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Cloning, in silico characterization, subcellular localization, and expression of a heat shock cognate 70 kDa protein/gene (EjHsc70-2) from Eriobotrya japonica

  • Shoukai Lin
  • Ti Wu
  • Ming Li
  • Xiaoming Huang
  • Yanqing Zhang
  • Linxiao Han
  • Bisha Wu
  • Yu Chen
  • Suying Lin
  • Dahe Lin
  • Minlan Wu
  • Jincheng WuEmail author
Original Article
  • 38 Downloads

Abstract

Heat shock protein 70 kDa proteins (Hsp70 s) are among the most important molecular chaperone groups and play a significant role in the stress responses and development of plants. In the present study, the full-length cDNA of the heat shock cognate 70 protein 2 gene EjHsc70-2, which encodes a loquat Hsp70 s member, was cloned and characterized, and its expression and subcellular localization were also investigated. The full-length cDNA of EjHsc70-2 consists of an open reading frame (ORF) of 1950 bp, a 5′-UTR of 103 bp, and a 3′-UTR of 62 bp, and the ORF encodes 649 amino acid residues. The structure of the loquat Hsc70-2 protein was analysed using several bioinformatics tools, and the results showed that the protein was, indeed, a member of the Hsp70 s. Phylogenetic tree analysis suggested that the genetic evolution of Hsc70-2 genes conformed well to the morphology based taxonomic classification of seed plants. BLAST and multiple alignment analyses determined that the Hsc70-2 genes and Hsc70-2 proteins were both highly conserved among loquat and other seed plants, suggesting that the functions of EjHsc70-2 might be similar to those of other Hsc70-2 genes. The bioinformatics and experimental subcellular localization analyses both supported that EjHsc70-2 was a cytoplasmic and/or nuclear protein. Quantitative real-time RT-PCR (RT-qPCR) suggested its conserved functions involved in loquat organ development. Moreover, EjHsc70-2 were also inducible, which may contribute to the low-temperature adaptation of loquat fruits in cold storage. These results provide new insights into the characteristics and functions of Hsp70 s in Eriobotrya japonica.

Keywords

Loquat EjHsc70-2 Cloning Characterization Subcellular localization Expression 

Notes

Acknowledgements

This research was supported by the Natural Science Foundation of Fujian Province (2017J01644, 2017J01645), the Education and Research Project of Young and Middle-aged Teachers of Fujian Province (JAT160434, JA15454), the College Outstanding Young Researchers Cultivation Program of Fujian education department, the Research and Innovation Special Foundation of Putian University (2016CX001, 2017081, 2018006, 2018064) and the Open Fund of Key laboratory of Loquat Germplasm Innovation and Utilization (Putian University), and Fujian Province University (2016001, 2017005). The English language of this manuscript was edited by American Journal Experts (AJE).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Supplementary material

11738_2019_2908_MOESM1_ESM.docx (108 kb)
Supplementary material 1 (DOCX 107 kb)

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Copyright information

© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2019

Authors and Affiliations

  • Shoukai Lin
    • 1
    • 3
  • Ti Wu
    • 2
    • 5
  • Ming Li
    • 1
    • 4
  • Xiaoming Huang
    • 1
  • Yanqing Zhang
    • 1
  • Linxiao Han
    • 1
  • Bisha Wu
    • 1
    • 3
  • Yu Chen
    • 1
    • 3
  • Suying Lin
    • 1
    • 3
  • Dahe Lin
    • 1
    • 3
  • Minlan Wu
    • 6
  • Jincheng Wu
    • 1
    • 3
    Email author
  1. 1.Key Laboratory of Loquat Germplasm Innovation and Utilization (Putian University)Fujian Province UniversityPutianChina
  2. 2.Faculty of AgricultureDalhousie UniversityTruroCanada
  3. 3.Fujian Provincial Key Laboratory of Ecology-toxicological Effects and Control for Emerging ContaminantsPutian UniversityPutianChina
  4. 4.Qinghai Academy of Agricultural and Forestry SciencesQinghai UniversityXiningChina
  5. 5.Overseas Education CollegeFujian Agriculture and Forestry UniversityFuzhouChina
  6. 6.Department of Garden and HorticultureZhangzhou City CollegeZhangzhouChina

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