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Çukurova Bölgesi Pamuk Alanlarında Bemisia tabaci (Gennadius)’de Aşırı Düzeyde Sitokrom P450 CYP6CM1 Gen Ekspresyonu

Yıl 2020, Cilt: 23 Sayı: 3, 634 - 640, 30.06.2020

Selçuk Ulusoy

https://doi.org/10.18016/ksutarimdoga.vi.605716

Öz

Çukurova bölgesi polikültür tarımın yapıldığı
bir bölgedir. Bu bölgede pamuk ekim alanlarında B. tabaci mücadelesinde neonikotinoid grubu insektisitler yoğun bir
şekilde kullanılmakta olup, direnç problemi ile karşılaşılmaktadır. Çalışmada
2018 yılı, Adana ili pamuk alanlarından toplanmış beş farklı B. tabaci popülasyonunda imidacloprid
dayanıklılığı göstergesi olan sitokrom P450 CYP6CM1 geni relatif expresyon
düzeyleri incelenmiştir. Ayrıca metabolik dayanıklılıkda etkili GST geni
relatif expresyon profili, sitokrom P450, karboksil esteraz(AcHE) ve Glutathion
S transferaz (GST) enzim etkinlikleri belirlenmiştir. En yüksek relatif
aktivite Meletmez populasyonunda CYP6CM1 geni 18.2 kat ve Gst geni 9 kat daha
fazla gen regülasyonu belirlenmiştir. Ayrıca en yüksek ve en düşük sitokrom
P450 enzim aktivitesi meletmez ve solaklı popülasyonlarında gözlemlenmiştir.
Yapılan analizlerde Çukurova bölgesi pamuk alanlarında B. tabaci popülasyonlarında yaygın imidacloprid ve neonikotinoid
dayanıklılığı durumundan bahsedilebilir. Sonuç olarak, aşırı gen expresyonu ve
yüksek düzeydeki enzim etkinlikleri, bölgede güçlendirilmiş monooksigenaz
etkinliğine dayanan direncin varlığını ortaya koymaktadır.

