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Plant Cell, Vol. 11, 865-874, May 1999, Copyright © 1999, American Society of Plant Physiologists

CHL1 Is a Dual-Affinity Nitrate Transporter of Arabidopsis Involved in Multiple Phases of Nitrate Uptake

Kun-Hsiang Liua,b, Chi-Ying Huangb, and Yi-Fang Tsaya
a Institute of Molecular Biology, Academia Sinica, Taipei, 11529 Taiwan
b Department of Botany, National Taiwan University, Taipei, 10660 Taiwan

Correspondence to: Yi-Fang Tsay, mbyftsay{at}ccvax.sinica.edu.tw (E-mail), 886-2-2782-6085 (fax)

Higher plants have both high- and low-affinity nitrate uptake systems. These systems are generally thought to be genetically distinct. Here, we demonstrate that a well-known low-affinity nitrate uptake mutant of Arabidopsis, chl1, is also defective in high-affinity nitrate uptake. Two to 3 hr after nitrate induction, uptake activities of various chl1 mutants at 250 µM nitrate (a high-affinity concentration) were only 18 to 30% of those of wild-type plants. In these mutants, both the inducible phase and the constitutive phase of high-affinity nitrate uptake activities were reduced, with the inducible phase being severely reduced. Expressing a CHL1 cDNA driven by the cauliflower mosaic virus 35S promoter in a transgenic chl1 plant effectively recovered the defect in high-affinity uptake for the constitutive phase but not for the induced phase, which is consistent with the constitutive level of CHL1 expression in the transgenic plant. Kinetic analysis of nitrate uptake by CHL1-injected Xenopus oocytes displayed a biphasic pattern with a Michaelis–Menten Km value of ~50 µM for the high-affinity phase and ~4 mM for the low-affinity phase. These results indicate that in addition to being a low-affinity nitrate transporter, as previously recognized, CHL1 is also involved in both the inducible and constitutive phases of high-affinity nitrate uptake in Arabidopsis.




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