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Controlled replication of 'Candidatus Liberibacter asiaticus' DNA in citrus leaf discs.
Microbial Biotechnology ( IF 4.8 ) Pub Date : 2020-01-20 , DOI: 10.1111/1751-7915.13531
Elham Attaran 1 , Anna Berim 2 , Nabil Killiny 3 , Haluk Beyenal 4 , David R Gang 2 , Anders Omsland 1
Affiliation  

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.

中文翻译:


柑橘叶盘中“Candidatus Liberibacter asiaticus”DNA 的受控复制。



Candidatus Liberibacter asiaticus ”是一种要求严格的细菌,也是柑橘绿化病(又名黄龙病,HLB)的假定病原体,柑橘绿化病是一种影响柑橘果实品质和树木健康的重要农业病害。在柑橘中,' Ca。 L. asiaticus ' 的韧皮部有限。缺乏研究“ Ca”的文化工具。 L. asiaticus的研究使这一重要生物体的分析变得更加复杂。提高对 ' Ca的理解。 L. asiaticus '-宿主相互作用,包括影响 ' Ca 的参数。亚洲乳杆菌的复制需要适合筛选病原体对理化和营养变量反应的方法。我们描述了一种基于叶盘的培养测定,可以高度选择性地测量Ca的变化。 L. asiaticus在特定理化和营养条件下培育的植物组织内的 DNA。针对假设基因 CD16-00155(A4 株)的 qPCR 分析允许对Ca进行选择性定量。 L. asiaticus受感染组织内的 DNA 含量。 ' Ca。在微需氧条件下观察到亚洲乳杆菌的 DNA 复制仅响应于葡萄糖,并且抗生素阿米卡星进一步增强了Ca。 L. asiaticus的 DNA 复制。代谢物分析显示 ' Ca具有中等影响。 L. asiaticus '对叶组织代谢和响应葡萄糖的能力的影响。
更新日期:2020-01-20
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