Anahtar Kelimeler

Bemisia tabaci Pamuk Sitokrom P450 Aşırı ekspresyon

Kaynakça

  • Ahmad, M., M. I. Arif & I. Denholm, 2003. High resistance of field populations of the cotton aphid Aphis gossypii Glover (Homoptera: Aphididae) to pyrethroid insecticides in Pakistan. Journal of Economic Entomology, 96: 875-878.
  • Anonymous, 2019. Türkiye istatistik kurumu. http://www.tuik.gov.tr (26.03.2019).
  • Bass, C. & L. M. Field, 2011. Gene amplification and insecticide resistance. Pest Management Science 67 (8): 886-890.
  • Bel-Kadhi, M.S., 2008. Molecular characterization of Bemisia tabaci biotypes in Southern Tunisia. Tunisian Journal of Plant Protection, 3(2): 79-85.
  • Bradford, M.M., 1976. A rapid and sensitive metod for the quantitation of microgram quantites of protein. utulizing the principle of protein dye binding. Analytical Biochemistry, 72:248-254.
  • Cahill, M., Gorman, K., Day, S., Denholm, I., Elbert, A. & Nauen, R., 1996. Baseline determination and detection of resistance to imidacloprid in Bemisia tabaci (Homoptera: Aleyrodidae). Bull. Entomol. Res. 86, 343–349.
  • Devonshire, A. L. & G. D. Moores, 1982. A carboxylesterase with broad substrate specify causes organophosphorus, carbamate and pyrethroid resistance in peach-potato aphids (Myzus persicae). Pesticide Biochemistry Physiology, 18: 235-246.
  • E. Roditakis, E. Morou, A. Tsagkarakou, M. Riga, R. Nauen, M. Paine, S. Morin & J. Vontas, 2011. Assessment of the Bemisia tabaci CYP6CM1vQ transcript and protein levels in laboratory and field-derived imidacloprid-resistant insects and cross-metabolism potential of the recombinant enzyme. Insect Science 18(1): 23–29.
  • Feyereisen, R., 2005. Insect Cytochrome P450. Comprehensive Molecular Insect Science: Biochemistry and Molecular Biology. Gilbert L.I., Iatrou K. And G. S.S. Amsterdam, Elsevier. 4: 1-77.
  • Field, L. M., R. L. Blackman, C. Tyler-Smith & A. L. Devonshire, 1999. Relationship between amount of esterase and gene copy number in insecticide resistant Myzus persicae (Sulzer). Biochemical Journal, 339 (3): 737-742.
  • Hemingway, J. & G. P. Georghiou, 1984. Differential suppression of organophosphorus resistance in Culex quinquefasciatus by the synergists IBP, DEF, and TPP. Pesticide Biochemistry and Physiology, 21 (1): 1-9.
  • Hsieh, C., C. Wang & C. Ko, 2006. Analysis of Bemisia tabaci (Hemiptera ) species complex and distribution in Eastern Asia based on mitochondrial DNA markers. Entomological Society of America, 1: 768– 775.
  • I. Karunker, E. Moroub, D. Nikoub, R. Nauen, R. Sertchook, B.J. Stevenson, M.J.I. Paine, S. Morin & J. Vontas, 2009. Structural model and functional characterization of the Bemisia tabaci CYP6CM1vQ, a cytochrome P450 associated with high levels of imidacloprid resistance. Insect Biochemical. Molecular Biology. 39:697–706.
  • Jeschke, P., R. Nauen, M. Schindler & A. Elbert, 2011. Overview of the status and global strategy for neonicotinoids. Journal of Agriculture Food Chemistry, 59: 2897-2908.
  • Kaygısız,H., 1976.Akdeniz Bölgesi Pamuklarında Zarar Yapan Beyazsinek (Bemisia tabaci Genn.)’nin Tanınması,Biyolojisi,Yayılış Alanları,Zararı,Konukçuları ve Mücadelesi Üzerinde Araştırmalar.Tarım ve Orman Bakanlığı Zirai Mücadele ve Zirai Karantina Genel Müdürlüğü,Adana Bölge Zirai Mücadele Araştırma Enstitüsü Müdürlüğü Yayınları,Araştırma Eserleri Serisi No : 45, 58.
  • Kranthi, K. R., 2005. Insecticide Resistance -Monitoring, Mechanisms and Management Manual. Central Institute for Cotton Research, Nagpur, 153: 75-77.
  • Karunker, I., Benting, J., Lueke, B., Ponge, T., Nauen, R., Rodikatis, E., Vontas, J., Gorman, K., Denholm, I. & Morin, S., 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 38:634– 644.
  • Li, j., Li, X., Bai, R., Shi, Y., Tang, Q., An, S., Song, Q. & Yan, F., 2014. RNA interference of the P450 CYP6CM1 gene has different efficacy in B and Q biotypes of Bemisia tabaci. DOI 10.1002/ps.3903.
