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Establishment of a gene function analysis system for the euhalophyte Salicornia europaea L.

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Abstract

Key message

A Salicornia europaea L. in vitro cell transformation system was developed and further applied to SeNHX1 function investigation.

Abstract

The exploration of salt-tolerant genes from halophyte has seriously been limited by the lack of self-dependent transformation system. Here, an Agrobacterium tumefaciens-mediated in vitro cell transformation system of euhalophyte Salicornia europaea L. was developed. Calli derived from hypocotyl of S. europaea were co-cultured for 3 days with Agrobacterium at OD600 ranging from 1.0 to 1.5 and then selected with 25 mg/L hygromycin (Hyg). The transformed cells were identified from Hyg positive calli by GUS assay and qRT-PCR, and the transformation efficiency was up to 74.4%. The practicality of this system was further tested via genetic manipulation of S. europaea Na+/H+ antiporter 1 (SeNHX1) gene by creating the overexpressing, silencing, and empty vector cells. Survival ratio and Na+ distribution under salt treatment showed obvious differences in SeNHX1-overexpressing, -silencing, and empty vector cells, indicating the feasibility of this system to analyze gene function. This investigation is enlightening for studies in other non-model plants lacking of self-dependent transformation system.

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Abbreviations

FDA:

Diacetate

GUS:

β-Glucuronidase

Hyg:

Hygromycin

Kan:

Kanamycin

LB:

Luria–Bertani

MS:

Murashige and Skoog

NAA:

a-Naphthaleneacetic acid

PI:

Propidium iodide

TDZ:

Thidiazuron

TMT:

Ticarcillin

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Acknowledgements

This work was supported by the National Research and Development Project of Transgenic Crops of China (Grant No. 2016ZX08009-003-002) and the National Natural Science Foundation of China (Grant No. 31270421). We would like to thank Professor Xuejun Hua from Institute of Botany, CAS for kindly providing plasmid pBISN1 with GUS gene.

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Authors and Affiliations

  1. Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, No. 20 Nanxincun, Xiangshan, Beijing, 100093, China

    Fang Tai, Sulian Lv, Ping Jiang, Jinhui Wang, Juanjuan Feng & Yinxin Li

  2. University of Chinese Academy of Sciences, Beijing, 100049, China

    Fang Tai & Jinhui Wang

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  1. Fang Tai
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  2. Sulian Lv
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  3. Ping Jiang
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  4. Jinhui Wang
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  5. Juanjuan Feng
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  6. Yinxin Li
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Correspondence to Yinxin Li.

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Communicated by Emmanuel Guiderdoni.

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Tai, F., Lv, S., Jiang, P. et al. Establishment of a gene function analysis system for the euhalophyte Salicornia europaea L.. Plant Cell Rep 36, 1251–1261 (2017). https://doi.org/10.1007/s00299-017-2150-z

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  • DOI: https://doi.org/10.1007/s00299-017-2150-z

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