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2020, v.45(24) 5958-5966

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华重楼种子休眠相关miRNA的鉴定和差异表达分析
Identification and differential expression of miRNA related to seed dormancy of Paris polyphylla var. chinensis

张成才;王红娟;梁晖辉;高真;罗丽娜;何超;向增旭;
ZHANG Cheng-cai;WANG Hong-juan;LIANG Hui-hui;GAO Zhen;LUO Li-na;HE Chao;XIANG Zeng-xu;College of Horticulture, Nanjing Agricultural University;

摘要(Abstract):

探究华重楼种胚休眠过程中miRNA的表达模式,为miRNA调控种子休眠与萌发机制提供参考资料。利用高通量测序技术、生物信息学分析和实时荧光定量PCR(qPCR)技术,分别对处于休眠和休眠解除时期的华重楼种胚的miRNA进行高通量测序、筛选和鉴定。结果显示华重楼种胚休眠、休眠解除时期4个样本文库中分别有23 811 977,24 276 695,20 611 876,20 601 811条unique序列。miRNA主要分布在21~24 nt,其中在休眠状态长度为24 nt的miRNA数量最多,在休眠解除后的生长发育阶段19 nt的最多。共鉴定出31个已知的miRNA,属于13个不同的家族,并利用生物信息学软件预测到了93个新的miRNA,比较分析休眠维持到休眠释放过程中差异表达的miRNA,筛选出10个差异表达的miRNA(miR156a-5p,miR160a-5p,miR160h-1,miR169a-5p,miR157d,miR159a-1,miR395-3,miR156f-5p,miR156-2,miR171a-3p)。该研究共鉴定出10个与华重楼种子休眠解除相关的miRNAs,GO与KEGG功能富集分析表明,差异表达的miRNA靶基因主要通过碳水化合物代谢、植物激素信号转导、细胞分裂和生长等途径调控种子休眠和萌发,qRT-PCR验证结果显示测序结果和实际结果一致。
The purpose of this study was to explore the expression pattern of miRNA in the process of embryo dormancy and provide a reference for the mechanism of regulating seed dormancy and germination by miRNA. We used high-throughput sequencing technology, bioinformatics analysis and real-time fluorescent quantitative PCR(qPCR) technology to sequence, screen and identify miRNAs of dormant and dormant embryos. The results showed that there were 23 811 977, 24 276 695, 20 611 876 and 20 601 811 unique sequences in the four sample libraries during the period of dormancy and dormancy release. MiRNAs are mainly distributed between 21 and 24 nt, among which the length of 24 nt occurred most frequently. A total of 31 known miRNAs were identified, belonging to 13 different families. 93 new miRNAs were predicted by bioinformatics software. Ten miRNAs(mir156 a-5 p, mir160 a-5 p, mir160 h-1, mir169 a-5 p, mir157 d, mir159 a-1, mir395-3, mir156 f-5 p, mir156-2 and mir171 a-3 p) were screened out. In this study, 10 miRNAs related to seed dormancy release were identified. The target genes mainly involved carbohydrate metabolism, plant hormone signal transduction, cell division and growth. The results of qRT-PCR showed that the sequencing results were consistent with the actual results.

关键词(KeyWords): 华重楼;休眠;miRNA;生物信息学;qPCR
Paris polyphylla var.chinensis;dormancy;miRNA;bioinformatics;qPCR

Abstract:

Keywords:

基金项目(Foundation): 现代农业产业技术体系建设专项(CARS-21);; 中央高校基本科研业务费基金项目(Z52911218)

作者(Author): 张成才;王红娟;梁晖辉;高真;罗丽娜;何超;向增旭;
ZHANG Cheng-cai;WANG Hong-juan;LIANG Hui-hui;GAO Zhen;LUO Li-na;HE Chao;XIANG Zeng-xu;College of Horticulture, Nanjing Agricultural University;

Email:

DOI: 10.19540/j.cnki.cjcmm.20200922.102

参考文献(References):

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