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 x 10(8) cfu ml(-1), 1 mm acetosyringone, 25-d-old mycelia at 0.2 g ml(-1), 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. (c) 2008 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Keywords

Activation tagging; Agrobacterium tumefaciens; Antrodia cinnamoma; Fungus; Transformation

Published in

Mycological Research
2009, Volume: 113, number: 3, pages: 290-297
Publisher: ELSEVIER SCI LTD

UKÄ Subject classification

Microbiology
Biochemistry and Molecular Biology


Publication identifier

DOI: https://doi.org/10.1016/j.mycres.2008.11.007

Permanent link to this page (URI)

https://res.slu.se/id/publ/90709