International Journal of Research in Pharmacy and Biosciences Volume 7, Issue 1, 2020, PP 10-15 ISSN 2394-5885 (Print) and ISSN 2394-5893 (Online)

The Exceptional Endophytic Fungi, Emericella (Berk.) and (Sacc.) Genera

Waill A. Elkhateeb*, Ghoson M. Daba Chemistry of Natural and Microbial Products Department, Pharmaceutical Industries Researches Division, National Research Centre, El Buhouth St., Dokki, 12311, Giza, Egypt *Corresponding Author: Waill A. Elkhateeb, Chemistry of Natural and Microbial Products Department, Pharmaceutical Industries Researches Division, National Research Centre, El Buhouth St., Dokki, 12311, Giza, Egypt. Email: [email protected]

ABSTRACT Fungi are rich sources of biologically active natural compounds, which are used in the manufacturing of wide range of clinically important drugs. Fungi produce important antibiotics such as the beta-lactam antibiotics members; penicillin and cephalosporin, which and their derivatives are dominating the most important antibiotic market until now. Endophytic fungi attracted attention due to their production of bioactive and chemically novel compounds that have medical, agriculture and industrial applications. This review focused on Emericella (Berk.) and Phoma (Sacc.) as models of endophytic fungi rich in therapeutic agents that have known medicinal application. Moreover, encourage further studies of metabolites produced by those potent fungi to discover additional applications. Keywords: Phoma; Emericella; biological activities; endophytic fungi; metabolites.

INTRODUCTION Emericella (Berk.) Description and Ecology The application of microbial secondary metabolites Aspergillus section Nidulantes includes species in various fields of biotechnology has attracted with striking morphological characters, such as the interests of researchers. Endophytic fungi biseriate conidiophores with brown-pigmented are the fungi residing symbiotically inside the stipes, and if present, the production of plant without causing any harmful effect to the ascomata embedded in masses of Hülle cells with often reddish brown ascospores. Based on plant. Phoma is an endophytic polyphyletic genus this approach, section Nidulantes is subdivided belonging to the phylum , class in seven clades and 65 species, and 10 species. , order , and family Emericella (Eidam) Vuill. Anamorph: Aspergillus Didymellaceae. Phoma (Sacc.) is a worldwide forms white cleistothecia surrounded by Hülle , which has been reported from plants, cells (Thick walled refractile cells like chlamy- soil and air. The genus Phoma was introduced doconidia) and produces purple ascospores. more than 170 years ago. More than 2000 Conidiophores usually have short brown stipes, species have been studied (Montel et al., 1991; bear both metulae and phialides, and produce Elkhateeb, 2005; Elkhateeb et al., 2016; Bennett columns of dark green conidia. et al., 2018; Daba et al., 2018). On the other Only one species, Emericella nidulans, is hand, the genus Emericella Berk. Belonging also common in foods. Emericella nidulans (Eidam) to the phylum Ascomycota, is one of the sexual Vuill. Anamorph: Aspergillus nidulans (Eidam) stages associated with Aspergillus. G. Winter Emericella nidulans, colonies may be The Endophytic Emericella is an important dark grass green if conidial heads are abundantly formed, or creamish if cleistothecia and Hülle source of natural products, exhibiting a wide cells are present. Violet or pink soluble pigment range of biological activities (Tarawneh et al., may be produced, and the reverse of the 2013; Majik et al., 2014; Ha was et al., 2016). In colonies is usually brightly coloured in shades this review, description, ecology, applications of pink, orange, brown or violet brown. This and biological activities exhibited by those species grows rapidly at 37 °C. Emericella important endophytic fungi have been discussed. nidulans grows optimum at 35–37 °C (Elkhateeb,

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2005; Pitt and Hocking, 2009; Elkhateeb et al., of this group have important biological activities 2016; Chen et al., 2016; Elkhateeb and Daba, (Chiang et al., 2009; Nadumane et al., 2018; Daba et al., 2018). 2016).The genus Emericella fungi produces a remarkable diversity of secondary metabolites, Emericella Species Benefits and Uses such as indole alkaloids, prenylated polyketides, Emericella nidulans is known for its industrial benzophenones, and xanthones (Fig. 1), with and medical significance. Emericella nidulans interesting biological properties thus representing was the source of a novel polyketide potential leads for the developing of new asperfuranone (Fig. 1). Polyketides are one of pharmaceutical agents (Kralj et al., 2006; the major classes of natural products and many Figueroa et al., 2009).

