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Expression of Drosera rotundifolia Chitinase in Transgenic Tobacco Plants Enhanced Their Antifungal Potential.

Authors
  • Durechova, Dominika1
  • Jopcik, Martin1
  • Rajninec, Miroslav1
  • Moravcikova, Jana1
  • Libantova, Jana2
  • 1 Institute of Plant Genetics and Biotechnology, Plant Science Biodiversity Center, Slovak Academy of Sciences, Akademicka 2, P. O. Box 39A, 950 07, Nitra, Slovak Republic.
  • 2 Institute of Plant Genetics and Biotechnology, Plant Science Biodiversity Center, Slovak Academy of Sciences, Akademicka 2, P. O. Box 39A, 950 07, Nitra, Slovak Republic. [email protected]
Type
Published Article
Journal
Molecular biotechnology
Publication Date
Dec 01, 2019
Volume
61
Issue
12
Pages
916–928
Identifiers
DOI: 10.1007/s12033-019-00214-1
PMID: 31555964
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

In this study, a chitinase gene (DrChit) that plays a role in the carnivorous processes of Drosera rotundifolia L. was isolated from genomic DNA, linked to a double CaMV35S promoter and nos terminator in a pBinPlus plant binary vector, and used for Agrobacterium-mediated transformation of tobacco. RT-qPCR revealed that within 14 transgenic lines analysed in detail, 57% had DrChit transcript abundance comparable to or lower than level of a reference actin gene transcript. In contrast, the transgenic lines 9 and 14 exhibited 72 and 152 times higher expression level than actin. The protein extracts of these two lines exhibited five and eight times higher chitinolytic activity than non-transgenic controls when measured in a fluorimetric assay with FITC-chitin. Finally, the growth of Trichoderma viride was obviously suppressed when the pathogen was exposed to 100 μg of crude protein extract isolated from line 9 and line 14, with the area of mycelium growth reaching only 56.4% and 45.2%, of non-transgenic control, respectively. This is the first time a chitinase from a carnivorous plant with substrate specificity for long chitin polymers was tested in a transgenic plant with the aim of exploring its antifungal potential.

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