Project description:Similar to other plant-parasitic nematodes, root lesion nematodes possess an array of enzymes that are involved in degradation of the plant cell wall. Here we report the identification of a gene encoding a cell wall degrading enzyme, pectin methylesterase PME (EC 3.1.1.11), in the root lesion nematode Pratylenchus penetrans. Both genomic and coding sequences of the gene were cloned for this species, showing the presence of four introns that excluded a potential bacterial contamination. Expression of the Pp-pme gene was localized in the esophageal glands of P. penetrans as determined by in situ hybridization. Temporal expression of Pp-pme in planta was validated for early time points of infection. The possible function and activity of the gene were assessed by transient expression of Pp-pme in N. benthamiana plants via a Potato virus X-based vector. To our knowledge, this is the first report on identification and characterization of a PME gene within the phylum Nematoda.
Project description:The CpG depleted Mycoplasma penetrans harbors a CpG specific C5 methyltransferase. The aim of this experiment was to confirm the specificity of the methyltransferase in vivo and in vitro. Genomic DNA from Mycoplasma penetrans and Escherichia coli genomic DNA that either was or was not methylated in vitro by M.MpeI were subjected to Illumina MiSeq bisulfite sequencing.
Project description:Leishmania donovani WHO reference strain MHOM/IN/80/DD8 and Leptomonas seymouri isolates Ld 2001 and Ld39 were used for proteome analysis which were originally isolated from clinical cases of kala azar patients with different inherent antimonial sensitivities. Ld 2001 was Sb-S and Ld 39 was Sb-R. The genome sequencing of these isolates had confirmed co-infection with Leptomonas.
Project description:Candida lusitaniae is an emerging human opportunistic yeast, which can switch from yeast to pseudohyphae, and one of the rare Candida species capable of sexual reproduction. Its haploid genome and the genetic tools available make it a model of interest to study gene function. This study describes the consequences of DPP3 inactivation on cell morphology and mating, both altered in the dpp3Δ knock-out. Interestingly, reintroducing a wild-type copy of the DPP3 gene in the dpp3Δ mutant failed to restore the wild-type phenotypes. Proteomic analyses showed that about 150 proteins were statistically deregulated in the dpp3Δ mutant, and that most of them did not return to their wild-type level in the reconstituted DPP3 strain. The analysis of the segregation of the dpp3Δ mutation and the phenotypes in the progeny of a cross (between the dpp3Δ knock-out and a wild-type strain) showed that the phenotypes are not linked to dpp3Δ, but to a secondary mutation. Genome sequencing of the dpp3Δ mutant allowed us to identify this secondary mutation.
