МОЛЕКУЛЯРНА ІДЕНТИФІКАЦІЯ БІЛКА ПЕПТИДОГЛІКАНУ КОРОТКОГО ТИПУ, GMPGRP-SC ВІД GRAPHOLITHA MOLESTA
Ключові слова:
східна плодожерка, вроджений імунітет, Beauveria bassiana, біологічна боротьба зі шкідниками
Анотація
Білок розпізнавання пептидогліканів (PGRP) є важливим рецептором розпізнавання паттернів, який міститься як у безхребетних, так і у хребетних. Він відіграє важливу роль в антибактеріальному імунітеті, завдяки своїй помітній здатності виявляти та усувати збудника інфекції. Однак PGRP, в основному ідентифіковано з Drosophila melanogaster та Bombyx mori, а щодо інших сільськогосподарських комах повідомлень мало про їхні функції та механізм. У цьому дослідженні короткотиповий ген PGRP під назвою GmPGRP-SC був ідентифікований у Grapholitha molesta – східної плодожерки (OFM) – на основі аналізу бази даних груп транскрипції OFM з нашої лабораторії та Національного центру інформації з біотехнологій (https://www.ncbi.nlm.nih.gov/). GmPGRP-SC містить домен PGRP і має найтісніший генетичний зв'язок з геном PGRP Leguminivora glycinivorella, відповідно до послідовності та філогенетичного аналізу. Метод ПЛР у реальному часі було використано для аналізу картини його експресії на стадіях розвитку OFM та в різних тканинах личинки OFM. Наразі, відносно рівні експресії гена GmPGRP-SC в OFM були проаналізовані після інфікування Beauveria bassiana. Результати показали, що загальна кДНК від GmPGRP-SC становила 3221 bp (базових, основних пар), а кодуючі області – 2268 bp, що кодують 756 амінокислотних залишків. Рівень експресії GmPGRP-SC був найвищим у стадії лялечки OFM. Водночас, у різних тканинах OFM його відносна експресія була вищою в епітелії та гемоциті, тоді як іншим стадіям та тканинам притаманна порівняно нижча та з невеликою різницею. Рівень експресії GmPGRP-SC був істотно різним, коли суспензія спор B. bassiana становить 105 конідій/мкл, інфікованих через 48 годин. Тоді, коли суспензія спор B. bassiana становить 107 конідій/мкл, рівень експресії GmPGRP-SC також був різним. Усі ці результати закладуть основу для вивчення ролі та функцій GmPGRP-SC у вродженому імунітеті OFM, а також сприятимуть подальшому вивченню молекулярної взаємодії між OFM та B. bassiana. Результати досліджень можуть допомогти знайти потенційні молекули-мішені та забезпечити наукову основу для розробки нових біогенних пестицидів та реалізації Зеленої боротьби зі шкідниками (GPM).
Посилання
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2. Armenteros, J. J. A., Tsirigos, K. D., Sonderby, C. K., Petersen, T. N., Winther, O., Brunak, S., Von Heijne, G. & Nielsen, H. (2019). SignalP 5.0 improves signal peptide predictions using deep neural networks. Nature Biotechnology, 37(4), 420 doi: 10.1038/s41587-019-0036-z
3. Blanco, G.A., Malchiodi, E.L. & De Marzi, M.C. (2008). Cellular clot formation in a sipunculan worm: entrapment of foreign particles, cell death and identification of a PGRP-related protein. Journal of invertebrate pathology, 99(2), 156–165.
4. Benelli, G., Lucchi, A., Thomson, D. & Loriatti, C. (2019). Sex pheromone aerosol devices for mating disruption: challenges for a brighter future. Insects, 10, 308.
5. Beutler, B. (2004). Innate immunity: an overview. Molecular Immunology, 40(12), 845–859. doi: 10.1016/j. molimm.2003.10.005
6. Bustin, S. A., Benes, V., Garson, J.A., Jan, H. Jim, H. Mikael, K. Reinhold, M. Tania, N. Pfaffl, M. W. & Shipley, G. L. (2009). The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem, 55(4), 611–622.
