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Convolutional neural network modelling: advancing identification of true mRNA cleavage sites

ABSTRACT: Degradome sequencing is commonly used to generate high-throughput information on mRNA cleavages by small RNAs. Here we developed an extension module based on a deep learning convolutional neural network (CNN) in a machine learning environment to discriminate false from true cleavage sites applied on datasets from potato (Solanum tuberosum, St) and the oomycete pathogen Phytophthora infestans (Pi). The core of the CNN module is a stochastic gradient descent optimizer with cyclical learning rate (CLR) which together with Bayesian optimization scored a validation accuracy of 100%. To verify the recognition of cleavage sites we applied the module on Arabidopsis thaliana microRNA cleavages. Our module managed to recognize all cleavages, confirming the reliability of the module. When applying this new model to evaluate our data, 7.3% of all cleavage windows represented true cleavages distributed as 214 sites in P. infestans and 444 sites in potato. The sRNA landscape of the potato-P. infestans interaction is complex with uneven sRNA production and cleavage regions. In total, 222 endogenous Pi-sRNAs, 565 endogenous St-sRNAs, 91 trans-acting Pi-sRNAs and 14 trans-acting St-sRNAs were discovered from our datasets. Groups of self-regulatory sRNAs, and sRNAs generated from effector sequences like RXLR and Crinklers suggest dual effector functions. In the potato genome, resistance genes were most targeted mainly as self-regulation but also as results of a trans-action event. Our new analytic model is freely accessible for anyone working on complex biological systems.

ORGANISM(S): Phytophthora infestans Solanum tuberosum

PROVIDER: GSE163382 | GEO | 2021/05/05

REPOSITORIES: GEO

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Phytophthora infestans miRNA translocation facilitates infection in potato

Project description:Phytophthora spp. encode large sets of effector proteins and distinct populations of small RNAs (sRNAs). Reports suggest that pathogen-derived sRNAs can modulate the expression of plant defense genes. The experiments reported here were designed to shed light on impact of sRNAs in the potato-P. infestans interaction. We used the Argonaute or Ago1 from P. infestans tagged with GFP transformed into the 88069 strain to infect potato cv. Bintje plants. Collected leaf materials were used in co-immunoprecipitation experiments together with P. infestans harboring GFP (control GFP) and P. infestans mycelia grown on media (control mycelia). These three materials were sequenced at a Ion Proton platform. The reads length of 8-38 nt were adaptor-trimmed and mapped to the P. infestans genome and the Solanom tuberosum genome v4.04. Both P. infestans-associated and potato derived sRNAs were identified.

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Project description:Background: The oomycete Phytophthora infestans possesses active RNA silencing pathways, which presumably enable this plant pathogen to control the large numbers of transposable elements present in its 240 Mb genome. Small RNAs (sRNAs), central molecules in RNA silencing, are known to also play key roles in this organism, notably in regulation of critical effector genes needed for infection of its potato host. Results: To identify additional classes of sRNAs in oomycetes, we mapped deep sequencing reads to transfer RNAs (tRNAs) thereby revealing the presence of 19-40 nt tRNA-derived RNA fragments (tRFs). Northern blot analysis identified abundant tRFs corresponding to half tRNA molecules. Some tRFs accumulated differentially during infection, as seen by examining sRNAs sequenced from P. infestans-potato interaction libraries. The putative connection between tRF biogenesis and the canonical RNA silencing pathways was investigated by employing hairpin RNA-mediated RNAi to silence the genes encoding P. infestans Argonaute (PiAgo) and Dicer (PiDcl) endoribonucleases. By sRNA sequencing we show that tRF accumulation is PiDcl1-independent, while Northern hybridizations detected reduced levels of specific tRNA-derived species in the PiAgo1 knockdown line. Conclusions: Our findings extend the sRNA diversity in oomycetes to include fragments derived from non-protein-coding RNA transcripts and identify tRFs with elevated levels during infection of potato by P. infestans. Small RNA sequence data from Phytophthora infestans-infected potato leaf tissue and P. infestans mycelium tissue. Three infection stage time-points. Two P. infestans lines: 88089 (wild-type) and PiDcl1 (transformant PiDcl1 knock-down). No replicates. Total number of samples: 8.

2014-12-12 | E-GEOD-63292 | biostudies-arrayexpress

Fragmentation of tRNA in Phytophthora infestans asexual life cycle stages and during host plant infection [host plant infection]

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