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BcNRT1, a plasma membrane-localized nitrate transporter from non-heading Chinese cabbage

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Abstract

A nitrate transporter, BcNRT1, was isolated from non-heading Chinese cabbage (Brassica campestris ssp. chinensis Makino) cultivar ‘Suzhouqing’. The full-length cDNA was obtained using the rapid amplification of cDNA ends technique and contains an open reading frame of 1,770 bp that predicts a protein of 589 acid residues that possesses 12 putative transmembrane domains. Using the GUS marker gene driven by the BcNRT1 promoter, we found BcNRT1 expression to be concentrated in primary and lateral root tips and in shoots of transgenic Arabidopsis plants. The YFP fused to BcNRT1 and transformed into cabbage protoplasts indicated that BcNRT1 was localized to the plasma membrane. The expression of BcNRT1 in roots was induced by exposure to 25 mM nitrate, and the BcNRT1 cRNA heterologously expressed in Xenopus laevis oocytes showed nitrate conductance when nitrate was included in the medium. Moreover, mutant chl1-5 plants harboring 35S::BcNRT1 showed sensitivity to chlorate treatment and exhibited restored nitrate uptake. In conclusion, the results indicate that BcNRT1 functions as a low affinity nitrate transporter in non-heading Chinese cabbage.

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Acknowledgments

We are grateful to Prof. Guohua Xu and Pro. Xiaorong Fan (College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing, China) for helping with experiment on Xenopus laevis oocytes, to Shanhua Lin (Institute of Molecular Biology, Academia Sinica, Taiwan, ROC) for kindly providing the mutants chl1-5 and identifying membrane proteins. We also thank Prof. F. R. Gu (College of Foreign Studies, NAU) for language editing of this manuscript. This work was supported by the earmarked fund for Modern Agro-industry Technology Research System (Nycytx-35-gw12) and the National Science and Technology Support Program (2009BADB8B03-1).

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Authors and Affiliations

  1. College of Horticulture, Nanjing Agricultural University, Nanjing, 210095, China

    Xuedong Yang, Aisheng Xiong, Feng Wang, Min Kong, Qian Wang, Jinyan Wang, Wei Dai & Xilin Hou

  2. Nanjing Institute of Vegetable Science, Nanjing, China

    Feifei Sun

  3. The College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing, China

    Xiudong Xia

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  1. Xuedong Yang
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Correspondence to Xilin Hou.

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Yang, X., Sun, F., Xiong, A. et al. BcNRT1, a plasma membrane-localized nitrate transporter from non-heading Chinese cabbage. Mol Biol Rep 39, 7997–8006 (2012). https://doi.org/10.1007/s11033-012-1646-4

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