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芥蓝转录组组装及多个组织中芥子油苷代谢相关基因的全局表达分析

Transcriptome Assembly of Chinese Kale and Global Expression Analysis of Genes Involved in Glucosinolate Metabolism in Multiple Tissues.

作者信息

Wu Shuanghua, Lei Jianjun, Chen Guoju, Chen Hancai, Cao Bihao, Chen Changming

机构信息

Department of Vegetable Science, College of Horticulture, South China Agricultural University Guangzhou, China.

Vegetable Research Institute, Guangdong Academy of Agricultural Sciences Guangzhou, China.

出版信息

Front Plant Sci. 2017 Feb 8;8:92. doi: 10.3389/fpls.2017.00092. eCollection 2017.

Abstract

Chinese kale, a vegetable of the cruciferous family, is a popular crop in southern China and Southeast Asia due to its high glucosinolate content and nutritional qualities. However, there is little research on the molecular genetics and genes involved in glucosinolate metabolism and its regulation in Chinese kale. In this study, we sequenced and characterized the transcriptomes and expression profiles of genes expressed in 11 tissues of Chinese kale. A total of 216 million 150-bp clean reads were generated using RNA-sequencing technology. From the sequences, 98,180 unigenes were assembled for the whole plant, and 49,582~98,423 unigenes were assembled for each tissue. Blast analysis indicated that a total of 80,688 (82.18%) unigenes exhibited similarity to known proteins. The functional annotation and classification tools used in this study suggested that genes principally expressed in Chinese kale, were mostly involved in fundamental processes, such as cellular and molecular functions, the signal transduction, and biosynthesis of secondary metabolites. The expression levels of all unigenes were analyzed in various tissues of Chinese kale. A large number of candidate genes involved in glucosinolate metabolism and its regulation were identified, and the expression patterns of these genes were analyzed. We found that most of the genes involved in glucosinolate biosynthesis were highly expressed in the root, petiole, and in senescent leaves. The expression patterns of ten glucosinolate biosynthetic genes from RNA-seq were validated by quantitative RT-PCR in different tissues. These results provided an initial and global overview of Chinese kale gene functions and expression activities in different tissues.

摘要

芥蓝是十字花科蔬菜,因其高含量的芥子油苷和营养品质,在中国南方和东南亚地区是一种受欢迎的作物。然而,关于芥蓝中芥子油苷代谢及其调控所涉及的分子遗传学和基因的研究很少。在本研究中,我们对芥蓝11个组织中表达的基因进行了转录组测序和特征分析。使用RNA测序技术共生成了2.16亿条150碱基对的clean reads。从这些序列中,为整个植株组装了98180个单基因,为每个组织组装了49582至98423个单基因。Blast分析表明,共有80688个(82.18%)单基因与已知蛋白质具有相似性。本研究中使用的功能注释和分类工具表明,主要在芥蓝中表达的基因大多参与基本过程,如细胞和分子功能、信号转导以及次生代谢物的生物合成。对芥蓝不同组织中所有单基因的表达水平进行了分析。鉴定出大量参与芥子油苷代谢及其调控的候选基因,并分析了这些基因的表达模式。我们发现,大多数参与芥子油苷生物合成的基因在根、叶柄和衰老叶片中高表达。通过定量RT-PCR在不同组织中验证了来自RNA-seq的10个芥子油苷生物合成基因的表达模式。这些结果提供了芥蓝基因功能和不同组织中表达活性的初步全局概述。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5e6b/5296335/32f409bef9fe/fpls-08-00092-g0001.jpg

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