@article{28e84faaf2e34aa1a693fbb67e3d9f3e,
title = "Host-free biofilm culture of “Candidatus Liberibacter asiaticus,” the bacterium associated with Huanglongbing",
abstract = "Inability to culture the phloem-restricted alpha-proteobacterium “Candidatus Liberibacter asiaticus” (“Ca. L. asiaticus”) or the closely related species (“Candidatus Liberibacter americanus” and “Candidatus Liberibacter africanus”) that are associated with Huanglongbing (HLB) hampers the development of effective long-term control strategies for this devastating disease. Here we report successful establishment and long-term maintenance of host-free “Ca. L. asiaticus” cultures, with the bacterium growing within cultured biofilms derived from infected citrus tissue. The biofilms were grown in a newly designed growth medium under specific conditions. The initial biofilm-based culture has been successfully maintained for over two years and has undergone over a dozen subcultures. Multiple independent cultures have been established and maintained in a biofilm reactor system, opening the door to the development of pure culture of “Ca. L. asiaticus” and the use of genetics-based methods to understand and mitigate the spread of HLB.",
keywords = "Biofilm, Candidatus Liberibacter asiaticus, Host-free culture, Huanglongbing, Isolation",
author = "Ha, {Phuc T.} and Ruifeng He and Nabil Killiny and Brown, {Judith K.} and Anders Omsland and Gang, {David R.} and Haluk Beyenal",
note = "Funding Information: We thank Julie Thayer for assistance with citrus tree growth in the Institute of Biological Chemistry (IBC) greenhouses. The authors would like to acknowledge the staff at the Franceschi Microscopy and Imaging Center (Washington State University) for their assistance and for providing the facilities for imaging analysis. This work was supported by the Citrus Disease Research and Extension Program of the Specialty Crop Research Initiative (grant no. 2016-70016-24824 ) of the USDA National Institute of Food and Agriculture . Any opinions, findings, conclusions, or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect the views of the U.S. Department of Agriculture. Funding Information: We thank Julie Thayer for assistance with citrus tree growth in the Institute of Biological Chemistry (IBC) greenhouses. The authors would like to acknowledge the staff at the Franceschi Microscopy and Imaging Center (Washington State University) for their assistance and for providing the facilities for imaging analysis. This work was supported by the Citrus Disease Research and Extension Program of the Specialty Crop Research Initiative (grant no. 2016-70016-24824) of the USDA National Institute of Food and Agriculture. Any opinions, findings, conclusions, or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect the views of the U.S. Department of Agriculture. Publisher Copyright: {\textcopyright} 2019 The Authors",
year = "2019",
month = dec,
doi = "10.1016/j.bioflm.2019.100005",
language = "English (US)",
volume = "1",
journal = "Biofilm",
issn = "2590-2075",
publisher = "Elsevier BV",
}