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[1]苏立遥 陈旭 黄倏祺 蒋梦琦 厉雪 张梓浩 赖钟雄 林玉玲**.eTM、microRNA319及其调控靶标在龙眼体胚发生早期的表达模式*[J].应用与环境生物学报,2020,26(03):1.[doi:10.19675/j.cnki.1006-687x.2019.07062]
 SU Liyao,CHEN Xu,HUANG Shuqi,et al.The expression profiles of eTM, microRNA319 and their targets in early longan somatic embryogenesis *[J].Chinese Journal of Applied & Environmental Biology,2020,26(03):1.[doi:10.19675/j.cnki.1006-687x.2019.07062]
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eTM、microRNA319及其调控靶标在龙眼体胚发生早期的表达模式*()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
26卷
期数:
2020年03期
页码:
1
栏目:
研究论文
出版日期:
2020-06-25

文章信息/Info

Title:
The expression profiles of eTM, microRNA319 and their targets in early longan somatic embryogenesis *
作者:
苏立遥 陈旭 黄倏祺 蒋梦琦 厉雪 张梓浩 赖钟雄 林玉玲**
福建农林大学园艺植物生物工程研究所 福州 350002
Author(s):
SU Liyao;? CHEN Xu;? HUANG Shuqi;? JIANG Mengqi;? LI Xue;? ZHANG Zhihao;? LAI Zhongxiong & LIN Yuling**
Institute of Horticultural Biotechnology, Fujian Agriculture and Forestry University, Fuzhou 350002, China
关键词:
龙眼;?miR319;?内源诱捕靶标;?靶基因;?体胚发生
Keywords:
Dimocarpus longan;? microRNA319;? target gene;? Endogenous Target Mimics;? somatic embryogenesis
DOI:
10.19675/j.cnki.1006-687x.2019.07062
摘要:
MicroRNA319作为最保守的miRNA家族之一,其在植物器官形态建成、激素信号转导、抵抗外界胁迫等途径中均有重要作用。为了解miR319在植物体胚中的调控途径,利用龙眼转录组数据库筛选miR319成熟体(miR319)和前体序列(pre-miR319),并通过DNAMAN 6.0和Mfold分析miR319的定位和pre-miR319的二级结构;进而采用TAPIR和psRNAtarget预测选定的miR319成员的内源诱捕靶标(eTM,Endogenous Target Mimics)及候选靶标,并通过RLM-RACE验证靶标裂解位点,最后分析eTM、miR319及其靶标在龙眼体胚发生早期的表达模式。结果显示,龙眼4个miR319成员有1个定位于miR319a scafflod517,有3个定位于miR319a scaffold225。因此,以miR319a scaffold225为研究对象,将其命名为pre-miR319a,其上的成熟体分别命名为miR319a-3p.1、miR319a-5p.1和miR319a-5p.2。裂解位点显示龙眼miR319a 3个成员中,miR319a-3p.1的靶基因为TCP2和TCP4 (Dlo_022819.1,Dlo_026743.1),miR319a-5p.1的靶基因为吲哚-3-甘油磷酸合成酶(IGS)和ABI3-5抑制因子,miR319a-5p.2未验证到靶基因。qPCR显示,在龙眼早期体胚发生过程中,miR319a 3个成员的表达模式基本一致,呈显着上调趋势。另外,除eTM8637外,其余eTM的表达模式与其相应的miR319a成员基本呈现为负相关趋势,均为下调趋势。miR319a-3p.1与其靶基因基本呈现为负调控趋势,miR319a-5p.1与ABI3-5抑制因子在龙眼早期体胚发生后期呈现负调控,与IGS没有明显负相关。本试验结果初步验证了“eTM-miR319a-靶基因”信号途径可能参与到龙眼早期体胚发生的形态建成。
Abstract:
MicroRNA319 (miR319) is one of the most conserved miRNA families, which plays an important role in plants organ morphogenesis, hormone signal transduction, stress resistance and so on. To understand the function of miR319a in plants somatic embryogenesis (SE), we extracted mature and precursor of miR319 from longan transcriptome. The location of miR319 members were analyzed by DNAMAN 6.0 and the secondary stem-loop structure of pre-miR319 were performed by Mfold. The TAPIR software was used to search the endogenous target mimics (eTM). The candidate target of longan miR319a was predicted by psRNAtarget software. The cleavage site of candidate targets were verified by modified RLM-RACE (RNA ligase-mediated amplification of cDNA ends). Then, quantitative real time-PCR (qPCR) was performed to identify the expression patterns of eTMs, miR319a and its targers in longan early SE. In this study, 1 of the 4 longan miR319 members located in miR319a scafflod517, and 3 located in miR319a scaffold225. Therefore, miR319a scaffold225 was selected and named pre-miR319a, and the miR319s were named miR319a-3p.1, miR319a-5p.1 and miR319a-5p.2, respectively. The cleavage sites suggested that miR319a-3p.1 cleaved TCP2 and TCP4 (Dlo_022819.1, Dlo_026743.1), miR319a-5p.1 cleaved indole-3-glycerol phosphate synthase (IGS) and suppressor of ABI3-5, miR319a-5p.2 had no target genes. qPCR analysis showed that three members of longan miR319a were significantly up-regulated and shared the same expression pattern during lonagn early SE. Except eTM8637, all eTMs were down-regulated in longan early SE and showed a inversely proportional to corresponding longan miR319a members. The expression patterns of miR319a-3p.1 and their targets were basically opposite. miR319a-5p.1 and suppressor of ABI3-5 were negatively regulated in day 6 to 12. And miR319a-5p.1 had no significant relative with IGS during longan early SE.The results indicated that "eTM-miR319a-target" signaling pathway was initially verified to participate in the morphogenesis of longan early SE.

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备注/Memo

备注/Memo:
收稿日期 Received: 2019-07-29 接受日期 Accepted: 2019-08-30
*国家自然科学基金项目(31672127,31572088)、福建省高原学科建设经费(102/71201801101)、福建农林大学园艺学院优秀硕士学位论文资助基金(2018S02)和福建农林大学科技创新专项基金(CXZX2017187)资助
**通讯作者Correspondence author (E-mail: buliang84@163.com)
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更新日期/Last Update: 2019-09-04