Figure1. Structure of the some compounds produced by Emericella Emericella Species as Antimicrobial of a epipolythio-dioxopiperazines (ETPs) group that exhibited cytototoxic activity against T47D, Emericella nidulans can exert inhibitory activity HeLa and WiDr cells. against the mycelial growth and spore production of F. oxysporum. It was reported that Hawas et al., (2012) revealed that the ethyl acetate methanolic extract of E. nidulans is more extract of culture broth of the endophytic effective than n-hexane and ethyl acetate crude Emericella nidulans possessed promising anti- extracts in inhibiting both mycelial growth and HCV protease activity and selective anticancer spore production (Sibounnavong et al., 2009; activity against liver cancer cell lines, which Khalil et al., 2014). In another study, crude may be attributed to the presence of adenosine extracted from endophytic fungus Emericella class compounds. Cultivation of this fungus in qaudrilineata possessing antibacterial activity Czapek’s peptone media led to the isolation of was subjected to bioassay guided fractionation five known metabolites: sterigmatocystin, in which toluene fraction exhibited maximum emericellin, cordycepin, ergosterol peroxide, antibacterial activity against Staphylococcus aureus and myristic acid (Fig. 1) from the ethyl acetate (gram positive) and Aeromonas hydrophilla (gram extract of the culture broth (Hawas et al., 2012). negative) respectively (Goutam et al., 2016). Emericella nidulans fungal strains were isolated and purified from ten Egyptian medicinal plants Emericella Species as Anticancer and Antiviral and their culture broth extracts were explored Nursid et al., (2011) reported extraction of an for HCV protease inhibitory activity and epipolythio-dioxypiperazine (emestrin A) from cytotoxicity. The ethyl acetate extracts of mycelium of Emericella nidulans. Emestrin A Emericella nidulans RPL-21showed the most (Fig. 1) exhibited anticancer properties against potent inhibition of HCV NS3/4A protease. The several cancer cell lines. Nursid et al., (2015) extracts of Emericella nidulans RSL24, mentioned that marine fungi Emericella Emericella nidulans RPL-21, exhibited strong nidulans produced emestrin B (Fig.1), a member cytotoxic activity against human breast cancer

