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Related Experiment Videos

GFLV replication in electroporated grapevine protoplasts.

Valat1, Toutain, Courtois

  • 1Laboratoire de Biologie et Physiologie Végétales (URVVC, UPRES EA 2069), URCA, BP1039, F-51687, Reims, France

Plant Science : an International Journal of Experimental Plant Biology
|May 18, 2000
PubMed
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Researchers developed a new method for directly inoculating grapevine plants with Grapevine fanleaf virus (GFLV) using protoplast electroporation. This technique allows for efficient virus replication studies and screening for GFLV resistance in grapevine biotechnology.

Area of Science:

  • Plant Virology
  • Plant Biotechnology
  • Molecular Biology

Background:

  • Grapevine fanleaf virus (GFLV) causes the economically significant court-noué disease in grapevines.
  • Natural transmission of GFLV occurs exclusively via Xiphinema nematodes.

Purpose of the Study:

  • To establish a method for direct inoculation of GFLV into grapevine protoplasts.
  • To investigate GFLV replication in grapevine protoplasts.
  • To assess the potential for grapevine biotechnology applications.

Main Methods:

  • Isolation of protoplasts from in vitro-grown grapevine mesophyll and embryogenic cell suspensions.
  • Optimization of protoplast permeation using calcein uptake and low salt poration medium.
  • Electroporation for direct inoculation of GFLV (isolate F13) and Arabis mosaic virus (ArMV).

Related Experiment Videos

  • Detection of viral replication via anti-P38 Western blotting.

    • Main Results:

    • Successful direct inoculation and replication of GFLV were achieved in both grapevine protoplast types.
    • Viral RNA and particles effectively initiated GFLV replication.
    • The method also worked for another GFLV isolate and the related nepovirus, ArMV.

    Conclusions:

    • Protoplast electroporation provides a viable method for direct GFLV inoculation and replication studies.
    • This technique is valuable for grapevine biotechnology, including screening for GFLV resistance and biorisk evaluation.