Plant Molecular Biology Reporter

, Volume 29, Issue 3, pp 582–596 | Cite as

Expression and Cellular Localization of ZIP1 Transporter Under Zinc Deficiency in Wild Emmer Wheat

  • Emel Durmaz
  • Ceyda Coruh
  • Gizem Dinler
  • Micheal A. Grusak
  • Zvika Peleg
  • Yashua Saranga
  • Tzion Fahima
  • Atilla Yazici
  • Levent Ozturk
  • Ismail Cakmak
  • Hikmet Budak
Article

Abstract

Zinc deficiency is a common problem leading to severe decreases in grain yield and has detrimental effects on nutritional quality in cereals. Wild emmer wheat, Triticum turgidum ssp. dicoccoides, exhibits a potential genetic resource for wheat improvement due to its compatibility with modern wheat. In this study, Zn deficiency response of wild progenitors and modern wheat were examined using molecular and physiological approaches with plants grown under various Zn concentrations. The results revealed wide variation in response to Zn deficiency between wild emmer accessions. Among the wild emmer accessions studied, accession MM 5/4 was found to be most tolerant and accession 19–36 was the most sensitive to Zn deficiency. To better understand Zn transport mechanisms in wild emmer wheat, we analyzed the expression patterns of a ZRT/IRT-like gene, Zrt-, Irt-like protein (ZIP)1, in the roots and shoots of several accessions that were maintained on different concentrations of Zn. Quantitative real-time polymerase chain reaction results revealed that ZIP1 transcript levels are elevated with decreasing Zn supply in all accessions. Particularly, ZIP1 transcript accumulation was lower in the roots of accession MM 5/4 while the susceptible, 19–36 accession, has elevated levels of ZIP1 transcript, revealing a Zn deficiency response for this genotype. We also identified and cloned a full-length ZIP1 transporter, named TdZIP1, and further analyzed the corresponding protein sequence for structural attributes. Under Zn deficiency, deleting the last 20 amino acids from the last transmembrane domain of TdZIP1 and tagging with GFP resulted in endoplasmic reticulum localization. Functional expression of the isolated TdZIP1 using Zn-uptake defective Saccharomyces cerevisiae strains on limiting Zn media showed that it could indeed transport Zn. However, overexpression of this transporter causes excess accumulation of Zn in the cells, thus generating a toxic environment. Overall, our results indicate the possibility of using Triticum dicoccoides for the genetic improvement of zinc deficiency tolerance in wheat.

Keywords

Triticum turgidum ssp. dicoccoides Zinc transport Heterologous expression ZRT/IRT-like proteins 

Supplementary material

11105_2010_264_MOESM1_ESM.doc (78 kb)
Supp. Fig. 1Flow cytometry analysis of GFP expression in ZHY3 strain. a Transformed with pPR3 only, b transformed with pPR3/GFP. (DOC 78 kb)
11105_2010_264_MOESM2_ESM.doc (68 kb)
Supp. Fig. 2In-frame cloning strategy of TdZIP1 from the 19 to 36 accession into pPR3/GFP. (DOC 68 kb)
11105_2010_264_MOESM3_ESM.doc (38 kb)
Supp. Fig. 3Severity of leaf symptoms as the development of response to Zn deficiency for wild and modern wheat. Severity levels were measured by the occurrence of chlorosis and necrosis as 1 demonstrates the least affected and 5 is for the most damaged leaves. Gray dashed and black solid bars are for T. dicoccoides and T. durum wheat, respectively. (DOC 38 kb)
11105_2010_264_MOESM4_ESM.doc (217 kb)
Supp. Fig. 4Complementation of ZHY3 yeast mutants by TdZIP1 obtained from susceptible and tolerant wild emmer wheat genotypes under complex media. A 10-fold serial dilutions from overnight cultures of ZHY3 mutant cells transformed with (a) the empty vector (pPR3-C), (b) TdZIP1 from the susceptible genotype (19–36) of wild wheat, (c) TdZIP1 from the susceptible genotype (24–39) of wild wheat. (DOC 217 kb)
11105_2010_264_MOESM5_ESM.docx (11 kb)
Supplementary Table 1Characterization of isolated ZIP1 genes from the genotypes used in this study. All the listed ZIP1s represent the coding sequences of the corresponding genes. (DOCX 11 kb)

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Emel Durmaz
    • 1
  • Ceyda Coruh
    • 1
  • Gizem Dinler
    • 1
    • 5
  • Micheal A. Grusak
    • 2
  • Zvika Peleg
    • 3
    • 6
  • Yashua Saranga
    • 3
  • Tzion Fahima
    • 4
  • Atilla Yazici
    • 1
  • Levent Ozturk
    • 1
  • Ismail Cakmak
    • 1
  • Hikmet Budak
    • 1
  1. 1.Faculty of Engineering and Natural Sciences, Biological Sciences and Bioengineering ProgramSabanci UniversityIstanbulTurkey
  2. 2.USDA-ARS Children’s Nutrition Research Center, Department of PediatricsBaylor College of MedicineHoustonUSA
  3. 3.The Robert H. Smith Institute of Plant Sciences and Genetics in AgricultureThe Hebrew University of JerusalemRehovotIsrael
  4. 4.Department of Evolutionary and Environmental Biology and the Institute of EvolutionUniversity of HaifaHaifaIsrael
  5. 5.Istanbul Technical UniversityIstanbulTurkey
  6. 6.Department of Plant SciencesUniversity of California, DavisDavisUSA

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