Abstract
The green mirid bug Cyrtorhinus lividipennis (Hemiptera: Miridae), known as a predator, plays an important role in modulating the population density of planthoppers and leafhoppers in rice ecosystem. To understand the molecular mechanisms underlying many physiological processes of C. lividipennis research on gene expression are required, and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) is the most commonly used method to quantify the expression of target genes. However, there are no previous reports on reliable reference genes in C. lividipennis. Here, we evaluated the stability of eight candidate reference genes (RPL32, β-Tub, SDHA, RPL9, GAPDH, EF1α, β-Act and α-Act) under different conditions with five algorithms (ΔCt method, BestKeeper, GeNorm, NormFinder and RefFinder). Results have indicated that RPL32 and RPL9 are suitable reference genes across nymphal stages, RPL32 and RPL9 are the suggested reference genes in different developmental stages of adult females, RPL32, RPL9 and β-Tub are suitable reference genes for all developmental stages, β-Tub, GAPDH, RPL9 and β-Act are reliable reference genes in different tissues of 5th instar females, EF1α, RPL9, RPL32 and β-Tub are stable reference genes in different tissues of one-day-old adult females, β-Tub, EF1α, RPL9, RPL32 and β-Act are suitable reference genes in all samples. These results highlight the importance of identifying reference genes under different experimental treatments. Our study will facilitate further phylogenetic and physiological mechanism research at the transcriptional level in C. lividipennis.
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Abbreviations
- 18S :
-
rRNA 18 S ribosomal RNA.
- Act :
-
Actin.
- cDNA:
-
Complementary deoxyribonucleic acid.
- ClJHAMT :
-
Cyrtorhinus lividipennis juvenile hormone acid methyl transferase.
- ClJHE :
-
Cyrtorhinus lividipennis juvenile hormone esterase.
- ClKr-h1 :
-
Cyrtorhinus lividipennis Krüppel homolog 1.
- ClSPATA13 :
-
Cyrtorhinus lividipennis spermatogenesis-associated protein 13.
- ClVg :
-
Cyrtorhinus lividipennis vitellogenin.
- Ct:
-
Threshold cycle.
- ddH2O:
-
Double distilled water.
- EF1α :
-
Eukaryotic elongation factor 1-alph.
- GAPDH :
-
Glyceraldehyde-3-phosphate dehydrogenase.
- NCBI:
-
National Center for Biotechnology Information.
- qPCR:
-
Quantitative polymerase chain reaction.
- RH:
-
Relative humidity.
- RNA:
-
Ribonucleic Acid.
- RPL32 :
-
Ribosomal protein L32.
- RPL9 :
-
Ribosomal protein L9.
- RPS15 :
-
Ribosomal protein S15.
- RT-qPCR:
-
Reverse transcription-quantitative polymerase chain reaction.
- SD:
-
Standard deviation.
- SDHA :
-
Succinate dehydrogenase complex flavoprotein subunit A.
- SV:
-
Stability value.
- TOR:
-
target of rapamycin.
- α-Act :
-
alph-Actin.
- β-Act :
-
beta-Actin.
- β-Tub :
-
beta-Tubin.
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This work was funded by the National Natural Science Foundation of China (No. 32001894 and 31872928).
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Hu, K., Chen, K., Fu, B. et al. Reliable reference genes for qPCR normalization in females of the mirid predator, Cyrtorhinus lividipennis (Hemiptera: Miridae). Biologia 77, 2383–2392 (2022). https://doi.org/10.1007/s11756-022-01093-9
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DOI: https://doi.org/10.1007/s11756-022-01093-9