[논문]살진균제인 캡탄 처리 후 갈색거저리의 해독효소 유전자 발현

장호암; 백형선; 김보배; 알리 모하마디 코줄 마리암; 패트나익 바랫 부산; 조용훈; 한연수

doi:10.5656/ksae.2022.02.0.011

살진균제인 캡탄 처리 후 갈색거저리의 해독효소 유전자 발현
Gene Expression of Detoxification Enzymes in Tenebrio molitor after Fungicide Captan Exposure 원문보기

한국응용곤충학회지 = Korean journal of applied entomology, v.61 no.1, 2022년, pp.155 - 163

장호암 (전남대학교 응용생물학과) , 백형선 (전남대학교 응용생물학과) , 김보배 (전남대학교 응용생물학과) , 알리 모하마디 코줄 마리암 (전남대학교 응용생물학과) , 패트나익 바랫 부산 (파키르 모한 대학교 생명공학과) , 조용훈 (전남대학교 응용생물학과) , 한연수 (전남대학교 응용생물학과)

초록
AI-Helper

최근 살진균제는 세계 식량 안보에 없어서는 안될 필수 요소이며, 그 사용량은 증가하고 있다. 살진균제는 직접적 또는 간접적으로 곤충에 영향을 미쳐 유전자 및 분자 수준의 변화를 일으킨다. 곤충은 다양한 해독 매커니즘을 통해 살진균제를 포함한 농약으로부터 유발되는 활성산소(ROS) 독성을 제거한다. 본 연구는 살진균제 캡탄의 비치명적 투여량(0.2, 2, and 20 ㎍/µL)을 주입 후 갈색거저리의 유충에서 해독효소의 mRNA 발현량을 분석했다. 갈색거저리의 전사체 분석을 통해 해독 매커니즘 관련 유전자인 퍼옥시다제(POX), 카탈라제(CAT), 슈퍼옥사이드 디스뮤타제(SOD) 및 글루타티온-S-트랜스퍼라제(GST)를 발굴하였다. 처리 24시간 후 TmPOX5 mRNA가 유의하게 증가한 것으로 나타났다. 처리 3 시간 후 TmSOD4의 mRNA가 유사하게 증가하였다. 또한 2 ㎍/µL 처리 24시간 후 TmCAT2의 mRNA 가 유의하게 증가하였다. 캡탄 노출 후 TmGST1 및 TmGST3의 mRNA 발현량도 증가하였다. 결론적으로, TmPOX5 및 TmSOD4 유전자는 갈색거저리에서 캡탄 노출에 대한 바이오마커 또는 생체이물 센서로 작용할 수 있음을 시사한다.

Abstract ▼ AI-Helper

The application of fungicides is indispensable to global food security, and their use has increased in recent times. Fungicides, directly or indirectly, have impacted insects, leading to genetic and molecular-level changes. Various detoxification mechanisms allow insects to eliminate reactive oxygen species (ROS) toxicity induced by agrochemicals including fungicides. In the present study, we analyzed the mRNA expression levels of detoxifying enzymes in Tenebrio molitor larvae following exposure to non-lethal doses (0.2, 2, and 20 ㎍/µL) of a fungicide captan. Transcripts of peroxidases (POXs), catalases (CATs), superoxide dismutases (SODs), and glutathione-s-transferases (GSTs) were screened from the T. molitor transcriptome database. RT-qPCR analysis showed that TmPOX5 mRNA increased significantly 24 h post-captan exposure. A similar increase was noticed for TmSOD4 mRNA 3 h post-captan exposure. Moreover, the expression of TmCAT2 mRNA increased significantly 24 h post-treatment with 2 ㎍/µL captan. TmGST1 and TmGST3 mRNA expression also increased noticeably after captan exposure. Taken together, these results suggest that TmPOX5 and TmSOD4 mRNA can be used as biomarkers or xenobiotics sensors for captan exposure in T. molitor, while other detoxifying enzymes showed differential expression.

주제어

표/그림 (5)

표 Table 1. Primers used in this study
그림 Fig. 1. Effect of captan (at 0.2, 2, and 20 μg/μL) on the relative mRNA expression of glutathione-S-transferases (GSTs) in the whole body of Tenebrio molitor larvae. Expression levels of TmGSTs mRNA in the negative control group were normalized to 1. Data are shown as the mean ± standard error (three biological replicates). Significant differences between the experimental and control groups are indicated by asterisks (* p < 0.05).
그림 Fig. 2. Effect of captan (at 0.2, 2, and 20 μg/μL) on the relative mRNA expression of peroxidases (POXs) in the whole body of Tenebrio molitor. Expression levels of TmPOXs mRNA in the negative control group were normalized to 1. Data are shown as the mean ±standard error (three biological replicates). Significant differences between the experimental and control groups are indicated by asterisks (* p < 0.05).
그림 Fig. 3. Effect of captan (at 0.2, 2, and 20 μg/μL) on the relative mRNA expression of catalases (CATs) in the whole body of Tenebrio molitor. Expression levels of TmCATs mRNA in the negative control group were normalized to 1. Data are shown as the mean ± standard error (three biological replicates). Significant differences between the experimental and control groups are indicated by asterisks (* p < 0.05).
그림 Fig. 4. Effect of captan (at 0.2, 2, and 20 μg/μL) on the relative mRNA expression of superoxide dismutases (SODs) in the whole body of Tenebrio molitor. Expression levels of TmSODs mRNA in the negative control group were normalized to 1. Data are shown as the mean ± standard error (three biological replicates). Significant differences between the experimental and control groups are indicated by asterisks (* p < 0.05).