7. Cao, J. J. (2015) The expression and functional study of ecdysone signal transduction pathway genes in Grapholitha molesta. [Doctor thesis].
8. Dziarski, R. (2003). Recognition of bacterial peptidoglycan by the innate immune system. Cellular and Molecular Life Sciences, 60, 1793–1804.
9. Dziarski, R. & Gupta, D. (2006) The peptidoglycan recognition proteins (PGRPs). Genome Biology, 7(8), 232. doi: 10.1186/gb-2006-7-8-232
10. Du, S.X., Wu, C., Sue, Y.H. Yang, M. & Nathalie, B. (2011). Effects on mass rearing on the olfactory of an invasive fruit month species, Grapholita molesta (Busck). [Master thesis].
11. Du, J., Guo, J.T., Zhang, Y.S. & Wu, J.X. (2009). Effect of Temperature on Development and Reproduction of Grapholitha molesta (Busck) (Lepidoptera: Tortricidae). Acta Agriculturae Boreali-occidentalis Sinica, 18(6), 314–318.
12. Fischer, B. & Siedler, F. (2004). Using the Agilent 2100 bioanalyzer for quality control of protein samples prior to MS-analysis. Agilent Technologies.
13. Gao, Y.F. (2013). Immune Function Analysis of PGRP-SC and Cathepsin L Genes in Musca domestica. [Master thesis].
14. Geer, L.Y., Marchler-Bauer, A., Geer, R.C., Han, L.Y., He, J., He, S.Q., Liu, C.L., Shi, W.Y. & Bryant, S.H. (2010). The NCBI BioSystems database. Nucleic Acids Research, 38, 492–496. doi: 10.1093/nar/gkp858
15. Gerardo, N.M., Altincicek, B. Anselme, C., Atamian, H.S., Barribeau, S.M., Martin de, V., Duncan, E.J., Evans, J., Gabaldón, T., Ghanim, M., Heddi, A., Kaloshian, I., Latorre, A., Moya, A., Nakabachi, A., Perez-Brocal, V., Pignatelli, M., Rahbe, Y., Ramsey, J.S. Spragg, C., Tamames, J., Tamarit, D., Tamborindeguy, C. & Vincent-Monegat, C. (2010). Immunity and other defenses in pea aphids, Acyrthosiphon pisum. Genome Biology, 11(2), R21. doi: 10.1186/gb-2010-11-2-r21
16. Gottar, M., Gobert, V., Michel, T. Belvin, M., Duyk, G., Hoffmann, J.A., Ferrandon, D. & Royet, J. (2002). The Drosophila immune response against Gram-negative bacteria is mediated by a peptidoglycan recognition protein. Nature, 416(6881), 640–643.
17. Hultmark, D. (2003). Drosophila immunity: paths and patterns. Current Opinion in Immunology, 15(1), 12–19. doi: doi: 10.1016/S0952-7915(02)00005-5
18. Ivica, L. & Peer, B. (2017). 20 years of the SMART protein domain annotation resource. Nucleic Acids Research, 46(1), 493–496. doi: 10.1093/nar/gkx922
19. Kang, D., Liu, G., Lundström, A., Gelius, E. & Steiner, H. (1998). A peptidoglycan recognition protein in innate immunity conserved from insects to humans. Proceedings of The National Academy of Sciences of The United States of America, 95(17), 10078–10082. doi: 10.1073/pnas.95.17.10078
20. Kanga, L. H. B., Pree, D. J., van Lier, J. L. & Walker, G. M. (2003). Management of insecticide resistance in oriental fruit moth (Grapholita molesta; Lepidoptera: Tortricidae) populations from Ontario. Pest Management Science, 59, 921–927.
