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雌雄异株葫芦科植物大籽栝楼花芽的转录组分析及性别分化相关基因的发现

Transcriptome profiling of the floral buds and discovery of genes related to sex-differentiation in the dioecious cucurbit Coccinia grandis (L.) Voigt.

作者信息

Mohanty Jatindra Nath, Nayak Sanghamitra, Jha Sumita, Joshi Raj Kumar

机构信息

Functional Genomics Laboratory, Centre of Biotechnology, Siksha O Anusandhan University, Kalinga Nagar, Ghatikia, Bhubaneswar 751003, India.

Centre of Advanced Study, Dept. of Botany, University of Calcutta, 35 Ballygunge Circular Road, Kolkata 700019, India.

出版信息

Gene. 2017 Aug 30;626:395-406. doi: 10.1016/j.gene.2017.05.058. Epub 2017 May 31.

Abstract

Dioecious species offer an inclusive structure to study the molecular basis of sexual dimorphism in angiosperms. Despite having a small genome and heteromorphic sex chromosomes, Coccinia grandis is a highly neglected dioecious species with little information available on its physical state, genetic orientation and key sex-defining elements. In the present study, we performed RNA-Seq and DGE analysis of male (MB) and female (FB) buds in C. grandis to gain insights into the molecular basis of sex determination in this plant. De novo assembly of 75 million clean reads resulted in 72,479 unigenes for male library and 63,308 unigenes for female library with a mean length of 736bp. 61,458 (85.57%) unigenes displayed significant similarity with protein sequences from publicly available databases. Comparative transcriptome analyses revealed 1410 unigenes as differentially expressed (DEGs) between MB and FB samples. A consistent correlation between the expression levels of DEGs was observed for the RNA-Seq pattern and qRT-PCR validation. Functional annotation showed high enrichment of DEGs involved in phytohormone biosynthesis, hormone signaling and transduction, transcriptional regulation and methyltransferase activity. High induction of hormone responsive genes such as ARF6, ACC synthase1, SNRK2 and BRI1-associated receptor kinase 1 (BAK1) suggest that multiple phytohormones and their signaling crosstalk play crucial role in sex determination in this species. Beside, the transcription factors such as zinc fingers, homeodomain leucine zippers and MYBs were identified as major determinants of male specific expression. Moreover, the detection of multiple DEGs as the miRNA target site implies that a small RNA mediated gene silencing cascade may also be regulating gender differentiation in C. grandis. Overall, the present transcriptome resources provide us a large number of DEGs involved in sex expression and could form the groundwork for unravelling the molecular mechanism of sex determination in C. grandis.

摘要

雌雄异株物种为研究被子植物性二态性的分子基础提供了一个全面的结构。尽管大孢儿瓜有一个小基因组和异形性染色体,但它是一个被高度忽视的雌雄异株物种,关于其物理状态、遗传取向和关键性别决定因素的信息很少。在本研究中,我们对大孢儿瓜的雄性(MB)和雌性(FB)芽进行了RNA测序和数字基因表达分析,以深入了解该植物性别决定的分子基础。对7500万个clean reads进行从头组装,雄性文库得到72479个单基因,雌性文库得到63308个单基因,平均长度为736bp。61458个(85.57%)单基因与来自公开数据库的蛋白质序列显示出显著相似性。比较转录组分析显示,MB和FB样本之间有1410个单基因差异表达(DEGs)。在RNA测序模式和qRT-PCR验证中,观察到DEGs表达水平之间存在一致的相关性。功能注释显示,参与植物激素生物合成、激素信号传导和转导、转录调控和甲基转移酶活性的DEGs高度富集。激素响应基因如ARF6、ACC合酶1、SNRK2和BRI1相关受体激酶1(BAK1)的高诱导表明,多种植物激素及其信号串扰在该物种的性别决定中起关键作用。此外,锌指、同源域亮氨酸拉链和MYB等转录因子被确定为雄性特异性表达的主要决定因素。此外,检测到多个DEGs作为miRNA靶位点,这意味着小RNA介导的基因沉默级联反应也可能在大孢儿瓜的性别分化中发挥作用。总体而言,目前的转录组资源为我们提供了大量参与性别表达的DEGs,并可为阐明大孢儿瓜性别决定的分子机制奠定基础。

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