A modified method for transformation of Fusarium graminearum
Publish place: Journal of Crop Protection، Vol: 2، Issue: 3
Publish Year: 1392
نوع سند: مقاله ژورنالی
زبان: English
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شناسه ملی سند علمی:
JR_JCP-2-3_005
تاریخ نمایه سازی: 13 آبان 1402
Abstract:
One of the important technical obstacles in the study of many filamentous fungi is the development of efficient transformation system. Transformation of filamentous fungi is difficult because they have a cell wall and for most frequently used approaches preparing of protoplast is necessary. Protoplast preparation is batch-dependent and often frustrating work. In this study, a modified method was introduced for gene transfer to the plant pathogenic fungus, Fusarium graminearum, the major causal agent of Fusarium Head Blight disease in small grains. This protocol was based on protoplast-PEG method. Age of mycelia, enzymes exposure time and mycelium/enzyme ratio were optimized for the purpose of protoplast preparation. The outcome showed that the best result for protoplast preparation was obtained when ۱.۵ × ۱۰۵ spores were let germinate for ۶ h then exposed to ۱۰ ml of enzyme solution for ۳ h. The effect of other parameters that might enhance transformation yields including PEG concentration, DNA quantity and number of protoplasts was also examined. The most efficient condition for transformation involved the use of ۱۰۶ protoplasts, ۲۰ µg DNA and ۳۰% PEG (w/v). In the course of this study, a simple and appropriate modified protocol for transformation of F. graminearum was established. The method introduced is also more economical and faster than other current methods.
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Authors
Sohrab Moradi
Plant Biotechnology Department, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
Forough Sanjarian
Plant Biotechnology Department, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
Naser Safaie
Plant Pathology Department, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran.
Amir Mousavi
Plant Biotechnology Department, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
Gholam Reza Bakhshi Khaniki
Faculty of Agricultural Sciences , Payame Noor University, Tehran, Iran.