Barros, D.R.

Batista, D.

Chaves, I.

Guerra-Guimarães, L.

Miguel, C.

Ricardo, C.P.

Várzea, V.

English

Guerra-Guimarães, Leonor

UTL Repository

www.alphavisa.com/asic/2021Poster presentations - Poster presentations Session 2: Plant pathology & protection5528th Conference of Association for the Science and Information on Coffee28 June - 1 July 2021 Montpellier, FranceBook of abstractsS2-PO-02Small-RNA characterization of Coffee Leaf Rust races having different virulence profiles Chaves Inês1, 2 (ichaves@itqb.unl.pt), Barros Danielle Ribeiro3, 4, Batista Dora3, 5, Miguel Célia5, Ricardo Cândido Pinto1, Várzea Vitor3, Guerra-Guimarães Leonor*31 ITQB NOVA, Universidade NOVA de Lisboa, Oeiras, Portugal ; 2 Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal ; 3 CIFC, LEAF, Instituto Superior de Agronomia, Universidade de Lisboa, Lisboa, Portugal ; 4 Departamento de Fitossanidade, Universidade Federal de Pelotas, Pelotas, Rio Grande do Sul, Brazil ; 5 cE3c, Faculdade de Ciências, Universidade de Lisboa, Lisboa, Portugal*Corresponding author: leonorguimaraes@edu.ulisboa.ptRATIONALEThe obligate biotrophic rust fungus Hemileia vastatrix (causal agent of Coffee Leaf Rust), which is the most widespread pathogen of Arabica coffee, causes severe yield losses. More than 50 rust physiological races have been detected resorting to 27 coffee differentials, since their molecular identification has not yet been possible. In the present study we have identified small-RNAs from urediniospores of three rust races with different spectra for virulence: race VI (v?-unknown) non-pathogenic to all known C. arabica genotypes and races II (v5) and XXIV (v2,4,5) pathogenic to the majority of C. arabica genotypes.METHODSUrediniospores of the three races were collected in triplicate samples that were immediately frozen in liquid nitrogen. Total RNA was isolated using the NOGEN KIT Plant/Fungi Total RNA Purification Kit and then sent to small-RNA-sequencing. The genome of H. vastatrix race XXXIII (Porto et al. 2019) and miRBase v22.0 were used to analyze the small-RNA-seq data by means of the miRPursuit pipeline. RESULTSThe three races of H. vastatrix have a predominance of reads with 22 and 29 nt length, but only in race XXIV the relative abundance of these small-RNA species is highly differentiated. The PCA analysis revealed that the profiles of small-RNAs among the three races are completely different with the first two components explaining 90 % of the variance. The majority of the miRNAs present in H. vastatrix was not yet reported in miRBase and so the number of sequences annotated as conserved miRNAs is low, with only about 10 conserved miRNAs being found.CONCLUSION AND PERSPECTIVESUnlike what has been described for the small-RNA profile in plants, in which the main species abundance has 21 or 24 nt length, H. vastatrix small-RNA profile revealed a higher abundance of species with 22 and 29 nt. With this approach we have identified differences between the three races that will potentially contribute to unveil the virulence divergence observed among H. vastatrix races.Acknowledgements: This work was supported by Portuguese Funds through FCT - Fundação para a Ciência e a Tecnologia, I.P., under the project PTDC/AGR-GPL/109990/2009 and co-funded by programs Lisboa 2020, Portugal 2020 and European Union, through FEDER under project PTDC/ASP-PLA/29189/2017, and the R&D Units “GREEN-IT - Bioresources for Sustainability, UIDB/Multi/04551/2020” and “LEAF - UID/AGR/04129/2019”. I. Chaves acknowledge DL 57/2016/CP1351/CT0003 grant.References:• Porto et al. 2019. PLoS ONE.doi:10.1371/journal.pone.0215598

Small-RNA characterization of Coffee Leaf Rust races having different virulence profiles

https://www.repository.utl.pt/bitstream/10400.5/23333/1/Chaves%20et%20al_Asic2021.pdf

Small-RNA characterization of Coffee Leaf Rust races having different virulence profiles

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