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Establishment and characterization of a Satureja khuzistanica Jamzad (Lamiaceae) cell suspension culture: a new in vitro source of rosmarinic acid

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Abstract

An in vitro approach to the production of rosmarinic acid (RA), a medicinally important caffeic acid ester, in a cell suspension culture (CSC) of Satureja khuzistanica Jamzad (Lamiaceae) has been investigated for the first time. The CSC was established from friable calli derived from shoot tip explants in Gamborg’s B5 liquid medium supplemented with 30 g/L sucrose, 20 mg/L l-glutamine, 200 mg/L casein hydrolysate, 5 mg/L benzyladenine (BA) and 1 mg/L indole-3-butyric acid (IBA). The effect of nitrogen source (KNO3 and (NH4)2SO4) and their different concentrations on the fresh and dry weight (g/L), as well as RA content (mg/g dry weight) were measured. CSC growth measurements indicated a maximum specific cell growth rate of 1.5/day, a doubling time of 7.6 days and a high percentage of cell viability (96.4 %) throughout the growth cycle. Maximum cell fresh weight (353.5 g/L), dry weight (19.7 g/L) and RA production (180.0 mg/g) were attained at day 21 of culture. Cell growth and RA content were affected by nitrogen deficiency. Media containing 8.3 mM of total nitrogen (¼ of B5 standard medium) led to a minimum cell fresh weight (243.0 g/L), dry weight (17.4 g/L) and RA content (38.0 mg/g) after 21 days. The established CSC provided useful material for further optimization experiments aimed at a large-scale production of RA.

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Abbreviations

RA:

Rosmarinic acid

CSC:

Cell suspension culture

BA:

Benzyladenine

IBA:

Indole-3-butyric acid

FW:

Fresh weight

DW:

Dry weight

SGR:

Specific growth rate

PGRs:

Plant growth regulators

HPLC:

High performance liquid chromatography

PVP:

Polyvinylpyrrolidone

CRD:

Completely randomized design

MS:

Mean square

DF:

Degrees of freedom

CV:

Coefficient of variation

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Acknowledgment

The authors thank the Shahid Beheshti University Research Council, the University of Tehran, Iran and the University of Salamanca, Spain for their financial support of this project. We also wish to thank Dr. Javad Hadian (Medicinal Plants research Institute, Shahid Beheshti University) for his kind help in plant material collection. This work is part of Amir Sahraroo PhD’s thesis.

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Author notes

  1. Amir Sahraroo

    Present address: Department of Horticulture, Faculty of Agriculture, University of Guilan, Rasht, Iran

Authors and Affiliations

  1. Department of Horticulture, Faculty of Agriculture, University of Tehran, Karaj, Iran

    Amir Sahraroo, Mesbah Babalar & Mohammad Reza Fattahi Moghadam

  2. Department of Agriculture, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University G. C., Tehran, Iran

    Mohammad Hossein Mirjalili

  3. Department of Plant Physiology, Campus Miguel de Unamuno, University of Salamanca, 37007, Salamanca, Spain

    Purificación Corchete

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  1. Amir Sahraroo
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  2. Mohammad Hossein Mirjalili
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  4. Mesbah Babalar
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  5. Mohammad Reza Fattahi Moghadam
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Correspondence to Mohammad Hossein Mirjalili.

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Sahraroo, A., Mirjalili, M.H., Corchete, P. et al. Establishment and characterization of a Satureja khuzistanica Jamzad (Lamiaceae) cell suspension culture: a new in vitro source of rosmarinic acid. Cytotechnology 68, 1415–1424 (2016). https://doi.org/10.1007/s10616-015-9901-x

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