A Soil‐free Plant Growth System to Facilitate Analysis of Plant Pathogen Interactions in Roots

Gunning, Tiffany; Cahill, David M.

doi:10.1111/j.1439-0434.2008.01503.x

Cited by 18 publications

(9 citation statements)

References 13 publications

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“…However, routine tissue culture systems do not provide a soil-like growth substrate for microbial colonization. Furthermore, agar-based systems are notorious for non-uniform nutrient and O2 delivery over time 9 . Hydroponic and aeroponic systems can alleviate issues with nutrient uniformity and O2-delivery by agitation and media replenishment, but such systems still do not provide a soil-like growth substrate for microbial colonization 10 .…”

Section: Existing Growth Methodology For Plant Microbiome Researchmentioning

confidence: 99%

FlowPot axenic plant growth system for microbiota research

Kremer

Rhodes

et al. 2018

Preprint

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The presence of resident microbiota on and inside plants is hypothesized to influence many phenotypic attributes of the host. Likewise, host factors and microbe-microbe interactions are believed to influence microbial community assembly. Rigorous testing of these hypotheses necessitates the ability to grow plants in the absence or presence of resident or defined microbiota. To enable such experiments, we developed the scalable and inexpensive FlowPot growth platform. FlowPots have a sterile peat substrate amenable to colonization by microbiota, and the platform supports growth of the model plant Arabidopsis thaliana in the absence or presence of soil-derived microbial communities. Mechanically, the FlowPot system is unique in that it allows for total-saturation of the sterile substrate by "flushing" with water and/or nutrient solution via an irrigation port. The irrigation port also facilitates passive drainage of the substrate, preventing root anoxia. Materials to construct an individual FlowPot total ~$2.A simple experiment with 12 FlowPots requires ~4.5 h of labor following peat and seed sterilization. Plants are grown on FlowPots within a standard tissue culture microbox after inoculation, thus the Flowpot system is modular and does not require a sterile growth chamber. Here, we provide a detailed assembly and microbiota inoculation protocol for the FlowPot system. Collectively, this standardized suite of tools and colonization protocols empowers the plant microbiome research community to conduct harmonized experiments to elucidate the rules microbial community assembly, the impact of microbiota on host phenotypes, and mechanisms by which host factors influence the structure and function of plant microbiota.3

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Section: Existing Growth Methodology For Plant Microbiome Researchmentioning

confidence: 99%

FlowPot axenic plant growth system for microbiota research

Kremer

Rhodes

et al. 2018

Preprint

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show abstract

“…The filter paper in each Petri dish was replaced each week with a fresh piece that was moistened with sdH 2 O. The germinated seeds were then transferred to a soil‐free plant growth system (SPS), 20 plants in each module, as described in Gunning & Cahill () where roots could grow vertically through a moistened filter paper support. The SPS system allowed the production of roots free from soil and enabled roots to be inoculated with minimal disturbance.…”

Section: Methodsmentioning

confidence: 99%

Transcriptome analysis, using RNA‐Seq of Lomandra longifolia roots infected with Phytophthora cinnamomi reveals the complexity of the resistance response

et al. 2017

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The plant pathogen Phytophthora cinnamon the causal agent of disease in numerous species, is a major threat to natural vegetation and has economic impacts in agriculture. The pathogen principally invades the root system, which, in susceptible species, is rapidly colonised and functionally destroyed. Few species are resistant, however, where resistance is expressed the pathogen is restricted to small, localised lesions. The molecular mechanisms that underpin this response in resistant species are not well understood. Lomandra longifolia, an Australian native species, is highly resistant to P. cinnamomi. In an earlier study, we showed induction of resistance-related components such as callose, lignin and hydrogen peroxide (H O ) in L. longifolia roots that had been inoculated with P. cinnamomi. Here, in order to further identify, during the very early stages of infection, the molecular components and regulatory networks that may trigger resistance, a comprehensive root transcriptome analysis was performed using next generation sequencing. Overall, 18 cDNA libraries were produced generating 52.8 GB 126 base pair reads, which were de novo assembled into contigs. Differentially expressed genes (DEGs) were identified allowing the identification of infection-responsive candidate genes that were putatively related to resistance, and from this set ten were selected for qRT-PCR to validate the RNA-Seq expression value. Further analysis of individual candidates revealed that many were involved in PAMP-triggered immunity (PTI; pattern recognition receptors, glutathione S-transferase, callose synthases, pathogenesis-related protein-1, mitogen activated protein kinases) and effector-triggered immunity (ETI) (NBS-LRR, signalling genes, transcription factors and anti-pathogenic compound synthase genes). As these candidate genes or mediated components activate different defence signalling systems, they may have potential for investigation of novel approaches to disease control and in transgenic approaches for improvement, in susceptible species, of resistance to P. cinnamomi.

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“…Seeds of Z. mays (‘Early Leaming’, Eden seeds, Lower Beechmont, QLD, Australia) and L. angustifolius (var. wonga , Naracoorte, SA, Australia) were germinated and grown on filter paper in a soil‐free plant growth system (SPS) as previously described (Gunning and Cahill 2009) within temperature‐controlled growth cabinets under 3 × 600 watt high‐pressure sodium lights with a 16/8 h photoperiod at 21°C at 200 μmol/m 2 /s. Seedlings were watered daily with a pressurized spray bottle containing 1/3 strength nutrient solution (Total Horticultural Concentrate, Excel Distributors, Doncaster East, Vic., Australia), and the filter paper and nutrient solution were replaced every 5 days.…”

Section: Methodsmentioning

confidence: 99%

Defining Plant Resistance to Phytophthora cinnamomi: A Standardized Approach to Assessment

Allardyce¹,

Rookes²,

Cahill³

2012

Journal of Phytopathology

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Phytophthora cinnamomi is a soil‐borne plant pathogen that causes devastating disease in agricultural and natural systems worldwide. While a small number of species survive infection by the pathogen without producing disease symptoms, the nature of resistance, especially under controlled conditions, remains poorly understood. At present, there are no standardized criteria by which resistance or susceptibility to P. cinnamomi can be assessed, and we have used five parameters consisting of plant fresh weight, root growth, lesion length, relative chlorophyll content of leaves and pathogen colonization of roots to analyse responses to the pathogen. The parameters were tested using two plant species, Zea mays and Lupinus angustifolius, through a time course study of the interactions and resistance and susceptibility defined 7 days after inoculation. A scoring system was devised to enable differentiation of these responses. In the resistant interaction with Z. mays, there was no significant difference in fresh weight, root length and relative chlorophyll content in inoculated compared with control plants. Both lesion size and pathogen colonization of root tissues were limited to the site of inoculation. Following inoculation L. angustifolius showed a significant reduction in plant fresh weight and relative leaf chlorophyll content, cessation of root growth and increased lesion lengths and pathogen colonization. We propose that this technique provides a standardized method for plant–P. cinnamomi interactions that could be widely used to differentiate resistant from susceptible species.

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A Soil‐free Plant Growth System to Facilitate Analysis of Plant Pathogen Interactions in Roots

Cited by 18 publications

References 13 publications

FlowPot axenic plant growth system for microbiota research

FlowPot axenic plant growth system for microbiota research

Transcriptome analysis, using RNA‐Seq of Lomandra longifolia roots infected with Phytophthora cinnamomi reveals the complexity of the resistance response

Defining Plant Resistance to Phytophthora cinnamomi: A Standardized Approach to Assessment

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