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Temporary immersion systems for Amaryllidaceae alkaloids biosynthesis by Pancratium maritimum L. shoot culture

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Abstract

The alkaloid patterns of sea daffodil (Pancratium maritimum L.) shoot culture, cultivated in a temporary immersion cultivation system were investigated. The shoots accumulated maximal amounts of biomass (0.8 g dry biomass/L and Growth Index = 1.6) at immersion frequency with 15 min flooding and 12 h stand-by periods. At this regime P. maritimum shoots achieved the highest degree of utilization of carbon source. Twenty-two alkaloids, belonging to narciclasine, galanthamine, haemanthamine, lycorine, montanine, tazettine, homolycorine and tyramine types were identified in intracellular and extracellular alkaloid extracts. The immersion frequency affected strongly the capacity of alkaloid biosynthesis in P. maritimum shoots and at the optimum conditions of cultivation, the total intracellular alkaloid content reached up to 3,469 μg/g dry biomass. The main biosynthesized alkaloids were haemanthamine (900.1 μg/g) and lycorine (799.9 μg/g). The obtained results proved that temporary immersion technology, as a cultivation approach, and P. maritimum shoots, as a biological system, are prospective for producing wide range bioactive alkaloids.

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Abbreviations

M&S:

Murashige and Skoog medium

NAA:

1-naphthaleneacetic acid

BAP:

6-benzylaminopurine

ddH2O:

Double-distilled water

GC-MS:

Gas chromatography–mass spectrometry

ADB:

Accumulated dry biomass

GIDW:

Growth indexes dry weight

AChE:

acetylcholinesterase

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Acknowledgments

This research has been supported by National Science Fund of Bulgaria under contract number TK-B-1605/2006. The authors acknowledge Ivaila Dincheva (AgroBioInstitute, Sofia, Bulgaria) for performing GC-MS runs.

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Authors and Affiliations

  1. Laboratory of Applied Biotechnologies, The Stephan Angeloff Institute of Microbiology, Bulgarian Academy of Sciences, 139 “Ruski” Blvd., 4000, Plovdiv, Bulgaria

    Vasil Georgiev, Ivan Ivanov & Atanas Pavlov

  2. Center for Viticulture and Small Fruit Research, Florida A & M University, 6505 Mahan Drive, Tallahassee, FL, 32317, USA

    Vasil Georgiev

  3. AgroBioInstitute, 8 Dragan Tzankov Blvd., 1164, Sofia, Bulgaria

    Strahil Berkov

  4. Department of Organic Chemistry, University of Food Technologies, 26 Maritza Blvd, 4002, Plovdiv, Bulgaria

    Ivan Ivanov & Atanas Pavlov

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  1. Vasil Georgiev
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  2. Ivan Ivanov
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  3. Strahil Berkov
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  4. Atanas Pavlov
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Correspondence to Vasil Georgiev.

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Georgiev, V., Ivanov, I., Berkov, S. et al. Temporary immersion systems for Amaryllidaceae alkaloids biosynthesis by Pancratium maritimum L. shoot culture. J. Plant Biochem. Biotechnol. 23, 389–398 (2014). https://doi.org/10.1007/s13562-013-0222-x

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