REVIEW PAPER 2010-2
Investigating RNA silencing as a method of controlling Fusarium verticillioides
infections
Score for this proposal – Good
Objectives and approaches
1. Identify how siRNAs enter cells of Fusarium verticillioides.
Fusarium verticillioides strain M‐3125 that has been modified to express green fluorescent
protein (GFP) will be exposed to small double stranded RNAs (dsRNA) on potato dextrose agar
(PDA). Controls will be exposed to buffer without dsRNA. Changes in gene expression due to
this exposure will be analyzed using suppression subtraction hybridization (SSH). Collected
DNA from the fungus will be sequenced to identify differentially expressed genes. Restriction
enzyme‐mediated integration (REMI) will be used to make Fusarium verticillioides mutants that
do not take up dsRNA and therefore do not undergo RNA silencing. Similar tests will be carried
out on these mutants.
2. Identify target genes that are effective in controlling the growth of Fusarium
verticillioides.
Genes involved in metabolism and growth of this fungus will be retrieved from the genomic
sequence database of Fusarium verticillioides. These genes will be compared with those
expressed when Fusarium verticillioides was grown on PDA in the presence of ground and
sterilized corn.
The objectives were clearly and concisely outlined. Approaches and expectations for each
objective were clearly stated and discussed. The proposal had a well explained timeline and
future directions. The author indicated his/her past training in this field which shows that he/she
is competent to carry out this study.
On the other hand, the author did not include any pitfalls in the approaches or alternative
approaches for the pitfalls. He/she also had few references to support the proposed study. This
makes it hard to believe that this study will be successful. The limited number of references
could be an indicator of author’s limited knowledge concerning other studies that have been
previously carried out in this field. This could potentially result in proposing a study that has
narrower impacts. Additionally, there was no justification for the money requested to be spent on
items listed in the budget.
I scored this proposal as good because it gave a good general picture of what the author hopes to
achieve from the proposed study. This study is capable of having good impact on plant-pathogen
related studies. Nevertheless, the proposal had a weak project summary and introduction. There
was a lack of connection between most statements. This plus the many grammatical errors and
wrong sentence construction made it hard to follow and understand what information the author
was trying to relay.