In Vitro- Exploration of Fungal Endophytes of Egyptian Cynara Scolymus L
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Global NEST Journal, Vol 21, No 3, pp 296-308 Copyright© 2019 Global NEST Printed in Greece. All rights reserved In vitro- exploration of fungal endophytes of Egyptian Cynara scolymus L. (artichoke) and investigation of some their bioactive potentials Nermien H.S.1,2,*, Thabet A.3,4, Markeb A.A.3, Sayed D.M.5 and El-Maali N.A.3 1Department of Botany and Microbiology, Faculty of Science, Assiut University Assiut 71516, Egypt 2College of Applied Medical Sciences – Jubail, Imam Abdulrahman Bin Faisal University, Dammam, KSA 3Department of Chemistry, Faculty of Science, Assiut University, Assiut 71516, Egypt 4Department of Chemistry, Faculty of Science, Sana’a University, Sana’a, Yemen 5Department of Oncological Clinical Pathology, South Egypt Cancer Institute, Assiut University, Assiut 71516, Egypt Received: 23/07/2018, Accepted: 09/11/2018, Available online: 21/11/2018 *to whom all correspondence should be addressed: e-mail: [email protected], [email protected] https://doi.org/10.30955/gnj.002836 Abstract prevention and treatment of different diseases and thereby avoid the adverse effects of organically Plant-associated microorganisms, especially endophytic synthesized drugs as well as the high-cost drug therapy fungi, represent an untapped resource for the discovery of biologically active natural products. The objectives of this (Roghani-Dehkordi and Kamkhah, 2009). Medicinal plants study were to isolate, identify endophytic fungi, and are the richest bio-resource of drugs from traditional produce their bioactive metabolites from the leaves of systems and modern medicines, nutraceuticals, food two varieties of Egyptian artichoke namely: French supplements, folk medicines, pharmaceutical Hyrious and Egyptian Baladi. In addition, assess of their intermediates and chemical entities for synthetic drugs. total antioxidant capacity (TAC), total phenolic content World Health Organization (WHO) encourages, advises (TPC) and total flavonoid content (TFC). The results of this and promotes herbal remedies in national health care novel study show a total of 35 endophytic fungal species programmers due to the low cost of these drugs (Amita belonging to 14 genera were isolated from both artichoke Pandey, 2014). Recently, medicinal plants have received leaves with gross total counts of colonizing endophytic the attention of the pharmaceutical and scientific fungi ranged from 71 to 123 cfu which is matching 78.89% societies. The biomolecules of pharmaceutical and to 136.67% of colonization frequency. All taxa recovered nutraceutical interest are mainly represented by were assigned to Ascomycetes. In addition, there is high flavonoids and phenolic acids, particularly caffeic acid and species richness and diversity indices of endophytic its derivates mono- and bi-caffeoylquinic acids (Saez et al., filamentous fungi in the leaves Baladi Artichoke 2013). Recent studies have reported that bioactive as compared to its French rival. Alternaria alternata phenolic constituents from plants and endophytic microbe were found to be the most frequently isolated have a wide variety of biological activities, such as dominant species. The TAC, TPC and TFC of the fungal antioxidants, hepatoprotective, anti-inflammatory, cultures ranged from 163 to 681 mgAAE/gDW, 10.38 to antifibrosis, and antiviral activities, and anticarcinogenic 40.30 mgGAE/gDW, and 13.92 to 173.55 mgQE/gDW, effects against different cancer cell line (Chutulo and respectively. Furthermore, LC-ESI-MS/MS confirmed Chalannavar, 2018; Gebhardt and Fausel, 1997; Liang the presence of 1,3-dicaffeoylquinic acid and et al., 2012; Llorach et al., 2002; Sihem DABBOU et al., 1,5-dicaffeoylquinic acid in the methanolic extract of A. 2017; Simona Spînu et al., 2017; Wang et al., 2014). Also, alternata. Hence, this novel study suggested that the Plant-derived bioactive compounds have achieved an metabolites produced by endophytic fungi associated with extensive interest due to their versatile applications and Egyptian artichoke could be explored as an economic and beneficial effects; for instance, antioxidant activity, potential natural resources with diverse pharmaceutical digestive stimulation action, anti-inflammatory, and biological activities. antimicrobial, hypolipidemic, antimutagenic effects and Keywords: Artichoke, fungal endophytes, antioxidant anticarcinogenic anticarcinogenic potential (Jacociunas capacity, total flavonoids, total phenolics contents. et al., 2013; Wojdylo et al., 2007), on health promotion because the low side effects in comparison to the 1. Introduction synthetic drugs (Farag et al., 2013; Ncube and Afolayan, As an alternative to drug therapy, it is desirable to use 2008). Th. Thus, many publications