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ACTA HORTICULTURAE SINICA ›› 2019, Vol. 46 ›› Issue (10): 1907-1918.doi: 10.16420/j.issn.0513-353x.2018-0845

• Research Papers • Previous Articles     Next Articles

Reference Gene Selection and Genes Expression Analysis During Adventitious Root Formation in Walnut

SONG Xiaobo1,CHANG Yingying1,LIU Hao1,2,XU Huimin3,and PEI Dong1,*   

  1. 1State 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;2Tianjin Research Institute of Forestry and Pomology,Tianjin 300112,China;3Forestry Bureau of Luoning County,Luoyang,Henan 471700,China
  • Online:2019-10-25 Published:2019-10-25

Abstract: The expression stability of eight candidate internal reference genes(18S,ACT,ACT2,TUB,H2B,UBI,EF-2α and 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.

Key words: walnut, adventitious root, reference gene selection, expression analysis

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