Induction and Resuscitation of the Viable but Non-culturable (VBNC) State in Acidovorax citrulli, the Causal Agent of Bacterial Fruit Blotch of Cucurbitaceous Crops

Yumin Kan, Na Jiang, Xin Xu, Qingyang Lyu, Vinoj Gopalakrishnan, Ronald Walcott, Saul Burdman, Jianqiang Li, Laixin Luo*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

21 Scopus citations


Acidovorax citrulli is a gram-negative bacterium that infects a wide range of cucurbits causing bacterial fruit blotch (BFB) disease. Copper-based compounds are the most widely-used chemicals for managing BFB and other bacterial diseases in the field. Many bacteria can enter a viable but non-culturable (VBNC) state in response to stress, including exposure to copper, and recover the culturability when favorable conditions return. The present study demonstrates that A. citrulli strain AAC00-1 is able to enter into the VBNC state by treatment with different concentrations of copper sulfate. It took 3 h, 5 and 15 days for all viable cells to lose culturability upon exposure to copper sulfate concentrations of 50, 10, and 5 μM, respectively. The VBNC A. citrulli cells regained culturability when the Cu2+ ions were removed by chelation with EDTA or by transfer of cells to LB broth, a cell-free supernatant from a suspension of AAC00-1, oligotrophic media amended with casein hydrolysate or watermelon seedling juice. We also found that the VBNC cells induced by Cu2+ were unable to colonize or infect watermelon seedlings directly, but the resuscitated cells recovered full virulence equivalent to untreated bacterial cells in the log phase. To the best of our knowledge, this is the first report on the VBNC state in A. citrulli and the factors that facilitate resuscitation and restoration of pathogenicity.

Original languageAmerican English
Article number1081
JournalFrontiers in Microbiology
Issue numberMAY
StatePublished - 2019

Bibliographical note

Funding Information:
This work was supported financially by the National Key Research and Development Program of China (Grant No.

Publisher Copyright:
Copyright © 2019 Kan, Jiang, Xu, Lyu, Gopalakrishnan, Walcott, Burdman, Li and Luo. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.


  • Acidovorax citrulli
  • Copper sulfate
  • Induction
  • Pathogenicity
  • Resuscitation
  • Viable but non-culturable


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