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JXB Advance Access originally published online on February 26, 2009
Journal of Experimental Botany 2009 60(6):1569-1577; doi:10.1093/jxb/erp022
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© 2009 The Author(s).
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. This paper is available online free of all access charges (see http://jxb.oxfordjournals.org/open_access.html for further details)

RESEARCH PAPER

NtCP56, a new cysteine protease in Nicotiana tabacum L., involved in pollen grain development

Xiao-mei Zhang, Ying Wang, Xiao-meng Lv, Hui Li, Peng Sun, Hai Lu* and Feng-lan Li

College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, PR China

* To whom correspondence should be addressed: luhai1974{at}yahoo.com

Proteinases play a critical role in developmental homeostasis and in response to environ-mental stimuli. Our present research reports that a new cysteine protease, NtCP56, is involved in the development of pollen grains in Nicotiana tabacum L. The NtCP56 gene, which encodes a protein of 361 amino acid residues with a calculated molecular mass of 40 kDa, is strongly expressed in anthers. The recombinant NtCP56 showed a high activity towards casein. Kinetic analysis revealed a Km of 2.20 mg ml–1 and Vmax of 11.07 µg ml–1 min–1. The recombinant NtCP56 retained more than 50% of its maximum enzymatic activity from 20 °C to 60 °C with an optimum Tm range of 30–50 °C. The enzyme had a maximum activity at approximately pH 6.5. Suppression of the NtCP56 gene in anti-sense transgenic tobaccos resulted in the sterility of pollen grains. Our data indicated that, as a cysteine protease, NtCP56 might play an important role in pollen development.

Key words: Cysteine protease, NtCP56, pollen, tobacco

Received 3 November 2008; Revised 2 January 2009 Accepted 20 January 2009


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