The Cultured Microbiome of Pollinated Maize Silks Shifts after Infection with Fusarium graminearum and Varies by Distance from the Site of Pathogen Inoculation

Thompson, Michelle E. H.; Shrestha, Anuja; Rinne, Jeffrey; Limay-Rios, Victor; Reid, Lana; Raizada, Manish N.

doi:10.3390/pathogens12111322

Cited by 6 publications

(1 citation statement)

References 164 publications

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“…Isolate R22 was also observed densely colonizing Fg hyphae, including near transition points where Fg was dying (turning from green to red). Isolate R22 contained genes for phenazine biosynthesis, acetoin biosynthesis, and colicin V biosynthesis (Thompson, 2023).…”

Section: Discussionmentioning

confidence: 99%

Protocols to enable fluorescence microscopy of microbial interactions on living maize silks (style tissue) that balance the requirements for containment, biological relevance, viability, and experimental accessibility

Thompson,

Raizada

2023

Preprint

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There is interest in studying microbes that colonize maize silks (style tissue, critical for reproduction) including the fungal pathogenFusarium graminearum(Fg) and its interactions with the microbiome and biocontrol agents.In plantaimaging of these interactions on living silks using confocal fluorescence microscopy would provide key insights. However, newly discovered microbes have unknown effects of on human health, and there are regulatory requirements to prevent the release of fluorescently-tagged microbes into the environment.Therefore, the microbe infection, colonization, and interaction stages on silks prior to microscopy must be contained. At the same time, silk viability must be maintained and experiments conducted that are biologically relevant (e.g. silks should remain attached to the cob), yet the silk tissue must be accessible to the researcher (i.e. not within husk leaves) and allow for multiple replicates. Here we present methods that meet these five contrasting criteria. We tested these methods usingFgand four silk-derived bacterial endophytes. The endophytes were previously known to have anti-Fgactivityin vitro, butin plantaobservations were lacking. In Method 1, a portion of the tip of a cob was dissected, and silks remained attached to the cob in a Petri dish. The cob was placed on a water agar disc to maintain hydration. DsRed-tagged bacteria and GFP-taggedFgwere inoculated onto the silks and incubated, allowing the two microbes to grow towards one another before staining with propidium iodide for confocal microscopy. A variation of the protocol was presented in Method 2, where detached silk segments were placed directly on water agar where they were inoculated with bacteria andFgto promote dense colonization, and to allow for many replicates and interventions such as silk wounding. The bacterial endophytes were successfully observed colonizingFghyphae, silk trichomes, and entering silks via cut ends and wounds. These protocols can be used to study other silk-associated microbes including several globally important fungal pathogens that enter maize grain through silks.

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Section: Discussionmentioning

confidence: 99%

Protocols to enable fluorescence microscopy of microbial interactions on living maize silks (style tissue) that balance the requirements for containment, biological relevance, viability, and experimental accessibility

Thompson,

Raizada

2023

Preprint

View full text Add to dashboard Cite

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Protocols to enable fluorescence microscopy of microbial interactions on living maize silks (style tissue)

Thompson,

Raizada

2024

Journal of Microbiological Methods

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Genome-wide identification and expression analysis of CASPL gene family in Zea mays (L.)

Xue,

Liang,

et al. 2024

Front. Plant Sci.

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Casparian strip membrane domain proteins like (CASPL), exhibit profound associations with root development, stress responsiveness and mineral element uptake in plants. Nonetheless, a comprehensive bioinformatics analysis of the ZmCASPL gene family in maize remains unreported. In the study, we have identified 47 ZmCASPL members at the whole-genome level, systematically classifying them into six distinct groups. Furthermore, our analysis revealed that the same group of ZmCASPL contains similar gene structures and conserved motifs. Duplication events showed whole genome duplication (WGD) and tandem duplication (TD) contribute to the generation of the ZmCASPL gene family together in maize, but the former plays a more prominent role. Furthermore, we observed that most ZmCASPL genes contain MYB-binding sites (CAACCA), which are associated with the Casparian strip. Utilizing RNA-seq data, we found that ZmCASPL21 and ZmCASPL47 are specifically highly expressed only in the roots. This finding implies that ZmCASPL21 and ZmCASPL47 may be involved in the Casparian strip development. Additionally, RNA-seq analysis illuminated that drought, salt, heat, cold stresses, low nitrogen and phosphorus conditions, as well as pathogen infection, significantly impact the expression patterns of ZmCASPL genes. RT-qPCR revealed that ZmCASPL 5/13/25/44 genes showed different expression patterns under PEG and NaCl treatments. Collectively, these findings provide a robust theoretical foundation for further investigations into the functional roles of the ZmCASPL gene family in maize.

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The Cultured Microbiome of Pollinated Maize Silks Shifts after Infection with Fusarium graminearum and Varies by Distance from the Site of Pathogen Inoculation

Cited by 6 publications

References 164 publications

Protocols to enable fluorescence microscopy of microbial interactions on living maize silks (style tissue) that balance the requirements for containment, biological relevance, viability, and experimental accessibility

Protocols to enable fluorescence microscopy of microbial interactions on living maize silks (style tissue) that balance the requirements for containment, biological relevance, viability, and experimental accessibility

Protocols to enable fluorescence microscopy of microbial interactions on living maize silks (style tissue)

Genome-wide identification and expression analysis of CASPL gene family in Zea mays (L.)

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