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Applied and Environmental Microbiology, September 2005, p. 5124-5128, Vol. 71, No. 9
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.9.5124-5128.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Improvement of the Fatty Acid Composition of an Oil-Producing Filamentous Fungus, Mortierella alpina 1S-4, through RNA Interference with {Delta}12-Desaturase Gene Expression

Seiki Takeno,1 Eiji Sakuradani,1 Akiko Tomi,1 Misa Inohara-Ochiai,2 Hiroshi Kawashima,3 Toshihiko Ashikari,2 and Sakayu Shimizu1*

Laboratory of Fermentation Physiology and Applied Microbiology, Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan,1 Institute for Advanced Technology, Suntory Ltd., Osaka 618-0001, Japan,2 Institute for Health Care Science, Suntory Ltd., Osaka 618-0001, Japan3

Received 12 October 2004/ Accepted 6 April 2005

An oleaginous fungus, Mortierella alpina 1S-4, is used commercially for arachidonic acid production. {Delta}12-Desaturase, which desaturates oleic acid (18:1n-9) to linoleic acid (18:2n-6), is a key enzyme in the arachidonic acid biosynthetic pathway. To determine if RNA interference (RNAi) by double-stranded RNA occurs in M. alpina 1S-4, we silenced the {Delta}12-desaturase gene. The silenced strains accumulate 18:2n-9, 20:2n-9, and Mead acid (20:3n-9), which are not detected in either the control strain or wild type strain 1S-4. The fatty acid composition of stable transformants was similar to that of {Delta}12-desaturation-defective mutants previously identified. Thus, RNAi occurs in M. alpina and could be used to alter the types and relative amounts of fatty acids produced by commercial strains of this fungus without mutagenesis or other permanent changes in the genetic background of the producing strains.

* Corresponding author. Mailing address: Laboratory of Fermentation Physiology and Applied Microbiology, Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan. Phone: 81-75-753-6115. Fax: 81-75-753-6128. E-mail: sim{at}kais.kyoto-u.ac.jp.

Applied and Environmental Microbiology, September 2005, p. 5124-5128, Vol. 71, No. 9
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.9.5124-5128.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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