Identification of suitable reference genes for miRNA quantitation in bumblebee (Hymenoptera: Apidae) response to reproduction
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The precise quantification of microRNAs (miRNAs) expression level is a critical factor in mastering its functions. We evaluate the suitability of two common genes and ten miRNAs as normalizers for miRNA quantification in the head and ovary at different reproductive status of bumblebees, Bombus lantschouensis by using four different algorithms and one consensus rank approach. For the head and ovary combination, miR-275 was the best candidate. For different tissues, miR-275 was the most stable candidate in the head, while the candidate for the ovary was miR-277. To test the best candidate accuracy, miR-315 was demonstrated to be downregulated based on miR-275 normalization in ovipositor bumblebees. The miR-275 and miR-277 combination is identified to be the most reliable and suitable reference genes for the head and ovary of bumblebees.
Keywordsbumblebee microRNAs reference gene real-time quantification normalization
The authors would like to thank Prof. Jiandong An and Dr. Tolera Kumsa Gemeda for their invaluable comments.
JD and JH conceived this research and designed experiments; JH, JL, and JW participated in the design and interpretation of the data; JD performed experiments and analysis; JD and JH wrote the paper and participated in the revisions of it. All authors read and approved the final manuscript.
This work was funded by China Agriculture Research System (CARS-44), the Agricultural Science and Technology Innovation Program (CAASASTIP-2018-IAR), and the Natural Science Foundation of China (U1603108).
Compliance with ethical standards
Conflicts of interest
The authors declare that they have no conflict of interest.
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