin</i>), <i>ribosomal protein 49</i> (<i>RP49</i>), <i>glyceraldehyde-3-phosphate dehydrogenase</i> (<i>GAPDH</i>), <i>nicotinamide adenine dinucleotide</i> (<i>NADH</i>), and <i>elongation factor 2</i> (<i>EF2</i>) of <i>T. hagenowii</i>. By analyzing expression stability with four algorithms (Delta Ct, geNorm, NormFinder, and BestKeeper), as well as comprehensive ranking with RefFinder, we identified <i>α-TUB</i> as the most stable reference gene for the larval, pupal, female adult, and male adult stages. Subsequently, we estimated the transcript levels of vitellogenin (<i>Vg</i>) and cuticle protein (<i>CP</i>) after normalization with <i>α-TUB</i> across various developmental stages. Significantly higher expression levels of <i>CP</i> and <i>Vg</i> were observed in pupae and female adults, respectively, consistent with previous findings in other insects. This study offers a reliable reference gene for normalizing transcription levels of <i>T. hagenowii</i> genes.https://www.mdpi.com/2075-4450/15/9/668<i>Tetrastichus hagenowii</i> (Ratzeburg)reference genesgene expression analysisRT-qPCR normalization |
| spellingShingle | Renke Dong Fengming Cao Jincong Yu Yuan Yuan Jiahui Wang Zining Li Chunxue Zhu Sheng Li Na Li Selection and Validation of Reference Genes for Quantitative Real-Time PCR Analysis in Cockroach Parasitoid <i>Tetrastichus hagenowii</i> (Ratzeburg) Insects <i>Tetrastichus hagenowii</i> (Ratzeburg) reference genes gene expression analysis RT-qPCR normalization |
| title | Selection and Validation of Reference Genes for Quantitative Real-Time PCR Analysis in Cockroach Parasitoid <i>Tetrastichus hagenowii</i> (Ratzeburg) |
| title_full | Selection and Validation of Reference Genes for Quantitative Real-Time PCR Analysis in Cockroach Parasitoid <i>Tetrastichus hagenowii</i> (Ratzeburg) |
| title_fullStr | Selection and Validation of Reference Genes for Quantitative Real-Time PCR Analysis in Cockroach Parasitoid <i>Tetrastichus hagenowii</i> (Ratzeburg) |
| title_full_unstemmed | Selection and Validation of Reference Genes for Quantitative Real-Time PCR Analysis in Cockroach Parasitoid <i>Tetrastichus hagenowii</i> (Ratzeburg) |
| title_short | Selection and Validation of Reference Genes for Quantitative Real-Time PCR Analysis in Cockroach Parasitoid <i>Tetrastichus hagenowii</i> (Ratzeburg) |
| title_sort | selection and validation of reference genes for quantitative real time pcr analysis in cockroach parasitoid i tetrastichus hagenowii i ratzeburg |
| topic | <i>Tetrastichus hagenowii</i> (Ratzeburg) reference genes gene expression analysis RT-qPCR normalization |
| url | https://www.mdpi.com/2075-4450/15/9/668 |
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