ARAKAKI, J. E.

BORRA, R. C.

FAVERO, A. P.

MALAGO JUNIOR, W.

MUDADU, M. de A.

SANTOS, P. M.

VIGNA, B. B. Z.

English

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Proceedings X-Meeting 2018Editor: AB3COctober 2018X-meeting • October 2018 • São PedroANALYSIS OF PASPALUMNOTATUM TRANSCRIPTOMEUNDER DROUGHT CONDITIONJoyce Etsuko Arakaki, Wilson Malagó Junior, MAURICIO DE ALVARENGAMUDADU, Patricia Menezes Santos, Alessandra Pereira Fávero, Ricardo CarneiroBorra, Bianca Baccili Zanotto VignaAbstractPaspalum notatum Flüggé is a perennial grass widely used as forage and as turf in manycountries as Argentina, Australia, United States and in the South of Brazil. The species presentdiploid and tetraploid citotypes with x=10 chromosomes. Transcriptomic and differential geneexpression (DGE) studies in plants, under water deficit, are used to investigate the molecularmechanisms of transcriptional regulation and potential genes involved in the response to drought,making possible the genetic modification and/or gene editing of Paspalum and other plantsof commercial interest, as already done in bahiagrass and maize. The objective of this study isto evaluate the gene expression profile of Paspalum notatum Flüggé in response to drought.The tetraploid accession P. notatum BGP 216 conserved at the germplasm bank of EmbrapaPecuária Sudeste (São Carlos, Brazil) was selected for a drought stress experiment based onthe climate characteristics of its site of collection, on the importance of the species and on theability of the species to tolerate drought. The experiment was conducted in greenhouse duringBrazilian spring (Nov/2016), in triplicates, and leaf samples of each biological replicate werecollected for mRNA extraction in two conditions, without drought stress and after five daysunder drought stress (4% of relative soil moisture). The samples were sequenced using NextGeneration Sequencing (NGS) technology on Illumina HiSeq 2500 equipment with HiSeq-V4 kitand 2x100 bp paired-end reads. Bioinformatics tools were used to characterize the quality ofthe de novo transcriptome assembly, to analyze DGE between plants with and without droughtstress and to perform the functional annotation of the genes. Around 80 millions of reads weresequenced per sample and Trinity software was used for de novo assembly, annotation and DGEstudies of the transcriptome. Around 244 million bases were assembled, 168.509 transcripts wereidentified from 56.718 predicted genes and the transcriptome had a N50 stats of 2.226 bp withmedian contig length of 1.083 bp. The alignment of the reads to the assembly was performedby Bowtie2 and 91.61% were mapped as proper pairs which is better than the usually expected70-80%. Diamond Blast determined that 17.381 of 47.514 proteins were covered by more than90% of their protein lengths. The latter result, additionally to the results of ExN50 stats, FastQCand Transrate (score of 0.3701) softwares showed satisfactory quality of the reads and assembly.BUSCO3 was used to search a dataset of ortholog conserved genes and returned a good resultof 86.9% of transcriptome completeness against the embryophyta dataset from OrthoDB v9database. DGE analyses were performed using EdgeR software and identified 1.978 differentiallyexpressed genes (DEG) (p-value < 0.001 and Fold Change > 2). The Trinotate software suite wasused for automatic functional annotation of the transcriptome. Initial searches of the top tenDEG in UniProt databases and PubMed showed relation to biological process and molecularfunctions. The annotated genes ARF9, ARF24, DJC76 and THI1-2 have been related to plant androot development, biotic and abiotic stresses responses (including drought) in other organismssuch as Arabidopsis thaliana, Gossypium barbadense, Solanum lycopersicum and Oryza sativa.The characterization of this accession as to their strategy of tolerance to drought and the betterunderstanding of the molecular and physiological mechanisms involved may contribute for210

Analysis of paspalum notatum transcriptome under drought condition.

http://www.alice.cnptia.embrapa.br/bitstream/doc/1101437/1/XMeetingAnalysisPaspalum.pdf

Analysis of paspalum notatum transcriptome under drought condition.

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