Project description:During a compatible interaction, root-knot nematodes (Meloidogyne spp.) induce the redifferentiation of root cells into multinucleate nematode feeding cells giant cells. These hypertrophied cells result from repeated nuclear divisions without cytokinesis, are metabolically active and present features typical of transfer cells. Hyperplasia of the surrounding cells leads to formation of the typical root gall. We investigate here the plant response to root-knot nematodes.
Project description:In this study a comparison was made between the local transcriptional changes at two time points upon root knot (Meloidogyne graminicola) and migratory nematode (Hirschmanniella oryzae) infection in rice. Using mRNA-Seq we have characterized specific and general responses of the root challenged with these endoparastic root nematodes with very different modes of action. Root knot nematodes induce major developmental reprogramming of the root tip, where they force the cortical cells to form multinucleate giant cells, resulting in gall-development. Our results show that root knot nematodes force the plant to produce and transfer nutrients, like sugars and amino acids, to this tissue. Migratory nematodes, on the other hand, induce the expression of proteins involved in plant death and oxidative stress, and obstruct the normal metabolic activity of the root. While migratory nematode infection also causes upregulation of biotic stress-related genes early in the infection, the root knot nematodes seem to actively suppress the local defence of the plant root. This is exemplified by a downregulation of genes involved in the salicylic acid and ethylene pathways. Interestingly, hormone pathways usually involved in plant development, were strongly induced (auxin and gibberellin) or repressed (cytokinin) in the galls. In addition, thousands of novel transcriptionally active regions as well as highly expressed nematode transcripts were detected in the infected root tissues. These results uncover previously unrecognized nematode-specific expression profiles and provide an interesting starting point to study the physiological function of many yet unannotated transcripts potentially targeted by these nematodes.
Project description:The global imperative to enhance crop protection while preserving the environment has increased interest in the application of biological pesticides. Bacillus thuringiensis (Bt) is a Gramm-positive bacterium that can produce nematicidal proteins and accumulate them in parasporal crystals. Root-knot nematodes are obligate root plant parasitic which are distributed worldwide, causing severe damages to the infested plants and, consequently, large yield reductions. In this work, we have evaluated the toxicity of the crystal proteins Cry5, Cry21, App6, and Xpp55 against two root-knot nematodes belonging to the Meloidogyne genus (M. incognita and M. javanica). The results show that all four proteins, when solubilized, were highly toxic for both nematode species. To check the potential of using Bt strains producing nematicidal crystal proteins as biopesticides to control plant parasitic nematodes in the field, in planta assays were conducted, using two wild Bt strains which produced Cry5 or a combination of App6 and Cry5 proteins. The tests were carried out with cucumber or with tomato plants infested with M. javanica J2, subjected to irrigation with spore+cristal mixtures of the respective strains. The results showed that the efficacy of the nematicidal activity was plant-dependent, as Bt was able to reduce emerged J2 in tomato plants but not in cucumber plants. In addition, the toxicity observed in the in planta assays was much lower than expected, highlighting the challenge of the crystal proteins to exert their toxicity. This emphasizes the delivery of the Bt proteins as crucial for its use to control root-knot nematodes.
Project description:In this study a comparison was made between the local transcriptional changes at two time points upon root knot (Meloidogyne graminicola) and migratory nematode (Hirschmanniella oryzae) infection in rice. Using mRNA-Seq we have characterized specific and general responses of the root challenged with these endoparastic root nematodes with very different modes of action. Root knot nematodes induce major developmental reprogramming of the root tip, where they force the cortical cells to form multinucleate giant cells, resulting in gall-development. Our results show that root knot nematodes force the plant to produce and transfer nutrients, like sugars and amino acids, to this tissue. Migratory nematodes, on the other hand, induce the expression of proteins involved in plant death and oxidative stress, and obstruct the normal metabolic activity of the root. While migratory nematode infection also causes upregulation of biotic stress-related genes early in the infection, the root knot nematodes seem to actively suppress the local defence of the plant root. This is exemplified by a downregulation of genes involved in the salicylic acid and ethylene pathways. Interestingly, hormone pathways usually involved in plant development, were strongly induced (auxin and gibberellin) or repressed (cytokinin) in the galls. In addition, thousands of novel transcriptionally active regions as well as highly expressed nematode transcripts were detected in the infected root tissues. These results uncover previously unrecognized nematode-specific expression profiles and provide an interesting starting point to study the physiological function of many yet unannotated transcripts potentially targeted by these nematodes. 2 or 3 biological replicates of nematode infected roots and root tips and their respective controls were sampled at two time points (1 biological replicate contains pooled tissue from 6 plants)
Project description:Plant-parasitic root-knot nematodes (Meloidogyne species) are highly polyphagous parasites that infect a wide variety of plants, including economically important crops. These parasites modulate cellular identity of terminally differentiated root cells to induce the formation of giant-cells and knot-like structures (galls), creating new organs in infected plant roots. In this study, we generated single cell RNA-seq data of galls and neighboring root tissues at two distinct stages of M. incognita infection of tomato plants.
Project description:Root-knot nematodes (RKNs) induce inside the vascular cylinder the giant cells (GCs) imbibed into a gall. Gene-repression in early developing GCs could be facilitated by small RNAs (sRNA) as miRNAs. 24nt-sRNAs, rasiRNAs and 21-22nt-sRNAs can also mediate epigenetic mechanisms. Three sRNA libraries from 3dpi galls and three from uninfected root segments were sequenced following Illumina-Solexa technology.
Project description:Conclusions: MiEFF12 overexpression modifies the expression of genes important for the ontogenesis of giant cells, notably those involved in microtubule cytoskeleton reorganization and cell cycle regulation, strongly suggesting that MiEFF12-mediated SmD1 inactivation in plants leads to a loss of susceptibility to root-knot nematodes.
Project description:Ascorbic acid (AA) is known to play a vital role in plant growth and detoxification of reactive oxygen species, however little is known about the significance of AA oxidation in plant defence against pathogens. • The role of ascorbate oxidation in rice defence against root-knot nematodes, Meloidogyne graminicola, was tested with application of AA, ascorbate oxidase (AO), dehydroascorbic acid (DHA), biosynthesis inhibitors and use of mutants. Transcriptome analysis was done on AO treated plants, and hormone measurements were executed to confirm the results. Biochemical analyses were used to study oxidative stress markers, including accumulation of H2O2, , malondialdehyde and AA/DHA.
Project description:Root-knot nematodes (RKNs) induce inside the vascular cylinder the giant cells (GCs) imbibed into a gall. Gene-repression in early developing GCs could be facilitated by small RNAs (sRNA) as miRNAs. 24nt-sRNAs, rasiRNAs and 21-22nt-sRNAs can also mediate epigenetic mechanisms. Three sRNA libraries from 3dpi galls and three from uninfected root segments were sequenced following Illumina-Solexa technology. Three sRNA libraries from 3dpi galls and three from uninfected root segments were sequenced