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Identification of an abscisic acid transporter by functional screening using the receptor complex as a sensor.

ABSTRACT: Movement of the plant hormone abscisic acid (ABA) within plants has been documented; however, the molecular mechanisms that regulate ABA transport are not fully understood. By using a modified yeast two-hybrid system, we screened Arabidopsis cDNAs capable of inducing interactions between the ABA receptor PYR/PYL/RCAR and PP2C protein phosphatase under low ABA concentrations. By using this approach, we identified four members of the NRT1/PTR family as candidates for ABA importers. Transport assays in yeast and insect cells demonstrated that at least one of the candidates ABA-IMPORTING TRANSPORTER (AIT) 1, which had been characterized as the low-affinity nitrate transporter NRT1.2, mediates cellular ABA uptake. Compared with WT, the ait1/nrt1.2 mutants were less sensitive to exogenously applied ABA during seed germination and/or postgermination growth, whereas overexpression of AIT1/NRT1.2 resulted in ABA hypersensitivity in the same conditions. Interestingly, the inflorescence stems of ait1/nrt1.2 had a lower surface temperature than those of the WT because of excess water loss from open stomata. We detected promoter activities of AIT1/NRT1.2 around vascular tissues in inflorescence stems, leaves, and roots. These data suggest that the function of AIT1/NRT1.2 as an ABA importer at the site of ABA biosynthesis is important for the regulation of stomatal aperture in inflorescence stems.

SUBMITTER: Kanno Y

PROVIDER: S-EPMC3386071 | biostudies-literature | 2012 Jun

REPOSITORIES: biostudies-literature

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Identification of an abscisic acid transporter by functional screening using the receptor complex as a sensor.

Kanno Yuri Y Hanada Atsushi A Chiba Yasutaka Y Ichikawa Takanari T Nakazawa Miki M Matsui Minami M Koshiba Tomokazu T Kamiya Yuji Y Seo Mitsunori M

Proceedings of the National Academy of Sciences of the United States of America 20120529 24

Movement of the plant hormone abscisic acid (ABA) within plants has been documented; however, the molecular mechanisms that regulate ABA transport are not fully understood. By using a modified yeast two-hybrid system, we screened Arabidopsis cDNAs capable of inducing interactions between the ABA receptor PYR/PYL/RCAR and PP2C protein phosphatase under low ABA concentrations. By using this approach, we identified four members of the NRT1/PTR family as candidates for ABA importers. Transport assay ...[more]

PMID: 22645333

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Project description:Abscisic acid (ABA) is a plant hormone, which plays an important role in plant growth, crop cultivation and modern agricultural engineering management. Accordingly, the detection of ABA content combined with new techniques and methods has become a more and more popular problem in the field of agricultural engineering. In this work, a SERRS and fluorescence dual-function sensor based on the fluorescence quenching and Raman enhancement properties of gold nanorods (AuNRs) was developed, and applied to the detection of plant hormone ABA. The dual-function reporter molecule Rhodamine isothiocyanate (RBITC) and complementary DNA (cDNA) were modified on AuNRs (AuNRs@RBITC@cDNA) as signal probes and aptamer modified magnetic nanoparticles (Fe3O4MNPs@Apt) as capture probes. Through the specific recognition of ABA aptamer and its complementary chains, an dual-function aptamer sensor based on SERRS and fluorescence was constructed. When ABA molecules were present in the detection system, the signal probes were detached from the capture probes due to the preferential binding between aptamer and ABA molecules. SERS signal of the reporter molecules appeared in the supernatant after magnetic separation, and it increased with the increase of ABA concentration. If the etching agent that can etch AuNRs was added to the supernatant, the AuNRs was etching disappeared, then the signal molecules fall off from the AuNRs, and the fluorescence signal intensity would recovered. The intensity of fluorescence signal also increased with the increase of ABA concentration. Thus, the quantitative relationship between ABA concentration and SERRS intensity and fluorescence intensity of signal molecules was established. The linear range of SERRS detection was 100 fM–0.1 nM, the detection limit was 38 fM; The linear range of fluorescence detection was 1 pM–100 nM, the detection limit is 0.33 p.m. The constructed dual-effect sensor was used in the recovery laboratory of real ABA samples, the recovery rate was up to 85–108%.

Dataset Information

Identification of an abscisic acid transporter by functional screening using the receptor complex as a sensor.

Publications

Identification of an abscisic acid transporter by functional screening using the receptor complex as a sensor.

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