recorded its medicinal foods, food components, and/or medicinal plants for the values as natural compounds. Nermien H.S., Thabet A., Markeb A.A., Sayed D.M. and El-Maali N.A. (2019), In vitro- exploration of fungal endophytes of Egyptian Cynara scolymus L. (artichoke) and investigation of some their bioactive potentials, Global NEST Journal, 21(3), 297-309. IN VITRO- EXPLORATION OF FUNGAL ENDOPHYTES OF EGYPTIAN CYNARA SCOLYMUS L. (ARTICHOKE) 297 Artichoke (Cynara scolymus L.) is an ancient herbaceous more economically to produce (Chen et al., 2010). perennial plant species belonging to the Asteraceae Metabolites isolated from the fungal endophytes are good family, genus Cynara (Farag et al., 2013; Xia et al., 2014), sources of novel secondary metabolic products possess originating from the Mediterranean region, and it is now diverse and unique structural groups (Chutulo and cultivated worldwide. According to FAO estimation, Italy Chalannavar, 2018; Schulz et al., 2002; Sharma et al., was the world’s largest artichoke producer in 2016 2016). Numerous work has been focused on the bioactive followed by Egypt, Spain Peru (FAO, 2016). The artichoke potential of endophytes, such as antiviral, antibacterial, was used as food and medicine by ancient Egyptians, antifungal, anticancer, antiviral, insecticide, antidiabetic Greeks, and Romans. Three Cynara scolymus (artichoke) and immunosuppressive activities, but few is known about cultivars: American Green Globe, French Hyrious, and their antioxidant capacity (Yadav et al., 2014). Shukla et al. Egyptian Baladi have been cultivated in the Northern part was isolating endophytic fungi from Ocimum sanctum and of Egypt (Farag et al., 2013). Moreover, the artichoke is showing their antioxidant activity. A strong antioxidant one of the oldest medicinal plants with multiple health activity was found in the endophytic fungus of Phyllosticta benefits (Xia et al., 2014). Artichoke has many natural sp. isolated from Guazumato mentos plant. Therefore, by antioxidants and multiple pharmacological actions. fermentation an endophytic fungus isolated from The antioxidant properties of artichoke are thought Artichoke (Cynara scolymus L.) can produce bioactive to be related to its abundant phenolic composition compounds such as chlorogenic acid, caffeoylquinic acids, (El Morsy and Kamel, 2014). Recent studies have reported caffeic acid and its derivatives (apigenin and luteolin) just that Artichoke has antitoxic activity (Heidarian like its host plant. This will protect the plant from and Rafieian-Kopaei, 2013), cholesterol-reducing effect extinction. Thus, an immense opportunity to discover (Kusku-Kiraz et al., 2010) anticarcinogenic, antigenotoxic, novel natural products from hitherto less investigated hepatoprotective, bile expelling, diuretic, and anti- fungal endophytes that reside in hitherto assumed inflammatory, as well as anti-HIV, antifungal, and medicinal plants. Also, microbial natural products antibacterial (Abu-Reidah et al., 2013). Additionally, its represent a huge and largely important resource of leaves are used in traditional medicine as an herbal unique chemical structures and probably play an medicament for the treatment of hepatitis, choleretic, important role for adaptation as a response to habitat diuretic, hyperlipidemia, obesity and dyspeptic disorders (Gunatilaka, 2006). (Nassar et al., 2013). Artichoke was ranked fourth out of Based on this theory, the aim of this study is: (1) to isolate more than 1000 food products in antioxidant content, and and identify each endophytic fungus from two artichoke the first among several selected vegetable crops (Yoo cultivars (French Hyrious and Egyptian Baladi), (2) to et al., 2012). Artichoke by-products (leaves, external investigate the total polyphenols and the total flavonoids bracts and stems) represent an enormous amount of content and (3) to evaluate the antioxidant activities using discarded materials (representing 80−85% of the biomass) in vitro study of the Methanolic crude extracts of (Lattanzio et al., 2009). Recently, the possibility to recover C. scolymus L. and its fungal endophytes. To the best of them has been proposed for economic, environmental our knowledge, no previous work on fungal endophytes concerns, adding value to agro-industrial by-products and its metabolites investigated from Artichoke plant, so (Ruiz-Cano this research considered a new record on this et al., 2014). investigation. By implementation of our present work, it Endophytic fungi residing internally in the healthy living becomes easier, cheaper and time consuming to get the plants tissues without causing any immediate harm to bioactive compounds (like flavonoids, phenolic their host (Yadav et al., 2014). All higher plants host one compound, anti-microbial or anti-cancer compounds) of or more