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DNA METHYLATION OF GENES RELATED TO HONEY BEE WORKER REPRODUCTIVE PHYSIOLOGY |
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| รหัสดีโอไอ | |
| Title | DNA METHYLATION OF GENES RELATED TO HONEY BEE WORKER REPRODUCTIVE PHYSIOLOGY |
| Creator | Manlika Kilaso |
| Contributor | Chanpen Chanchao, Benjamin P. Oldroyd |
| Publisher | Chulalongkorn University |
| Publication Year | 2558 |
| Keyword | Bees -- Reproduction, DNA, ผึ้ง -- การสืบพันธุ์, ดีเอ็นเอ |
| Abstract | Venom gland and ovary are the important organs related to a reproductive system in honey bee workers. The aim of this study is to investigate whether DNA methylation is involved in the regulation of function role of both organs in workers. In the first part, the frequency of DNA methylation is determined in four Thai native honey bee species which are A. andreniformis, A. florea, A. cerana indica and A. dorsata by focusing on venom phospholipase A2 (PLA2). Reverse transcriptase polymerase chain reaction (RT-PCR) revealed that the highest expression of PLA2 is found in house bees of all four Apis spp. while the high expression of PLA2 is observed in foragers of A. florea and A. dorsata. In contrast, expression of PLA2 is undetectable in pupae from all Apis spp. The PLA2 activity and specific activity from crude venom extract shows the higher level in house bees than in black-eyed pupae in all studied bees. DNA methylation level of pupae is higher than that of house bees and foragers of A. florea and A. dorsata but it is not the case in A. andreniformis and A. cerana indica. In the second part, under the queenless condition, workers are divided into control and experimental (CO2 narcosis) cages. Using methylation-sensitive amplified fragment length polymorphism (MS-AFLP) assay, there is no difference detectable in a pattern of genome-wide DNA methylation between workers with non-active and active ovaries in any comparisons. Similarly, non-treated and CO2-treated workers are not significantly different in DNA methylation pattern. For the last part, DNA methylation at target region of Krüppel homolog-1 (Kr-h1) is determined in 7-day old queenless workers. Bisulfite sequencing shows 12 methylated CpG sites found across amplified region of Kr-h1 in examined workers. The overall DNA methylation level of Kr-h1 is significantly higher in non-active ovaries than active ovaries. CO2 narcosis does not have a significant effect to the DNA methylation change in Kr-h1 in worker ovaries. In addition, it is found that there is one deletion site specific to one site of CpG methylation. |
| URL Website | cuir.car.chula.ac.th |
