Determination of Optimum Incubation Time for the Formation and Quantification of Xanthomonas axonopodis pv. citri Biofilms on Abiotic Surface

Author: Ganesuni Lakshmi Prasanna, K. Gopal, Ch. Ruth, Y. Sireesha, Syed Sadarunnisa and V.V. Padmaja

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Abstract

Xanthomonas axonopodis pv. citri (Xac) is the most devastating pathogen to Citrus spp. as well as acid lime and are able to form a biofilm, making it difficult to manage canker disease. Biofilm formation is one of the mechanisms that bacterial communities use to adapt to unfavourable environmental conditions and to survive and colonize in host plants. Hence, the present study aims to investigate the ability of thirty-two isolates of Xac to form biofilms on abiotics surface at different intervals and quantification of biofilm has done. The results revealed that, the biofilms formed strongly adhered to the plates on days one, three, five, seven and nine. Interestingly on day two (48 h) of incubation most of the isolates showed the highest biofilms formation. However, moderate biofilm formation onto the microtiter plates by the isolates were observed on days five and seven, but non-adherence was observed on days one. These strains were grouped into four categories. The isolate which does not formed any biofilm (Xac-23), the isolates with week biofilm formation viz., Xac-15 and Xac-26, the isolates with moderately formed biofilm viz., Xac-2 and Xac-32, and the remaining isolates viz., Xac-1, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 16, 17, 18, 19, 20, 21, 22, 24, 25, 26, 27, 28, 29, 30, 31, formed strong biofilm on the walls of microtiter plate compared to negative control. The results concluded that 36-48h is the optimum cultivation period for Xac to form a strong biofilm in microtiter plate and could be used to study biofilm inhibitor studies further.

Keywords

Biofilm, Crystal violet, Ethanol, Microtiter plate, Quantification and Xanthomonas axonopodis pv. citri

Conclusion

Plant pathogenic bacteria can use biofilm formation as a crucial survival tactic in their natural state. To locate and stop the systemic spread of these bacteria through abiotic surfaces, the adherence characteristic of these bacteria must be detected. In this study, we assessed the ability of Xanthomonas axonopodis pv. citri, a plant-associated bacterium, to form biofilm on an abiotic surface, such as a microtiter plate, and we timed the development of mature biofilms. After 48 hours of incubation, we discovered that Xac strains successfully formed the highest amount of biofilm on a 96-well microtiter plate using nutrient broth, and Xac then started inhibiting the growth of biofilm.

References

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How to cite this article

Ganesuni Lakshmi Prasanna, K. Gopal, Ch. Ruth, Y. Sireesha, Syed Sadarunnisa and V.V. Padmaja (2023). Determination of Optimum Incubation Time for the Formation and Quantification of Xanthomonas axonopodis pv. citri Biofilms on Abiotic Surface. Biological Forum – An International Journal, 15(5): 681-687.