Journal of Crop Protection
Journal of Crop Protection
JCP
Agriculture
http://jcp.modares.ac.ir
1
admin
2251-9041
2251-905X
en
jalali
1392
6
1
gregorian
2013
9
1
2
3
online
1
fulltext
en
A modified method for transformation of Fusarium graminearum
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, <em>Fusarium graminearum</em>, 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 1.5 × 10<sup>5 </sup>spores were let germinate for 6 h then exposed to 10 ml of enzyme solution for 3 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 10<sup>6</sup> protoplasts, 20 µg DNA and 30% PEG (w/v). In the course of this study, a simple and appropriate modified protocol for transformation of <em>F. graminearum</em> was established. The method introduced is also more economical and faster than other current methods.
Filamentous fungi,Fusarim graminearum,PEG,Protoplast,Transformation
297
304
http://jcp.modares.ac.ir/browse.php?a_code=A-3-1000-6083&slc_lang=en&sid=3
Sohrab
Moradi
Sohrab
Moradi
100319475328460040326
100319475328460040326
No
Plant Biotechnology Department, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
Plant Biotechnology Department, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
Forough
Sanjarian
Forough
Sanjarian
100319475328460040325
100319475328460040325
Yes
Plant Biotechnology Department, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
Plant Biotechnology Department, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
Naser
Safaie
Naser
Safaie
100319475328460040328
100319475328460040328
No
Plant Pathology Department, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran.
Plant Pathology Department, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran.
Amir
Mousavi
Amir
Mousavi
100319475328460040327
100319475328460040327
No
Plant Biotechnology Department, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
Plant Biotechnology Department, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
Gholam Reza
Bakhshi Khaniki
Gholam Reza
Bakhshi Khaniki
100319475328460040329
100319475328460040329
No
Faculty of Agricultural Sciences , Payame Noor University, Tehran, Iran.
Faculty of Agricultural Sciences , Payame Noor University, Tehran, Iran.