Hi, I am using a TRV-based virus expression system to express...

YW
Ykn Wong
Apr 13, 2019
protocol Protocol: Plant Tissue Trypan Blue Staining During Phytopathogen Infection
Hi, I am using a TRV-based virus expression system to express in Nicotiana benthamiana. Necrosis can be clearly seen. But according to the protocol, necrosis area cannot be dyed blue.How to solve this problem? Thank you!
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Marta Berrocal-Lobo Author Answered Apr 16, 2019

Centro de Biotecnología y Genómica de Plantas, Universidad Politécnica de Madrid (UPM) – Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Parque Científico y Tecnológico, UPM, Campus de Montegancedo, Spain

Dear Sr,
This protocol has been used several times with Arabidopsis leaves infected with different fungi, but some users working with tobacco infected with virus showed that trypan blue solution needs higher concentrations for staining necrosis areas. Probably virus infect vascular tissue areas more difficult to stain than necrotic leave areas.
I would suggest to stain leaves of younger plants or stain leaves during longer periods and at higher concentrations.
Please let me know about your results in order to add them to notes about the article.
I never had problems, but with Arabidopsis leaves.
Hoping to be useful
Sincerely
Marta
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