Bemisia tabaci (Gennadius)’de İki Nikotinik Asetilkolin Reseptör Genin Klonlanması

Nikotinik asetilkolin reseptörleri (nAChR), böcek sinir sisteminde hızlı kolinerjik sinaptik geçişe aracılık ederler. Neonikotinoid grubu insektisitler, bu reseptörleri hedefleyerek böceklerde ölüme neden olurlar. Bu çalışmanın amacı, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) nAChR alt birim genlerinin cDNA klonlarını izole etmektir. Bu genlerin elde edebilmesi için RACE (cDNA Ends'in hızlı amplifikasyonu) tekniği uygulanmıştır. Bu çalışmada, B. tabaci (Bt)’den sırayla iki kısmi nAChR alt birim geni β1 ve α8 (Btβ1-1168 bp ve Btα8-755 bp) elde edilmiştir. Bu çalışmayla Bemisia tabaci nAChR α8 alt birimi, klonlanarak ilk kez elde edilmiştir. Btβ1 ve Btα8'in nAChR alt birimlerinin tipik ve birbirinden farklı özelliklerine sahip olduğunu belirlenmiştir. Filogenetik ağaç sonuçları, B. tabaci’nin Btβ1 ve Btα8 alt birim genlerinin, farklı böcek türlerine ait ortolog gen bölgeleriyle kümelendiğini göstermektedir.

Cloning of Two Nicotinic Acetylcholine Receptor Genes from Bemisia tabaci (Gennadius)

Nicotinic acetylcholine receptors (nAChRs) mediate fast cholinergic synaptic transmission in theinsect nervous system. Neonicotinoid insecticides exhibit insecticidal activities by targeting thesereceptors. The aim of this study was to isolate cDNA clones of nAChR subunit genes of Bemisiatabaci (Gennadius) (Hemiptera: Aleyrodidae). RACEs (Rapid Amplification of cDNA Ends) wereapplied to obtain partial-length cDNA sequences. We identified two partial cDNA clones encodingβ1 and α8 subunit genes (Btβ1-1168 bp and Btα8-755 bp), respectively, from Bemisia tabaci (Bt).This is the first report of isolation of α8 from B. tabaci by cloning. Btβ1 and Btα8 possesscharacteristics that are typical of nAChR subunits. Phylogenetic analysis showed that Btβ1 and Btα8clustered with the orthologous genes of other insect species.

