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Schlueter, T.

Publications and source records attributed to Schlueter, T..

2 recordsLinked to original sources

Contrasting and conserved roles of NPR proteins in diverged land plant lineages

The NPR proteins function as salicylic acid (SA) receptors in Arabidopsis thaliana. AtNPR1 plays a central role in SA-induced transcriptional reprogramming whereby positively regulates SA-mediated defense. NPRs are found in the genomes of nearly all land plants. However, we know little about the molecular functions and physiological roles of NPRs in most plant species. Our phylogenetic and alignment analyses show that Brassicaceae NPR1-like proteins have characteristically gained or lost functional residues or motifs identified in AtNPRs, pointing to the possibility of a unique evolutionary trajectory for the Brassicaceae NPR1-like proteins that has resulted in peculiar functions. In line with this observation, we find that the only NPR in Marchantia polymorpha, MpNPR, is not the master regulator of SA-induced transcriptional reprogramming and negatively regulates bacterial resistance in this species. Interspecies complementation analysis indicated that the molecular properties of AtNPR1 and MpNPR are partially conserved, implying the diversification of NPR-associated pathways contributed to distinct roles of NPR in different species. The Mpnpr transcriptome suggested potential roles of MpNPR in heat and far-red light responses. We identify both Mpnpr and Atnpr1-1 display enhanced thermomorphogenesis. NPRs and NPR-associated pathways clearly have evolved distinctively in diverged land plant lineages to cope with different terrestrial environments.

plant biology↗

A versatile Tn7 transposon-based bioluminescence tagging tool for quantitative and spatial detection of bacteria in plants

Investigation of plant-bacteria interactions requires quantification of in planta bacterial titers by means of colony counting assays. However, colony counting assays are cumbersome and time-consuming, and are unable to detect spatial patterns of bacterial colonization in plants. Here, to overcome these shortcomings, we devised a broadly applicable genetic engineering tool for bioluminescence-based quantitative and spatial detection of bacteria in plants. We developed plasmid vectors that have broad host ranges and enable Tn7 transposon-mediated integration of the luxCDABE luciferase operon into a specific genomic location ubiquitously found across bacterial phyla. These vectors allowed for generation of bioluminescent transformants of various plant pathogenic bacteria belonging to the genera Pseudomonas, Rhizobium (Agrobacterium), and Ralstonia. The bioluminescent transformant of Pseudomonas syringae pv. tomato DC3000 (Pto-lux) was as virulent in Arabidopsis thaliana as its parental strain. Direct luminescence measurements of Pto-lux-inoculated plant tissues reported bacterial titers in A. thaliana, Solanum lycopersicum, Nicotiana benthamiana, and Marchantia polymorpha as accurately as conventional colony counting assays. We further showed the utility of our vectors for converting the previously generated Pto derivatives to isogenic bioluminescent strains. Importantly, quantitative bioluminescence assays using these Pto-lux strains accurately reported the effects of plant immunity and bacterial effectors on bacterial growth with a dynamic range of 4 orders of magnitude. Moreover, macroscopic bioluminescence imaging illuminated spatial colonization patterns of the Pto-lux in/on inoculated plant tissues. Taken together, our vectors offer untapped opportunities for developing bioluminescence-based quantitative and spatial analysis of bacterial growth in a variety of plant-bacteria interactions. SIGNIFICANCE STATEMENTWe developed broad-host-range plasmid vectors that integrate the luciferase operon, luxCDABE, into a specific genomic location ubiquitously found across bacterial phyla. Using these vectors, we established a high-throughput method for bioluminescence-based quantitative assays of in planta bacterial growth with a dynamic range of 4 orders of magnitude and visualized spatiotemporal patterns of bacterial colonization in/on inoculated plant tissues.

plant biology↗