참고문헌 (45)

Abdollahi, M., Ranjbar, A., Shadnia, S., Nikfar, S., Rezaie, A., 2004. Pesticides and oxidative stress: a review. Med. Sci. Monit. 10, Ra141-147.
Barbehenn, R.V., 2002. Gut-based antioxidant enzymes in a polyphagous and a graminivorous grasshopper. J. Chem. Ecol. 28, 1329-1347.

상세보기
Barreda, M., LOpez, F.J., Villarroya, M., Beltran, J., GarciaBaudin, J.M., HernAndez, F., 2006. Residue determination of captan and folpet in vegetable samples by gas chromatography/negative chemical Ionization mass spectrometry. J. AOAC Int. 89, 1080-1087.

상세보기
Brucker, R.M., Funkhouser, L.J., Setia, S., Pauly, R., Bordenstein, S.R., 2012. Insect innate immunity database (IIID): an annotation tool for identifying immune genes in insect genomes. PLoS ONE 7, e45125.

상세보기
Caverzan, A., Piasecki, C., Chavarria, G., Stewart, C.N., Jr., Vargas, L., 2019. Defenses against ROS in crops and weeds: the effects of interference and herbicides. Int. J. Mol. Sci. 20, 1086.

상세보기
Edosa, T.T., Jo, Y.H., Keshavarz, M., Bae, Y.M., Kim, D.H., Lee, Y.S., Han, Y.S., 2020. TmSpz6 is essential for regulating the Immune response to Escherichia coli and Staphylococcus aureus infection in Tenebrio molitor. Insects 11, 105.

상세보기
Everich, R., Schiller, C., Whitehead, J., Beavers, M., Barrett, K., 2009. Effects of captan on Apis mellifera brood development under field conditions in California almond orchards. J. Econ. Entomol. 102, 20-29.

상세보기
Felton, G.W., Summers, C.B., 1995. Antioxidant systems in insects. Arch. Insect Biochem. Physiol. 29, 187-197.

상세보기
Freyre, E.O., Valencia, A.T., Guzman, D.D., Maldonado, I.C., Ledezma, L.E.B., Carrillo, M.F.Z., Escorza, M.A.Q., 2021. Oxidative stress as a molecular mechanism of exposure to organophosphorus pesticides: a review. Curr. Protein Pept. Sci. 22, 890-897.

상세보기
Fridovich, I., 1975. Superoxide dismutases. Annu. Rev. Biochem. 44, 147-159.

상세보기
Han, J.B., Li, G.Q., Wan, P.J., Zhu, T.T., Meng, Q.W., 2016. Identification of glutathione S-transferase genes in Leptinotarsa decemlineata and their expression patterns under stress of three insecticides. Pestic. Biochem. Physiol. 133, 26-34.

상세보기
He, B., Liu, Z., Wang, Y., Cheng, L., Qing, Q., Duan, J., Xu, J., Dang, X., Zhou, Z., Li, Z., 2021. Imidacloprid activates ROS and causes mortality in honey bees (Apis mellifera) by inducing iron overload. Ecotoxicol. Environ. Saf. 228, 112709.

상세보기
He, Q.K., Xu, C.L., Li, Y.P., Xu, Z.R., Luo, Y.S., Zhao, S.C., Wang, H.L., Qi, Z.Q., Liu, Y., 2022. Captan exposure disrupts ovarian homeostasis and affects oocytes quality via mitochondrial dysfunction induced apoptosis. Chemosphere 286, 131625.

상세보기
Houbraken, M., Spranghers, T., De Clercq, P., Cooreman-Algoed, M., Couchement, T., De Clercq, G., Verbeke, S., Spanoghe, P., 2016. Pesticide contamination of Tenebrio molitor (Coleoptera: Tenebrionidae) for human consumption. Food Chem. 201, 264-269.

상세보기
Irato, P., Santovito, G., 2021. Enzymatic and non-enzymatic molecules with antioxidant function. Antioxidants 10, 579.

