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...
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Frontiers Media S.A.
2019-05-01
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Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2019.01081/full |
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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. |
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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 |
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