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Scientia Silvae Sinicae ›› 1999, Vol. 35 ›› Issue (3): 10-15.

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CLONING OF ACC SYNTHASE cDNA AND ITS INHIBITION OF ETHYLENE SYNTHESIS BY ITS ANTISENSE RNA IN TRANSGENIC POPULUS DELTOIDES

Mingliang Li1,Yifan Han1,Deyou Qiu1,Ning Li2,Yingchuan Tian3   

  1. 1. The Institute of Forestry, Chinese Academy of Forestry Beijing 100091
    2. Department of Biology, the Hong Kong University of Science and Technology Hong Kong
    3. The Institute of Microbiology, Chinese Academy of Sciences Beijing 100080
  • Received:1998-08-12 Online:1999-05-25 Published:1999-05-25

Abstract:

A 1.5 kb fragment of ACC synthase cDNA fragment prepared from total RNA of soybean was amplified by polymerase chain reaction (PCR) and cloned into pGEM®-T vector. The cloned ACC synthase gene was further inserted into a binary vector, pBin438, in an inverted orientation between the CaMV 35S promoter and Nos 3'termination sequence (pBACS). Transgenic poplar plants were obtained by regeneration of Agrobacterium-mediated transformation of leaves of Populus deltoides. PCR and Southern blotting analyses confirmed the integration of a single antisense ACC synthase gene in transformed poplar genome. The results form reverse transcription PCR (RT-PCR) of RNAs isolated from transgenic poplar leaves confimed that the antisense RNA of ACC synthase presented in these transgenic plants. The amount of ethylene released from transgenic poplar was reduced significantly to about 22% of that released from non-transformed control poplars.

Key words: ACC(l-aminocyclopropane-l-carboxylate)synthase, Gene, Antisense RNA, Populus deltoides