Steczkiewicz, K. | bioRxiv Science

bioRxiv2025

The NLR immune receptor Roq1 recognizes the Pseudomonas syringae HopAG1 effector via its Nudix domain

O_LIPlant nucleotide-binding leucine-rich repeat receptors (NLRs) sense directly or indirectly effectors - proteins employed by pathogens during infection. While indirect recognition can be initiated by unrelated effectors targeting the same cellular component, direct recognition is restricted by conformational constraints of the pathogen protein and corresponding receptor. C_LIO_LIRecognition of XopQ (Roq1) receptor confers resistance to bacteria expressing members of the highly conserved effector family comprising XopQ/HopQ1/RipB. The structure of the activated Roq1 was solved previously, revealing that LRR and C-JID domains of Roq1 are involved in XopQ binding. C_LIO_LIIn this study, we found that Roq1 detects HopAG1, a member of a structurally different effector family. Presence of HopAG1 in Pseudomonas syringae reduced bacterial growth and symptoms development in wild-type but not in roq1 Nicotiana benthamiana plants. Moreover, HopAG1 co-immunoprecipitated with Roq1 indicating that both proteins associate. Based on deletion and structural analyses, we postulate that this interaction occurs via the Nudix domain of HopAG1 that binds to a similar receptor interface like XopQ. C_LIO_LINudix fold is predicted in multiple pathogen effectors suggesting that Roq1 may recognize effector families other than XopQ and HopAG1. Accordingly, it is also conceivable that other TNLs may detect a broader range of effector families than supposed. C_LI

plant biology↗

bioRxiv2020

Metabolic potential, ecology and presence of endohyphal bacteria is reflected in genomic diversity of Mucoromycotina

We describe the genomes of six Mucoromycotina fungi representing distant saprotrophic lineages within the subphylum (i.e. Umbelopsidales and Mucorales). We selected two Umbelopsis isolates from soil (i.e. U. isabellina, U. vinacea), two soil-derived Mucor isolates (i.e. M. circinatus, M. plumbeus), and two Mucorales representatives with extended proteolytic activity (i.e. Thamnidium elegans and Mucor saturninus). We complement genome analyses with a description of their digestive capabilities, their cell wall carbohydrate composition, and total lipid profiles. Finally, we link the presence of endohyphal bacteria with observed characteristics. One of the genomes, Thamnidium elegans, harbours a complete genome of an associated bacterium classified to Paenibacillus sp. This fungus displays multiple altered traits compared to remaining isolates regardless of their evolutionary distance. T. elegans has expanded carbon assimilation capabilities particularly efficiently degrades carboxylic acids, has a higher diacylglycerol: triacylglycerol ratio and phospholipid composition suggesting a more rigid cellular membrane. Comparison of early-diverging Umbelopsidales with evolutionary younger Mucorales points at several differences particularly in their carbon source preferences and encoded carbohydrate repertoire. All tested Mucoromycotina shares features including the ability to produce 18:3 gamma-linoleic acid and fucose as a cell wall component. Author SummaryIn our paper, we report on the genomic sequences of six Mucoromycotina strains and an associated bacterium from Paenibacillus genus. Mucoromycotina are often studied in pathogenic context albeit their basic biology remains understudied. This manuscript expands on the collection of currently sequenced Mucorales and Umbelopsidales, including the first sequenced Thamnidium isolate, which was sequenced together with a Paenibacillus bacterium. The interaction with a bacterial partner alters the metabolism, cell membrane composition but not the exoskeleton of the fungus. The associated bacterium provided multiple enzymes that significantly expanded the digestive capabilities of the fungal host. Parallel sequencing and phenotyping of Mucorales and Umbelopsidales enabled us to look at the differences of both lineages within Mucoromycotina. We demonstrate that the predicted digestive capabilities are in line with experimental validation. Based on the cell wall composition data and genomic underpinnings of carbohydrate metabolism we were able to confirm the universal presence of fucose in Mucoromycotina cell walls. Fatty acid, phospholipid and acylglycerol composition support the usage of 18:3 gamma-linoleic acid as a chemotaxonomic marker of Mucoromycotina and corroborate TAG as a dominant storage lipid in these organisms. Genomic features, digestive capabilities, fatty acid composition differ between Mucorales and Ubelopsidales pointing at subtle but significant changes in the course of Mucoromycotina radiation.

genomics↗

bioRxiv2020

Utilization of cobalamin is ubiquitous in early-branching fungal phyla

Cobalamin is a cofactor present in essential metabolic pathways in animals and one of the water-soluble vitamins. It is a complex compound synthesized solely by prokaryotes. Cobalamin dependence is scattered across the tree of life. In particular, fungi and plants were deemed devoid of cobalamin. We demonstrate that cobalamin is utilized by all fungal lineages, except for Dikarya. This observation is supported by the genomic presence of both B12 dependent enzymes and cobalamin modifying enzymes. Moreover, the genes identified are actively transcribed in many taxa. Most fungal cobalamin dependent enzymes and cobalamin metabolism proteins are highly similar to their animal homologs. Phylogenetic analyses support a scenario of vertical inheritance of the cobalamin trait with several losses. Cobalamin usage was probably lost in Mucorinae and at the base of Dikarya which groups most of the model organisms which hindered B12-dependent metabolism discovery in fungi. Our results indicate that cobalamin dependence was a widely distributed trait at least in Opisthokonta, across diverse microbial eukaryotes and likely in the LECA.

evolutionary biology↗