铁皮石斛CLE基因家族鉴定与功能分析  

Identification and functional analysis of CLE gene family in Dendrobium officinale Kimura et Migo

作　　者：诸燕[1] 丁兰 陈忆乾 黄秀静 姜伟伟 陈东红 ZHU Yan;DING Lan;CHEN Yiqian;HUANG Xiujing;JIANG Weiwei;CHEN Donghong(The State Key Laboratory of Subtropical Silviculture,Zhejiang A&F University,Hangzhou 311300 China;Agricultural and Forestry Technology Extension Center of Lin an District,Hangzhou 311300,China)

机构地区：[1]浙江农林大学省部共建亚热带森林培育国家重点实验室,浙江杭州311300 [2]临安区农林技术推广中心,浙江杭州311300

出　　处：《浙江农业学报》2024年第7期1583-1590,共8页Acta Agriculturae Zhejiangensis

基　　金：浙江省农业新品种选育重大科技专项(2021C02074)。

摘　　要：CLE多肽是一类小分子分泌蛋白,参与植物的生长发育和细胞间通讯,在维持干细胞的增殖与分化中发挥关键调控作用。为研究兰科药用植物铁皮石斛(Dendrobium officinale Kimura et Migo)中CLE基因家族成员的功能,从铁皮石斛叶、根、茎、花苞、果实的组织中提取RNA,反转录成cDNA,通过半定量PCR检测CLE家族基因成员在各组织中的表达情况。结果表明:在铁皮石斛基因组中鉴定出17个CLE成员,它们均拥有保守的12氨基酸CLE基序。半定量RT-PCR结果表明,CLE基因家族成员在根、茎、叶、花、果实中展示不同的组织表达谱,尤其是CLE 19635在花苞和果实中特异性表达,CLE 22175在果实中特异性表达。用体外合成的CLE多肽处理拟南芥,结果显示,CLE合成肽参与拟南芥根的发育,其中,CLE05351对根生长有促进作用,CLE18468对根影响不明显,其他CLE多肽均对根的生长有抑制作用。mPS-PI染色并未发现根尖分生组织结构模式受到CLE合成肽的影响。此外,CLE 02038过表达的拟南芥T 2代纯合系具有短根表型,与CLE合成肽处理结果一致。研究结果为进一步挖掘CLE家族在铁皮石斛附生根中的作用及其信号调控网络奠定了基础,并为铁皮石斛理想根系的育种提供了潜在靶基因。CLE peptide are a class of small secreted proteins that widely participate in plant development and cell-to-cell communication,playing a key regulatory role in maintaining the proliferation and differentiation of stem cells.In order to study the function of CLE family member in Dendrobium officinale Kimura et Migo,RNA was extracted from the leaves,roots,stems,buds and fruits of D.officinale and reverse transcribed into cDNA,and the expression of CLE family genes in each tissue was detected by semi-quantitative RT-PCR.The results showed that 17 CLE members were identified in D.officinale,all of which conserved 12-amino acid CLE motifs.Semi-quantitative RT-PCR performed on roots,stems,leaves,flowers and fruits showed that these CLE members had different tissue expression profiles,particularly CLE 19635 with specifical expression in flower buds and fruits,and CLE 22175 with specifical expression in fruits.In vitro synthesis of CLE peptides for treatment of Arabidopsis thaliana revealed that CLE peptides impacted the growth of A.thaliana roots,CLE05351 promoted root elongation,CLE18468 had no obvious effect,but the other CLE peptides all inhibited root elongation.mPS-PI staining did not find that the cell pattern of root-tip meristem was affected by CLE peptides.In addition,T 2 homozygotes of A.thaliana overexpressing CLE02038 showed a short root phenotype,which was consistent with the result of CLE peptides treatment.The results laid a foundation for the further exploring the role of CLE family in the epiphytic root of D.officinale and its signal regulation network,and provided potential target genes for breeding of ideal root systems in D.officinale.

关 键 词：铁皮石斛 CLE基因家族 肽激素 根发育 拟南芥 

分 类 号：S567.239[农业科学—中草药栽培]