Plant and Cell Physiology, 2001, Vol. 42, No. 3 308-313
© 2001 Oxford University Press
Molecular Cloning and Characterization of a cDNA for a Novel Ethylene Receptor, NT-ERS1, of Tobacco (Nicotiana tabacum L.)
1 Laboratory of Bio-adaptation, Graduate School of Agricultural Sciences, Tohoku University, Tsutsumidori-amamiyamachi 1-1, Aoba-ku, Sendai, 981-8555 Japan 2 Laboratory of Plant Pathology, Graduate School of Agricultural Sciences, Tohoku University, Tsutsumidori-amamiyamachi 1-1, Aoba-ku, Sendai, 981-8555 Japan
The cDNA encoding a novel member (NT-ERS1) of ethylene receptor family of tobacco (Nicotiana tabacum L.) was obtained by a combination of RT-PCR and 5'-/3'-RACE cloning. The cDNA was 2,092 nucleotides long and had an open reading frame of 1,911 bp encoding 637 amino acids. The deduced polypeptide lacked a response regulator domain, indicating that the ethylene receptor belongs to an ERS-group. The amino acid sequence was similar to respective members of the tobacco ethylene receptor family: 67.8% to NT-ETR1, 39.1% to NTHK1 and 31.1% to NTHK2. Comparison of amino acid sequence suggested that NT-ERS1 is the counterpart of Nr in the ethylene receptor family of tomato, which belongs to Solanaceae as does tobacco. Northern blot analysis showed that mRNA of NT-ERS1 was present in leaf, shoot and root tissues, and accumulated in leaves treated with exogenous ethylene. A mutated NT-ERS1 cDNA transgene, obtained by introducing one nucleotide substitution into NT-ETR1 cDNA, conferred ethylene insensitivity in tobacco plants, indicating that the translation product of the cDNA actually functioned in the plants.
3 Present address: YT, Kyushu National Agricultural Experiment Station; SF, Kagawa Prefectural Agricultural Research Center.
4 Corresponding author: E-mail, ssatoh@ bios.tohoku.ac.jp; Fax, +81-22-717-8834.
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