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AhDMT1, a Fe2+ transporter, is involved in improving iron nutrition and N2 fixation in nodules of peanut intercropped with maize in calcareous soils

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Abstract

Peanut (Arachis hypogaea L.) is an important legume providing edible proteins and N2 fixation. However, iron deficiency severely reduces peanut growth in calcareous soils. The maize/peanut intercropping effectively improves iron nutrition and N2 fixation of peanut under pot and field conditions on calcareous soils. However, little was known of how intercropping regulates iron transporters in peanut. We identified AhDMT1 as a Fe2+ transporter which was highly expressed in mature nodules with stronger N2 fixation capacity. Promoter expression analysis indicated that AhDMT1 was localized in the vascular tissues of both roots and nodules in peanut. Short-term Fe-deficiency temporarily induced an AhDmt1 expression in mature nodules in contrast to roots. However, analysis of the correlation between the complex regulation pattern of AhDmt1 expression and iron nutrition status indicated that sufficient iron supply for long term was a prerequisite for keeping AhDmt1 at a high expression level in both, peanut roots and mature nodules. The AhDmt1 expression in peanut intercropped with maize under 3 years greenhouse experiments was similar to that of peanut supplied with sufficient iron in laboratory experiments. Thus, the positive interspecific effect of intercropping may supply sufficient iron to enhance the expression of AhDmt1 in peanut roots and mature nodules to improve the iron nutrition and N2 fixation in nodules. This study may also serve as a paradigm in which functionally important genes and their ecological significance in intercropping were characterized using a candidate gene approach.

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Abbreviations

BPDS:

Bathophenanthrolinedisulfonic acid

DMT:

Divalent metal ion transporter genes

hiTAIL-PCR:

High-efficiency thermal asymmetric interlaced PCR

IP:

Intercropping peanut

IRT:

Iron-regulated transporter genes

MP:

Monocropping peanut

NAS:

Nicotianamin synthase genes

NRAMP:

Natural resistance-associated macrophage protein

PBM:

Peribacteroid membrane

PLACE:

Plant cis-acting regulatory DNA elements

SNF:

Symbiotic N2 fixation

TM:

Transmembrane domains

YSL:

Yellow stripe-like genes

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Acknowledgments

We thank Dr. Jiping Liu (USDA-ARS, Cornell University), Dr. Takanori Kobayashi (Graduate School of Agricultural and Life Sciences, The University of Tokyo) for reading and comments of the manuscript. This work was supported by the National Natural Science Foundation of China (Grant No. 31272223, 31071840) and the innovative group of NSFC (Grant No. 31121062).

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Authors and Affiliations

  1. Key Laboratory of Plant-Soil Interactions, MOE, Centre for Resource, Environment and Food Surity, College of Resources and Environmental Sciences, China Agricultural University, No. 2, Yuanmingyuan West Road, Beijing, 100193, China

    Hongyun Shen, Hongchun Xiong, Xiaotong Guo, Pengfei Wang, Penggen Duan, Lixia Zhang, Fusuo Zhang & Yuanmei Zuo

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  1. Hongyun Shen
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  2. Hongchun Xiong
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Correspondence to Yuanmei Zuo.

Additional information

H. Shen and H. Xiong contributed equally to this work.

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Supporting information

Additional supporting information can be found in the online version of this article:

Suppl. Fig. S1 Sequence analysis of nucleotide and deduced amino acid sequence of AhDmt1 (TIFF 6642 kb)

Suppl. Fig. S2 Analysis of promoter sequence of AhDmt1 and DNA structure of GmDmt1 (TIFF 1758 kb)

Suppl. Fig. S3 Functional analysis of AhDmt1 in yeast strains with expression vector pDR195 (TIFF 2122 kb)

425_2014_2033_MOESM4_ESM.tif

Suppl. Fig. S4 Subcellular localization of GFP (a–c) and AhDmt1-GFP fusion (d–f) proteins in onion epidermal cells with the expression vector pEZS-NL (TIFF 632 kb)

425_2014_2033_MOESM5_ESM.tif

Suppl. Fig. S5 Semi-quantitative RT-PCR analysis of mRNA abundance of AhDmt1 in peanut plant tissues in response to iron deficiency under the control of housekeeping gene 18S and 25S ribosomal RNA, respectively (TIFF 36 kb)

425_2014_2033_MOESM6_ESM.tif

Suppl. Fig. S6 The active iron concentration in peanut primary and mature nodules under the different iron nutrition conditions (TIFF 92 kb)

Supplementary material 7 (DOCX 19 kb)

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Shen, H., Xiong, H., Guo, X. et al. AhDMT1, a Fe2+ transporter, is involved in improving iron nutrition and N2 fixation in nodules of peanut intercropped with maize in calcareous soils. Planta 239, 1065–1077 (2014). https://doi.org/10.1007/s00425-014-2033-2

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