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Acta Agron Sin ›› 2017, Vol. 43 ›› Issue (06): 839-848.doi: 10.3724/SP.J.1006.2017.00839


Functional Analysis of 1-Aminocyclopropane-1-carboxylate Oxidase Gene’s Promoter in Mulberry

YU Jian,LIU Chang-Ying,ZHAO Ai-Chun,WANG Chuan-Hong,CAI Yu-Xiang,YU Mao-De*   

  1. College of Biotechnology, Southwest University / State Key Laboratory of Silkworm Genome Biology, Chongqing 400715, China?
  • Received:2016-07-25 Revised:2017-01-21 Online:2017-06-12 Published:2017-02-17
  • Contact: Yu maode, E-mail: yumd@163.com, Tel: 023-68250191 E-mail:yujian1949@163.com
  • Supported by:

    This study was supported by China Special Fund for Agro-scientific Research in the Public Interest (201403064), Graduate Research and Innovation Projects of Chongqing (CYS2015070), and the China Agriculture Research System (CARS-22).


1-Aminocyclopropane-1-carboxylate oxidase (ACO) as a key enzyme catalyzes the reaction from 1-Aminocyclopropane- 1-carboxylate (ACC) to ethylene. In order to explore the function of MnACO in mulberry growth and development and resisting external stresses, we constructed pMnACO::GUS fusion and transformed it into Arabidopsis thaliana, and used GUS histochemical staining to identify GUS activities of transgenic Arabidopsis thaliana treated by different stresses in different growth stages . MnACO1 and MnACO2 promoter fragments obtained by PCR were 1518 bp and 1429 bp, respectively. There were lots of cis-acting elements such as TATA-box, CAAT-box and others responded to external stimuli. MnACO promoter could drive GUS to express in Arabidopsis thaliana; MnACO1 promoter expressed in the root, leaf, flower petal, anther, filament, stigma and silique of Arabidopsis thaliana and their activities were higher than those of MnACO2; but there was no expression of MnACO2 promoter in silique. The pMnACO1::GUS and pMnACO2::GUS transgenetic plants had different activities in different treatments. The GUS activities of the pMnACO1::GUS transgenetic plants were weakened, while those of the pMnACO2::GUS transgenetic plants were strengthened with the elongation of stress treatment time. Detection of MaACO expression in stress treatment of 2-week-old seedlings by qRT-PCRindicated that the trends of expression pattern of MaACO and its GUS activity changes were almost the same. This research indicated that MnACO is inducible promoter, MnACO1 has the characteristic of constitutive promoter, and MnACO2 has the characteristic of tissue-specific promoter. MnACO1 had more ability to respond to stresses in the transgenic plant, indicating it can be used to regulate the target gene of improving mulberry stress resistance. MaACO2 might be relevant to fruit maturation, thus its promoter could be used as fruit-specific promoter to improve the quality of mulberry fruit.

Key words: Mulberry, Ethylene, ACO, Promoter, GUS, Functional Analysis

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