상세보기
Jang, H.A., Patnaik, B.B., Ali Mohammadie Kojour, M., Kim, B.B., Bae, Y.M., Park, K.B., Lee, Y.S., Jo, Y.H., Han, Y.S., 2021. TmSpz-like plays a fundamental role in response to E. coli but not S. aureus or C. albican Infection in Tenebrio molitor via regulation of antimicrobial peptide production. Int. J. Mol. Sci. 22, 10888.

상세보기
Jia, Z., Misra, H.P., 2007. Reactive oxygen species in in vitro pesticide-induced neuronal cell (SH-SY5Y) cytotoxicity: role of NFkappaB and caspase-3. Free Radic. Biol. Med. 42, 288-298.

상세보기
Jo, Y.H., Patnaik, B.B., Hwang, J., Park, K.B., Ko, H.J., Kim, C.E., Bae, Y.M., Jung, W.J., Lee, Y.S., Han, Y.S., 2019. Regulation of the expression of nine antimicrobial peptide genes by TmIMD confers resistance against Gram-negative bacteria. Sci. Rep. 9, 10138.

상세보기
Johnson, R.M., Dahlgren, L., Siegfried, B.D., Ellis, M.D., 2013. Acaricide, fungicide and drug interactions in honey bees (Apis mellifera). PLoS ONE 8, e54092.

상세보기
Kim, S.G., Jo, Y.H., Seong, J.H., Park, K.B., Noh, M.Y., Cho, J.H., Ko, H.J., Kim, C.E., Tindwa, H., Patnaik, B.B., Bang, I.S., Lee, Y.S., Han, Y.S., 2017. TmSR-C, scavenger receptor class C, plays a pivotal role in antifungal and antibacterial immunity in the coleopteran insect Tenebrio molitor. Insect Biochem. Mol. Biol. 89, 31-42.

상세보기
Kobayashi, Y., Nojima, Y., Sakamoto, T., Iwabuchi, K., Nakazato, T., Bono, H., Toyoda, A., Fujiyama, A., Kanost, M.R., Tabunoki, H., 2019. Comparative analysis of seven types of superoxide dismutases for their ability to respond to oxidative stress in Bombyx mori. Sci. Rep. 9, 2170.

상세보기
Kolawole, A.O., Kolawole, A.N., 2014. Insecticides and bio-insecticides modulate the glutathione-related antioxidant defense system of cowpea storage bruchid (Callosobruchus maculatus). Int. J. Insect Sci. 6, IJIS.S18029.
Kolawole, A.O., Olajuyigbe, F.M., Ajele, J.O., Adedire, C.O., 2014. Activity of the antioxidant defense system in a typical bioinsecticide-and synthetic insecticide-treated cowpea storage beetle Callosobrochus maculatus F. (Coleoptera: Chrysomelidae). Int. J. Insect Sci. 6, IJIS.S19434.
Kostaropoulos, I., Mantzari, A.E., Papadopoulos, A.I., 1996. Alterations of some glutathione S-transferase characteristics during the development of Tenebrio molitor (Insecta: Coleoptera). Insect Biochem. Mol. Biol 26, 963-969.

상세보기
Kostaropoulos, I., Papadopoulos, A.I., Metaxakis, A., Boukouvala, E., Papadopoulou-Mourkidou, E., 2001. The role of glutathione S-transferases in the detoxification of some organophosphorus insecticides in larvae and pupae of the yellow mealworm, Tenebrio molitor (Coleoptera: Tenebrionidae). Pest Manag. Sci. 57, 501-508.

상세보기
Lu, Q., Sun, Y., Ares, I., Anadon, A., Martinez, M., Martinez-Larranaga, M.R., Yuan, Z., Wang, X., Martinez, M.A., 2019. Deltamethrin toxicity: a review of oxidative stress and metabolism. Environ. Res. 170, 260-281.

상세보기
Nazir, A., Mukhopadhyay, I., Saxena, D.K., Siddiqui, M.S., Chowdhuri, D.K., 2003. Evaluation of toxic potential of captan: Induction of hsp70 and tissue damage in transgenic Drosophila melanogaster (hsp70-lacZ) Bg9. J. Biochem. Mol. Toxicol. 17, 98-107.

상세보기
Piechowicz, B., Sienko, J., Mytych, J., Grodzicki, P., Podbielska, M., Szpyrka, E., Zareba, L., Piechowicz, I., Sadlo, S., 2021. Assessment of risk to honey bees and honey consumers resulting from the insect exposure to captan, thiacloprid, penthiopyrad, and λ-cyhalothrin used in a commercial apple orchard. Environ. Monit. Assess. 193, 129.

