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

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-cultura...

Full description

Bibliographic Details
Main Authors: Yumin Kan, Na Jiang, Xin Xu, Qingyang Lyu, Vinoj Gopalakrishnan, Ronald Walcott, Saul Burdman, Jianqiang Li, Laixin Luo
Format: Article
Language:English
Published: Frontiers Media S.A. 2019-05-01
Series:Frontiers in Microbiology
Subjects:
Online Access:https://www.frontiersin.org/article/10.3389/fmicb.2019.01081/full
_version_ 1819070624739360768
author Yumin Kan
Na Jiang
Xin Xu
Qingyang Lyu
Vinoj Gopalakrishnan
Ronald Walcott
Saul Burdman
Jianqiang Li
Laixin Luo
author_facet Yumin Kan
Na Jiang
Xin Xu
Qingyang Lyu
Vinoj Gopalakrishnan
Ronald Walcott
Saul Burdman
Jianqiang Li
Laixin Luo
author_sort Yumin Kan
collection DOAJ
description 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.
first_indexed 2024-12-21T17:08:54Z
format Article
id doaj.art-7d4c9ee790d24a2d8d89c2ba63ec421b
institution Directory Open Access Journal
issn 1664-302X
language English
last_indexed 2024-12-21T17:08:54Z
publishDate 2019-05-01
publisher Frontiers Media S.A.
record_format Article
series Frontiers in Microbiology
spelling doaj.art-7d4c9ee790d24a2d8d89c2ba63ec421b2022-12-21T18:56:27ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2019-05-011010.3389/fmicb.2019.01081454323Induction and Resuscitation of the Viable but Non-culturable (VBNC) State in Acidovorax citrulli, the Causal Agent of Bacterial Fruit Blotch of Cucurbitaceous CropsYumin Kan0Na Jiang1Xin Xu2Qingyang Lyu3Vinoj Gopalakrishnan4Ronald Walcott5Saul Burdman6Jianqiang Li7Laixin Luo8Department of Plant Pathology, China Agricultural University, Beijing Key Laboratory of Seed Disease Testing and Control, Beijing, ChinaDepartment of Plant Pathology, China Agricultural University, Beijing Key Laboratory of Seed Disease Testing and Control, Beijing, ChinaDepartment of Plant Pathology, China Agricultural University, Beijing Key Laboratory of Seed Disease Testing and Control, Beijing, ChinaDepartment of Plant Pathology, China Agricultural University, Beijing Key Laboratory of Seed Disease Testing and Control, Beijing, ChinaDepartment of Plant Pathology and Microbiology, The Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot, IsraelDepartment of Plant Pathology, University of Georgia, Athens, GA, United StatesDepartment of Plant Pathology and Microbiology, The Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot, IsraelDepartment of Plant Pathology, China Agricultural University, Beijing Key Laboratory of Seed Disease Testing and Control, Beijing, ChinaDepartment of Plant Pathology, China Agricultural University, Beijing Key Laboratory of Seed Disease Testing and Control, Beijing, ChinaAcidovorax 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.https://www.frontiersin.org/article/10.3389/fmicb.2019.01081/fullAcidovorax citrulliviable but non-culturablecopper sulfateinductionresuscitationpathogenicity
spellingShingle Yumin Kan
Na Jiang
Xin Xu
Qingyang Lyu
Vinoj Gopalakrishnan
Ronald Walcott
Saul Burdman
Jianqiang Li
Laixin Luo
Induction and Resuscitation of the Viable but Non-culturable (VBNC) State in Acidovorax citrulli, the Causal Agent of Bacterial Fruit Blotch of Cucurbitaceous Crops
Frontiers in Microbiology
Acidovorax citrulli
viable but non-culturable
copper sulfate
induction
resuscitation
pathogenicity
title Induction and Resuscitation of the Viable but Non-culturable (VBNC) State in Acidovorax citrulli, the Causal Agent of Bacterial Fruit Blotch of Cucurbitaceous Crops
title_full Induction and Resuscitation of the Viable but Non-culturable (VBNC) State in Acidovorax citrulli, the Causal Agent of Bacterial Fruit Blotch of Cucurbitaceous Crops
title_fullStr Induction and Resuscitation of the Viable but Non-culturable (VBNC) State in Acidovorax citrulli, the Causal Agent of Bacterial Fruit Blotch of Cucurbitaceous Crops
title_full_unstemmed Induction and Resuscitation of the Viable but Non-culturable (VBNC) State in Acidovorax citrulli, the Causal Agent of Bacterial Fruit Blotch of Cucurbitaceous Crops
title_short Induction and Resuscitation of the Viable but Non-culturable (VBNC) State in Acidovorax citrulli, the Causal Agent of Bacterial Fruit Blotch of Cucurbitaceous Crops
title_sort induction and resuscitation of the viable but non culturable vbnc state in acidovorax citrulli the causal agent of bacterial fruit blotch of cucurbitaceous crops
topic Acidovorax citrulli
viable but non-culturable
copper sulfate
induction
resuscitation
pathogenicity
url https://www.frontiersin.org/article/10.3389/fmicb.2019.01081/full
work_keys_str_mv AT yuminkan inductionandresuscitationoftheviablebutnonculturablevbncstateinacidovoraxcitrullithecausalagentofbacterialfruitblotchofcucurbitaceouscrops
AT najiang inductionandresuscitationoftheviablebutnonculturablevbncstateinacidovoraxcitrullithecausalagentofbacterialfruitblotchofcucurbitaceouscrops
AT xinxu inductionandresuscitationoftheviablebutnonculturablevbncstateinacidovoraxcitrullithecausalagentofbacterialfruitblotchofcucurbitaceouscrops
AT qingyanglyu inductionandresuscitationoftheviablebutnonculturablevbncstateinacidovoraxcitrullithecausalagentofbacterialfruitblotchofcucurbitaceouscrops
AT vinojgopalakrishnan inductionandresuscitationoftheviablebutnonculturablevbncstateinacidovoraxcitrullithecausalagentofbacterialfruitblotchofcucurbitaceouscrops
AT ronaldwalcott inductionandresuscitationoftheviablebutnonculturablevbncstateinacidovoraxcitrullithecausalagentofbacterialfruitblotchofcucurbitaceouscrops
AT saulburdman inductionandresuscitationoftheviablebutnonculturablevbncstateinacidovoraxcitrullithecausalagentofbacterialfruitblotchofcucurbitaceouscrops
AT jianqiangli inductionandresuscitationoftheviablebutnonculturablevbncstateinacidovoraxcitrullithecausalagentofbacterialfruitblotchofcucurbitaceouscrops
AT laixinluo inductionandresuscitationoftheviablebutnonculturablevbncstateinacidovoraxcitrullithecausalagentofbacterialfruitblotchofcucurbitaceouscrops