Abstract
In situ hybridization (ISH) associated to immunohistochemistry has become a powerful tool for the examination of gene expression. Application of these techniques in grapevine tissues is limited mainly because of technical difficulties with this plant material. Here we present detailed protocols for ISH and immunohistochemistry, recommended controls and troubleshooting, along with examples of several applications to grapevine tissues.
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Abbreviations
- AP:
-
Alkaline phosphatase
- DIG:
-
Digoxigenine
- GS primer:
-
Gene specific primer
- ISH:
-
In situ hybridization
- NTE:
-
NaCl-Tris-EDTA buffer
- PAL:
-
Phenylalanine ammonia-lyase
- PBS:
-
Phosphate buffer saline
- Rib:
-
26S ribosomic RNA
- SSC:
-
Saline sodium citrate
- STS:
-
Stilbene synthase
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Acknowledgments
The authors thank the EMOA laboratory UMR INRA FARE 614 Reims for giving us access to their confocal equipment.
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Colas, S. et al. (2010). Expression Analysis in Grapevine by In Situ Hybridization and Immunohistochemistry. In: Delrot, S., Medrano, H., Or, E., Bavaresco, L., Grando, S. (eds) Methodologies and Results in Grapevine Research. Springer, Dordrecht. https://doi.org/10.1007/978-90-481-9283-0_26
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DOI: https://doi.org/10.1007/978-90-481-9283-0_26
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