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Divalent Cations (Ca2+, Mg2+, Mn2+, Fe2+, Ni2+, and Zn2+) Enhance Growth of Xanthomonas citri and X. campestris by Reducing Generation Time

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Author(s):
Merighi, Davi Gabriel Salustiano ; Goncalves, Caue Augusto Boneto ; Ferrari, Anielle Salviano de Almeida ; Andrade, Maxuel de Oliveira ; Guzzo, Cristiane Rodrigues
Total Authors: 5
Document type: Journal article
Source: ACS OMEGA; v. 10, n. 32, p. 15-pg., 2025-08-06.
Abstract

Bacterial ion homeostasis is critical for adaptation and survival. In this study, we show that supplementing 2xTY medium with divalent cations (Ca2+, Mg2+, Mn2+, Fe2+, Ni2+, and Zn2+) from 50 to 1,000 mu M significantly enhances the growth of Xanthomonas species by reducing generation time, while monovalent ions (Na+, K+, Li+, and Cl-) had no significant effect. Among them, Ca2+ effectively boosted X. citri fitness, increasing the specific growth rate by up to 223%, shortening the cell cycle, and enhancing adhesion to abiotic surfaces up to 8-fold. Calcium also induced a more uniform and smaller cell morphology, suggesting improved cell division and metabolic efficiency. Colony forming unit counts confirmed a higher viability, excluding dead cell accumulation. These effects were conserved in other Xanthomonadaceae members (X. campestris and S. maltophilia) but not in nonrelated species. Based on these findings, we propose an optimized growth medium named 2xTY-Ca (16 g/L tryptone, 10 g/L yeast extract, 5 g/L NaCl, and 1 mM CaCl2), which significantly improves Xanthomonas spp. growth kinetics in vitro, enhancing X. citri and X. campestris cultivation for laboratory and industrial applications. Though the precise molecular mechanism behind these phenotypes has yet to be determined, we highlight X. citri as a promising organism to understand the role of divalent cations in bacterial physiology. (AU)

FAPESP's process: 20/04680-0 - Design of different fragments of SARS-CoV-2 surface protein spike for the development of fast diagnostic test and vaccine
Grantee:Cristiane Rodrigues Guzzo Carvalho
Support Opportunities: Regular Research Grants
FAPESP's process: 21/10577-0 - Biology of Bacteria and Bacteriophages Research Center
Grantee:Shaker Chuck Farah
Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC
FAPESP's process: 22/04601-8 - CryoEM study of the adsorption of a phage to a type IV pili
Grantee:Davi Gabriel Salustiano Merighi
Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
FAPESP's process: 23/13894-1 - CryoEM study of the adsorption of a phage to a type IV pilus
Grantee:Davi Gabriel Salustiano Merighi
Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
FAPESP's process: 23/07366-2 - Heterologous Expression of a PilZ Homolog for Studying c-di-GMP Signaling in Leptospira interrogans
Grantee:Cauê Augusto Boneto Gonçalves
Support Opportunities: Scholarships in Brazil - Scientific Initiation
FAPESP's process: 22/08730-7 - Protein structural biology applied in the study of c-di-GMP signaling and bacterial secretion systems
Grantee:Cristiane Rodrigues Guzzo Carvalho
Support Opportunities: Regular Research Grants
FAPESP's process: 17/17303-7 - Structure and function of bacterial secretion systems
Grantee:Shaker Chuck Farah
Support Opportunities: Research Projects - Thematic Grants