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Lueck, S.

Publications and source records attributed to Lueck, S..

3 recordsLinked to original sources

Deep phenotyping platform for microscopic plant-pathogen interactions

1O_LIThe initial phases of plant-pathogen interactions are critical since they are often decisive for the successful infection. However, these early stages of interaction are typically microscopic, making it challenging to study on a large scale. C_LIO_LIFor this reason, using the powdery mildew fungi of cereals as a model, we have developed an automated microscopy pipeline coupled with deep learning-based image analysis for the high-throughput phenotyping of plant-pathogen interactions. C_LIO_LIThe system can quantify fungal microcolony count and density, the precise area of the secondary hyphae of each colony, and different morphological parameters. Moreover, the high throughput and sensitivity allow quantifying rare microscopic phenotypes in a large sample size. One of these phenotypes is the cryptic infection of non-adapted pathogens, marking the hidden transition stages of pathogen adaptation and breaking the nonhost barrier. Thus, our tool opens the nonhost resistance phenomenon to genetics and genomics studies. C_LIO_LIWe have developed an open-source high-throughput automated microscopy system for phenotyping the initial stages of plant-pathogen interactions, extendable to other microscopic phenotypes and hardware platforms. Furthermore, we have validated the systems performance in disease resistance screens of genetically diverse barley material and performed Genome-wide associations scans (GWAS), discovering several resistance-associated loci, including conferring nonhost resistance. C_LI

plant biology↗

'Macrobot' - an automated segmentation-based system for powdery mildew disease quantification

Plant diseases, as one of the perpetual problems in agriculture, is increasingly difficult to manage due to intensifying of the field production, global trafficking, reduction of genetic variability of crops, climatic changes-driven expansion of pests, redraw and loss of effectiveness of pesticides and rapid breakdown of the disease resistance in the field. The substantial progress in genomics of both plants and pathogens, achieved in the last decades has the potential to counteract this negative trend, however, only when the genomic data is supported by relevant phenotypic data that allows linking the genomic information to specific traits. In this respect, phenotyping is and will remain an essential element of any comprehensive functional genomics study. We have developed a set of methods and equipment and combined them into a "Macrophenomics pipeline". The pipeline has been optimized for the quantification of powdery mildew infection symptoms on wheat and barley but it can be adapted to other diseases and host plants. The Macrophenomics pipeline scores the visible disease symptoms, typically 5-7 days after inoculation (dai) in a highly automated manner. The system can precisely and reproducibly quantify the percentage of the infected leaf area with a throughput of the image acquisition module of up to 10 000 individual samples per day, making it appropriate for phenotyping of large germplasms collections and crossing populations.

plant biology↗

The partial duplication of an E3-ligase gene in Triticeaespecies mediates resistance to powdery mildew fungi

In plant-pathogen interactions, components of the plant ubiquitination machinery are preferred targets of pathogen-encoded effectors suppressing defense responses or co-opting host cellular functions for accommodation. Here, we employed transient and stable gene silencing-and over-expression systems in Hordeum vulgare (barley) to study the function of HvARM1 (for H. vulgare Armadillo 1), a partial gene duplicate of the U-box/armadillo-repeat E3 ligase HvPUB15 (for H. vulgare Plant U-Box 15). The partial ARM1 gene was derived from an ancient gene-duplication event in a common ancestor of the Triticeae tribe of grasses comprising the major crop species H. vulgare, Triticum aestivum and Secale cereale. The barley gene HvARM1 contributed to quantitative host as well as nonhost resistance to the biotrophic powdery mildew fungus Blumeria graminis, and allelic variants were found to be associated with powdery mildew-disease severity. Both HvPUB15 and HvARM1 proteins interacted in yeast and plant cells with the susceptibility-related, plastid-localized barley homologs of THF1 (for Thylakoid formation 1) and of ClpS1 (for Clp-protease adaptor S1) of Arabidopsis thaliana. The results suggest a neo-functionalization HvARM1 to increase resistance against powdery mildew and provide a link to plastid function in susceptibility to biotrophic pathogen attack.

plant biology↗