Transcriptome assembly and expression analysis in Colletotrichum gloeosporioides -tolerant Rubus glaucus Benth.

ABSTRACT Andean blackberry (Rubus glaucus Benth.) is an important crop of the Andean region affected by Colletotrichum gloeosporioides. In Colombia, tolerant plant material has been detected, but it has not been completely characterized. The objective of this research was oriented to analyze de novo...

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Autores principales: Arias,Juliana, Rincón,Juan C., López,Ana M., Marulanda,Marta L.
Lenguaje:English
Publicado: Instituto de Investigaciones Agropecuarias, INIA 2019
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spelling oai:scielo:S0718-583920190004005652019-11-19Transcriptome assembly and expression analysis in Colletotrichum gloeosporioides -tolerant Rubus glaucus Benth.Arias,JulianaRincón,Juan C.López,Ana M.Marulanda,Marta L. Blackberry fungal tolerance molecular markers RNA-seq tolerance-related genes ABSTRACT Andean blackberry (Rubus glaucus Benth.) is an important crop of the Andean region affected by Colletotrichum gloeosporioides. In Colombia, tolerant plant material has been detected, but it has not been completely characterized. The objective of this research was oriented to analyze de novo transcriptome assembly of R. glaucus, and the comparison of the assembly with different reference genomes to further complete differential expression analysis of R. glaucus tolerant to C. gloespoiorides attack. To achieve this, three groups were used: infected tolerant material, infected susceptible material, and a susceptible group without inoculation. The RNA-seq sequencing was achieved through Illumina Hi-seq 2000. De novo assembly (Trinity, CD-HIT, TopHat) and functional annotation of sequences were carried out, additionally, mapping with reference genomes belonging to Rosaceae families was conducted (Bowtie2, TopHat). Subsequently, the differential expression was quantified (Cuffdiff) and analyzed through EdgeR. Variant analysis was made using MISA and SAMtools. After editing and assembly, 43579 consensus sequences were obtained (N50 = 489 bp; GC = 44.6%), annotation detected 35824 and 35602 sequences in Nt (partially non-redundant nucleotide sequences) and Nr (non-redundant protein sequences) databases, respectively. The 85% of Nr sequences was linked to members of Rosaceae family, mainly strawberry (67.6%). A total of 3570 simple sequence repeat (SSR) markers and 38791 single nucleotide polymorphisms (SNP) were found. The transcriptome of tolerant plants exhibited less SNPs. Finally, differentially expressed genes were found, including RPM1, MAPKBP1, CKX2. This research represents a contribution for future understanding of R. glaucus transcriptome, since there is little information available, and it would help further tolerance-related analysis.info:eu-repo/semantics/openAccessInstituto de Investigaciones Agropecuarias, INIAChilean journal of agricultural research v.79 n.4 20192019-12-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0718-58392019000400565en10.4067/S0718-58392019000400565
institution Scielo Chile
collection Scielo Chile
language English
topic Blackberry
fungal tolerance
molecular markers
RNA-seq
tolerance-related genes
spellingShingle Blackberry
fungal tolerance
molecular markers
RNA-seq
tolerance-related genes
Arias,Juliana
Rincón,Juan C.
López,Ana M.
Marulanda,Marta L.
Transcriptome assembly and expression analysis in Colletotrichum gloeosporioides -tolerant Rubus glaucus Benth.
description ABSTRACT Andean blackberry (Rubus glaucus Benth.) is an important crop of the Andean region affected by Colletotrichum gloeosporioides. In Colombia, tolerant plant material has been detected, but it has not been completely characterized. The objective of this research was oriented to analyze de novo transcriptome assembly of R. glaucus, and the comparison of the assembly with different reference genomes to further complete differential expression analysis of R. glaucus tolerant to C. gloespoiorides attack. To achieve this, three groups were used: infected tolerant material, infected susceptible material, and a susceptible group without inoculation. The RNA-seq sequencing was achieved through Illumina Hi-seq 2000. De novo assembly (Trinity, CD-HIT, TopHat) and functional annotation of sequences were carried out, additionally, mapping with reference genomes belonging to Rosaceae families was conducted (Bowtie2, TopHat). Subsequently, the differential expression was quantified (Cuffdiff) and analyzed through EdgeR. Variant analysis was made using MISA and SAMtools. After editing and assembly, 43579 consensus sequences were obtained (N50 = 489 bp; GC = 44.6%), annotation detected 35824 and 35602 sequences in Nt (partially non-redundant nucleotide sequences) and Nr (non-redundant protein sequences) databases, respectively. The 85% of Nr sequences was linked to members of Rosaceae family, mainly strawberry (67.6%). A total of 3570 simple sequence repeat (SSR) markers and 38791 single nucleotide polymorphisms (SNP) were found. The transcriptome of tolerant plants exhibited less SNPs. Finally, differentially expressed genes were found, including RPM1, MAPKBP1, CKX2. This research represents a contribution for future understanding of R. glaucus transcriptome, since there is little information available, and it would help further tolerance-related analysis.
author Arias,Juliana
Rincón,Juan C.
López,Ana M.
Marulanda,Marta L.
author_facet Arias,Juliana
Rincón,Juan C.
López,Ana M.
Marulanda,Marta L.
author_sort Arias,Juliana
title Transcriptome assembly and expression analysis in Colletotrichum gloeosporioides -tolerant Rubus glaucus Benth.
title_short Transcriptome assembly and expression analysis in Colletotrichum gloeosporioides -tolerant Rubus glaucus Benth.
title_full Transcriptome assembly and expression analysis in Colletotrichum gloeosporioides -tolerant Rubus glaucus Benth.
title_fullStr Transcriptome assembly and expression analysis in Colletotrichum gloeosporioides -tolerant Rubus glaucus Benth.
title_full_unstemmed Transcriptome assembly and expression analysis in Colletotrichum gloeosporioides -tolerant Rubus glaucus Benth.
title_sort transcriptome assembly and expression analysis in colletotrichum gloeosporioides -tolerant rubus glaucus benth.
publisher Instituto de Investigaciones Agropecuarias, INIA
publishDate 2019
url http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0718-58392019000400565
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