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Plant Physiol, March 2000, Vol. 122, pp. 887-894
Analysis of Reductant Supply Systems for Ferredoxin-Dependent
Sulfite Reductase in Photosynthetic and Nonphotosynthetic Organs of
Maize1
Keiko
Yonekura-Sakakibara,*
Yayoi
Onda,
Toshihiko
Ashikari,
Yoshikazu
Tanaka,
Takaaki
Kusumi, and
Toshiharu
Hase
Institute for Fundamental Research, Suntory Ltd., Wakayamadai,
Shimamoto, Mishima, Osaka, 618-8503 Japan (K.Y.-S., T.A., Y.T., T.K.);
and Institute for Protein Research, Osaka University, Yamada-oka,
Suita, Osaka, 565-0871 Japan (Y.O., T.H.)
Sulfite reductase (SiR) catalyzes the
reduction of sulfite to sulfide in chloroplasts and root plastids using
ferredoxin (Fd) as an electron donor. Using purified maize (Zea
mays L.) SiR and isoproteins of Fd and Fd-NADP+
reductase (FNR), we reconstituted illuminated thylakoid membrane- and
NADPH-dependent sulfite reduction systems. Fd I and L-FNR were
distributed in leaves and Fd III and R-FNR in roots. The stromal
concentrations of SiR and Fd I were estimated at 1.2 and 37 µM, respectively. The molar ratio of Fd III to SiR in
root plastids was approximately 3:1. Photoreduced Fd I and Fd III
showed a comparable ability to donate electrons to SiR. In contrast, when being reduced with NADPH via FNRs, Fd III showed a several-fold higher activity than Fd I. Fd III and R-FNR showed the highest rate of
sulfite reduction among all combinations tested. NADP+
decreased the rate of sulfite reduction in a dose-dependent manner. These results demonstrate that the participation of Fd III and high
NADPH/NADP+ ratio are crucial for non-photosynthetic
sulfite reduction. In accordance with this view, a cysteine-auxotrophic
Escherichia coli mutant defective for NADPH-dependent
SiR was rescued by co-expression of maize SiR with Fd III but not with
Fd I.
1
This work was supported in part by Grants-in-Aid
for Research on Priority Areas (nos. 09274101 and 09274103 to T.H.)
from the Ministry of Education, Science and Culture of Japan.
*
Corresponding author; e-mail Keiko_Sakakibara{at}suntory.co.jp;
fax 81-75-962-8807.
© 2000 American Society of Plant Physiologists
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