Enhancement of lignan biosynthesis in suspension cultures of Linum nodiflorum by coronalon, indanoyl-isoleucine and methyl jasmonate
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The effect of the two synthetic elicitors coronalon and indanoyl-isoleucine and of methyl jasmonate (MeJA) on the accumulation and biosynthesis of lignans by cell suspension cultures of Linum nodiflorum (Linaceae) was investigated. The production of 6-methoxypodophyllotoxin (MPTOX) could be increased more than tenfold, the maximal content reaching up to over 2.5% of the cell dry weight. The highest yield was achieved by administering 50 μM of the synthetic elicitors on the fourth day and extracting the products on the tenth day of the culture period. An additional lignan accumulated in elicitor-treated cultures. Its structure was elucidated by extensive 1D and 2D NMR measurements, revealing its identity as 5′-demethoxy-MPTOX (5′-dMPTOX). Its average content amounted up to over 5% of the cell dry weight. Growth was only slightly affected by the addition of the elicitors. Methyl jasmonate exerted a moderate stimulating effect on the L. nodiflorum cells with MPTOX and 5′-dMPTOX contents going up to 1.4 and 2.1% of the cell dry weight, respectively. The activities of deoxypodophyllotoxin 6-hydroxylase and β-peltatin 6-O-methyltransferase, two enzymes involved in MPTOX biosynthesis, were increased up to 21.9-fold and 14.6-fold, respectively, in the treated cultures.
KeywordsCell cultures Coronalon Elicitation Linum nodiflorum (Linaceae) 6-Methoxypodophyllotoxin 5′-Demethoxy-6-methoxypodophyllotoxin
We are thankful to Prof. Dr. A.W. Alfermann (Düsseldorf, Germany) for the suspension cultures of Linum nodiflorum established in his laboratory as well as for a sample of MPTOX. β-Peltatin and β-peltatin-A methylether were kindly donated by Prof. Dr. M. Medarde (Salamanca, Spain). We also thank I. Klaiber (Hohenheim, Germany) for recording MS spectra. The financial support by the Deutsche Forschungsgemeinschaft is gratefully acknowledged.
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