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JXB Advance Access published online on June 4, 2004
Journal of Experimental Botany, doi:10.1093/jxb/erh182
© 2004 by Oxford University Press
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Received February 19, 2004; accepted April 21, 2004
© 2004 Society for Experimental Biology

GENE NOTE

Differential expression of the nitrite reductase gene family in tobacco as revealed by quantitative competitive RT-PCR

Chiharu Kato 1, Misa Takahashi 1, Atsushi Sakamoto 1, and Hiromichi Morikawa 2*

1 Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Higashi-Hiroshima 739-8526, Japan
2 Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Higashi-Hiroshima 739-8526, Japan; Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), Kawaguchi, Saitama 332-0012, Japan

* To whom correspondence should be addressed. E-mail: hmorikaw{at}sci.hiroshima-u.ac.jp.


  Abstract

Tobacco (Nicotiana tabacum L. cv. Xanthi XHFD8) possesses four nitrite reductase (NiR) genes: nii1, nii2, nii3, and nii4. Their differential expression in leaves and roots was investigated by quantitative competitive RT-PCR using gene-specific primer pairs. These results appear to contradict existing views on the expression of these NiR genes: (i) the mRNA of each of the four NiR genes was distinguishable both in leaves and roots; (ii) nitrate treatment increased nii1 and nii3 mRNA in leaves and roots by at least 4-fold (at least 5-fold in nii2 and nii4 mRNA); and (iii) the steady-state levels of nii1 and nii3 mRNA were almost the same in leaves (6-7x105 and about 3x106 copies µg-1 of total RNA before and after nitrate treatment, respectively) and in roots (3-4x104 and 3-6x105 copies µg-1 of total RNA before and after nitrate treatment, respectively). Very similar relationships were obtained for the steady-state levels of nii2 and nii4 mRNA in roots (2-4x105 and 8x106 copies µg-1 of total RNA before and after nitrate treatment, respectively), and in leaves (5-9x104 and 4x105 copies µg-1 of total RNA before and after nitrate treatment, respectively). These results demonstrate that nii1 and nii3 transcripts are a dominating, but not exclusive, NiR mRNA in leaves, and the same is true for nii2 and nii4 transcripts in roots.

Key words: Differential gene expression, nitrite reductase (NiR), quantitative competitive RT-PCR, tobacco.


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