Molecular Characterization of SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) Gene Family in Betula luminifera.
Molecular Characterization of SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) Gene Family in Betula luminifera.
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光皮桦鳞状启动子结合蛋白样 (SPL) 基因家族的分子特征
DOI：
10.3389/fpls.2018.00608
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发表时间：
2018
影响因子：
5.6
通讯作者：
Zhang JH
中科院分区：
文献类型：
--
作者：
Li XY;Lin EP;Huang HH;Niu MY;Tong ZK;Zhang JH
鳞状病毒启动子结合蛋白类基因(SPL)是植物特有的转录因子家族，在植物的生长发育和逆境响应中发挥着重要的调控作用。在本研究中，从光皮桦中鉴定和克隆了18个SPL基因。所有BlSPLs的SBP结构域中均存在两个锌指结构和一个核定位信号(NLS)片段。系统发育分析表明，这些基因聚为9个组(第I-IX组)。内含子/外显子结构和基序组成在同一组内高度保守。在18个BlSPL中有12个被实验证实为miR156的靶标，并且在这些miR156靶向的BlSPL基因中检测到两个切割位点。在BlSpls的启动子区域发现了许多与光、胁迫和植物激素反应相关的顺式元件，提示BlSPL基因可能参与了重要的生理过程和发育事件。组织特异性表达分析表明，miR156靶向的BlSPL呈现出更强的差异性表达模式，而大多数miR156非靶向的BlSPL倾向于结构性表达，这表明miR156靶向和非靶向的BlSPL在光皮藻的发育和生长中具有不同的作用。进一步的表达分析表明，以miR156为靶标的Blspl随着年龄的增长而显著上调，而成熟BlmiR156的表达水平随着年龄的增长而明显下降，这表明miR156/SPL模块在光皮桦营养阶段的变化中发挥了重要作用。此外，酵母双杂交实验表明，一些miR156靶向和非靶向的BlSPLs可以与两种Della蛋白(BlRGA和BlRGL)相互作用，这表明某些BlSpls通过与Della蛋白的相互作用参与了GA的调控过程。这些结果为进一步研究BlSpls在光皮藻中的生物学功能提供了重要依据。
As a major family of plant-specific transcription factors, SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) genes play vital regulatory roles in plant growth, development and stress responses. In this study, 18 SPL genes were identified and cloned from Betula luminifera. Two zinc finger-like structures and a nuclear location signal (NLS) segments were existed in the SBP domains of all BlSPLs. Phylogenetic analysis showed that these genes were clustered into nine groups (group I-IX). The intron/exon structure and motif composition were highly conserved within the same group. 12 of the 18 BlSPLs were experimentally verified as the targets of miR156, and two cleavage sites were detected in these miR156-targeted BlSPL genes. Many putative cis-elements, associated with light, stresses and phytohormones response, were identified in the promoter regions of BlSPLs, suggesting that BlSPL genes are probably involved in important physiological processes and developmental events. Tissue-specific expression analysis showed that miR156-targeted BlSPLs exhibited a more differential expression pattern, while most miR156-nontargeted BlSPLs tended to be constitutively expressed, suggesting the distinct roles of miR156-targeted and nontargeted BlSPLs in development and growth of B. luminifera. Further expression analysis revealed that miR156-targeted BlSPLs were dramatically up-regulated with age, whereas mature BlmiR156 level was apparently declined with age, indicating that miR156/SPL module plays important roles in vegetative phase change of B. luminifera. Moreover, yeast two-hybrid assay indicated that several miR156-targeted and nontargeted BlSPLs could interact with two DELLA proteins (BlRGA and BlRGL), which suggests that certain BlSPLs take part in the GA regulated processes through protein interaction with DELLA proteins. All these results provide an important basis for further exploring the biological functions of BlSPLs in B. luminifera.
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