转录组测序揭示棘胸蛙棘刺季节性变化的分子机制  

Transcriptome Sequencing Reveals the Molecular Mechanism of Seasonal Changes in Spines of Quasipaa spinosa

作　　者：胡一斌 蒋泽元[1] 周奕言 赵晓蕊 章书声 郑荣泉[1,4] HU Yi-Bin;JIANG Ze-Yuan;ZHOU Yi-Yan;ZHAO Xiao-Rui;ZHANG Shu-Sheng;ZHENG Rong-Quan(Provincial Key Laboratory of Wildlife Conservation and Utilization Technology,Zhejiang Normal University,Jinhua 321004,China;College of Life Sciences,Beijing Normal University,Beijing 100875,China;The Management Center of Wuyanling National Natural Reserve in Zhejiang,Wenzhou 325500,China;Xingzhi College,Zhejiang Normal University,Jinhua 321004,China)

机构地区：[1]浙江师范大学野生动物生物技术与保护利用省重点实验室,金华321004 [2]北京师范大学生命科学学院,北京100875 [3]浙江乌岩岭国家级自然保护区管理中心,温州325500 [4]浙江师范大学行知学院,金华321004

出　　处：《农业生物技术学报》2022年第9期1783-1796,共14页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(31772482,31472015);浙江省自然科学基金(LZ17C030001);浙江省万人计划(2018R52028)。

摘　　要：棘胸蛙(Quasipaa spinosa)作为我国特有的经济两栖动物,兼具食用和药用价值。雄性棘胸蛙胸部的棘刺作为蛙类的性二态特征,对于提高抱对成功率有着重要意义,同时也是棘蛙类群的重要鉴别特征,对其产生和变化的分子机制研究,有助于阐明两栖动物棘蛙类群的系统进化关系。为了探究棘胸蛙棘刺季节性变化的分子机制,并挖掘棘刺形成的相关功能基因,本研究对雄性棘胸蛙繁殖期和繁殖后期胸部皮肤以及繁殖期腹部皮肤3组样本进行了高通量转录组测序,结果共获得100312条Unigenes,进行注释后发现多种角质化蛋白基因在繁殖期雄性棘胸蛙的胸部皮肤中表达量显著升高。在繁殖期与繁殖后期胸部棘刺皮肤的差异表达基因(differentially expressd genes,DEGs)中含有与细胞凋亡、细胞增殖、性激素合成等相关的基因,KEGG通路富集分析显示,多个差异表达基因富集于皮肤角质层胞间脂质代谢相关通路中。推测繁殖期角蛋白基因表达增加是棘刺形成的主要原因之一,皮肤角质层中的脂类物质合成减少以及细胞增殖减慢会促使棘刺形态变小。另外,性激素合成相关的基因在繁殖期到繁殖后期中表达减少,说明棘胸蛙棘刺的季节性变化可能受激素变化的调节。通过qPCR技术验证15个差异表达基因,结果表明转录组结果可靠。本研究阐明了棘胸蛙棘刺形成和季节性变化的分子机制,研究结果为棘蛙类群分类提供依据,相关候选基因和代谢通路为其他脊椎动物皮肤衍生物的季节性变化和性二态特征的分子机制研究提供参考。Quasipaa spinosa,a characteristic economic amphibian in China,have both gastronomic and medicinal values.As a sexual dimorphic feature of frogs,the spines of Q.spinosa skin derivatives are of great significance to improve the success rate of holding and also an important classification and identification feature of Tribe Paini.The molecular mechanism of its generation and changes will help to clarify the phylogenetic relationship of the amphibian Tribe Paini.In order to explore the molecular mechanism of the seasonal changes in spines of the Q.spinosa,this study performed transcriptome sequencing on 3 sets of samples from the breast skin of the male Q.spinosa during the breeding period and post-breeding period and the abdomen skin of the male Q.spinosa during the breeding period.A total of 100312 Unigenes were obtained.After annotation,the result showed that the expression of multiple keratinizing protein genes significantly increased in the breast skin of male Q.spinosa during the breeding period.Genes related to apoptosis,cell proliferation and sex hormone synthesis were found in the differentially expressed genes of chest spiny skin in breeding period and post-breeding period.KEGG analysis showed that multiple differentially expressed genes were found to be enriched in the intercellular lipid metabolism of the skin stratum corneum.It was speculated that the increased keratin gene expression during the breeding period was one of the direct causes of the formation of spines.The reduction of lipid synthesis in the stratum corneum of the skin and the slowdown of cell proliferation might cause the spines to become smaller.In addition,the expression of genes related to sex hormone synthesis decreased during the breedding period to post-breeding period,indicating that the seasonal changes in spines of the Q.spinosa might be regulated by hormone changes.The 15 differentially expressed genes were verified by q PCR,the results showed that the transcriptome results were reliable.This study elucidates the molecular mechan

关 键 词：棘胸蛙 棘刺 转录组测序 差异表达基因 

分 类 号：S917.1[农业科学—水产科学]