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辣椒果实发育过程中的转录组动态变化。

Dynamics of the chili pepper transcriptome during fruit development.

作者信息

Martínez-López Luis A, Ochoa-Alejo Neftalí, Martínez Octavio

机构信息

Laboratorio Nacional de Genómica para la Biodiversidad (LANGEBIO), Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (Cinvestav), 36821 Irapuato, Guanajuato, México.

出版信息

BMC Genomics. 2014 Feb 21;15:143. doi: 10.1186/1471-2164-15-143.

Abstract

BACKGROUND

The 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.

RESULTS

RNA-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.

CONCLUSIONS

In 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.

摘要

背景

细胞中存在的所有mRNA分子构成转录组。转录组因细胞类型而异,并且在发育过程中会响应内部和外部刺激而发生变化。在此,我们展示了一项关于辣椒果实发育和成熟过程中转录组变化的研究。

结果

利用RNA测序技术获取了在开花后10、20、40和60天(DAA)采集的整个塞拉诺型辣椒果实(辣椒属;“坦皮克尼奥74”)的转录组。15,550,468条Illumina MiSeq读数被从头组装成34,066个辣椒基因。我们使用统计标准对单个基因以及归类到生物过程本体和代谢途径类别的基因的表达模式进行了分类。对于基因组分析,我们加入了单个基因的加权表达。该方法在解释表达变化的总体模式方面有效,并提高了分析的统计效力。我们还估计了辣椒发育过程中转录组多样性和特异性的变化。大约17%的基因在至少一个采样间隔中表现出显著的表达变化。相比之下,在至少一个间隔中检测到大约63%的生物过程和80%的代谢途径存在显著差异。正如之前报道所证实的,与辣椒素和抗坏血酸生物合成相关的基因在开花后20天显著上调,而与类胡萝卜素生物合成相关的基因在果实成熟的最后阶段(40 - 60天)高度表达。我们通过qRT - PCR检测参与类胡萝卜素生物合成的九个基因的表达,对我们的RNA测序数据进行了验证。

结论

总体而言,在10至20天和40至60天的间隔中观察到辣椒果实转录组有更深刻的变化。最后一个间隔,即40至60天,包含了所有检测到的显著变化的49%,其主要特征是基因表达整体下降。这个时期标志着辣椒果实成熟的结束和衰老的开始。开花后60天的转录组是所采样的四个状态中最具特异性且多样性最低的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ba44/3936869/ff3aac00d2fd/1471-2164-15-143-1.jpg

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