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|標題:||Identify the Novel Innate Immunity Genes against Virus in Plants
Identifying the novel innate immunity genes in plants is one of the important research topics in studying the interaction between pathogens and plants. These genes may play potential roles for plants in defending the invading pathogens. Identifying these potential and novel innate immunity genes in plant will be very important not only for basic research but also for the application in the future. Previously, we have used the cDNA-amplified fragment length polymorphism (cDNA-AFLP) in identifying 90 differentially expressed cDNA fragments with 8 selective primers on Bamboo mosaic virus (BaMV)-infected N. benthamiana plants. In this project we are going to focus on 31 of those differential expressed genes whose identifies are related to cell rescue, death, defense, ageing, and energy in plants. In preliminary experiments, we had knocked down the expression levels by virus-induced gene silencing (VIGS) for some of these genes and showed the characters as the potential immunity genes that enhances the viral accumulation in the knockdown plants. These results indicted that they may suppress the virus accumulation when they are normally expressed. To identify these potential immunity genes, we are planning to screen these genes by knocking down the expression levels with VIGS, and following by virus inoculation to see if virus accumulation levels are increased. Besides BaMV inoculation in the experiment, we are planning to inoculate Potato virus X and Cucumber mosaic virus to see whether the effect is specific on certain type of virus such as BaMV, or common to all plant viruses. Once we get the potential innate immunity gene candidates, we will clone the full-length of these genes by 3- and 5-RACE. Cloning the full-length of these genes will be the most important step in this project that we can use them not only in studying the basic research such as how do these genes defend the invading pathogens, but also in improving the plant defense capability by over-expressing their products in transgenic plants. Another part of this project is related to the known innate immunity genes in the pathway of autophagy. One of the early genes in the autophagy pathway, Beclin-1, was shown upregulated during BaMV infection in our preliminary experiment. We are wondering whether the autophagy used to be reported involving in hypersensitive response (HR) of pathogen invading is also involved in none-HR pathogen such as BaMV infection in N. benthamiana. We are going to inspect a few genes in the autophagy pathway to make sure this pathway is truly involved. Following up the confirmation of the genes in the pathway do involve in defending BaMV, we are going to inspect the autophagosome formation by using confocal microscopy and study how the autophagosome to remove the viral products.
|Appears in Collections:||生物科技學研究所|
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