title: Viability of 'Candidatus Liberibacter asiaticus' prolonged by addition of citrus juice to culture medium.
abstract: Huanglongbing, or citrus greening disease, is associated with infection by the phloem-limited bacterium 'Candidatus Liberibacter asiaticus'. Infection with 'Ca. L. asiaticus' is incurable; therefore, knowledge regarding 'Ca. L. asiaticus' biology and pathogenesis is essential to develop a treatment. However, 'Ca. L. asiaticus' cannot currently be successfully cultured, limiting its study. To gain insight into the conditions conducive for growth of 'Ca. L. asiaticus' in vitro, 'Ca. L. asiaticus' inoculum obtained from seed of fruit from infected pomelo trees (Citrus maxima 'Mato Buntan') was added to different media, and cell viability was monitored for up to 2 months using quantitative polymerase chain reaction in conjunction with ethidium monoazide. Media tested included one-third King's B (K), K with 50% juice from the infected fruit, K with 50% commercially available grapefruit juice, and 100% commercially available grapefruit juice. Results show that juice-containing media dramatically prolong viability compared with K in experiments reproduced during 2 years using different juice sources. Furthermore, biofilm formed at the air-liquid interface of juice cultures contained 'Ca. L. asiaticus' cells, though next-generation sequencing indicated that other bacterial genera were predominant. Chemical characterization of the media was conducted to discuss possible factors sustaining 'Ca. L. asiaticus' viability in vitro, which will contribute to future development of a culture medium for 'Ca. L. asiaticus'.
authors.forename: Jennifer K
authors.initials: JK
authors.lastname: Parker
authors.forename: Sarah R
authors.initials: SR
authors.lastname: Wisotsky
authors.forename: Evan G
authors.initials: EG
authors.lastname: Johnson
authors.forename: Faraj M
authors.initials: FM
authors.lastname: Hijaz
authors.forename: Nabil
authors.initials: N
authors.lastname: Killiny
authors.forename: Mark E
authors.initials: ME
authors.lastname: Hilf
authors.forename: Leonardo
authors.initials: L
authors.lastname: De La Fuente
mesh_terms: D001628:Beverages; D032083:Citrus paradisi / Q000737:chemistry* / Q000382:microbiology; D003470:Culture Media / Q000494:pharmacology; D010935:Plant Diseases / Q000382:microbiology*; D060888:Real-Time Polymerase Chain Reaction; D012230:Rhizobiaceae / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development; D012639:Seeds / Q000737:chemistry* / Q000382:microbiology
chemical_list: D003470:Culture Media