  • Liang, P., Guo, Y., Zhou, X. & Gao, X., 2014. Expression Profiling in Bemisia tabaci under Insecticide Treatment: Indicating the Necessity for Custom Reference Gene Selection.. PLoS ONE 9(1): e87514. doi:10.1371/journal.pone.0087514.
  • Livak, K.J. & T.D. Schmittgen, 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2[-Δ Δ C[T]] method]. Methods, 25:402-408.
  • Moya, A, P. Guirao, D. Cifuentes, F. Beitia & J.L. Cenis, 2001. Genetic diversity of Iberian populations of Bemisia tabaci (Hemiptera: Aleyrodidae) based on random amplified polymorphic DNA-polymerase chain reaction. Molecular Ecology, 10(4): 891–897.
  • NAUEN R., STUMPF N. & ELBERT A., 2002.- Toxicological and mechanistic studies on neonicotinoid cross resistance in Q type Bemisia tabaci (Hemiptera: Aleyrodidae).- Pest Management Science, 58 (9): 868-875.
  • Nauen R., 2007. Insecticide resistance in disease vectors of public health importance. Pest Manag. Sci. 63: 628-633.
  • Nauen, R., P. Jeschke & L. Copping, 2008. In Focus: Neonicotinoid insecticides Editorial. Pest Management Science 64 (11): 1081-1081.
  • P.J. De Barro, S.S. Liu, L.M. Boykin & A.B. Dinsdale, 2011. Bemisia tabaci: a statement of species status, Annual. Review. Entomology. 56: 1–19.
  • Puinean, A.M., Foster, S.P., Oliphant, L., Denholm, I., Field, L.M., Millar, N.S., Williamson, M.S. & Bass, C., 2010. Amplification of a Cytochrome P450 Gene Is Associated with Resistance to Neonicotinoid Insecticides in the Aphid Myzus persicae. PLoS Genet 6(6): e1000999. doi:10.1371/journal.pgen.1000999.
  • Qiong, R., Yong-hua X., Chen, L., Hong-yu, Z., Jones, C.M., Devine, G.J., Gorman & K., Denholm, I., 2012. Characterisation of Neonicotinoid and Pymetrozine Resistance in Strains of Bemisia tabaci (Hemiptera: Aleyrodidae) from China. Journal of Integrative Agriculture . 11(2): 321-326.
  • Rauch, N., & R. Nauen, 2003. Identification of biochemical markers linked to neonicotinoid cross resistance in Bemisia tabaci (Hemiptera: Aleyrodidae). Archives of Insect Biochemistry and Physiology. 54(4): 165-176.
  • Rauch, N. & R. Nauen, 2003. Identification of biochemical markers linked to neonicotinoid cross resistance in Bemisia tabaci (Hemiptera: Aleyrodidae). Archives of Insect Biochemistry and Physiology, 54 (4): 165-176.
  • Satar, G., Ulusoy, R.M., 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.Scott, J.G., 1999. Cytochromes P450 and insecticide resistance. Insect Biochemistry and Molecular Biology, 29: 757–777.
  • Sparks T.C. & Nauen R., 2015. IRAC: Mode of action classification and insecticide resistance management. Pesticide Biochemistry and Physiology 121 (2015) 122–128.Şekeroğlu, E., K. Karut, S. Yıldız & C. Kazak, 1998. Spatial distribution of preimaginal Bemisia tabaci (Homoptera: Aleyrodidae) in cotton. Proceedings of the World Cotton Research Conference 2, Athens, Greece pp. 644-647.
  • Ulusoy, M.R., A. Sarı, C. Can & N. Uygun, 1996. Pamuk beyazsineği, Bemisia tabaci (Gennadius)'nin farklı kültür bitkileri üzerindeki gelişmesinin saptanması, 186- 191. Türkiye 3. Entomoloji Kongresi (24-28 Eylül 1996, Ankara) Bildirileri, 716 s.
  • Ulusoy, S., Atakan, E. & Dinçer, S., 2018. Neonicotinoid resistance of Aphis gossypii Glover, 1877 (Hemiptera: Aphididae) in cotton fields of Çukurova Region, Turkey. Turkish Entomolgy, 42 (1): 23-31.
  • Wang, K. I., Q. L. Guo, X. M. Xia, H. Y. Wang & T. X. Liu, 2007. Resistance of Aphis gossypii (Homoptera: Aphididae) to selected insecticides on cotton from five cotton production regions in Shandong, China. Journal of Pesticide Science, 32 (4): 372-378.
  • Wang, Z., M. Yao & Y. Wu, 2009. Cross-resistance, inheritance and biochemical mechanisms of imidacloprid resistance in B-biotype Bemisia tabaci. Pest Management Science, 65 (11):1189-94.
  • Yang, X., He, C., Xie, W., Liu, Y., Xia, J., Yang, Z., Guo, L., Wen, Y., Wang, S., Wu, Q., Yang, F., Zhou, X., Zhang, Y., 2014. Glutathione S-transferases are involved in thiamethoxam resistance in the field whitefly Bemisia tabaci Q (Hemiptera: Aleyrodidae). doi: 10.1016/j.pestbp.2016.04.003

