Open AccessControlled replication of ‘ Candidatus Liberibacter asiaticus ‘ DNA in citrus leaf discs
Author(s) -
Attaran Elham,
Berim Anna,
Killiny Nabil,
Beyenal Haluk,
Gang David R.,
Omsland Anders
Publication year - 2020
Publication title -
microbial biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.287
H-Index - 74
ISSN - 1751-7915
DOI - 10.1111/1751-7915.13531
Subject(s) - biology , bacteria , dna , diaphorina citri , fastidious organism , microbiology and biotechnology , botany , genetics , hemiptera
Summary ‘ Candidatus Liberibacter asiaticus ’ is a fastidious bacterium and a putative agent of citrus greening disease (a.k.a., huanglongbing, HLB), a significant agricultural disease that affects citrus fruit quality and tree health. In citrus, ‘ Ca. L. asiaticus ’ is phloem limited. Lack of culture tools to study ‘ Ca. L. asiaticus ’ complicates analysis of this important organism. To improve understanding of ‘ Ca. L. asiaticus ’–host interactions including parameters that affect ‘ Ca. L. asiaticus ’ replication, methods suitable for screening pathogen responses to physicochemical and nutritional variables are needed. We describe a leaf disc‐based culture assay that allows highly selective measurement of changes in ‘ Ca. L. asiaticus ’ DNA within plant tissue incubated under specific physicochemical and nutritional conditions. qPCR analysis targeting the hypothetical gene CD16‐00155 (strain A4) allowed selective quantification of ‘ Ca. L. asiaticus ’ DNA content within infected tissue. ‘ Ca. L. asiaticus ’ DNA replication was observed in response to glucose exclusively under microaerobic conditions, and the antibiotic amikacin further enhanced ‘ Ca. L. asiaticus ’ DNA replication. Metabolite profiling revealed a moderate impact of ‘ Ca. L. asiaticus ’ on the ability of leaf tissue to metabolize and respond to glucose.