Abstract

Purpose: To study the effect of palmarumycin C13, an elicitor from the endophytic fungus Berkleasmium sp. Dzf12, on growth and diosgenin production in plantlet or cell cultures of its host plant, Dioscorea zingiberensis.Methods: Palmarumycin C13 was isolated from the ethyl acetate extract of the endophytic fungus Berkleasmium sp. Dzf12 using a combination of high-speed counter-current chromatography (HSCCC), Sephadex LH-20 chromatography and preparative high performance liquid chromatography (HPLC). The biomass of the plantlet and cell cultures of D. zingiberensis as well as their diosgenin content and yield were analyzed after treatment with palmarumycin C13.Results: Optimal elicitation of diosgenin production by palmarumycin C13 in D. zingiberensis plantlet and cell cultures was achieved when palmarumycin C13 was added to the medium at a concentration of 60 mg/L (for plantlet culture) at the beginning of culturing or 10 mg/L (for cell culture) on day 25 after inoculation. By using these optimal concentrations, the diosgenin yield of the cultured plantlets reached its maximum of 6.44 mg/L, that is, > 1.4-fold increase, while diosgenin yield of the cultured cells reached a maximum of 10.73 mg/L, which is an > 8.0-fold increase.Conclusion: Addition of palmarumycin C13 from the endophytic fungus, Berkleasmium sp. Dzf12, is a potentially effective strategy for enhancing diosgenin production in D. zingiberensis plantlet and cell cultures.Keywords: Endophytic fungus, Berkleasmium sp. Dzf12, Spirobisnaphthalene, Palmarumycin C13, Dioscorea zingiberensis, Diosgenin, Elicitation, Cell culture, Plantlet culture

Highlights

  • IntroductionThe latent phytopathogens or mutualistic symbionts presenting inter- and/or intra-cellularly in the normal tissue of host plants, have been proved to be a promising source of novel structures and/or strong bioactivities [5]

  • The results verified that day 32 was a suitable time for harvesting plantlet cultures for diosgenin production by treatment with palmarumycin C13 at 60 mg/L

  • The highest diosgenin yield (10.73 mg/L), which was 9.02-fold of control (1.19 mg/L), was obtained when palmarumycin C13 was added on the 25th day at 10 mg/L

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Summary

Introduction

The latent phytopathogens or mutualistic symbionts presenting inter- and/or intra-cellularly in the normal tissue of host plants, have been proved to be a promising source of novel structures and/or strong bioactivities [5]. Dzf was an endophytic and spirobisnapthalenes-producing fungus isolated from the rhizomes of D. zingiberensis in our previous studies [6,7,8]. This fungus was found to be a high producer of palmarumycin C13 [9,10,11], which has been shown to have obvious antibacterial and antifungal activity [6,12], antitumor activity and inhibitory activity on phospholipase D (PLD) [13]

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