Evaluation of Reference Genes for Real-Time Quantitative PCR Analysis in Tissues from Bumble Bees ( Bombus Terrestris ) of Different Lines

• Published in: International journal of molecular sciences 2022-11, Vol.23 (22), p.14371
• Main Authors: Sankar, Kathannan, Yoon, Hyung Joo, Lee, Young Bo, Lee, Kyeong Yong
• Format: Article
• Language: English
• Subjects: Actin; Actins - genetics; Animals; Arginine kinase; Bees; Bees - genetics; bumble bee; Dehydrogenases; Elongation; Fat body; Female; Gene expression; Genes; Glyceraldehyde-3-phosphate dehydrogenase; Glyceraldehyde-3-Phosphate Dehydrogenases; Kinases; Molecular modelling; Ovaries; Peptide Elongation Factor 1 - genetics; Phospholipase A2; Phospholipases A2; Pollinators; Polymerase chain reaction; Proteins; qRT-PCR; Real time; Real-Time Polymerase Chain Reaction - methods; reference gene; RefFinder; Ribosomal proteins; Stability analysis; Thorax
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms232214371

Bumble bees are important alternative pollinators and model insects due to their highly developed sociality and colony management. In order to better understand their molecular mechanisms, studies focusing on the genetic and molecular aspects of their development and behavior are needed. Although quantitative real-time polymerase chain reaction (qRT-PCR) can be used to quantify the relative expression of target genes, internal reference genes (which are stably expressed across different lines and tissues) must first be identified to ensure the accurate normalization of target genes. In order to contribute to molecular studies on bumble bees, we used to determine the expression stability of eight reference genes (β-actin (ACT), Arginine Kinase (AK), Phospholipase A2 (PLA2), Elongation factor 1 alpha (EF-1), Ribosomal proteins (S5, S18, S28) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH)) in five different lines and several tissues (ovary, thorax, fat body, and head) using RT-qPCR procedures and four analysis programs (RefFinder, NormFinder, BestKeeper, and geNorm). In general, the S28, S5, and S18 ribosomal protein genes and the PLA2 and EF-1 genes showed the highest stability and were therefore identified as suitable reference genes for the bumble bee species and their defined lines and tissues. Our results also emphasized the need to evaluate the stability of candidate reference genes for any differently designed lines and tissue conditions in bumble bee species.