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A two-component high-affinity nitrate uptake system in barley.

Yiping Tong¹, Jing-Jiang Zhou, Zhensheng Li

¹Crop Performance and Improvement Division, Harpenden, Hertfordshire, AL5 2JQ, UK.

The Plant Journal : for Cell and Molecular Biology

|January 22, 2005

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Plant nitrate transport requires a two-gene system, similar to algae. Functional analysis in Xenopus oocytes confirmed that barley nitrate transporter HvNRT2.1 needs a partner protein, nar2-like, for activity.

Area of Science:

Plant molecular biology
Nutrient transport mechanisms
Algal-biochemistry

Background:

Genome analysis reveals plant genes homologous to algal nitrate transport system components.
Algal high-affinity nitrate transporter (Chlamydomonas reinhardtii) requires two gene products: a carrier protein and a smaller protein (nar2).
Investigating the relevance of this two-component model in plant nitrate uptake.

Purpose of the Study:

To determine if plant nitrate transport also relies on a two-gene system.
To functionally characterize barley nitrate transporter HvNRT2.1 and its interaction with nar2-like proteins.
To understand the regulatory factors influencing this nitrate transport system.

Main Methods:

Genome database screening for plant genes related to algal nitrate transporters.

Related Experiment Videos

Functional expression of plant genes in Xenopus oocytes.

Development and application of a novel (15)N-enriched nitrate uptake assay for high-throughput screening.

Co-injection of candidate transporter and nar2-like mRNAs into oocytes.

Main Results:

The barley nitrate transporter HvNRT2.1 requires co-expression of a specific nar2-like gene product for functional nitrate transport in oocytes.
Out of three cloned barley nar2-like genes, only one facilitated HvNRT2.1 transport activity.
Nitrate transport mediated by this two-gene system is sensitive to external pH and cytoplasmic acidification.

Conclusions:

Plant high-affinity nitrate transport likely utilizes a conserved two-gene system, analogous to algae.
The specific interaction between transporter and nar2-like proteins is crucial for function.
Findings have significant implications for genetically engineering nitrate uptake in crop plants.