Determination of Farnesoic acid O-methyltransferase Gene Polymorphism in Caucasian Bee (Apis mellifera caucasica, G.)

Abstract

Honey bees contribute significantly to the balance and sustainability of the ecosystem through pollination. The Caucasian bee (Apis mellifera caucasica G.), which differs from other honey bee races with distinctly different morphological and physiological characteristics, stands out with its ability to easily adapt to low temperatures and superior wintering ability. Juvenile hormone (JH) plays a vital role in regulating physiological processes in adult bees. In this study, it was aimed to determine the DNA polymorphisms in the 6th, 7th, 4th and 1st exon regions of juvenile hormone esterase, juvenile hormone acid (Farnesoic acid) O-methyltransferase, methyl farnesoate epoxidase and farnesoal epoxidase genes by the PCR-RFLP method. Genomic DNA was isolated from samples from 30 colonies. HinfI was used to cut the regions amplified by PCR. Fragments of 50-55 bc in size were obtained due to cutting the 111 bc FAMET region of the Juvenile hormone acid (Farnesoic acid) O-methyltransferase gene amplified by PCR with the HinfI enzyme. It is thought that polymorphisms up to 5 base pairs in fragment size may be caused by SNP or insertion-deletion mutation. This study is a preliminary study showing that the difference detected in the Juvenile hormone acid (Farnesoic acid) O-methyltransferase gene may be effective on physiological and behavioural characteristics specific to the Caucasian bee, and may be among the molecular causes of the race-specific differences seen in complex processes such as social and seasonal behaviours, age-related task change and colony dynamics and more and detailed studies are needed on this subject.

Keywords

• Biosynthesis
• caucasian bee
• DNA polymorphism
• juvenile hormone
• PCR-RFLP

Kafkas Arısı’nda (Apis mellifera caucasica G.) Farnesoik asit O-metiltransferaz Gen Polimorfizminin Belirlenmesi

Abstract

Polinasyon aracılığıyla ekosistemin dengesine ve sürdürülebilirliğine önemli katkılar sağlayan bal arıları, aynı zamanda tarımsal üretim açısından yüksek ekonomik değere sahiptir. Belirgin farklı morfolojik ve fizyolojik özellikler ile diğer bal arısı ırklarından ayrılan Kafkas arısı (Apis mellifera caucasica G.), düşük sıcaklıklara kolayca adapte olabilmesi ve üstün kışlama yeteneği ile öne çıkmaktadır. Endokrin hormonlardan biri olan Juvenil hormon (JH); bal arılarında başkalaşımın kontrol edilmesi, larval büyüme, ergin dişi böceklerde üreme gibi fizyolojik süreçlerin düzenlenmesinde önemli rol almaktadır. Bu çalışmada JH biyosentezinde rol oynayan juvenil hormon esteraz, juvenil hormon asit (Farnesoik asit) O-metiltransferaz, metil farnesoat epoksidaz ve farnesoal eposidaz genlerinin sırasıyla 6., 7., 4. ve 1. ekzon bölgelerindeki DNA polimorfizmlerinin PCR-RFLP yöntemi ile belirlenmesi amaçlanmıştır. Bu amaçla 30 farklı Kafkas arısı kolonisinden alınan örneklerden genomik DNA izole edilmiştir. PCR ile çoğaltılan bölgelerin kesiminde HinfI restriksiyon endonükeazı kullanılmıştır. Kafkas arı ırkında PCR ile çoğaltılan Juvenil hormon asit (Farnesoik asit) O-metiltransferaz geninin 111 bç büyüklüğündeki FAMET bölgesinin HinfI enzimiyle kesilmesi sonucunda 50-55 bç büyüklüğünde fragmentler elde edilmiştir. Fragment büyüklüğündeki 5 baz çiftine kadar olan polimorfizmin SNP veya indel (insersiyon-delesyon) mutasyonundan kaynaklanabildiği düşünülmektedir. Bu çalışma, JH biyosentezinde rol alan Juvenil hormon asit (Farnesoik asit) O-metiltransferaz geninde tespit edilen farklılığın Kafkas arısına özgü fizyolojik ve davranışsal özellikler üzerine etkili olabileceğini, sosyal ve mevsimsel davranışlar, yaşa bağlı görev değişimi ve koloni dinamikleri gibi karmaşık süreçlerde görülen ırka özgü farklılığın da moleküler nedenleri arasında yer alabileceğini göstermiş olup, bu konuda daha fazla çalışmanın yapılmasına gereksinim bulunmaktadır.

