Abstract
Bacillus spp. can promote the growth of plants and reduce plant disease incidence or severity by triggering induced systemic resistance to plant pathogens. In addition, bacteria of this genus are chitinase enzyme producers. Chitinase inhibits the growth of fungi by breaking down the chitin-containing cell wall of plant pathogenic fungi. In this study, 270 native Bacillus spp., isolated from various habitats in Kayseri and Adana, Turkey, were screened by PCR for the chitinase gene and 66 of them were found to have this gene. Nine Bacillus thuringiensis (Bt) isolates showing high insecticidal activity against insect pests in our previous studies were selected from 66 isolates containing the chitinase gene. Chitinases with different molecular weights ranging from ~ 40 to 113 kDa were determined by SDS-PAGE. To determine the antagonistic effects against plant pathogenic fungi (Fusarium oxysporum f.sp. niveum, Verticillium dahliae and Aspergillus niger), a dual culture assay was used with nine native and two standard strains of Bt, and radial growth inhibition was calculated as a percentage. Of all the tested isolates, SY33.3 showed the strongest antagonistic activity and thus, can be used as an effective biological control agent against plant pathogenic fungi.
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Abbreviations
- SY:
-
Semih Yilmaz
- U:
-
Ugur Azizoglu
- Bt :
-
Bacillus thuringiensis
- chi :
-
chitinase
- PCR:
-
Polymerase Chain Reaction
- PIRG:
-
Percentage Inhibition of Radial Growth
- LB:
-
Luria-Bertani
- mM:
-
mili Molar
- TAE:
-
Tris-Acetate-EDTA
- L:
-
Liter
- rpm:
-
revolutions per minute
- SDS-PAGE:
-
Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis
- Tm:
-
melting temperature
- kb:
-
kilobase
- F:
-
Forward
- R:
-
Reverse
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Acknowledgements
Special thanks to Dr. Sibel Derviş for supplying phytopathogenic fungi. The authors would like to express special thanks to Dr. Hatice Korkmaz Güvenmez for supplying chitin. This study is the master thesis entitled ‘Screening of chitinase genes from local Bacillus isolates and testing their antifungal activity’ of Zehra Busra Azizoglu and presented on Ecology 2017 International Symposium held on 11–13 May 2017 in Kayseri, Turkey.
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This project was funded by Erciyes University Scientific Project Unit under the codes of FBD-08-540.
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ZBA performed bioassays and molecular studies, and contributed to the planning of the study. SY contributed to the material supply, molecular studies, manuscript preparation and data analysis. UA participated in experimental studies, manuscript preparation and molecular studies. SK carried out statistical analysis of the data and contributed to the figures. RT contributed to experimental studies. AA conceived of the study, contributed to the design and coordination and helped to draft the manuscript. All authors read and approved the manuscript.
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Azizoglu, Z.B., Yilmaz, S., Azizoglu, U. et al. Molecular characterization of the chitinase genes of native Bacillus thuringiensis isolates and their antagonistic activity against three important phytopathogenic fungi. Biologia 76, 2745–2755 (2021). https://doi.org/10.1007/s11756-021-00802-0
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DOI: https://doi.org/10.1007/s11756-021-00802-0