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[1]赖瑞联,林玉玲,赖钟雄.龙眼生长素受体基因TIR1的克隆及其与miR393互作关系[J].应用与环境生物学报,2016,22(01):95-102.[doi:10.3724/SP.J.1145.2015.05051]
 LAI Ruilian,LIN Yuling & LAI Zhongxiong**.Cloning of auxin receptor gene TIR1 and its interaction with miR393 in Dimocarpus longan Lour.[J].Chinese Journal of Applied & Environmental Biology,2016,22(01):95-102.[doi:10.3724/SP.J.1145.2015.05051]
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龙眼生长素受体基因TIR1的克隆及其与miR393互作关系()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
22卷
期数:
2016年01期
页码:
95-102
栏目:
研究论文
出版日期:
2016-02-25

文章信息/Info

Title:
Cloning of auxin receptor gene TIR1 and its interaction with miR393 in Dimocarpus longan Lour.
作者:
赖瑞联 林玉玲 赖钟雄
福建农林大学园艺植物生物工程研究所 福州 350002
Author(s):
LAI Ruilian LIN Yuling & LAI Zhongxiong**
Institute of Horticultural Biotechnology, Fujian Agriculture and Forestry University, Fuzhou 350002, China
关键词:
龙眼体胚发生生长素受体启动子miR393激素定量PCR
Keywords:
Dimocarpus longan Lour. somatic embryogenesis auxin receptor promoter miR393 hormone qPCR
分类号:
S667.203 : Q786
DOI:
10.3724/SP.J.1145.2015.05051
摘要:
为了解龙眼生长素受体基因TIR1的功能,以龙眼胚性愈伤组织为材料,在转录水平上对TIR1(命名为DlTIR1-3)及其启动子进行克隆和分析,并在转录后水平上研究TIR1与miR393的相互调控关系. 结果表明,DlTIR1-3 cDNA全长2 196 bp,包含ORF 1 716 bp,编码571个氨基酸(GenBank登录号:KR558761);生物信息学分析发现,DlTIR1-3属于不稳定蛋白,不含有信号肽,含有跨膜螺旋结构,具有F-box保守结构域和富含异亮氨酸重复结构,且DlTIR1-3与碧桃TIR1保持较高的同源性. 染色体步移法克隆获得DlTIR1-3启动子序列2909 bp(GenBank登录号:KR558763),该启动子不存在CpG岛但含有大量光反应、激素应答、组织特异性调控、胁迫响应以及其他生长发育相关的作用元件. psRNAtarget结合改良RLM-RACE验证表明miR393能够裂解DlTIR1-3从而抑制其mRNA转录;qPCR结果显示,DlTIR1-3与miR393相互平衡能够调节龙眼体胚形态建成及响应激素调控. 由此可见,DlTIR1-3通过转录水平的调控作用在龙眼生长发育、激素应答、组织分化及胁迫响应等过程中发挥重要功能,而在转录后水平上miR393通过裂解DlTIR1-3参与相关调控过程. (图7 表1 参32)
Abstract:
This study aimed to investigate the functions of TIR1 in longan. The TIR1 (named DlTIR1-3) and its promoter were cloned and analyzed at the transcriptional level in embryogenic callus of Dimocarpus longan Lour., then the relationship between TIR1 and miR393 was researched at the post-transcriptional level. The results showed the cDNA sequence length of DlTIR1-3 as 2196 bp, containing ORF sequence 1716 bp, encoding 571 amino acids (GenBank accession number KR558761). Bioinformatics analysis showed that DlTIR1-3 was unstable protein, with transmembrane structure and signal peptide, F-box conserved domain and leucine-rich repeats. In addition, DlTIR1-3 shared higher homology with TIR1 of Prunus persica. By using the Genome Walking method, 2909 bp of DlTIR1-3 promoter was amplified (GenBank accession number KR558763). Without an CpG island, DlTIR1-3 had multiple cis-elements correlated to plant growth and development, including light response, hormone response, tissue specific regulatory and stress response. According to the result of psRNAtarget and RLM-RACE, DlTIR1-3 was regulated by miR393 in longan. The qPCR results revealed that DlTIR1-3 and miR393 kept a mutual equilibrium to regulate the morphological?formation of somatic?embryogenesis and responses to hormone in longan. Therefore, DlTIR1-3 plays an important role in the growth, hormone response, tissue differentiation and stress response of longan at the transcriptional level, and miR393 may crack DlTIR1-3 to involve in the regulation at the post-transcriptional level.

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更新日期/Last Update: 2016-02-25