蜡梅CpWOX13基因的表达及功能

doi:10.11707/j.1001-7488.20190708

摘要/Abstract

摘要： [目的]WOX13基因属于WOX（WUSCHEL related homeobox）转录因子家族中古老进化支成员之一，主要参与成花转化与根的发育过程。本文通过对蜡梅CpWOX13基因表达模式的分析和过表达拟南芥植株表型的观察，分析CpWOX13在蜡梅生长发育中的作用，为了解进化地位极低的物种中WOX13基因的作用机制奠定基础。[方法]基于前期‘罄口’蜡梅露瓣、盛开和衰败3个时期转录组数据，克隆蜡梅CpWOX13基因并通过邻接法构建系统进化树。利用实时定量RT-PCR （qRT-PCR）技术分析CpWOX13基因在4月、5月、7月和9月花芽（4FB、5FB、7FB、9FB）中的相对表达量；对35S∷CpWOX13/Columbia-0拟南芥植株和对照植株的莲座叶、不定根和株高等进行形态观察和生长指标测定，并通过qRT-PCR检测过表达拟南芥株系内源开花相关基因的表达差异，分析过表达CpWOX13对拟南芥开花时间、不定根发育和株高的影响。[结果]蜡梅CpWOX13与葡萄和拟南芥的WOX13聚在一起，同属于WOX家族古老进化支。qRT-PCR结果表明CpWOX13基因在4FB、5FB、7FB和9FB中均有表达，在5FB时期表达量最高，是4FB中表达量的6.5倍，分别是7FB和9FB的3.8倍和3.3倍。35S∷CpWOX13/Columbia-0过表达T₂代拟南芥株系，莲座叶数量少于野生型，开花较野生型提前；根长明显长于野生型，且侧根分支明显增多，株高远高于野生型，最大相差可达12 cm。[结论]蜡梅WOX家族同源基因CpWOX13在不同花发育阶段均有表达，在5月份雌雄蕊原基分化阶段的花芽中表达量最高，过表达CpWOX13会促进拟南芥早花并促进其侧根的发育。

关键词: 蜡梅, CpWOX13基因, 花发育, 早花, 功能分析

Abstract: [Objective] WOX13 gene is a member of the ancient clade of the transcription factor of WOX (WUSCHEL related homeobox),which is mainly involved in the regulation of floral transition and root development. In this study,the expression patterns and overexpression phenotypic traits in Arabidopsis thaliana were analyzed to explore the gene function of CpWOX13 and also to provide a basis for comprehensive research on the mechanism of WOX13 genes in different species with low phylogenetic status.[Method] Based on the RNA-seq analysis at three different developmental stages (petal appears,flowering peak and declining) in Chimonanthus praecox ‘grandiflorus’,CpWOX13 gene was obtained by Reverse Transcription Polymerase Chain Reaction (RT-PCR). Phylogenetic tree was constructed by Neighbor-joining (NJ) method. CpWOX13 expression analysis in flower bud in April,May,July and September (4FB, 5FB, 7FB, 9FB) was carried out using quantitative real-time RT-PCR (qRT-PCR). The genetic effects of CpWOX13 on plant development were explored by studying the phenotypes, including rosette leaves,length of axial and lateral root and plant height between the 35S∷CpWOX13/Col-0 Arabidopsis lines and the wild type.Relative gene expression of CpWOX13 and endogenous gene from Arabidopsis was also carried out in three overexpression lines and WT by qRT-PCR.[Result] CpWOX13 clustered closely to WOX13 in Vitis vinifera and Arabidopsis, belonging to ancient clade of WUS family. qRT-PCR indicated that CpWOX13 transcripts were found at different developmental stages,4FB,5FB,7FB and 9FB.The highest CpWOX13 expression was observed in 5FB, nearly 6.5-fold higher than that of 4FB,and about 3.8-and 3.3-fold of 7FB and 9FB,respectively. Phenotypes were observed in different 35S∷CpWOX13/Col-0 lines compared with the Col-0. 35S∷CpWOX13 overexpression (OE) lines (OE6#,OE3# and OE8#)induced early flowering,in which the number of rosette leaves was slightly less than that of Arabidopsis Col-0. In addition,the length of axial root and the number of lateral root of transgenic lines increased significantly,the plant height was much higher than that of the wild type,with a maximum of 12 cm.[Conclusion] CpWOX13,one of the WOX family homologous genes,expressed in different developmental stages of C. praecox. The highest expression was detected in flower bud during the differentiation of stamen and pistil primordia in May.Overexpression of CpWOX13 affects the development of lateral root and results in early flowering in transgenic Arabidopsis.

Key words: Chimonanthus praecox, CpWOX13 gene, flower development, early flowering, functional analysis

中图分类号:

S718.46

李志能, 江英杰, 陈竟, 李婷, 眭顺照, 李名扬. 蜡梅CpWOX13基因的表达及功能[J]. 林业科学, 2019, 55(7): 77-85.

Li Zhineng, Jiang Yingjie, Chen Jing, Li Ting, Sui Shunzhao, Li Mingyang. Expression and Function of CpWOX13 Gene in Chimonanthus praecox[J]. Scientia Silvae Sinicae, 2019, 55(7): 77-85.

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