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Developmental stages and secondary compound biosynthesis of mycobiont and whole thallus cultures of Buellia subsororioides

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Abstract

Lichen species have unique culture media preferences, and established cultures are known for the synthesis of secondary metabolites. This paper reports observations on the developmental stages and secondary compound biosynthesis by the mycobiont and whole thallus cultures of Buellia subsororioides. It also investigates the suitable media compositions for the culture growth, the role (nutrient or stressor) of sucrose concentrations on the growth stages, biomass, secondary compound profiles, and the quantity of biosynthesized known compounds/g of culture biomass for each treatment using mycobiont cultures. The ascospore-derived mycobiont cultures and thallus macerate-derived whole thallus cultures of B. subsororioides were established and grown using malt yeast extract (MY) medium. Mycobiont cultures were subcultured in MY medium supplemented with sucrose and its concentrations ranging from 0 to 30 % (with 2 % increment between treatments) for 120 days. The molecular identity of cultures was confirmed using nuclear ribosomal Internal transcribed spacer (ITS) DNA sequences obtained from the cultured mycobiont and from the natural thallus. The ITS DNA sequences of the mycobiont showed 99 % similarity with the sequences of the natural thallus. The mycobiont cultures under varying sucrose concentrations initiated as white cottony stages and transformed to brown compact mycelia, with optimum biomass and biosynthesis of nine secondary compounds in MY 10 %. The number of compounds (1–9) varies according to treatments. The whole thallus cultures (MY 0 %) showed a profile of secondary compounds similar to that of the natural thalli along with a trace of one unknown compound. The obtained results are encouraging for the synthesis of the desired quantities of lichen secondary compounds through cultures for relevant applications.

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Abbreviations

NT:

Natural thallus

MY:

Malt yeast extract medium

MYC:

Mycobiont culture

WTC:

Whole thallus culture

UV:

Ultraviolet wavelength

BA:

Baeomycesic acid

NA:

Norstictic acid

AT:

Atranorin

HPTLC:

High performance thin layer chromatography

ITS:

Internal transcribed spacer

SE:

Standard error

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Acknowledgments

We thank Prof. M.S. Swaminathan, Founder Chairman, and Dr. Ajay Parida, Executive Director, M.S. Swaminathan Research Foundation, for their constant encouragement and support; Dr Brian Coppins, Royal Botanic Garden Edinburgh, for correcting the language, Dr. G. Ganesan, C. Divya for suggestions, Mr. G.K. Dayanandham, Senior Secretary for his immense help in the final version of manuscript preparation and Mr. E. Siva for his help in the laboratory and Department of Biotechnology, Government of India, for the financial assistance.

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Authors and Affiliations

  1. Lichen Ecology and Bioprospecting Laboratory, M. S. Swaminathan Research Foundation, Taramani, Chennai, 600 113, India

    Karthik Shanmugam, Muthukumar Srinivasan & G. N. Hariharan

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  1. Karthik Shanmugam
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  2. Muthukumar Srinivasan
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  3. G. N. Hariharan
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Correspondence to G. N. Hariharan.

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Section Editior: Gerhard Rambold

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Shanmugam, K., Srinivasan, M. & Hariharan, G.N. Developmental stages and secondary compound biosynthesis of mycobiont and whole thallus cultures of Buellia subsororioides . Mycol Progress 15, 41 (2016). https://doi.org/10.1007/s11557-016-1184-x

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