Cloning and functional characterization of a constitutively expressed nitrate transporter gene, OsNRT1, from rice.
Elucidating how rice (Oryza sativa) takes up nitrate at the molecular level could help improve the low recovery rate (<50%) of nitrogen fertilizer in rice paddies. As a first step toward that goal, we have cloned a nitrate transporter gene from rice called OsNRT1. OsNRT1 is a new member of a growing transporter family called PTR, which consists not only of nitrate transporters from higher plants that are homologs of the Arabidopsis CHL1 (AtNRT1) protein, but also peptide transporters from a wide variety of genera including animals, plants, fungi, and bacteria. However, despite the fact that OsNRT1 shares a higher degree of sequence identity with the two peptide transporters from plants (approximately 50%) than with the nitrate transporters (approximately 40%) of the PTR family, no peptide transport activity was observed when OsNRT1 was expressed in either Xenopus oocytes or yeast. Furthermore, contrasting the dual-affinity nitrate transport activity of CHL1, OsNRT1 displayed only low-affinity nitrate transport activity in Xenopus oocytes, with a K(m) value of approximately 9 mM. Northern-blot and in situ hybridization analysis indicated that OsNRT1 is constitutively expressed in the most external layer of the root, epidermis and root hair. These data strongly indicate that OsNRT1 encodes a constitutive component of a low-affinity nitrate uptake system for rice.
Lin
C M
CM
Department of Life Science, School of Life Science, National Tsing Hua University, 30043, Hsin-Chu, Taiwan.
Koh
S
S
Stacey
G
G
Yu
S M
SM
Lin
T Y
TY
Tsay
Y F
YF
D000595:Amino Acid Sequence; D000818:Animals; D027321:Anion Transport Proteins; D029681:Arabidopsis Proteins; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary; D006863:Hydrogen-Ion Concentration; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D009566:Nitrates / Q000378:metabolism*; D012275:Oryza / Q000235:genetics*; D010940:Plant Proteins; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D014981:Xenopus
D027321:Anion Transport Proteins; D029681:Arabidopsis Proteins; D002352:Carrier Proteins; D018076:DNA, Complementary; C101492:NRT1.1 protein, Arabidopsis; D009566:Nitrates; D010940:Plant Proteins; D012333:RNA, Messenger