21. Kong, W. N., Wang, Y., Guo, Y. F., Chai, X. H., Li, J. & Ma, R. Y. (2020). Behavioral effects of different attractants on adult male and female oriental fruit moths, Grapholita molesta. Pest Management Science, 76(9), 3225–3235. doi: 10.1002/ps.5878
22. Leone, P., Bischoff, V., Kellenberger, C., Hetru, C., Royetd, J. & Roussel, A. (2008). Crystal structure of Drosophila PGRP-SD suggests binding to DAP-type but not lysine-type peptidoglycan. Molecular Immunology, 45, 2521–2530. doi: 10.1016/j.molimm.2008.01.015
23. Lin, C. Q. & Yao, B. (2012). Recent Advance in Digital PCR. Progress in Chemistry, 24(12), 2415–2423
24. Livak, K. J. & Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2 (–Delta Delta C(T)) Method. Methods, 25, 402–408.
25. Liu, Y., Shen, D. X., Zhou, F., Wang, G. R. & An, C. J. (2014). Identification of Immunity-Related Genes in Ostrinia furnacalis against Entomopathogenic Fungi by RNA-Seq Analysis, Plos One, 9(1), e86436. doi: 10.1371/journal.pone.0086436
26. Liu W. (2019). Immune response of intestinal reactive oxygen species and peptidoglycan recognition protein genes in Antheraea pernyi. [Doctor thesis].
27. Lu, Y. Z., Su, F. H., Li, Q.L., Zhang, J., Li, Y. J., Tang, T., Hu, Q. H. & Yu, X. Q. (2020). Pattern recognition receptors in Drosophila immune responses. Developmental and Comparative Immunology, 102, 9. doi: 10.1016/j.dci.2019.103468
28. Myllymaki, H., Valanne, S. & Ramet, M., (2014). The Drosophila imd signaling pathway. The Journal of Immunology, 192, 3455–3462
29. Natale, D., Mattiacci, L., Hern, A., Pasqualini, E. & Dorn, S. (2003). Response of female Cydia molesta (Lepidoptera: Tortricidae) to plant derived volatiles. Bulletin of Entomological Research, 93, 335–342. doi: 10.1079/BER2003250
30. Nolan, T., Hands, R.E. & Bustin, S.A. (2006). Quantification of mRNA using real-time RT-PCR. Nature Protocol, 1, 1559–1582.
31. Ochiai, M. & Ashida, M. (1999). A pattern recognition protein for peptidoglycan. Cloning the cDNA and the gene of the silkworm, Bombyx mori. Journal of Biological Chemistry, 274(17), 11854–11858. doi: https://doi.org/10.1074/jbc.274.17.11854
32. Pavsic, J., Zel, J. & Milavec, M (2016). Assessment of the real-time PCR and different digital PCR platforms for DNA quantification. Analytical and Bioanalytical Chemistry, 408(1), 107–121.
33. Pattyn, F., Speleman, F., De Paepe, A. & Vandesompele, J. (2003). RTPrimerDB: the real-time PCR primer and probe database. Nucleic Acids Research, 31(1), 122–123. doi: 10.1093/nar/gkg011
34. Rufer, A. (2010) Therapeutic protein analysis with the Agilent 2100 Bioanalyzer. Biotechniques, 49(3), 669–671.
35. Saranraj, P. & Jayaprakash, A. (2017). A grobeneficial entomopathogenic fungi–Beauveria bassiana: A Review. Asian Journal of Multidisciplinary Research, 3(2), 1051–1087. doi: 10.22192/iajmr.2017.3.2.4
36. Sarker, S., Woo, Y.H. & Lim, U.T. (2020). Laboratory Evaluation of Beauveria bassiana ARP14 Against Grapholita molesta (Lepidoptera: Tortricidae), Current Microbiology, 77(9), 2365–2373. doi: 10.1007/s00284-020-02012-4
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Опубліковано
2022-02-21
Як цитувати
Цао, Ж., ЦaoЦ., Чжу, Х., & Власенко, В. А. (2022). МОЛЕКУЛЯРНА ІДЕНТИФІКАЦІЯ БІЛКА ПЕПТИДОГЛІКАНУ КОРОТКОГО ТИПУ, GMPGRP-SC ВІД GRAPHOLITHA MOLESTA. Вісник Сумського національного аграрного університету. Серія: Агрономія і біологія, 45(3), 52-63. https://doi.org/10.32845/agrobio.2021.3.7
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