Plant Molecular Biology Reporter

, Volume 33, Issue 3, pp 401–413 | Cite as

Tobacco PIC1 Mediates Iron Transport and Regulates Chloroplast Development

  • Xun Gong
  • Changhong Guo
  • Toru Terachi
  • Hongsheng Cai
  • Diansi Yu
Original Paper
First Online:

Abstract

Iron is an essential element for plant growth and development, playing important roles in a variety of cellular activities including cell respiration, chlorophyll biosynthesis, and DNA synthesis. In chloroplasts, iron is essential for photosynthetic electron transport and functions as a cofactor for superoxide dismutases. Thus, iron homeostasis is critical for chloroplast and plant development. To better understand the mechanisms by which iron is transported into chloroplasts, we cloned and characterized the Nicotiana tabacum homolog of AtPIC1, which is a chloroplast iron transporter in Arabidopsis thaliana. NtPIC1 was expressed in many tissues of the tobacco plant, and the encoded protein was localized to the chloroplast envelope. Southern blotting indicated that there is a single copy of NtPIC1 in the tobacco genome. Moreover, yeast complementation assays suggested that NtPIC1 transports iron. We used RNA interference to downregulate NtPIC1 in transgenic plants, which resulted in albinism, dwarfism, iron-deficient chloroplasts, and chloroplast that exhibited ultrastructural defects. NtPIC1 overexpression resulted in deep-green leaves, elevated levels of chlorophyll, more densely packed chloroplasts, and the accumulation of iron and starch grains within chloroplasts. In addition, NtPIC1 influenced the expression of genes involved in iron transport, iron storage, iron oxidative stress, and the biogenesis of iron-sulfur proteins. These results suggest that NtPIC1 transports iron into chloroplasts, regulates iron homeostasis, and influences plant development.

Keywords

Tobacco Chloroplast NtPIC1 Iron transporter RNA interference 

Abbreviations

qRT-PCR

Quantitative real-time polymerase chain reaction

RT-PCR

Reverse transcriptase-polymerase chain reaction

GFP

Green fluorescent protein

RNAi

RNA interference

WT

Wild type

OX

Overexpression

IRT1

Iron regulated transporter1

NRAMP1

Natural resistance associated macrophage protein1

YSL1

Yellow stripe like1

FER

Ferritin

FSD

Fe superoxide dismutase

FD

Ferredoxin

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Notes

Acknowledgments

We thank Dr. Mikio Nozoe (Faculty of Engineering, Kyoto Sangyo University, Japan) for helping to clone NtPIC1. We also thank Dr. Yao Guang Liu (South China Agricultural University, China) for kindly providing the pRNAi vector, Prof. Shaojian Zheng (Zhejiang University) for providing the yeast strain BY4741, and Prof. Yuanmei Zuo (China Agricultural University) for providing the yeast strain DEY1453. This work was supported by MOST 863 project (2013AA102607-5), and the Graduate Innovation Fund of Harbin Normal University (HSDBSCX2012-05) and Natural Science Foundation of Heilongjiang Province of China (C201308C0601).

Supplementary material

11105_2014_758_MOESM1_ESM.tif (5.9 mb)
Fig. S1 Southern blot analysis of tobacco (N. tabacum cv. SR-1) genomic DNA digested with XbaI or DraI. An NtPIC1 probe was used (TIFF 6043 kb)
11105_2014_758_MOESM2_ESM.tif (11.4 mb)
Fig. S2 Southern blot analysis of NtPIC1 transgenic tobacco plants. Genomic DNA was isolated from leaves of WT or transgenic (OX1, OX11, OX13, OX19, OX30, and OX33) plants. DNA was digested with DraI and selection marker gene as probe (This result was previously done, using X-ray exposure imaging) (TIFF 11680 kb)

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Xun Gong
    • 1
  • Changhong Guo
    • 1
  • Toru Terachi
    • 2
  • Hongsheng Cai
    • 1
  • Diansi Yu
    • 1
  1. 1.Key Laboratory of Molecular Cytogenetics and Genetic Breeding of Heilongjiang Province, College of Life Science and TechnologyHarbin Normal UniversityHarbin CityChina
  2. 2.Department of Bioresource and Environmental Sciences, Faculty of Life SciencesKyoto Sangyo UniversityKita-kuJapan

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