摘要
通过荧光定量PCR技术和geNorm、NormFinder软件对8个候选内参基因(18S、ACT、ACT2、TUB、H2B、UBI、EF-2α和UBQ)在核桃(Juglans reiga)不定根发生过程中的表达稳定性进行评估。利用筛选出的最佳内参基因ACT2进行校正,对核桃不定根发生过程中的12个重要功能基因(YUC4、PIN6、AUX/IAA8、SAUR24、WOX11、WOX5、LBD16、WRKY14、SHR、SCL、CYCA和CYCD)的表达水平进行定量分析。结果显示,YUC4在扦插过程中持续显著下调表达,其余11个基因均在不同时期显著上调表达。结合基因功能推测:PIN6等基因介导的生长素极性运输对于不定根发生初期所需的高浓度IAA富集具有重要作用;SHR、WOX11、LBD16、WRKY14和SCL主要参与形成层细胞向根原基原始细胞的转化过程,SAUR24、WOX5、CYCA和CYCD主要参与根原基原始细胞向根原基细胞转变的调控;同时,细胞周期蛋白相关基因(CYCA、CYCD)可以作为插穗中根原基原始细胞向根原基细胞转变的标志基因。
The expression stability of eight candidate internal reference genes(18S,ACT,ACT2,TUB,H2B,UBI,EF-2αand UBQ)during adventitious root development was evaluated by real-time PCR and geNorm and NormFinder software.Using the best selected internal reference genes for calibration,12 important functional genes(YUC4,PIN6,AUX/IAA8,SAUR24,WOX11,WOX5,LBD16,WRKY14,SHR,SCL,CYCA and CYCD)were found,and the level of expression was quantified during the adventitious root formation of walnut.The results of stability analysis of internal reference gene expression showed that ACT2 gene could be used as the best reference gene in qRT-PCR study of walnut adventitious root formation.ACT2 was used as reference to quantitatively analyze the expression levels of 12 genes in adventitious root formation.The results showed:the YUC gene was down-regulated significantly during the cutting process,and the other 11 genes were significantly up-regulated during different periods of adventitious root development.It is speculated that PIN6 and other gene-mediated auxin polar transport plays an important role in the high concentration of IAA enrichment required in the early stage of adventitious root formation.The SHR,WOX11,LBD16,WRKY14 and SCL genes are mainly involved in the transformation of the stratified cells into the root primordial cells.The SAUR24,WOX5,CYCA and CYCD genes are mainly involved in the regulation of root primordial cells to root primordial cells.At the same time,cyclin-related genes(CYCA,CYCD)can serve as marker genes for the transformation of root primordial cells into root primordial cells.
作者
宋晓波
常英英
刘昊
徐慧敏
裴东
SONG Xiaobo;CHANG Yingying;LIU Hao;XU Huimin;PEI Dong(State Key Laboratory of Tree Genetics and Breeding,Key Laboratory of Tree Breeding and Cultivativation of State Forestry Administration,Research Institute of Forestry Chinese Academy of Forestry,Beijing 100091,China;Tianjin Research Institute of Forestry and Pomology,Tianjin 300112,China;Forestry Bureau of Luoning County,Luoyang,Henan 471700,China)
出处
《园艺学报》
CAS
CSCD
北大核心
2019年第10期1907-1918,共12页
Acta Horticulturae Sinica
基金
中央级公益性科研院所基本科研业务费专项(CAFYBB2017ZA004-2)
国家自然科学基金项目(31672126)
关键词
核桃
不定根
内参基因筛选
表达分析
walnut
adventitious root
reference gene selection
expression analysis
