Project description:Asterids is one of the major plant clades comprising of many commercially important medicinal species. One of the major concerns in medicinal plant industry is adulteration/contamination resulting from misidentification of herbal plants. This study reports the construction and validation of a microarray capable of fingerprinting medicinally important species from the Asterids clade.
Project description:Phytopathogenic Rathayibacter species are unique bacterial plant pathogens as they are obligately vectored by plant parasitic Anguinid nematodes to the developing seedheads of forage grasses and cereals. This understudied group of plant associated Actinomycetes includes the neurotoxigenic Plant Pathogen Select Agent, R. toxicus, which causes annual ryegrass toxicity in grazing livestock. The complexRathayibacter disease cycle requires intimate interactions with the nematode cuticle and plant hosts, which warrants an increased understanding of the secretory and surface-associated proteins that mediate these diverse eukaryotic interactions. Here we present the first comparative secretome analysis for this complex, nematode vectoredRathayibacter genera that compares three agronomically damaging Rathayibacterspecies, R. toxicus, R. iranicus, and R. tritici. The exoproteomic comparison identified 1,423 unique proteins between the three species using LC-MS/MS. Of the uniquely identified proteins, 94 homologous proteins were conserved between the three Rathayibacter exoproteomes and comprised between 43.4 to 58.6% of total protein abundance. Comparative analyses revealed both conserved and uniquely expressed extracellular proteins, which, interestingly, had more similarities to extracellular proteins commonly associated with bacterial animal pathogens than classical plant pathogens. This comparative exoproteome analysis will facilitate the characterization of proteins essential for vector and host colonization and assist in the development of diagnostic targets.
Project description:Molecular phylogenomics investigates evolutionary relationships based on genomic data. However, despite genomic sequence conservation, changes in protein interactions can occur relatively rapidly and may cause strong functional diversification. To investigate such functional evolution, we here combine phylogenomics with interaction proteomics. We develop this concept by investigating the molecular evolution of the shelterin complex, which protects telomeres, across 16 vertebrate species from zebrafish to humans covering 450 million years of evolution. Our phylointeractomics screen discovers previously unknown telomere-associated proteins and reveals how homologous proteins undergo functional evolution. For instance, we show that TERF1 evolved as a telomere-binding protein in the common stem lineage of marsupial and placental mammals. Phylointeractomics is a versatile and scalable approach to investigate evolutionary changes in protein function and thus can provide experimental evidence for phylogenomic relationships.
Project description:In plant-associated fungi, the role of the epigenome is increasingly recognized as an important regulator of genome structure and of the expression of genes involved in the interaction(s) with the host plant. Two closely-related phytopathogenic species, Leptosphaeria maculans ‘brassicae’ (Lmb) and L. maculans ‘lepidii’ (Lml) exhibit a large conservation of genome synteny but contrasted genome structure. Lmb has undergone massive invasion of its genome by transposable elements summing up one third of its genome and clustered in large TE-rich regions on chromosomal arms, while Lml genome has a low amount of repeats (3% of the genome). Previous information also showed that the TE-rich regions of Lmb host a few species-specific effector genes, expressed during plant infection, with main incidence on the adaptive potential of the fungus. The distinct genome organisation between Lmb and Lml thus provides us with a model of choice for the comparison of the epigenomic organization in two closely related phytopathogenic fungi, in order to investigate pathogenicity/effector gene landscape with respect to the chromatin landscape. To address this, we performed chromatin immunoprecipitation, targeting either histone modifications typical for heterochromatin or euchromatin, during axenic culture, combined with transcriptomic analysis to analyse the influence of the chromatin organisation on gene expression. Our data comfort in both species the postulate that facultative heterochromatin landscapes, associated with H3K27me3 domains, are enriched in genes with no annotation, including numerous candidate effector and species-specific genes. Notably orthologous genes located in the same H3K27me3 domains are enriched in genes encoding putative proteinaceous and metabolic effectors. These genes are mostly silenced in vegetative growth conditions and are likely to be involved in interaction with the host. Compared to other fungal species, including Lml, Lmb has the particularity to have large H3K9me3-domains within chromosomal arms, strongly associated to TEs, and hosting numerous species-specific effector-encoding genes. These two distinct heterochromatin landscapes hosting genes involved in interaction with the host now questions their involvement in regulation of pathogenicity, the dynamics of the domains during plant infection, and the selective advantage for the fungus to host effector genes in H3K9me3 or H3K27me3 domains.
Project description:This a reciprocal transplant experiment. Two hebivorous lepidoptera species (Ostrinia nubilalis and Ostrinia scapulalis) have been fed either their preferred plant (corn for O.nubilalis and Mugwort for O. scapulalis) or the reciprocal plant. At 4th larval instar, RNA was extracted from whole larvae, size selected to enrich in small RNAs and sequenced by Illumina single-end 50bp.