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JXB Advance Access originally published online on June 4, 2004
Journal of Experimental Botany 2004 55(403):1761-1763; doi:10.1093/jxb/erh182
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Journal of Experimental Botany, Vol. 55, No. 403, © Society for Experimental Biology 2004; all rights reserved

GENE NOTES

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

Chiharu Kato1, Misa Takahashi1, Atsushi Sakamoto1 and Hiromichi Morikawa1,2,*

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

* To whom correspondence should be addressed. Fax: +81 824 24 0749. 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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