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Phylogenetic and Expression Analysis of Mn-CDF Transporters in Pear (Pyrus bretschneideri Rehd.)

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Abstract

Metal tolerance proteins (MTPs) play important roles in heavy metal homeostasis and tolerance in plants. As a part of the MTP family, manganese-cation diffusion facilitator (Mn-CDF) subfamily members transport Mn and iron (Fe) in plants. However, the role of these Mn-CDFs in pear is not well established. In this study, seven members of the Mn-CDF subgroup were identified in Chinese white pear (Pyrus bretschneideri Rehd.), based on recently released draft genome of pear. These members were predicted to encode proteins ranging from 394 to 416 amino acids, including 6 putative transmembrane domains (TMDs), and predicted to localize in the Golgi or vacuole membrane. Furthermore, all the members contained a highly conserved signature sequence in TMD-II and TMD-V. Yeast functional complementation assays demonstrated that the expression of PbMTP8.1, PbMTP8.2, PbMTP9, and PbMTP10 in particular conferred Mn and Fe tolerance, while PbMTP11.1 and PbMTP11.2 only transport Mn, and PbMTP11.3 transports neither Mn nor any other ion that was used in the study. Quantitative real-time PCR analysis indicated that these genes were widely expressed in the root, stem, and leaf tissues of the pear seedlings, but the expression levels differed among the different tissues. The results provide us with systematic information about the Mn-CDF subgroup genes and their possible roles in the distribution and homeostasis of Mn and Fe in pear.

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Abbreviations

CAX:

cation exchanger

CDF:

cation diffusion facilitator

IRT:

iron-regulated transporter

MTP:

metal tolerance protein

NRAMP:

natural resistance–associated macrophage protein

SD-Ura:

synthetic drop-out uracil

TMD:

transmembrane domain

VIT:

vacuolar iron transporter

WT:

wild-type

YPD:

yeast extract peptone dextrose

YSL:

yellow stripe-like

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Funding

This work was supported by the National Natural Science Foundation of China (grant numbers 31301839, 51572131) and a project funded by the Fundamental Research Funds for the Central Universities (grant number KJZ201743).

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Authors and Affiliations

  1. Department of Biochemistry & Molecular Biology, College of Life Sciences, Nanjing Agricultural University, Nanjing, 210095, Jiangsu, China

    Lei Hou, Dongfang Gu, Yun Li, Jiyu Li, Jingjun Li, Xi Chen & Wei Zhang

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  1. Lei Hou
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  2. Dongfang Gu
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  4. Jiyu Li
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  5. Jingjun Li
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  7. Wei Zhang
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Correspondence to Xi Chen or Wei Zhang.

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Hou, L., Gu, D., Li, Y. et al. Phylogenetic and Expression Analysis of Mn-CDF Transporters in Pear (Pyrus bretschneideri Rehd.). Plant Mol Biol Rep 37, 98–110 (2019). https://doi.org/10.1007/s11105-019-01142-9

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