De novo assembly and functional annotation of Myrciaria dubia fruit transcriptome reveals multiple metabolic pathways for L-ascorbic acid biosynthesis

Myrciaria dubia is an Amazonian fruit shrub that produces numerous bioactive phytochemicals, but is best known by its high L-ascorbic acid (AsA) content in fruits. Pronounced variation in AsA content has been observed both within and among individuals, but the genetic factors responsible for this va...

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Autores principales: Castro Gómez, Juan Carlos, Maddox, J. Dylan, Cobos Ruiz, Marianela, Requena Anicama, David Ruben, Zimic Peralta, Mirko Juan, Bombarely Gomez, Aureliano, Imán Correa, Sixto Alfredo, Cerdeira Gutiérrez, Luis Alexander, Medina Bardales, Anderson E.
Formato: info:eu-repo/semantics/article
Lenguaje:Inglés
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://repositorio.inia.gob.pe/handle/20.500.12955/736
https://doi.org/10.1186/s12864-015-2225-6
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author Castro Gómez, Juan Carlos
Maddox, J. Dylan
Cobos Ruiz, Marianela
Requena Anicama, David Ruben
Zimic Peralta, Mirko Juan
Bombarely Gomez, Aureliano
Imán Correa, Sixto Alfredo
Cerdeira Gutiérrez, Luis Alexander
Medina Bardales, Anderson E.
author_browse Bombarely Gomez, Aureliano
Castro Gómez, Juan Carlos
Cerdeira Gutiérrez, Luis Alexander
Cobos Ruiz, Marianela
Imán Correa, Sixto Alfredo
Maddox, J. Dylan
Medina Bardales, Anderson E.
Requena Anicama, David Ruben
Zimic Peralta, Mirko Juan
author_facet Castro Gómez, Juan Carlos
Maddox, J. Dylan
Cobos Ruiz, Marianela
Requena Anicama, David Ruben
Zimic Peralta, Mirko Juan
Bombarely Gomez, Aureliano
Imán Correa, Sixto Alfredo
Cerdeira Gutiérrez, Luis Alexander
Medina Bardales, Anderson E.
author_sort Castro Gómez, Juan Carlos
collection Repositorio INIA
description Myrciaria dubia is an Amazonian fruit shrub that produces numerous bioactive phytochemicals, but is best known by its high L-ascorbic acid (AsA) content in fruits. Pronounced variation in AsA content has been observed both within and among individuals, but the genetic factors responsible for this variation are largely unknown. The goals of this research, therefore, were to assemble, characterize, and annotate the fruit transcriptome of M. dubia in order to reconstruct metabolic pathways and determine if multiple pathways contribute to AsA biosynthesis.RESULTS: In total 24,551,882 high-quality sequence reads were de novo assembled into 70,048 unigenes (mean length = 1150 bp, N50 = 1775 bp). Assembled sequences were annotated using BLASTX against public databases such as TAIR, GR-protein, FB, MGI, RGD, ZFIN, SGN, WB, TIGR_CMR, and JCVI-CMR with 75.2 % of unigenes having annotations. Of the three core GO annotation categories, biological processes comprised 53.6 % of the total assigned annotations, whereas cellular components and molecular functions comprised 23.3 and 23.1 %, respectively. Based on the KEGG pathway assignment of the functionally annotated transcripts, five metabolic pathways for AsA biosynthesis were identified: animal-like pathway, myo-inositol pathway, L-gulose pathway, D-mannose/L-galactose pathway, and uronic acid pathway. All transcripts coding enzymes involved in the ascorbate-glutathione cycle were also identified. Finally, we used the assembly to identified 6314 genic microsatellites and 23,481 high quality SNPs. CONCLUSIONS: This study describes the first next-generation sequencing effort and transcriptome annotation of a non-model Amazonian plant that is relevant for AsA production and other bioactive phytochemicals. Genes encoding key enzymes were successfully identified and metabolic pathways involved in biosynthesis of AsA, anthocyanins, and other metabolic pathways have been reconstructed. The identification of these genes and pathways is in agreement with the empirically observed capability of M. dubia to synthesize and accumulate AsA and other important molecules, and adds to our current knowledge of the molecular biology and biochemistry of their production in plants. By providing insights into the mechanisms underpinning these metabolic processes, these results can be used to direct efforts to genetically manipulate this organism in order to enhance the production of these bioactive phytochemicals.The accumulation of AsA precursor and discovery of genes associated with their biosynthesis and metabolism in M. dubia is intriguing and worthy of further investigation. The sequences and pathways produced here present the genetic framework required for further studies. Quantitative transcriptomics in concert with studies of the genome, proteome, and metabolome under conditions that stimulate production and accumulation of AsA and their precursors are needed to provide a more comprehensive view of how these pathways for AsA metabolism are regulated and linked in this species.
