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Combining Clearing and Fluorescence Microscopy for Visualising Changes in Gene Expression and Physiological Responses to Plasmodiophora brassicae
JoVE Journal
Biologia
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JoVE Journal Biologia
Combining Clearing and Fluorescence Microscopy for Visualising Changes in Gene Expression and Physiological Responses to Plasmodiophora brassicae

Combining Clearing and Fluorescence Microscopy for Visualising Changes in Gene Expression and Physiological Responses to Plasmodiophora brassicae

DOI:
10.3791/64297-v

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06:58 min

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August 05, 2022

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, , , , , ,
1Integrative Plant Biology Team,Institute of Plant Genetics Polish Academy of Sciences, 2ZMBP-Center for Plant Molecular Biology,University of Tübingen, 3Department of General Botany, Institute of Experimental Biology, Faculty of Biology,Adam Mickiewicz University

Capítulos

  • 00:05Introduction
  • 01:36Tissue Preparation and Fixation
  • 02:13Tissue Embedding and Sectioning
  • 03:03Vibratome Sectioning and Specimen Clearing
  • 04:03Staining Procedure and Microscopy
  • 04:49Results: Microscopic Analysis of the Biotrophic Interaction and Gene Expression Regulations Between Plasmodiophora brassicae and Arabidopsis Plants
  • 06:06Conclusion

Summary

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The present protocol describes an optimized method for the histological observation of galls induced by Plasmodiophora brassicae. Vibratome sections of hypocotyls are cleared before fluorescence imaging to study the involvement of transcription factors and phytohormones during disease progression. This protocol overcomes resin embedding limitations, enabling in planta visualization of fluorescent proteins.

Tags

Plasmodiophora BrassicaeClubrootGene ExpressionFluorescence MicroscopyClearingGallsPhytohormoneLignificationPlant-pathogen InteractionAgarose EmbeddingVibratome Sectioning

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