Gao Shuai, Yang Lu, Zeng Hou Qing, Zhou Zhao Sheng, Yang Zhi Min, Li Hua, Sun Di, Xie Fuliang, Zhang Baohong
Department of Biochemistry and Molecular Biology, College of Life Science, Nanjing Agricultural University, Nanjing 210095, China.
College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou 310036, China.
Sci Rep. 2016 Jan 27;6:19736. doi: 10.1038/srep19736.
The present study functionally identified a new microRNA (microRNA ovual line 5, miRNVL5) with its target gene GhCHR from cotton (Gossypium hirsutum). The sequence of miRNVL5 precursor is 104 nt long, with a well developed secondary structure. GhCHR contains two DC1 and three PHD Cys/His-rich domains, suggesting that GhCHR encodes a zinc-finger domain-containing transcription factor. miRNVL5 and GhCHR express at various developmental stages of cotton. Under salt stress (50-400 mM NaCl), miRNVL5 expression was repressed, with concomitant high expression of GhCHR in cotton seedlings. Ectopic expression of GhCHR in Arabidopsis conferred salt stress tolerance by reducing Na(+) accumulation in plants and improving primary root growth and biomass. Interestingly, Arabidopsis constitutively expressing miRNVL5 showed hypersensitivity to salt stress. A GhCHR orthorlous gene At2g44380 from Arabidopsis that can be cleaved by miRNVL5 was identified by degradome sequencing, but no confidential miRNVL5 homologs in Arabidopsis have been identified. Microarray analysis of miRNVL5 transgenic Arabidopsis showed six downstream genes (CBF1, CBF2, CBF3, ERF4, AT3G22920, and AT3G49200), which were induced by salt stress in wild-type but repressed in miRNVL5-expressing Arabidopsis. These results indicate that miRNVL5 is involved in regulation of plant response to salt stress.
本研究从棉花(陆地棉)中功能鉴定了一种新的微小RNA(微小RNA卵系5,miRNVL5)及其靶基因GhCHR。miRNVL5前体序列长104个核苷酸,具有发育良好的二级结构。GhCHR包含两个DC1和三个富含半胱氨酸/组氨酸的PHD结构域,这表明GhCHR编码一种含锌指结构域的转录因子。miRNVL5和GhCHR在棉花的各个发育阶段均有表达。在盐胁迫(50 - 400 mM NaCl)下,棉花幼苗中miRNVL5的表达受到抑制,同时GhCHR高表达。在拟南芥中异位表达GhCHR可通过减少植物体内Na(+)积累、促进主根生长和增加生物量来赋予盐胁迫耐受性。有趣的是,组成型表达miRNVL5的拟南芥对盐胁迫表现出超敏反应。通过降解组测序鉴定出拟南芥中一个可被miRNVL5切割的GhCHR直系同源基因At2g44380,但未在拟南芥中鉴定到miRNVL5的保守同源物。对miRNVL5转基因拟南芥的微阵列分析显示有六个下游基因(CBF1、CBF2、CBF3、ERF4、AT3G22920和AT3G49200),这些基因在野生型中受盐胁迫诱导,但在表达miRNVL5的拟南芥中受到抑制。这些结果表明miRNVL5参与调控植物对盐胁迫的反应。