DEVELOPMENT OF AN EST MICROARRAY FOR Nicotiana benthamiana

¹ Department of Genetics, Box 7614, North Carolina State University, Raleigh, NC 27965-7614 USA

² Department of Horticultural Science , Box 7609, North Carolina State University, Raleigh, NC 27695-7609

³ Department of Plant Pathology, Box 7616, North Carolina State University, Raleigh, NC 27695-7616

Indigenous to Australia, Nicotiana benthamiana is an amphiploid species with 38 chromosomes and is thought to be the result of the hybridization of N. suaveolens (n=16) and N. debneyi (n=24). It is a close relative to cultivated tobacco (N. tabacum) and a member of the Solanaceae family that includes important crops such as potatoes and tomatoes. N. benthamiana has proven to be an important model host for the study of plant disease interactions. More specifically, it is extremely valuable in the elucidation of plant-virus interactions, as well as the ongoing characterization of gene silencing. We have and continue to sequence deeply into three different N. benthamiana EST libraries constructed from both root tissue and expanded leaf tissue (normalized and non-normalized libraries). Approximately 60% of the leaf libraries have been sequenced with a return rate of 20% unique genes. Currently a test array of 800 clones has been designed and is in the construction phase. Ultimately, we will construct an array composed of 5,000 or more unigenes, which would be made available to the scientific community. Our group’s specific goal is to identify host genes involved in the host-pathogen interaction, with an emphasis on plant virus pathogenesis.