Abstract

BackgroundThe set of all mRNA molecules present in a cell constitute the transcriptome. The transcriptome varies depending on cell type as well as in response to internal and external stimuli during development. Here we present a study of the changes that occur in the transcriptome of chili pepper fruit during development and ripening.ResultsRNA-Seq was used to obtain transcriptomes of whole Serrano-type chili pepper fruits (Capsicum annuum L.; ‘Tampiqueño 74’) collected at 10, 20, 40 and 60 days after anthesis (DAA). 15,550,468 Illumina MiSeq reads were assembled de novo into 34,066 chili genes. We classified the expression patterns of individual genes as well as genes grouped into Biological Process ontologies and Metabolic Pathway categories using statistical criteria. For the analyses of gene groups we added the weighted expression of individual genes. This method was effective in interpreting general patterns of expression changes and increased the statistical power of the analyses. We also estimated the variation in diversity and specialization of the transcriptome during chili pepper development. Approximately 17% of genes exhibited a significant change of expression in at least one of the intervals sampled. In contrast, significant differences in approximately 63% of the Biological Processes and 80% of the Metabolic Pathways studied were detected in at least one interval. Confirming previous reports, genes related to capsaicinoid and ascorbic acid biosynthesis were significantly upregulated at 20 DAA while those related to carotenoid biosynthesis were highly expressed in the last period of fruit maturation (40–60 DAA). Our RNA-Seq data was validated by examining the expression of nine genes involved in carotenoid biosynthesis by qRT-PCR.ConclusionsIn general, more profound changes in the chili fruit transcriptome were observed in the intervals between 10 to 20 and 40 to 60 DAA. The last interval, between 40 to 60 DAA, included 49% of all significant changes detected, and was characterized predominantly by a global decrease in gene expression. This period signals the end of maturation and the beginning of senescence of chili pepper fruit. The transcriptome at 60 DAA was the most specialized and least diverse of the four states sampled.

Highlights

  • The set of all mRNA molecules present in a cell constitute the transcriptome

  • MRNA was purified and cDNA libraries created from two independent samplings of whole fruits

  • This study presents a detailed analysis of gene expression in chili pepper fruit at four stages of development

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Summary

Introduction

The set of all mRNA molecules present in a cell constitute the transcriptome. The set of all RNA molecules transcribed in an organ or tissue at a particular point of time under a given set of environmental conditions constitute the transcriptome. RNA-Seq [1] is a robust technology to obtain genomewide estimates of relative gene expression. Sequence fragments are mapped to a reference genome and the frequency of these alignments is used to estimate the expression of each gene [2]. When a published reference genome is not available, the cDNA reads can be assembled de novo in order to obtain a high quality reference to which the reads can be re-mapped [3]. RNA-Seq experiments demand a careful design including replicates that permit the estimation of statistical error or variation that is not explained by the experimental treatment [4]

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