桂花OfABFs基因克隆和表达分析

Cloning and expression analysis of OfABFs gene in Osmanthus fragrans

【目的】研究OfABFs基因家族成员在桂花‘堰虹桂’Osmanthus fragrans ‘Yanhonggui’不同组织和花开放进程中的表达模式,筛选参与调控花开放的关键成员。【方法】在桂花基因组数据库中筛选出相关OfABFs基因序列,克隆序列全长并对其进行生物信息学分析,通过荧光定量PCR分析OfABFs基因在‘堰虹桂’不同组织及花开放进程中的时空表达模式。【结果】筛选得到5条OfABFs基因序列。生物信息学分析得出:桂花OfABFs蛋白具有亲水性,较不稳定,无信号肽段;预测其蛋白二、三级结构具有相似的特性,无规则卷曲和α-螺旋为其结构组成的主要元件;5条OfABFs转录因子均含有bZIP转录因子家族保守结构域。亚细胞定位预测表明:5条OfABFs编码蛋白均定位在细胞核。荧光定量PCR结果表明:OfABF1、OfABF2、OfABF3、OfABF4和OfABF6在桂花中的表达具有组织特异性,其中OfABF1、OfABF4和OfABF6在花中有较高表达。OfABF1在顶壳期被转录激活,表达显著升高;铃梗期后表达水平虽呈下降趋势,但仍保持较高水平。OfABF2的表达在顶壳期到达峰值,且相对表达水平明显高于其他时期。OfABF3、OfABF4和OfABF6在花朵衰老期的相对表达量最高。【结论】OfABF1、OfABF2可能与桂花花开放进程有关,而OfABF3、OfABF4和OfABF6参与调控桂花花朵衰老的可能性较大。图10表4参32。

[Objective]The aim of this study is to explore the expression patterns of OfABF gene family members in different tissues and flower opening process of Osmanthus fragrans‘Yanhonggui’,and screen the key members involved in regulating flower opening.[Method]The related OfABFs gene sequence was screened from the O.fragrans genome database for the full-length sequence and bioinformatics analysis.The spatio-temporal expression mode of OfABFs gene in different tissues and flower opening process of O.fragrans‘Yanhonggui’were analyzed by fluorescence quantitative PCR.[Result]5 OfABFs sequences were screened.Bioinformatics analysis showed that O.fragrans OfABFs gene family protein was hydrophilic and unstable,and had no signal peptide.It was predicted that the protein secondary and tertiary structure had similar characteristics and irregular curl,andα-spiral was the main component of its structure.The 5 OfABFs transcription factors all contained the conserved domain of bZIP transcription factor family.Subcellular localization prediction showed that 5 OfABFs coding proteins were located in the nucleus.The results of fluorescence quantitative PCR showed that the expressions of OfABF1,OfABF2,OfABF3,OfABF4 and OfABF6 were tissue-specific,among which OfABF1,OfABF4 and OfABF6 were highly expressed in flowers.OfABF1 was transcriptionally activated in the apical shell stage,and its expression increased significantly.Although the expression level decreased after boll stem stage,it still maintained a high level.The expression of OfABF2 peaked in the apical shell stage,and the relative expression level was significantly higher than that in other stages.The relative expression levels of OfABF3,OfABF4 and OfABF6 were the highest in flower senescence stage.[Conclusion]OfABF1 and OfABF2 may be related to the blooming process of O.fragrans,while OfABF3,OfABF4 and OfABF6 are more likely to participate in the regulation of O.fragrans flower senescence.[Ch,10 fig.4 tab.32 ref.]