Overexpression of Cytochrome P450 CYP6CM1 Gene in Bemisia tabaci (Gennadius) in the Cotton Fields of the Çukurova Region, Turkey

Yıl 2020, Cilt: 23 Sayı: 3, 634 - 640, 30.06.2020

Selçuk Ulusoy

https://doi.org/10.18016/ksutarimdoga.vi.605716

Öz

In Cukurova region, neonicotinoid group
insecticides are widely used in cotton cultivation areas for controlling B. tabaci; as a result of this use,
there is a problem of resistance in these areas. In this study, cytochrome P450
CYP6CM1 relative expression levels, which are the indicators of imidacloprid
resistance, were analyzed. Five different B.
tabaci populations were collected from Adana city cotton cultivation areas
in 2018 and the indicators were observed. On the other hand, GST gene relative
expression profile, which is an efficient factor in metabolic resistance,
cytochrome P450, carboxylesterase (AcHE) and Glutathion S transferase (GST)
enzyme activities were determined. The highest cyp6cm1 and GST genes relative
activities were determined in Meletmez population; the activity level was 18, 2
and 9 times higher than susceptible population. On the other hand, the highest
and lowest cytochrome P450 enzyme activity was observed in Meletmez and Solaklı
populations, respectively. According to the analyses, it can be concluded that
there is extensive imidacloprid and neonicotinoid resistance of B. tabaci
populations in Çukurova Region cotton cultivation areas. As a result, excessive
gene expression and high-level enzyme activities indicate that there is a
resistance in the studied area based on enforced monooxygenases activity. 

