赤桉eca-miRn33及其靶基因EcaICE1的鉴定与表达分析  

Identification and expression analysis of eca⁃miRn33 and its target gene EcaICE1 in Eucalyptus camaldulensis

作　　者：刘艳 张紫阳 谭芷茵 周茜 林元震 LIU Yan;ZHANG Ziyang;TAN Zhiyin;ZHOU Qian;LIN Yuanzhen(College of Forestry and Landscape Architecture,South China Agricultural University,Guangzhou,Guangdong 510642,China)

机构地区：[1]华南农业大学林学与风景园林学院,广东广州510642

出　　处：《福建农林大学学报（自然科学版）》2023年第6期800-805,共6页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：国家自然科学基金项目(31470673)。

摘　　要：以赤桉组培苗为材料,通过PCR扩增eca-miRn33前体基因序列,结合5′RLM-RACE和烟草瞬时表达分析其对靶基因EcaICE1的剪切特征,并利用实时荧光定量PCR分析叶片和茎干中eca-miRn33与靶基因EcaICE1在低温胁迫下的表达模式。结果显示:赤桉eca-miRn33前体基因序列全长60 bp,可形成典型的发卡结构,eca-miRn33成熟序列位于其前体的5′端;5′RLM-RACE和农杆菌介导的烟草瞬时表达试验结果表明eca-miRn33可靶向剪切EcaICE1;eca-miRn33表达量与EcaICE1表达量之间呈显著负相关。The eca⁃miRn33 precursor gene was amplified by PCR from the tissue culture plantlets of Eucalyptus camaldulensis,and the cleavage effect of eca⁃miRn33 on the target gene EcaICE1 was analyzed by 5′RLM⁃RACE and tobacco transient expression sys⁃tem.The expression patterns of eca⁃miRn33 and EcaICE1 in leaves and stems under low temperature stress were analyzed by real⁃time quantitative PCR.The results showed that the eca⁃miRn33 precursor gene sequence was 60 bp in length and could generate a typical hairpin structure,and the mature eca⁃miRn33 was located at the 5′terminal of its precursor.The results of 5′RLM⁃RACE and Agrobacterium⁃mediated transient tobacco expression assays confirmed that eca⁃miRn33 could cleave EcaICE1,and its expression levels were significantly negatively correlated with those of EcaICE1.

关 键 词：赤桉 miRn33 ICE1 低温胁迫 5′RLM-RACE 

分 类 号：S718.46[农业科学—林学] Q943.2[生物学—植物学]