상세보기
Pita-Oliveira, M., Rodrigues-Soares, F., 2021. Influence of GSTM1, GSTT1, and GSTP1 genetic polymorphisms on disorders in transplant patients: a systematic review. Drug Metab. Pers. Ther. in press.
Radhakrishnan, R., Alqarawi, A.A., Abd Allah, E.F., 2018. Bioherbicides: current knowledge on weed control mechanism. Ecotoxicol. Environ. Saf. 158, 131-138.

상세보기
Rauf, A., Wilkins, R.M., 2021. Malathion-resistant Tribolium castaneum has enhanced response to oxidative stress, immunity, and fitness. bioRxiv, 2021.2011.2016.468861.
Semren, T., Zunec, S., Pizent, A., 2018. Oxidative stress in triazine pesticide toxicity: a review of the main biomarker findings. Arh. Hig. Rada Toksikol. 69, 109-125.

상세보기
Seo, G.W., Jo, Y.H., Seong, J.H., Park, K.B., Patnaik, B.B., Tindwa, H., Kim, S.A., Lee, Y.S., Kim, Y.J., Han, Y.S., 2016. The Silencing of a 14-3-3ɛ Homolog in Tenebrio molitor leads to increased antimicrobial activity in hemocyte and reduces larval survivability. Genes (Basel) 7, 53.

상세보기
Seong, J.H., Jo, Y.H., Seo, G.W., Park, S., Park, K.B., Cho, J.H., Ko, H.J., Kim, C.E., Patnaik, B.B., Jun, S.A., Choi, Y.S., Kim, Y.W., Bang, I.S., Lee, Y.S., Han, Y.S., 2018. Molecular cloning and effects of Tm14-3-3ζ-silencing on larval survivability against E. coli and C. albicans in Tenebrio molitor. Genes (Basel) 9, 330.

상세보기
Shakir, S.K., Irfan, S., Akhtar, B., Rehman, S.U., Daud, M.K., Taimur, N., Azizullah, A., 2018. Pesticide-induced oxidative stress and antioxidant responses in tomato (Solanum lycopersicum) seedlings. Ecotoxicology 27, 919-935.

상세보기
Thannickal, V.J., Fanburg, B.L., 2000. Reactive oxygen species in cell signaling. Am. J. Physiol. Lung Cell. Mol. Physiol. 279, L1005- 1028.

상세보기
Tindwa, H., Jo, Y.H., Patnaik, B.B., Lee, Y.S., Kang, S.S., Han, Y.S., 2015. Molecular cloning and characterization of autophagy-related gene TmATG8 in Listeria-invaded hemocytes of Tenebrio molitor. Dev. Comp. Immunol. 51, 88-98.

상세보기
Wang, H., Lu, Z., Li, M., Fang, Y., Qu, J., Mao, T., Chen, J., Li, F., Sun, H., Li, B., 2020. Responses of detoxification enzymes in the midgut of Bombyx mori after exposure to low-dose of acetamiprid. Chemosphere 251, 126438.

상세보기
Xin, S., Zhang, W., 2020. Construction and analysis of the protein-protein interaction network for the olfactory system of the silkworm Bombyx mori. Arch. Insect Biochem. Physiol. 105, e21737.

상세보기
Yu, L., Wan, F., Dutta, S., Welsh, S., Liu, Z., Freundt, E., Baehrecke, E.H., Lenardo, M., 2006. Autophagic programmed cell death by selective catalase degradation. Proc. Natl. Acad. Sci. U. S. A. 103, 4952-4957.

상세보기
Zhang, J., Lu, A., Kong, L., Zhang, Q., Ling, E., 2014. Functional analysis of insect molting fluid proteins on the protection and regulation of ecdysis. J. Biol. Chem. 289, 35891-35906.

상세보기
Zhao, H., Yi, X., Hu, Z., Hu, M., Chen, S., Muhammad, R.-u.-H., Dong, X., Gong, L., 2013. RNAi-mediated knockdown of catalase causes cell cycle arrest in SL-1 cells and results in low survival rate of Spodoptera litura (Fabricius). PLoS ONE 8, e59527.

상세보기
Zhou, C., Yang, H., Wang, Z., Long, G.Y., Jin, D.C., 2018. Protective and detoxifying enzyme activity and ABCG subfamily gene expression in Sogatella furcifera under insecticide stress. Front. Physiol. 9, 1890.

상세보기
Zhou, Y., Chen, X., Teng, M., Zhang, J., Wang, C., 2019. Toxicity effects of captan on different life stages of zebrafish (Danio rerio). Environ. Toxicol. Pharmacol. 69, 80-85.

상세보기
Zhu, W., Schmehl, D.R., Mullin, C.A., Frazier, J.L., 2014. Four common pesticides, their mixtures and a formulation solvent in the hive environment have high oral toxicity to honey bee larvae. PLoS ONE 9, e77547.

상세보기

저자의 다른 논문 :

표제어: PCR

동의어: Packet Collision Rate

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