Abstract
Due to the deleterious effect of chemical pesticides on human and environment, accelerated researchers to find a novel environmental friendly alternatives. Chitinase enzyme is a promising biocontrol agent against various crop pests including insects, nematodes and pathogenic fungi. In this context, here we elucidate the molecular toxicity of chitinase against the crop pest Corcyra cephalonica. Genotoxic effects of chitinase in C. cephalonica was evaluated and analyzed by estimation of Reactive Oxygen Species (ROS), Flow cytometry, DNA fragmentation and Comet assay. The results revealed that chitinase is able to induce oxidative stress followed by ROS generation (33,101.63 AU at the dose of 500 ppm), DNA damage and apoptosis (in live cell 70.09% and early apoptotic 28.07%). Moreover, by off target effects of chitinase on murine cell lines (L929) by cell viability method. The assay showed that chitinase doesn’t exhibit negative effects in L929 cells. Overall the present study conclude that chitinase cause genotoxic effects in the larvae of C. cephalonica. Therefore, chitinase can be utilized as a potent biocontrol agent against various crop pests.
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Abbreviations
- ROS:
-
Reactive Oxygen Species
- DNA:
-
Deoxyribonucleic acid
- AU:
-
Arbitrary unit
- ppm:
-
Parts per million
- DMEM:
-
Dulbecco’s modified Eagles’ medium
- FBS:
-
Fetal Bovine Serum
- PBS:
-
Phosphate Buffer Saline
- DMSO:
-
Dimethyl Sulfoxide
- NCCS:
-
National Centre for Cell Sciences
- MTT:
-
3-(4,5 Dimethyl-thiozole-2-yl)-2,5diphenyl tetrazolium bromide
- µL:
-
Microlitre
- TNAU:
-
Tamil Nadu Agriculture University
- RGR:
-
Relative growth rate
- RCR:
-
Relative consumption rate
- ECI:
-
Efficiency of conversion of ingested food
- FDI:
-
Feeding deterrence index and mortality percentage
- ACB:
-
Anticoagulant buffer
- mM:
-
Millimolar
- DCFH-DA:
-
2′-7′-Dichlorodihydrofluorescein diacetate
- DCF:
-
Dihydrodichlorofluorescein
- TCA:
-
Trichloroacetic acid
- DPA:
-
Diphenylamine
- FACS:
-
Fluorescence-activated cell sorting
- SOD:
-
Superoxide dismutase
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Acknowledgments
The authors acknowledge Karpagam Academy of Higher Education (KAHE) for provided the laboratory facilities to carry out this research work. The authors acknowledge the DST-FIST program (File No. SR/FST/LS-1/2018/187) for the sophisticated instrument facility.
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AYS designed the work, analyzed the data of genotoxicity studies and tailored the final manuscript; MN identification of Corcyra cephalonica, collection of samples for this studies; NV conducted the experiments and wrote the manuscript; LHF analyzed the data of ROS studies; HB analyzed the data of cytotoxicity studies; CR reviewed and modified the manuscript. The authors have read and approved the final manuscript.
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Vijayakumar, N., Nalini, M., Rajkuberan, C. et al. Genotoxic and cytotoxic effect of chitinase against Corcyra cephalonica larvae under laboratory conditions. Int J Trop Insect Sci 41, 2937–2946 (2021). https://doi.org/10.1007/s42690-021-00478-8
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DOI: https://doi.org/10.1007/s42690-021-00478-8