Anahtar Kelimeler

Bemisia tabaci Cotton Cytochrome P450 Overexpression

Kaynakça

  • Ahmad, M., M. I. Arif & I. Denholm, 2003. High resistance of field populations of the cotton aphid Aphis gossypii Glover (Homoptera: Aphididae) to pyrethroid insecticides in Pakistan. Journal of Economic Entomology, 96: 875-878.
  • Anonymous, 2019. Türkiye istatistik kurumu. http://www.tuik.gov.tr (26.03.2019).
  • Bass, C. & L. M. Field, 2011. Gene amplification and insecticide resistance. Pest Management Science 67 (8): 886-890.
  • Bel-Kadhi, M.S., 2008. Molecular characterization of Bemisia tabaci biotypes in Southern Tunisia. Tunisian Journal of Plant Protection, 3(2): 79-85.
  • Bradford, M.M., 1976. A rapid and sensitive metod for the quantitation of microgram quantites of protein. utulizing the principle of protein dye binding. Analytical Biochemistry, 72:248-254.
  • Cahill, M., Gorman, K., Day, S., Denholm, I., Elbert, A. & Nauen, R., 1996. Baseline determination and detection of resistance to imidacloprid in Bemisia tabaci (Homoptera: Aleyrodidae). Bull. Entomol. Res. 86, 343–349.
  • Devonshire, A. L. & G. D. Moores, 1982. A carboxylesterase with broad substrate specify causes organophosphorus, carbamate and pyrethroid resistance in peach-potato aphids (Myzus persicae). Pesticide Biochemistry Physiology, 18: 235-246.
  • E. Roditakis, E. Morou, A. Tsagkarakou, M. Riga, R. Nauen, M. Paine, S. Morin & J. Vontas, 2011. Assessment of the Bemisia tabaci CYP6CM1vQ transcript and protein levels in laboratory and field-derived imidacloprid-resistant insects and cross-metabolism potential of the recombinant enzyme. Insect Science 18(1): 23–29.
  • Feyereisen, R., 2005. Insect Cytochrome P450. Comprehensive Molecular Insect Science: Biochemistry and Molecular Biology. Gilbert L.I., Iatrou K. And G. S.S. Amsterdam, Elsevier. 4: 1-77.
  • Field, L. M., R. L. Blackman, C. Tyler-Smith & A. L. Devonshire, 1999. Relationship between amount of esterase and gene copy number in insecticide resistant Myzus persicae (Sulzer). Biochemical Journal, 339 (3): 737-742.
  • Hemingway, J. & G. P. Georghiou, 1984. Differential suppression of organophosphorus resistance in Culex quinquefasciatus by the synergists IBP, DEF, and TPP. Pesticide Biochemistry and Physiology, 21 (1): 1-9.
  • Hsieh, C., C. Wang & C. Ko, 2006. Analysis of Bemisia tabaci (Hemiptera ) species complex and distribution in Eastern Asia based on mitochondrial DNA markers. Entomological Society of America, 1: 768– 775.
  • I. Karunker, E. Moroub, D. Nikoub, R. Nauen, R. Sertchook, B.J. Stevenson, M.J.I. Paine, S. Morin & J. Vontas, 2009. Structural model and functional characterization of the Bemisia tabaci CYP6CM1vQ, a cytochrome P450 associated with high levels of imidacloprid resistance. Insect Biochemical. Molecular Biology. 39:697–706.
  • Jeschke, P., R. Nauen, M. Schindler & A. Elbert, 2011. Overview of the status and global strategy for neonicotinoids. Journal of Agriculture Food Chemistry, 59: 2897-2908.
  • Kaygısız,H., 1976.Akdeniz Bölgesi Pamuklarında Zarar Yapan Beyazsinek (Bemisia tabaci Genn.)’nin Tanınması,Biyolojisi,Yayılış Alanları,Zararı,Konukçuları ve Mücadelesi Üzerinde Araştırmalar.Tarım ve Orman Bakanlığı Zirai Mücadele ve Zirai Karantina Genel Müdürlüğü,Adana Bölge Zirai Mücadele Araştırma Enstitüsü Müdürlüğü Yayınları,Araştırma Eserleri Serisi No : 45, 58.
  • Kranthi, K. R., 2005. Insecticide Resistance -Monitoring, Mechanisms and Management Manual. Central Institute for Cotton Research, Nagpur, 153: 75-77.
  • Karunker, I., Benting, J., Lueke, B., Ponge, T., Nauen, R., Rodikatis, E., Vontas, J., Gorman, K., Denholm, I. & Morin, S., 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 38:634– 644.
  • Li, j., Li, X., Bai, R., Shi, Y., Tang, Q., An, S., Song, Q. & Yan, F., 2014. RNA interference of the P450 CYP6CM1 gene has different efficacy in B and Q biotypes of Bemisia tabaci. DOI 10.1002/ps.3903.
  • Liang, P., Guo, Y., Zhou, X. & Gao, X., 2014. Expression Profiling in Bemisia tabaci under Insecticide Treatment: Indicating the Necessity for Custom Reference Gene Selection.. PLoS ONE 9(1): e87514. doi:10.1371/journal.pone.0087514.
  • Livak, K.J. & T.D. Schmittgen, 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2[-Δ Δ C[T]] method]. Methods, 25:402-408.