Keywords

• Biyosentez
• DNA polimorfizmi
• juvenil hormon
• kafkas arısı
• PCR-RFLP

References

1. 1. Güler, A. (2017). Bal arısı (Apis mellifera L.) yetiştiriciliği hastalıkları ve ürünleri. Basım. Azim Matbaacılık. Ankara, 419.
2. 2. Karakaş, G. (2022). Pestisit Kullanımının Bal Verimi Üzerine Etkisi; Panel Veri Analizi. Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi, 25(5), 1163-1167. https://doi.org/10.18016/ksutarimdoga.vi.849861
3. 3. Luo, J., Liu, S., Hou, J., Chen, L., Li, H., Liao, S., ... & Li, X. (2021). The comparison of juvenile hormone and transcriptional changes between three different juvenile hormone analogs insecticides on honey bee worker larval’s development. Agronomy, 11(12), 2497. https://doi.org/10.3390/agronomy11122497
4. 4. Güler, A., Bek, Y., Genç, O., Nisbet, C., Konak, F., Hs, E., ... Öztürk, H.(2013). Change level of hygienic behaviour in the caucasian honey bee A M Caucasia G subspecies . 43. International Apicultural Congress, KİYEV, Ukraine
5. 5. Gül, A., & Nergiz, R. (2022). Kafkas Bal Arısı (Apis mellifera caucasia) gen merkezinin bozulmasına neden olan etmenler ve çözüm önerileri. Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi, 25(Ek Sayı 2), 545-554. https://doi.org/10.18016/ksutarimdoga.vi.1084184
6. 6. Li, W., Huang, Z. Y., Liu, F., Li, Z., Yan, L., Zhang, S., ... & Su, S. (2013). Molecular cloning and characterization of juvenile hormone acid methyltransferase in the honey bee, Apis mellifera, and its differential expression during caste differentiation. PloS one, 8(7), e68544. https://doi.org/10.1371/journal.pone.0068544
7. 7. Pandey, A., & Bloch, G. (2015). Juvenile hormone and ecdysteroids as major regulators of brain and behavior in bees. Current Opinion in Insect Science, 12, 26-37. https://doi.org/10.1016/j.cois.2015.09.006
8. 8. Santos, C. G., Humann, F. C., & Hartfelder, K. (2019). Juvenile hormone signaling in insect oogenesis. Current opinion in insect science, 31, 43-48. https://doi.org/10.1016/j.cois.2018.07.010

