Micropropagation and Quantification of Bioactive Compounds in Mertensia maritima (L.) Gray

Han Yong Park,Judy Gopal,Young-Soo Keum,Iyyakkannu Sivanesan,Doo Hwan Kim,Ramesh Kumar Saini

doi:10.3390/ijms20092141

Han Yong Park, Judy Gopal + Show 4 more

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https://doi.org/10.3390/ijms20092141

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Abstract

The goal of this study was to establish an efficient protocol for the large-scale propagation of Mertensia maritima (L.) Gray, and evaluate the carotenoid, fatty acid, and tocopherol contents in the leaves of in vitro regenerated shoots. Surface-disinfected node and shoot tip explants were placed on semisolid Murashige and Skoog (MS) medium with 0–16 µM N6-benzyladenine (BA), kinetin, (KN), and thidiazuron (TDZ) alone, or in combination with, 1 or 2 µM α-naphthaleneacetic acid (NAA). Of the three different cytokinins employed, TDZ elicited the best results for axillary shoot proliferation. A maximum frequency of shoot initiation above 84%, with a mean of 8.9 and 4.8 shoots per node and shoot tip, respectively, was achieved on the culture medium supplemented with 4 µM TDZ. A combination of TDZ + NAA significantly increased the percentage of multiple shoot formation and number of shoots per explant. The best shoot induction response occurred on MS medium with 4 µM TDZ and 1 µM NAA. On this medium, the node (93.8%) and shoot tip (95.9%) explants produced an average of 17.7 and 8.6 shoots, respectively. The highest root induction frequency (97.4%) and number of roots per shoot (25.4), as well as the greatest root length (4.2 cm), were obtained on half-strength MS medium supplemented with 4 µM indole-3-butyric acid (IBA). The presence of six carotenoids and α-tocopherol in the leaf tissues of M. maritima was confirmed by HPLC. Gas chromatography-mass spectrometry analysis confirmed the presence of 10 fatty acids, including γ-linolenic acid and stearidonic acid in the leaf tissues of M. maritima. All-E-lutein (18.49 μg g−1 fresh weight, FW), α-tocopherol (3.82 μg g−1 FW) and α-linolenic acid (30.37%) were found to be the significant compounds in M. maritima. For the first time, a successful protocol has been established for the mass propagation of M. maritima with promising prospects for harnessing its bioactive reserves.

Highlights

Micropropagation is widely used technique for germplasm conservation and extensive propagation of various plants
We investigated the bioactive reservoirs such as carotenoid, tocopherol and fatty acid contents of in vitro-derived shoots
Shoots were produced within a week from both node and shoot tip explants when they were cultured on semisolid Murashige and Skoog (MS) medium supplemented with N6-benzyladenine (BA), kinetin (KN), or thidiazuron (TDZ)

Summary

Introduction

Micropropagation is widely used technique for germplasm conservation and extensive propagation of various plants. This method has been employed in several biotechnological applications including the production of bioactive compounds [1]. Mertensia maritima (L.) Gray (Boraginaceae), commonly known as the oyster plant, is a perennial herb native to northern Europe, including the British Isles [10]. It inhabits shingle beaches, and rarely sandy beaches, and is difficult to cultivate in the garden because of stringent temperature requirements [11]. There are no reports on the content and composition of carotenoids, fatty acids and tocopherols from tissues and organs of M. maritima

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