Visualization of spatial gene expression in plants by modified RNAscope fluorescent<i>in situ</i>hybridization

Solanki, Shyam; Ameen, Gazala; Zhao, Jin; Flaten, Jordan; Borowicz, Paweł; Brueggeman, Robert

doi:10.1101/2020.04.01.020727

Cited by 2 publications

(2 citation statements)

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“…By exploiting the ability to nondestructively and spatially resolve infection spots, redox imaging can serve as an ideal tool for investigation of the molecular changes in tissues at different stages of infection. For example, the combination of redox imaging and spatial transcriptomics or mass spectrometry imaging (Dong et al, 2016; Giacomello et al, 2017; Petras et al, 2017; Schleyer et al, 2019; Solanki et al, 2020) has the potential to precisely tap into the genetic and metabolic processes underlying disease onset.…”

Section: Resultsmentioning

confidence: 99%

Early detection of late blight in potato by whole-plant redox imaging

Hipsch

Yaron

Lampl

et al. 2022

Preprint

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Late blight caused by the oomycete Phytophthora infestans is a most devastating disease of potatoes (Solanum tuberosum). Its early detection is crucial for suppressing disease spread. Necrotic lesions are normally seen in leaves at 4 dpi (days post inoculation) when colonized cells are dead, but early detection of the initial biotrophic growth stage, when the pathogen feeds on living cells, is challenging. Here, the biotrophic growth phase of P. infestans was detected by whole-plant redox imaging of potato plants expressing chloroplast-targeted reduction-oxidation sensitive green fluorescent protein (chl-roGFP2). Clear spots on potato leaves with a lower chl-roGFP2 oxidation state were detected as early as 2 dpi, before any visual symptoms were recorded. These spots were particularly evident during light-to-dark transitions and reflected mislocalization of chl-roGFP2 outside the chloroplasts, demonstrating perturbation of the chloroplast import system by the pathogen. Image analysis based on machine learning enabled systematic identification and quantification of spots and unbiased classification of infected and uninfected leaves in inoculated plants. Comparing redox to chlorophyll fluorescence imaging showed that infected leaf areas which exhibit mislocalized chl-roGFP2 also showed reduced non-photochemical quenching (NPQ) and enhanced quantum PSII yield (ΦPSII) compared to the surrounding leaf areas. The data suggest that mislocalization of chloroplast-targeted proteins is an efficient marker of late blight infection and demonstrate how it can be utilized for nondestructive monitoring of the disease biotrophic stage using whole-plant redox imaging.

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Section: Resultsmentioning

confidence: 99%

Early detection of late blight in potato by whole-plant redox imaging

Hipsch

Yaron

Lampl

et al. 2022

Preprint

View full text Add to dashboard Cite

show abstract

“…By exploiting the ability to non‐destructively and spatially resolve infection spots, redox imaging can serve as an ideal tool for investigating the molecular changes in tissues at different stages of infection. For example, the combination of redox imaging and spatial transcriptomics or mass spectrometry imaging (Dong et al., 2016; Giacomello et al., 2017; Petras et al., 2017; Schleyer et al., 2019; Solanki et al., 2020) has the potential to precisely tap into the genetic and metabolic processes underlying disease onset and assist in the discovery of early natural markers for P. infestans infection.…”

Section: Discussionmentioning

confidence: 99%

Early detection of late blight in potato by whole‐plant redox imaging

et al. 2023

View full text Add to dashboard Cite

Late blight caused by the oomycete Phytophthora infestans is a most devastating disease of potatoes (Solanum tuberosum). Its early detection is crucial for suppressing disease spread. Necrotic lesions are normally seen in leaves at 4 days post-inoculation (dpi) when colonized cells are dead, but early detection of the initial biotrophic growth stage, when the pathogen feeds on living cells, is challenging. Here, the biotrophic growth phase of P. infestans was detected by whole-plant redox imaging of potato plants expressing chloroplasttargeted reduction-oxidation sensitive green fluorescent protein (chl-roGFP2). Clear spots on potato leaves with a lower chl-roGFP2 oxidation state were detected as early as 2 dpi, before any visual symptoms were recorded. These spots were particularly evident during light-to-dark transitions, and reflected the mislocalization of chl-roGFP2 outside the chloroplasts. Image analysis based on machine learning enabled systematic identification and quantification of spots, and unbiased classification of infected and uninfected leaves in inoculated plants. Comparing redox with chlorophyll fluorescence imaging showed that infected leaf areas that exhibit mislocalized chl-roGFP2 also showed reduced non-photochemical quenching and enhanced quantum PSII yield (ΦPSII) compared with the surrounding leaf areas. The data suggest that mislocalization of chloroplast-targeted proteins is an efficient marker of late blight infection, and demonstrate how it can be utilized for non-destructive monitoring of the disease biotrophic stage using whole-plant redox imaging.

show abstract

Visualization of spatial gene expression in plants by modified RNAscope fluorescentin situhybridization

Abstract: 22 and semi-quantification of target RNAs which can be employed in other plants such as the widely utilized 40 model dicot plant Arabidopsis. 41 Keywords 42 RNAscope, multiplexing, barley, in situ hybridization, RNA, probe, HvGAPDH, Rpg1 43 44

Cited by 2 publications

References 21 publications

Early detection of late blight in potato by whole-plant redox imaging

Early detection of late blight in potato by whole-plant redox imaging

Early detection of late blight in potato by whole‐plant redox imaging

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