IDENTIFICATION AND CHARACTERIZATION OF PLANT COMPONENTS INVOLVED IN AGROBACTERIUM MEDIATED PLANT TRANSFORMATION
Various techniques have been used to identify plant factors required for Agrobacterium-mediated plant transformation (AGMT). The latest and most versatile, Virus Induced Gene Silencing (VIGS) has been successfully established and used in our laboratory to identify many unique genes not published before (Anand et al., MPMI 2007) and also confirm previously known genes during this interaction. Using this tool, a screen of an N. benthamiana cDNA library has yielded several new candidates.
We are using an experimental pipeline shown here
which will help determine the role of the gene in plant transformation as well as at what step it is involved. We are also actively looking for genes which have a negative regulatory role in plant transformation and which when silenced will lead to increased transformation. Some candidates are being analysed.
The other major undertaking is to identify candidates involved in the T-DNA integration step. A screen for role of DNA repair proteins in T-DNA integration has identified some interesting candidates. We are targeting the NHEJ machinery of proteins to determine their role in this process.
Publications
1.Zarir Vaghchhipawala , Ronald Bassüner , Kathryn Clayton , Kimberley Lewers , Randy Shoemaker , and Sally Mackenzie (2001). “Modulations in gene expression and Mapping of genes associated with cyst nematode infection of soybean" . Molecular Plant Microbe Interactions 14(1):42-54
2. Zarir E. Vaghchhipawala, Jessica A. Schlueter, Randy C. Shoemaker, and Sally A. Mackenzie (2004). Soybean FGAM synthase promoters direct ectopic nematode feeding site activity. Genome 47(2): 404-413
3. Chen, S.R*., Vaghchhipawala, Z.E.*, Li, W., Asard, H., and Dickman, M. (2004) “Tomato Phospholipid Hydroperoxide Glutathione Peroxidase Inhibits Cell Death Induced by Bax and Oxidative Stresses in Yeast and Plants”. Plant Physiology 135:1630-1641 (*Co-First Author)
4. Ajith Anand, Zarir Vaghchhipawala, Choong-Min Ryu, Li Kang, Keri Wang, Olga del-Pozo, Gregory B. Martin and Kirankumar S. Mysore. (2007) “Identification and Characterization of Plant Genes Involved in Agrobacterium-Mediated Plant Transformation by Virus-Induced Gene Silencing” MPMI Vol 20 (1):41-52.
