Metabolic profiling, in vitro propagation, and genetic assessment of the endangered rare plant Anarrhinum pubescens

Asmaa Abdelsalam,Ehab Mahran,Kamal Chowdhury,Arezue Boroujerdi

doi:10.1186/s43141-021-00210-6

Abstract

BackgroundAnarrhinum pubescens Fresen. (Plantaginaceae) is a rare plant, endemic to the Saint Catherine area, of South Sinai, Egypt. Earlier studies have reported the isolation of cytotoxic and anti-cholinesterase iridoid glucosides from the aerial parts of the plant. The present study aimed to investigate the chemical profiling of the wild plant shoots as well as establish efficient protocols for in vitro plant regeneration and proliferation with further assessment of the genetic stability of the in vitro regenerated plants. ResultsTwenty-seven metabolites have been identified in wild plant shoots using the Nuclear Magnetic Resonance (NMR) spectroscopy. The metabolites include alkaloids, amino acids, carbohydrates, organic acids, vitamins, and a phenol. In vitro propagation of the plant was carried out through nodal cutting-micropropagation and leaf segment-direct organogenesis. The best results were obtained when nodal cutting explants were cultured on Murashige and Skoog medium with Gamborg B5 vitamins supplemented with 6-benzylaminopurine (BAP) (1.0 mg/L) and naphthaleneacetic acid (NAA) (0.05 mg/L), which gave a shoot formation capacity of 100% and a mean number of shoots of 27.67 ± 1.4/explant. These shoots were successfully rooted and transferred to the greenhouse and the survival rate was 75%. Genetic fidelity evaluation of the micropropagated clones was carried out using random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) molecular markers. Jaccard’s similarity coefficient indicated a similarity as high as 98% and 95% from RAPD and ISSR markers, respectively. ConclusionsThis study provides the chemical profiling of the aerial part of Anarrhinum pubescens. Moreover, in vitro regeneration through different tissue culture techniques has been established for mass propagation of the plant, and the genetic fidelity of the in vitro regenerated plants was confirmed as well. Our work on the in vitro propagation of A. pubescens will be helpful in ex situ conservation and identification of bioactive metabolites.

Highlights

Anarrhinum pubescens Fresen. (Plantaginaceae) is a rare plant, endemic to the Saint Catherine area, of South Sinai, Egypt
We report the metabolic profiling, in vitro propagation, and callus induction for genus Anarrhinum, which could be an important basic reference for any further research concerned with chemical analysis and mass propagation of the A. pubescens plant or other species sharing the same genus
Metabolic profiling of the wild plant shoots The peaks from the 1H spectra were assigned using Chenomx software (8.6) and its 700 MHz database

Summary

Introduction

Anarrhinum pubescens Fresen. (Plantaginaceae) is a rare plant, endemic to the Saint Catherine area, of South Sinai, Egypt. (Plantaginaceae) is a rare plant, endemic to the Saint Catherine area, of South Sinai, Egypt. (Family: Plantaginaceae) is an endemic perennial plant that grows on granite rocks, in the Saint Catherine region of Egypt [1,2,3,4]. Phytochemical and biological analysis of the plant showed the presence of iridoid compounds which possess cytotoxic and acetylcholinesterase inhibitory activities [8, 9], suggesting the possible application of the plant in developing anticancer and Alzheimer disease relieving drugs. The genus Anarrhinum is characterized by the presence of iridoid glycoside compounds. These structurally diverse metabolites are well known for their biological activities, principally, the antimicrobial activities [10, 11]. Iridoid glycosides were isolated from Anarrhinum orientale and some of them showed a selective inhibition to hepatitis C virus protease [12, 13]

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