Tissue-specific transcriptome profiling of the citrus fruit epidermis and subepidermis using laser capture microdissection

Most studies of the biochemical and regulatory pathways that are associated with, and control, fruit expansion and ripening are based on homogenized bulk tissues, and do not take into consideration the multiplicity of different cell types from which the analytes, be they transcripts, proteins or met...

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Autores principales: Matas, Antonio J., Agustí, Javier, Tadeo, Francisco R., Talón, Manuel, Rose, Jocelyn K. C.
Formato: article
Lenguaje:Inglés
Publicado: 2017
Acceso en línea:http://hdl.handle.net/20.500.11939/5613
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author Matas, Antonio J.
Agustí, Javier
Tadeo, Francisco R.
Talón, Manuel
Rose, Jocelyn K. C.
author_browse Agustí, Javier
Matas, Antonio J.
Rose, Jocelyn K. C.
Tadeo, Francisco R.
Talón, Manuel
author_facet Matas, Antonio J.
Agustí, Javier
Tadeo, Francisco R.
Talón, Manuel
Rose, Jocelyn K. C.
author_sort Matas, Antonio J.
collection ReDivia
description Most studies of the biochemical and regulatory pathways that are associated with, and control, fruit expansion and ripening are based on homogenized bulk tissues, and do not take into consideration the multiplicity of different cell types from which the analytes, be they transcripts, proteins or metabolites, are extracted. Consequently, potentially valuable spatial information is lost and the lower abundance cellular components that are expressed only in certain cell types can be diluted below the level of detection. In this study, laser microdissection (LMD) was used to isolate epidermal and subepidermal cells from green, expanding Citrus clementina fruit and their transcriptomes were compared using a 20k citrus cDNA microarray and quantitative real-time PCR. The results show striking differences in gene expression profiles between the two cell types, revealing specific metabolic pathways that can be related to their respective organelle composition and cell wall specialization. Microscopy provided additional evidence of tissue specialization that could be associated with the transcript profiles with distinct differences in organelle and metabolite accumulation. Subepidermis predominant genes are primarily involved in photosynthesis- and energy-related processes, as well as cell wall biosynthesis and restructuring. By contrast, the most epidermis predominant genes are related to the biosynthesis of the cuticle, flavonoids, and defence responses. Furthermore, the epidermis transcript profile showed a high proportion of genes with no known function, supporting the original hypothesis that analysis at the tissue/cell specific levels can promote gene discovery and lead to a better understanding of the specialized contribution of each tissue to fruit physiology.
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spelling ReDivia56132025-04-25T14:43:25Z Tissue-specific transcriptome profiling of the citrus fruit epidermis and subepidermis using laser capture microdissection Matas, Antonio J. Agustí, Javier Tadeo, Francisco R. Talón, Manuel Rose, Jocelyn K. C. Most studies of the biochemical and regulatory pathways that are associated with, and control, fruit expansion and ripening are based on homogenized bulk tissues, and do not take into consideration the multiplicity of different cell types from which the analytes, be they transcripts, proteins or metabolites, are extracted. Consequently, potentially valuable spatial information is lost and the lower abundance cellular components that are expressed only in certain cell types can be diluted below the level of detection. In this study, laser microdissection (LMD) was used to isolate epidermal and subepidermal cells from green, expanding Citrus clementina fruit and their transcriptomes were compared using a 20k citrus cDNA microarray and quantitative real-time PCR. The results show striking differences in gene expression profiles between the two cell types, revealing specific metabolic pathways that can be related to their respective organelle composition and cell wall specialization. Microscopy provided additional evidence of tissue specialization that could be associated with the transcript profiles with distinct differences in organelle and metabolite accumulation. Subepidermis predominant genes are primarily involved in photosynthesis- and energy-related processes, as well as cell wall biosynthesis and restructuring. By contrast, the most epidermis predominant genes are related to the biosynthesis of the cuticle, flavonoids, and defence responses. Furthermore, the epidermis transcript profile showed a high proportion of genes with no known function, supporting the original hypothesis that analysis at the tissue/cell specific levels can promote gene discovery and lead to a better understanding of the specialized contribution of each tissue to fruit physiology. 2017-06-01T10:12:40Z 2017-06-01T10:12:40Z 2010 JUL 2010 article publishedVersion Matas, A. J., Agusti, J., Tadeo, F.R., Talón, M., Rose, Jocelyn K. C. (2010). Tissue-specific transcriptome profiling of the citrus fruit epidermis and subepidermis using laser capture microdissection. Journal of experimental botany, 61(12), 3321-3330. 0022-0957 http://hdl.handle.net/20.500.11939/5613 10.1093/jxb/erq153 en openAccess Impreso
spellingShingle Matas, Antonio J.
Agustí, Javier
Tadeo, Francisco R.
Talón, Manuel
Rose, Jocelyn K. C.
Tissue-specific transcriptome profiling of the citrus fruit epidermis and subepidermis using laser capture microdissection
title Tissue-specific transcriptome profiling of the citrus fruit epidermis and subepidermis using laser capture microdissection
title_full Tissue-specific transcriptome profiling of the citrus fruit epidermis and subepidermis using laser capture microdissection
title_fullStr Tissue-specific transcriptome profiling of the citrus fruit epidermis and subepidermis using laser capture microdissection
title_full_unstemmed Tissue-specific transcriptome profiling of the citrus fruit epidermis and subepidermis using laser capture microdissection
title_short Tissue-specific transcriptome profiling of the citrus fruit epidermis and subepidermis using laser capture microdissection
title_sort tissue specific transcriptome profiling of the citrus fruit epidermis and subepidermis using laser capture microdissection
url http://hdl.handle.net/20.500.11939/5613
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