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Area of Science:

  • Plant Biotechnology
  • Functional Genomics
  • Agricultural Science

Background:

  • Genetic transformation is crucial for plant improvement and gene function studies.
  • Existing grapevine transformation methods are inefficient, hindering functional genomics.
  • Vitis amurensis, a wild grape, possesses valuable cold tolerance but lacks an efficient transformation system.

Purpose of the Study:

  • To establish an efficient genetic transformation method for Vitis amurensis.
  • To facilitate the functional identification of cold stress-related genes in V. amurensis.
  • To enable functional analysis of agronomic traits in grapevines.

Main Methods:

  • Agrobacterium-mediated transformation of Vitis amurensis using petiole segments.
  • Optimization of transformation parameters including kanamycin concentration, Agrobacterium strains, bacterial densities, infection treatments, and co-cultivation time.
  • Simultaneous selection of transgenic calli using neomycin phosphotransferase II (NPTII) gene resistance and enhanced green fluorescent protein (eGFP) fluorescence.

Main Results:

  • Petiole segments showed superior callus differentiation capacity compared to leaf discs and stem segments.
  • Transgenic calli were successfully produced with up to 20% efficiency from petiole segments after 2 months.
  • Integration of the NPTII gene and expression of the eGFP gene were confirmed in transgenic calli.

Conclusions:

  • An efficient and rapid Agrobacterium-mediated transformation system for Vitis amurensis was successfully established.
  • This system significantly improves the potential for functional genomics research in V. amurensis.
  • The developed method will accelerate the study of cold tolerance and other agronomic traits in grapevines.