The tetrazole analogue of the auxin indole-3-acetic acid binds preferentially to TIR1 and not AFB5

Auxin is considered one of the cardinal hormones in plant growth and development. It regulates a wide range of processes throughout the plant. Synthetic auxins exploit the auxin-signalling pathway and are valuable as herbicidal agrochemicals. Currently, despite a diversity of chemical scaffolds all synthetic auxins have a carboxylic acid as the active core group. By applying bio-isosteric replacement we discovered that indole-3-tetrazole was active by surface plasmon resonance (SPR) spectrometry, showing that the tetrazole could initiate assembly of the TIR1 auxin co-receptor complex. We then tested the tetrazole’s efficacy in a range of whole plant physiological assays and in protoplast reporter assays which all confirmed auxin activity, albeit rather weak. We then tested indole-3-tetrazole against the AFB5 homologue of TIR1, finding that binding was selective against TIR1, absent with AFB5. The kinetics of binding to TIR1 are contrasted to those for the herbicide picloram, which shows the opposite receptor preference as it binds to AFB5 with far greater affinity than to TIR1. The basis of the preference of indole-3-tetrazole for TIR1 was revealed to be a single residue substitution using molecular docking, and assays using tir1 and afb5 mutant lines confirmed selectivity in vivo. Given the potential that a TIR1-selective auxin might have for unmasking receptor-specific actions, we followed a rational design, lead optimisation campaign and a set of chlorinated indole-3-tetrazoles was synthesised. Improved affinity for TIR1 and the preference for binding to TIR1 was maintained for 4- and 6-chloroindole-3-tetrazoles, coupled with improved efficacy in vivo. This work expands the range of auxin chemistry for the design of receptor-selective synthetic auxins

Transcriptional dynamics driving MAMP-triggered immunity and pathogen effector-mediated immunosuppression in Arabidopsis leaves following infection with Pseudomonas syringae pv tomato DC3000

Transcriptional reprogramming is integral to effective plant defense. Pathogen effectors act transcriptionally and posttranscriptionally to suppress defense responses. A major challenge to understanding disease and defense responses is discriminating between transcriptional reprogramming associated with microbial-associated molecular pattern (MAMP)-triggered immunity (MTI) and that orchestrated by effectors. A high-resolution time course of genome-wide expression changes following challenge with Pseudomonas syringae pv tomato DC3000 and the nonpathogenic mutant strain DC3000hrpA- allowed us to establish causal links between the activities of pathogen effectors and suppression of MTI and infer with high confidence a range of processes specifically targeted by effectors. Analysis of this information-rich data set with a range of computational tools provided insights into the earliest transcriptional events triggered by effector delivery, regulatory mechanisms recruited, and biological processes targeted. We show that the majority of genes contributing to disease or defense are induced within 6 h postinfection, significantly before pathogen multiplication. Suppression of chloroplast-associated genes is a rapid MAMP-triggered defense response, and suppression of genes involved in chromatin assembly and induction of ubiquitin-related genes coincide with pathogen-induced abscisic acid accumulation. Specific combinations of promoter motifs are engaged in fine-tuning the MTI response and active transcriptional suppression at specific promoter configurations by P. syringae

Identyfikacja cDNA genu RSH [RelA-SpoT homolog] zaangazowanego w odpowiedz Pharbitis nil na warunki stresowe

Stres środowiskowy jest z jednym z głównych czynników limitujących wzrost i rozwój roślin. Przeszukiwanie biblioteki cDNA Pharbitis nil z liścieni roślin zaindukowanych do kwitnienia sondą SOD3.1 kodującą manganową dysmutazę ponadtlenkową pszenicy, umożliwiło zidentyfikowanie sekwencji długości 798 pz. Sekwencja ta wykazuje najwyższą homologię do roślinnych genów RSH, homologicznych do bakteryjnych RelA i SpoT. Białka kodowane przez te geny uczestniczą w odpowiedzi ścisłej - wysoce konserwowanym mechanizmie odpowiedzi na stres.Environmental stress is one of the main factors limiting plant growth and development. The cDNA library constructed from cotyledons of Pharbitis nil plants photoinduced for flowering was screened by probe SOD3.1 coding wheat manganese superoxide dismutase. The cDNA sequence of 798 bp was isolated showing the highest identity to the plant RSH genes which are homologs of bacterial RelA and SpoT genes. Their protein products participate in the stringent response - a highly conserved mechanism of stress response