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Two AMT2-Type Ammonium Transporters from Pyrus betulaefolia Demonstrate Distinct Expression Characteristics

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Abstract

Ammonium transporters (AMTs) play dominant roles in ammonium transport and distribution across cellular membranes. However, the AMT2-type AMTs from pear remain poorly understood. To characterize ammonium transport pathways in Pyrus betulaefolia Bunge, one popular rootstock for pear and two new genes with high homology to AMT2-type AMTs, PbAMT2 and PbAMT3, were isolated from its seedlings. The analysis of the PbAMT expression patterns in different organs of P. betulaefolia showed that expression of a given transporter could be very specific and their regulatory mechanisms were distinct. Both responded to various nitrogen regimes, PbAMT2 was detected in all organs, and the highest transcription was found in roots, whereas PbAMT3 was restricted to leaves. In addition, transcript level of PbAMT2 was strongly affected by the diurnal cycle, and PbAMT3 was exclusively expressed in leaves after treatment with two phytohormones: abscisic acid and methyl jasmonate. A complementation experiment using the yeast strain 31019b revealed that both PbAMT2 and PbAMT3 encoded functional, high-affinity AMTs. Furthermore, uptake studies indicated that these two AMTs possessed disparate affinities for ammonium and had opposite responses to external pH change. In summary, the present study provides basic genomic, transcript information and nitrogen transporter features for the pear AMT2 subfamily; indeed, PbAMT2 and PbAMT3 demonstrated distinct expression characteristics, which will pave the way for deciphering the high-affinity ammonium transport system in pear.

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Abbreviations

ABA:

Abscisic acid

AMT:

Ammonium transporter

EST:

Expressing sequence tags

MEGA:

Molecular Evolutionary Genetics Analysis

MeJ:

Methyl jasmonate

N:

Nitrogen

NCBI:

The National Center for Biotechnology Information

NH3 :

Ammonia

NH4 + :

Ammonium

OD600 :

Absorbance value at 600 nm

ORF:

Open reading frame

qPCR:

Quantitative real-time PCR

RACE:

Rapid amplification of cDNA ends

TM:

Transmembrane domains

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Acknowledgments

This work was supported by the National Natural Sciences Foundation of China (No. 31372051).

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Authors and Affiliations

  1. Institute of Horticulture, Jiangsu Academy of Agricultural Sciences/Jiangsu Key Laboratory for Horticultural Crop Genetic Improvement, No. 50 Zhongling Street, 210014, Nanjing, Jiangsu, China

    Hui Li, You-hong Chang & Jing Lin

  2. State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences, No. 71 East Beijing Road, 210008, Nanjing, Jiangsu, China

    Yu Cong

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  1. Hui Li
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  2. Yu Cong
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  3. You-hong Chang
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  4. Jing Lin
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Correspondence to You-hong Chang.

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Li, H., Cong, Y., Chang, Yh. et al. Two AMT2-Type Ammonium Transporters from Pyrus betulaefolia Demonstrate Distinct Expression Characteristics. Plant Mol Biol Rep 34, 707–719 (2016). https://doi.org/10.1007/s11105-015-0957-8

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  • DOI: https://doi.org/10.1007/s11105-015-0957-8

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