9. 9. Brejcha, M., Prušáková, D., Sábová, M., Peska, V., Černý, J., Kodrík, D., ... & Frydrychová, R. Č. (2023). Seasonal changes in ultrastructure and gene expression in the fat body of worker honey bees. Journal of insect physiology, 146, 104504. https://doi.org/10.1016/j.jinsphys.2023.104504
10. 10. Ando, H., Ukena, K., & Nagata, S. (Eds.). (2021). Handbook of hormones: comparative endocrinology for basic and clinical research. Academic Press. Doi: 10.1016/C2013-0-15395-0
11. 11. Villalobos-Sambucaro, M. J., Nouzova, M., Ramirez, C. E., Eugenia Alzugaray, M., Fernandez-Lima, F., Ronderos, J. R., & Noriega, F. G. (2020). The juvenile hormone described in Rhodnius prolixus by Wigglesworth is juvenile hormone III skipped bisepoxide. Scientific Reports, 10(1), 3091. https://doi.org/10.1038/s41598-020-59495-1
12. 12. Barchuk, A. R., Cristino, A. S., Kucharski, R., Costa, L. F., Simões, Z. L., & Maleszka, R. (2007). Molecular determinants of caste differentiation in the highly eusocial honeybee Apis mellifera. BMC developmental biology, 7, 1-19. https://doi.org/10.1186/1471-213x-7-70
13. 13. Dominguez, C. V., & Maestro, J. L. (2018). Expression of juvenile hormone acid O‐methyltransferase and juvenile hormone synthesis in Blattella germanica. Insect science, 25(5), 787-796. https://doi.org/10.1111/1744-7917.12467
14. 14. Li, K., Jia, Q. Q., & Li, S. (2019). Juvenile hormone signaling–a mini review. Insect science, 26(4), 600-606. https://doi.org/10.1111/1744-7917.12614
15. 15. Nur Aliah, N. A., Ab-Rahim, S., Moore, H. E., & Heo, C. C. (2021). Juvenile hormone: Production, regulation, current application in vector control and its future applications. https://doi.org/10.47665/tb.38.3.066
16. 16. Tian, Z., Guo, S., Li, J. X., Zhu, F., Liu, W., & Wang, X. P. (2021). Juvenile hormone biosynthetic genes are critical for regulating reproductive diapause in the cabbage beetle. Insect Biochemistry and Molecular Biology, 139, 103654. https://doi.org/10.1016/j.ibmb.2021.103654
17. 17. Gunawardene, Y. S., Tobe, S. S., Bendena, W. G., Chow, B. K. C., Yagi, K. J., & Chan, S. M. (2002). Function and cellular localization of farnesoic acid O‐methyltransferase (FAMeT) in the shrimp, Metapenaeus ensis. European Journal of Biochemistry, 269(14), 3587-3595. https://doi.org/10.1046/j.1432-1033.2002.03048.x
18. 18. Burtenshaw, S. M., Su, P. P., Zhang, J. R., Tobe, S. S., Dayton, L., & Bendena, W. G. (2008). A putative farnesoic acid O-methyltransferase (FAMeT) orthologue in Drosophila melanogaster (CG10527): Relationship to juvenile hormone biosynthesis?. Peptides, 29(2), 242-251. https://doi.org/10.1016/j.peptides.2007.10.030
19. 19. Qu, Z., Bendena, W. G., Tobe, S. S., & Hui, J. H. (2018). Juvenile hormone and sesquiterpenoids in arthropods: biosynthesis, signaling, and role of MicroRNA. The Journal of Steroid Biochemistry and Molecular Biology, 184, 69-76. https://doi.org/10.1016/j.jsbmb.2018.01.013
20. 20. Hui, J. H., Hayward, A., Bendena, W. G., Takahashi, T., & Tobe, S. S. (2010). Evolution and functional divergence of enzymes involved in sesquiterpenoid hormone biosynthesis in crustaceans and insects. Peptides, 31(3), 451-455. https://doi.org/10.1016/j.peptides.2009.10.003
21. 21. Elekonich, M. M., Schulz, D. J., Bloch, G., & Robinson, G. E. (2001). Juvenile hormone levels in honey bee (Apis mellifera L.) foragers: foraging experience and diurnal variation. Journal of Insect Physiology, 47(10), 1119-1125. https://doi.org/10.1016/S0022-1910(01)00090-7
22. 22. Dostálková, S., Dobeš, P., Kunc, M., Hurychová, J., Škrabišová, M., Petřivalský, M., ... & Danihlík, J. (2021). Winter honeybee (Apis mellifera) populations show greater potential to induce immune responses than summer populations after immune stimuli. Journal of Experimental Biology, 224(3), jeb232595. https://doi.org/10.1242/jeb.232595
23. 23. Evans, J. D., Schwarz, R. S., Chen, Y. P., Budge, G., Cornman, R. S., De la Rua, P., ... & Pinto, M. A. (2013). Standard methods for molecular research in Apis mellifera. Journal of Apicultural Research, 52(4), 1-54. https://doi.org/10.3896/IBRA.1.52.4.11
24. 24. Untergasser, A., Cutcutache, I., Koressaar, T., Ye, J., Faircloth, B. C., Remm, M., & Rozen, S. G. (2012). Primer3—new capabilities and interfaces. Nucleic acids research, 40(15), e115-e115. https://doi.org/10.1093/nar/gks596
25. 25. Bomtorin, A. D., Mackert, A., Rosa, G. C. C., Moda, L. M., Martins, J. R., Bitondi, M. M. G., ... & Simões, Z. L. P. (2014). Juvenile hormone biosynthesis gene expression in the corpora allata of honey bee (Apis mellifera L.) female castes. PloS one, 9(1), e86923. https://doi.org/10.1371/journal.pone.0086923
26. 26. Yagi, K. J., Konz, K. G., Stay, B., & Tobe, S. S. (1991). Production and utilization of farnesoic acid in the juvenile hormone biosynthetic pathway by corpora allata of larval Diploptera punctata. General and comparative endocrinology, 81(2), 284-294. https://doi.org/10.1016/0016-6480(91)90013-V
27. 27. Luo, J., Yi, G., Liu, S., Mei, Y., Chen, W., Hou, J., ... & Li, X. (2022). Juvenile Hormone III R Stereoisomer Is Specifically Synthesized by Honeybees (Apis mellifera ligustica) and Shows a Higher Biological Activity in Regulating Their Social Behavior. Journal of Agricultural and Food Chemistry, 70(20), 6097-6107. https://doi.org/10.1021/acs.jafc.2c00762
28. 28. Ferreira, H. M., Di Pietro, V., Wenseleers, T., & Oi, C. A. (2023). Conserved role of juvenile hormone in regulating behavioural maturation and division of labour in a highly eusocial wasp. Animal Behaviour, 200, 59-69. https://doi.org/10.1016/j.anbehav.2023.03.013
29. 29. Huang, Z. Y., & Robinson, G. E. (1995). Seasonal changes in juvenile hormone titers and rates of biosynthesis in honey bees. Journal of Comparative Physiology B, 165, 18-28. https://doi.org/10.1007/BF00264682