Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
4357572 | Mycological Research | 2009 | 8 Pages |
Abstract
This study describes the development of an efficient and reliable activation tagging system for the medicinal fungus Antrodia cinnamomea. For successful Agrobacterium tumefaciens-mediated transformation, different parameters were considered. The Agrobacterium concentration of 5 Ã 108 cfu mlâ1, 1 mm acetosyringone, 25-d-old mycelia at 0.2 g mlâ1, and co-culture period of 6 d were found to be the most optimal conditions for enhancing the transformation efficiency. The mitotic stability of transferred DNA (T-DNA) was demonstrated by growing eight randomly selected putative transformants in malt extract agar medium for five subcultures. Insertion of T-DNA into the genome of transformants was confirmed by PCR and Southern hybridization. Results showed that 88 % of the mutants contained a single T-DNA insertion. Two of the mutants were observed with different triterpenoid profiles compared with the untransformed cultures. Our results suggest a new functional genomics approach to tag the triterpenoid biosynthesis genes in A. cinnamomea.
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Authors
Emily Chin-Fun Chen, Yu-Hsiu Su, Selvaraju Kanagarajan, Dinesh Chandra Agrawal, Hsin-Sheng Tsay,