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Abbick J. 1991. The biochemistry of imidacloprid. Pflanzenschutz-NACh. Richten Bayer, 44: 198–195. EPPO, 2004. Bemisia tabaci. Bulletin OEPP/EPPO Bulletin, 34: 281–288.
Arias HR. 1997. Topology of ligand binding sites on the nikotinik asetilkoline receptor. Brain Res. Brain Res. Rev., 25(2): 133–91.
Bass C, Puinean AM, Andrews M, Cutler P, Daniels M, Elias J, Paul VL, Crosswaite AJ, Denholm I, Field LM, Foster SP, Lind R, Williamson MS, Slater R. 2011. Mutation of a nicotinic acetylcholine receptor β subunit is associated with resistance to neonicotinoid insecticides in the aphid Myzus persicae. BMC Neuroscience, 12: 51.
Blom N, Gammeltoft S, Brunak S. 1999. Sequence and structure based prediction of eukaryotic protein phosphorylation sites. J. Mol. Biol., 294: 1351–1362
Campanella JJ, Bitincka L, Smalley J. 2003. MatGAT: An application that generates similarity/identity matrices using protein or DNA sequences. BMC Bioinformatics, 4. http:// www.biomedcentral.com/1471-2105/4/29.
Ilias A, Lagnel J, Kapantaidaki D, Roditakis E, Tsigenopoulos CS, Vontas J, Tsagkarakou A. 2015. Transcription analysis of neonicotinoid resistance in Mediterranean (MED) populations of B. tabaci reveal novel cytochrome. BMC Genomics, 16 (1): 939.
Jeschke P, Nauen R. 2008. Neonicotinoids–from zero to hero in insecticide chemistry. Pest Manag. Sci, 64: 1084–1098.939 (2015).
Jones AK, Satelle DB. 2010. Diversity of insect nicotinic acetylcholine receptor subunits. In: Steeve Hervé Thany (editor) Insect nicotinic acetylcholine receptors. New York NY, Springer. pp. 25-43. ISBN 978-1-4419-6445-8 (Online).
Karunker I, Juergen B, Bettina L, Tanja P, Nauen R, Emmanouil R, John V, Kevin G, Ian D, Shai M. 2008. Over-expression of cytochrome P450 CYP6CM1 is associated with high resistance to imidacloprid in the B and Q biotypes of Bemisia tabaci (Hemiptera: Aleyrodidae). Insect Biochemistry and Molecular Biology, 38(6): 634–44.
Millar NS, Denholm I. 2007. Nikotinik asetilkoline receptors: targets for commercially important insecticides. Invert. Neurosci, 7: 53–66.
Nauen R, Vontas J, Kaussmann M, Wolfel K. 2013. Pymetrozine is hydroxylated by CYP6CM1, a cytochrome P450 conferring neonicotinoid resistance in Bemisia tabaci. Pest Management Science, 69(4): 457–61.
NCBI, 2019. https://www.ncbi.nlm.nih.gov/nuccore/?term= Bemisia+tabaci+ Nicotinic+acetylcholine (18.12.2019).
Nicholas KB, Nicholas HBJ, Deerfıeld DW. 1997. GeneDoc: Analysis and Visualization of Genetic Variation. http://www.psc.edu/biomed/genedoc, EMBNEW.NEWS, 4: 14.
Rice P, Longden I, Bleasby A. 2000. EMBOSS: the European MolecularBiology Open Software Suite. Trends Genet., 16: 276–277
Roditakis E, Roditakis NE, Tsagkarakou A. 2005. Insecticide resistance in Bemisia tabaci (Homoptera: Aleyrodidae) populations from. Crete. Pest Manag Sci, 61: 577–582.
Roditakis E, Morou E, Tsagkarakou A, Riga M, Nauen R, Paine M, Vontas J. 2011. Assessment of the Bemisia tabaci CYP6CM1vQ transcript and protein levels in laboratory and field-derived imidacloprid-resistant insects and crossmetabolism potential of the recombinant enzyme. Insect Science, 18(1): 23–29.
Satar G, Ulusoy MR, Nauen R, Dong K. 2018. Neonicotinoid insecticide resistance among populations of Bemisia tabaci in the Mediterranean region of Turkey. Bulletin of Insectology, 71(2): 171–177.
Shi X, Zhu Y, Xia X, Qiao K, Wang H, Wang K. 2012. The mutation in nicotinic acetylcholine receptor β1 subunit may confer resistance to imidacloprid in Aphis gossypii (Glover). Journal of Food, Agriculture & Environment, 10: 1227–1230.
Stenersen J. 2004. Nicotinoids and Neonicotinoids Chemical pesticides. Mode of Action and Toxicology, pp. 135-136.
Tan J, Salgado VL, Hollingworth RW. 2008. Neural actions of imidacloprid and their involvement in resistance in the Colorado potato beetle, Leptinotarsa decemlineata (Say). Pest Manag. Sci, 64: 37–47.
Tomizawa M, Casida JE. 2003. Selective Toxicity of Neonicotinoids attributable to Specifıcity of Insect and Mammalian Nicotinik Receptors. Annu. Rev. Entomol., 48: 339–64.
Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. 1997. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res., 25: 4876–4882.
Wang W, Wang S, Han G, Du Y, Wang J 2017. Lack of crossresistance between neonicotinoids and sulfoxaflor in field strains of Q-biotype of whitefly, Bemisia tabaci, from eastern China. Pesticide Biochemistry and Physiology, 136: 46–51.
Wang R, Fang Y, Mu C, Qu C, Li F, Wang Z, Luo C. 2018. Baseline susceptibility and cross-resistance of cycloxaprid, a novel cis-nitromethylene neonicotinoid insecticide, in Bemisia tabaci MED from China. Crop Protection, 110: 283– 287.
Zaidi SSEA, Briddon RW, Mansoor S. 2017. Engineering dual begomovirus-Bemisia tabaci resistance in plants. Trends in Plant Science, 22(1): 6–8.
Zewen L, Zhaojun H, Yinchang W, Lingchun Z, Hongwei Z, Chengjun L. 2003. Selection for imidacloprid resistance in Nilaparvata lugens: cross-resistance patterns and possible mechanisms. Pest Manag. Science, 59: 1355–1359