文献类型：期刊文献

中文题名：杨树次生壁纤维素合酶的表达与互作模式分析

英文题名：Interaction and Expression of Secondary Wall CESAs in Populus

作者：魏凯莉[1] 周厚君[1] 江成[1] 赵岩秋[1] 宋学勤[1,2] 卢孟柱[1,2]

第一作者：魏凯莉

机构：[1]国家林业局林木培育重点实验室,中国林业科学研究院林业研究所,北京100091;[2]南方现代林业协同创新中心,南京林业大学,南京210037

年份：2017

卷号：30

期号：2

起止页码：245-253

中文期刊名：林业科学研究

外文期刊名：Forest Research

收录：CSTPCD;;Scopus;北大核心:【北大核心2014】；CSCD:【CSCD2017_2018】；

基金：国家重点基础研究发展计划(“973”计划)形成层干细胞维持、分化以及次生木质部发育的调控机制(2012CB114503)

语种：中文

中文关键词：纤维素合酶CESA;次生壁;表达分析;互作;激素;杨树

外文关键词：CESA ; Secondary cell wall ; Expression analysis ; Interaction ; Hormone ; Populus

分类号：S792.11

摘要：[目的]纤维素的合成在木材形成过程中具有重要的作用,纤维素合酶(Cellulose Synthase,CESA)是参与纤维素合成的关键酶。由于3种CESA形成一个有功能的纤维素合酶复合体,而杨树中包含CESA4、CESA7A、CESA7B、CESA8A和CESA8B5种次生壁CESA,本文从这5种CESA的表达模式与互作分析入手,探讨了其在次生壁纤维素合成中的工作模式。[方法]利用RNA-seq与基因芯片数据进行基因表达分析,揭示5种次生壁CESA在根、茎、叶组织的表达模式与次生维管再生过程中的表达变化。利用启动子驱动的GUS转基因材料GUS染色分析与实时定量PCR揭示5种次生壁CESA在各个组织的表达模式与在激素处理下的响应模式。利用荧光素酶互补实验揭示CESA7A、CESA7B、CESA8A与CESA8B之间的互作模式。[结果]基因表达分析表明,杨树5种次生壁CESA基因在杨树成熟茎中高表达,尤其在次生维管组织发育的后期高表达,表明其主要参与木材次生壁纤维素的合成。对启动子驱动的GUS转基因材料观察表明,CESA4、CESA7B、CESA8A和CESA8B的GUS信号在杨树茎和叶中较强,但这些次生壁CESA的表达模式存在一定的差异,这种差异在叶脉中表现地尤为明显。此外,在赤霉素GA3和细胞分裂素6-BA处理下,杨树次生壁CESA的表达量显著上调;在生长素NAA、油菜素内酯BR和乙烯处理下,杨树次生壁CESA的表达量下调,但不同的CESA对激素响应的表达量变化幅度存在差异。荧光素酶互补实验表明,杨树次生壁CESA7B和CESA8B、CESA7B和CESA8A、CESA7A与CESA8B之间存在互作,说明它们之间可以形成复合体。[结论]这些数据显示杨树5种次生壁CESA基因在不同组织与不同激素处理下的表达变化存在一定的差异,而它们之间均能互作,提示杨树5个次生壁CESA基因虽然具有同等的能力形成功能性的纤维素合酶复合体,但在不同组织、不同激素作用下可能有不同的组合方式,进而会导致木材成分的差异。
Cellulose synthesis is important for the wood formation of woody plant.CESAsplay important roles in the cellulose synthesis.Since that three kind of CESAs constitute a functional cellulose synthesis complex and five secondary wall CESAs existed in the poplar genome, we examined the expression pattern and interaction of these CESAs in Populus toreveal the work model of them in secondary wall cellulose synthesis.[Method]RNA-seq and microarray data analysis were used to examine the expression profile of the five secondary wall CESAsin root, stem and leaf, and in the regeneration of secondary vascular bundle, respectively. Promotor driving GUS expression assay and qRT-PCR were carried out to reveal the tissue expression pattern and hormone response of the five secondary wall CESAs. Firefly luciferase complementation assay was used to check the interaction between CESA7A, CESA7B, CESA8A and CESA8B.[Result]Expression analysis showed that all the five secondary wall CESAsexhibited high expression in the mature stem and preferred to express in the later stage of stem development, suggesting that they involved in the secondary wall synthesis during wood formation. Promotor driving GUS expression assay of CESA4,CESA7B, CESA8A and CESA8B demonstrated high expression of these secondary wall CESAs in stem and leaf, and some differences, however, were existed in the detail expression pattern, especially in leaf veins. In addition, the expression of secondary wall CESAswere upregulated during GA3 and 6-BA treatment, while down-regulated during NAA, BR and ethylene treatment. The expression variation of these CESAs in response to different hormones were different. Firefly luciferase complementation assayshowed that CESA7A and CESA7B could interact with CESA8A and CESA8B, indicating that they parallelly determine the final secondary wall cellulose synthesis complex.[Conclusion]The five secondary wall CESAsshowed different expression from one another in different tissue and under different hormone treatment. However, these CESAs could interact with each other.Thesedata suggest that poplar secondary wall CESAs could form the cellulose synthesis complex in equal possibility, but the complex might be varied in different tissues and in response to different hormones, which might have an effect on the wood chemical composition.