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spelling INIA7362023-11-28T15:32:49Z De novo assembly and functional annotation of Myrciaria dubia fruit transcriptome reveals multiple metabolic pathways for L-ascorbic acid biosynthesis Castro Gómez, Juan Carlos Maddox, J. Dylan Cobos Ruiz, Marianela Requena Anicama, David Ruben Zimic Peralta, Mirko Juan Bombarely Gomez, Aureliano Imán Correa, Sixto Alfredo Cerdeira Gutiérrez, Luis Alexander Medina Bardales, Anderson E. Camu-camu Metabolic pathway reconstruction Next-generation sequencing Plant vitamin C metabolism Biotecnología Agrícola y de alimentos Myrciaria dubia is an Amazonian fruit shrub that produces numerous bioactive phytochemicals, but is best known by its high L-ascorbic acid (AsA) content in fruits. Pronounced variation in AsA content has been observed both within and among individuals, but the genetic factors responsible for this variation are largely unknown. The goals of this research, therefore, were to assemble, characterize, and annotate the fruit transcriptome of M. dubia in order to reconstruct metabolic pathways and determine if multiple pathways contribute to AsA biosynthesis.RESULTS: In total 24,551,882 high-quality sequence reads were de novo assembled into 70,048 unigenes (mean length = 1150 bp, N50 = 1775 bp). Assembled sequences were annotated using BLASTX against public databases such as TAIR, GR-protein, FB, MGI, RGD, ZFIN, SGN, WB, TIGR_CMR, and JCVI-CMR with 75.2 % of unigenes having annotations. Of the three core GO annotation categories, biological processes comprised 53.6 % of the total assigned annotations, whereas cellular components and molecular functions comprised 23.3 and 23.1 %, respectively. Based on the KEGG pathway assignment of the functionally annotated transcripts, five metabolic pathways for AsA biosynthesis were identified: animal-like pathway, myo-inositol pathway, L-gulose pathway, D-mannose/L-galactose pathway, and uronic acid pathway. All transcripts coding enzymes involved in the ascorbate-glutathione cycle were also identified. Finally, we used the assembly to identified 6314 genic microsatellites and 23,481 high quality SNPs. CONCLUSIONS: This study describes the first next-generation sequencing effort and transcriptome annotation of a non-model Amazonian plant that is relevant for AsA production and other bioactive phytochemicals. Genes encoding key enzymes were successfully identified and metabolic pathways involved in biosynthesis of AsA, anthocyanins, and other metabolic pathways have been reconstructed. The identification of these genes and pathways is in agreement with the empirically observed capability of M. dubia to synthesize and accumulate AsA and other important molecules, and adds to our current knowledge of the molecular biology and biochemistry of their production in plants. By providing insights into the mechanisms underpinning these metabolic processes, these results can be used to direct efforts to genetically manipulate this organism in order to enhance the production of these bioactive phytochemicals.The accumulation of AsA precursor and discovery of genes associated with their biosynthesis and metabolism in M. dubia is intriguing and worthy of further investigation. The sequences and pathways produced here present the genetic framework required for further studies. Quantitative transcriptomics in concert with studies of the genome, proteome, and metabolome under conditions that stimulate production and accumulation of AsA and their precursors are needed to provide a more comprehensive view of how these pathways for AsA metabolism are regulated and linked in this species. Peer reviewed 2018-09-27T22:33:29Z 2018-09-27T22:33:29Z 2015-11-24 info:eu-repo/semantics/article Castro, J. C., Maddox, J. D., Cobos, M., Requena, D., Zimic, M., Bombarely, A., ... Medina, A. E. (2015). De novo assembly and functional annotation of Myrciaria dubia fruit transcriptome reveals multiple metabolic pathways for L-ascorbic acid biosynthesis. BMC genomics, 16(1): 997. doi: 10.1186/s12864-015-2225-6 1471-2164 https://repositorio.inia.gob.pe/handle/20.500.12955/736 BMC Genomics https://doi.org/10.1186/s12864-015-2225-6 eng BMC Genomics,16:997, 2015 https://doi.org/10.1186/s12864-015-2225-6 info:eu-repo/semantics/openAccess application/pdf application/pdf Amazonia Perú BioMed Central Reino Unido Instituto Nacional de Innovación Agraria Repositorio Institucional - INIA
spellingShingle Camu-camu
Metabolic pathway reconstruction
Next-generation sequencing
Plant vitamin C metabolism
Biotecnología Agrícola y de alimentos
Castro Gómez, Juan Carlos
Maddox, J. Dylan
Cobos Ruiz, Marianela
Requena Anicama, David Ruben
Zimic Peralta, Mirko Juan
Bombarely Gomez, Aureliano
Imán Correa, Sixto Alfredo
Cerdeira Gutiérrez, Luis Alexander
Medina Bardales, Anderson E.
De novo assembly and functional annotation of Myrciaria dubia fruit transcriptome reveals multiple metabolic pathways for L-ascorbic acid biosynthesis
title De novo assembly and functional annotation of Myrciaria dubia fruit transcriptome reveals multiple metabolic pathways for L-ascorbic acid biosynthesis
title_full De novo assembly and functional annotation of Myrciaria dubia fruit transcriptome reveals multiple metabolic pathways for L-ascorbic acid biosynthesis
title_fullStr De novo assembly and functional annotation of Myrciaria dubia fruit transcriptome reveals multiple metabolic pathways for L-ascorbic acid biosynthesis
title_full_unstemmed De novo assembly and functional annotation of Myrciaria dubia fruit transcriptome reveals multiple metabolic pathways for L-ascorbic acid biosynthesis
title_short De novo assembly and functional annotation of Myrciaria dubia fruit transcriptome reveals multiple metabolic pathways for L-ascorbic acid biosynthesis
title_sort de novo assembly and functional annotation of myrciaria dubia fruit transcriptome reveals multiple metabolic pathways for l ascorbic acid biosynthesis
topic Camu-camu
Metabolic pathway reconstruction
Next-generation sequencing
Plant vitamin C metabolism
Biotecnología Agrícola y de alimentos
url https://repositorio.inia.gob.pe/handle/20.500.12955/736
https://doi.org/10.1186/s12864-015-2225-6
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