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Cloning of ACO gene and inhibition of ethylene evolution in tomatoes with RNAi

Published online by Cambridge University Press:  13 February 2008

Chen Yin-Hua ,
Ouyang Bo ,
Li Han-Xia  and
Ye Zhi-Biao
Chen Yin-Hua
Affiliation:
National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, China Key Laboratory for Tropical Biological Resources, Ministry of Education, Haikou 570228, China College of Life Science and Agriculture, Hainan University, Haikou 570228, China
Ouyang Bo
Affiliation:
National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, China
Li Han-Xia
Affiliation:
National Center for Vegetable Improvement (Central China), Huazhong Agricultural University, Wuhan 430070, China
Ye Zhi-Biao*
Affiliation:
National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, China National Center for Vegetable Improvement (Central China), Huazhong Agricultural University, Wuhan 430070, China
*
*Corresponding author. E-mail: zbye@mail.hzau.edu.cn
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Abstract

A 1018 bp fragment of the ACO gene cDNA sequence was cloned from tomato (Lycopersicon esculentum) leaves incubated with a pathogen mixture using reverse transcriptase-polymerase chain reaction (RT-PCR) with two PCR primers designed according to the sequence of a tomato cDNA clone (E11). A BLAST search showed the sequence presenting a very high match with the ACO genes in other plants, with 83–99% homology. Using this sequence, an RNA interference (RNAi) transformation vector (pD311) was constructed and transformed into tomato. Twenty-seven regenerated plants with kanamycin resistance were obtained, showing that the transgene was integrated into the tomato genome; this was confirmed by PCR and Southern blotting. Ethylene production by the RNAi transgenic tomato plants was measured by gas chromatography, and showed that ethylene evolution was specifically inhibited in leaves and fruits of the transgenic plants.

Keywords

ACC oxidase geneRNA interferencetomatotransformation
Type
Research Article
Information
Chinese Journal of Agricultural Biotechnology , Volume 4 , Issue 3 , December 2007 , pp. 193 - 197
Copyright
Copyright © China Agricultural University and Cambridge University Press 2007

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Footnotes

First published in Journal of Agricultural Biotechnology 2007, 15(3): 464–468

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