  • Moya, A, P. Guirao, D. Cifuentes, F. Beitia & J.L. Cenis, 2001. Genetic diversity of Iberian populations of Bemisia tabaci (Hemiptera: Aleyrodidae) based on random amplified polymorphic DNA-polymerase chain reaction. Molecular Ecology, 10(4): 891–897.
  • NAUEN R., STUMPF N. & ELBERT A., 2002.- Toxicological and mechanistic studies on neonicotinoid cross resistance in Q type Bemisia tabaci (Hemiptera: Aleyrodidae).- Pest Management Science, 58 (9): 868-875.
  • Nauen R., 2007. Insecticide resistance in disease vectors of public health importance. Pest Manag. Sci. 63: 628-633.
  • Nauen, R., P. Jeschke & L. Copping, 2008. In Focus: Neonicotinoid insecticides Editorial. Pest Management Science 64 (11): 1081-1081.
  • P.J. De Barro, S.S. Liu, L.M. Boykin & A.B. Dinsdale, 2011. Bemisia tabaci: a statement of species status, Annual. Review. Entomology. 56: 1–19.
  • Puinean, A.M., Foster, S.P., Oliphant, L., Denholm, I., Field, L.M., Millar, N.S., Williamson, M.S. & Bass, C., 2010. Amplification of a Cytochrome P450 Gene Is Associated with Resistance to Neonicotinoid Insecticides in the Aphid Myzus persicae. PLoS Genet 6(6): e1000999. doi:10.1371/journal.pgen.1000999.
  • Qiong, R., Yong-hua X., Chen, L., Hong-yu, Z., Jones, C.M., Devine, G.J., Gorman & K., Denholm, I., 2012. Characterisation of Neonicotinoid and Pymetrozine Resistance in Strains of Bemisia tabaci (Hemiptera: Aleyrodidae) from China. Journal of Integrative Agriculture . 11(2): 321-326.
  • Rauch, N., & R. Nauen, 2003. Identification of biochemical markers linked to neonicotinoid cross resistance in Bemisia tabaci (Hemiptera: Aleyrodidae). Archives of Insect Biochemistry and Physiology. 54(4): 165-176.
  • Rauch, N. & R. Nauen, 2003. Identification of biochemical markers linked to neonicotinoid cross resistance in Bemisia tabaci (Hemiptera: Aleyrodidae). Archives of Insect Biochemistry and Physiology, 54 (4): 165-176.
  • Satar, G., Ulusoy, R.M., 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.Scott, J.G., 1999. Cytochromes P450 and insecticide resistance. Insect Biochemistry and Molecular Biology, 29: 757–777.
  • Sparks T.C. & Nauen R., 2015. IRAC: Mode of action classification and insecticide resistance management. Pesticide Biochemistry and Physiology 121 (2015) 122–128.Şekeroğlu, E., K. Karut, S. Yıldız & C. Kazak, 1998. Spatial distribution of preimaginal Bemisia tabaci (Homoptera: Aleyrodidae) in cotton. Proceedings of the World Cotton Research Conference 2, Athens, Greece pp. 644-647.
  • Ulusoy, M.R., A. Sarı, C. Can & N. Uygun, 1996. Pamuk beyazsineği, Bemisia tabaci (Gennadius)'nin farklı kültür bitkileri üzerindeki gelişmesinin saptanması, 186- 191. Türkiye 3. Entomoloji Kongresi (24-28 Eylül 1996, Ankara) Bildirileri, 716 s.
  • Ulusoy, S., Atakan, E. & Dinçer, S., 2018. Neonicotinoid resistance of Aphis gossypii Glover, 1877 (Hemiptera: Aphididae) in cotton fields of Çukurova Region, Turkey. Turkish Entomolgy, 42 (1): 23-31.
  • Wang, K. I., Q. L. Guo, X. M. Xia, H. Y. Wang & T. X. Liu, 2007. Resistance of Aphis gossypii (Homoptera: Aphididae) to selected insecticides on cotton from five cotton production regions in Shandong, China. Journal of Pesticide Science, 32 (4): 372-378.
  • Wang, Z., M. Yao & Y. Wu, 2009. Cross-resistance, inheritance and biochemical mechanisms of imidacloprid resistance in B-biotype Bemisia tabaci. Pest Management Science, 65 (11):1189-94.
  • Yang, X., He, C., Xie, W., Liu, Y., Xia, J., Yang, Z., Guo, L., Wen, Y., Wang, S., Wu, Q., Yang, F., Zhou, X., Zhang, Y., 2014. Glutathione S-transferases are involved in thiamethoxam resistance in the field whitefly Bemisia tabaci Q (Hemiptera: Aleyrodidae). doi: 10.1016/j.pestbp.2016.04.003
Toplam 36 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Konular Ziraat, Veterinerlik ve Gıda Bilimleri
Bölüm ARAŞTIRMA MAKALESİ (Research Article)
Yazarlar

Selçuk Ulusoy 0000-0002-2278-8471

Yayımlanma Tarihi 30 Haziran 2020
Gönderilme Tarihi 16 Ağustos 2019
Kabul Tarihi 21 Ocak 2020
Yayımlandığı Sayı Yıl 2020Cilt: 23 Sayı: 3

Kaynak Göster

APA Ulusoy, S. (2020). Overexpression of Cytochrome P450 CYP6CM1 Gene in Bemisia tabaci (Gennadius) in the Cotton Fields of the Çukurova Region, Turkey. Kahramanmaraş Sütçü İmam Üniversitesi Tarım Ve Doğa Dergisi, 23(3), 634-640. https://doi.org/10.18016/ksutarimdoga.vi.605716
 Kapak Resmi İndir

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