11 International Journal of Research in Pharmacy and Biosciences V7 ● I1 ● 2020 The Exceptional Endophytic Fungi, Emericella (Berk.) and Phoma (Sacc.) Genera cell lines (MCF-7). Emericella nidulans RSS- (Rai et al., 2009).Fungi of the genus Phoma are 22, Emericella nidulans RSL24, displayed a reported from different plants as phytopathogens potent cytotoxic effect on human liver cancer (Kövics et al., 1999), as saprophytes from soils cell lines (HEP-G2). Emericellin, shamixanthone, (Elkhateeb et al., 2016). arugosin C were isolated from the ethyl acetate Phoma (Sacc.) Benefits and Uses extract of Emericella nidulans RPL-21 (El- Kassem et al., 2019). In recent years, the main goal of both pharmaceutical and agrochemical industries is Phoma (Sacc.) Description and Ecology the discovery of new natural products, which Phoma species colonies are greenish gray to can be used as medicine as well as environmental brown; some species may develop red or pinkish friendly agrochemicals. There are many secondary pigmentation. The growth rate varies widely metabolites like: phomodione, cercosporamide depending on the species. Dark, globose to Phomenon, Phomin, and Cytochalasins, subglobose to pyriform pycnidia with single or Dehydrophomin secreted by Phomaspp, which multiple ostioles occur on or immersed in the have already demonstrated antiviral, antifungal, agar and may form singly or in aggregates. The antibacterial, antiprotozoal, anticancer and hyaline conidiogenous cells lining the inner weedicide activities, and therefore, can be joined walls of the pycnidia produce their conidia by for the biological control of harmful weeds and percurrent proliferation, which are then extruded also against different microbes. in slimy masses. The variably shaped conidia Phoma Species as Bioherbicide Activities are unicellular and hyaline. Alternarioid chlamydoconidia are seen in some species Many microorganisms can be used for (Boerema et al., 2004; Elkhateeb, 2005; Aveskamp bioherbicide production, including fungi from et al., 2008). Phoma Spp. identification determined the genera Phoma sp. (Klaic et al., 2013; Klaic by morphological characteristics, such as the et al., 2017). Parra et al., (2005) optimized the formation of conidia (asexual spores), pycnidia production of secondary metabolites of Phoma (asexual fruiting bodies), and chlamydospores sp. with herbicide activity. Bailey et al., (2011) (enlarged, thick-walled vegetative cells within evaluated the action of Phoma macro stoma as hyphae or at hyphal tips) (Guégan et al., 2016; bioherbicide. Various species of the genus Choi and Kim, 2017). Phoma produce numerous important secondary metabolites, which includes phytotoxins, There are two major groups of Phoma, viz., antimicrobials, and mycoherbicides. Different terrestrial and marine. Recently researchers all phytotoxins like Deoxaphomin, Phomenon, over the world have focused on marine-derived Phomin, Cytochalasins, Dehydrophomin and p- Phoma for their bioactive compounds. The hydroxybenzaldehyde produced by Phoma spp. marine Phoma are very rich sources for novel were found to be effective herbicidal agents. bioactive secondary metabolites, which could The potential of these biocontrol agents have potentially be used as drugs. Recently, a large been studied by Cimmino et al., (2008). number of structurally unique metabolites with potential biological and pharmacological Phoma Species as Antimicrobial Activities activities have been isolated from the marine An endophytic Phoma species from guinea Phoma species (Fig. 2), mainly Phoma (Saurauia scaberrinae) showed to produce herbarum, P. sorghina, and P. tropica. These important metabolites such as usnic acid, metabolites mainly include diterpenes, enolides, phomodione, and cercosporamide in the culture lactones, quinine, phthalate, and anthraquinone. broth. These compounds demonstrated remarkable Most of these compounds possess antimicrobial, activity against Staphylococcus aureus, Pythium anticancer, radical scavenging, and cytotoxic ultimum, Sclerotinia sclerotiorum, and properties (Rai et al., 2018). Rhizoctonia solani (Hoffman et al., 2008). Some species of Phoma like P. herbarum, P. Antibacterial activity was tested by Bhimba et exigua var. exigua, P. glomerata, Phoma al., (2012), using ethyl acetate extract from macdonaldii, Phoma tracheiphila, Phoma Phoma herbarum, give maximum activity in proboscis, P. multirostrata, and Phoma foveata 100ul concentration against Micrococcus secrete phytotoxin and anthraquinone pigments leuteus and Vibreo cholerae. Thus secondary as secondary metabolites, which have great metabolites of the marine derived fungi Phoma potential for the biological control of weeds, and herbarum have a promising potential of can be exploited for the production of antibacterial activity to be included in drug mycopesticides, agrophytochemicals, and dyes. discovery. The endophytic Phoma species

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ZJWCFOO6 isolated from Arisaema erubescens activities, while cercosporaminde, β – Sitosterol in China produce different metabolites that trichodermin possess broad spectrum of possess strong and moderate antifungal and antifungal and antitumor activities (Wang et al., cytotoxic activities. A compound (3 S)-3, 6, 7- 2012). trihydroxy-α-tetralone, showed antifungal

Figure2. Structure of the some compounds produced by Phoma Phoma Species as Anticancer Activities support/replace nowadays spreading diseases. Endophytic fungi are promising candidates for Cytochalasin derivatives and cytochalasin B such mission specially the two genera Phoma were isolated from Phoma sp. recovered from and Emericella. Further studies are required to jellyfish (Nemopilema nomurai) which showed uncover the real therapeutic potency as well as cytotoxic activity against human solid tumor industrial applications of such powerful fungi. cell lines (A549, SKOV-3, SK-MEL-2, XF 498, and HCT15) and HeLa cell line, respectively CONFLICT OF INTEREST (Kim et al., 2012).Phoma sensu lato is currently The authors declare that they have no competing considered as one of the most prominent groups of endophytes and abundant source of novel interests bioactive compounds with potential for REFERENCES application in medicine, and industry (Nisa et al., 2015). [1] Aveskamp MM, De Gruyter J, Crous PW. (2008). Biology and recent developments in the Mycologists predict that less than 10% of all systematics of Phoma, a complex genus of fungal species have been isolated so far, major quarantine significance. Fungal indicating a huge potential for further discovery Diversity, 31: 1-18.‏ of novel bioactive chemical scaffolds. The [2] Bailey KL, Pitt WM, Falk S, Derby J. (2011). cytochalasins are produced by many fungal The effects of Phoma macrostoma on nontarget genera including Aspergillus, Emericella, plant and target weed species. Biological Hypoxylon, Metarrhizium, Zygosporium, Control, 58: 379–386. Hypocrella and Phoma. Many of the [3] Bennett A, Ponder MM, Garcia-Diaz J. (2018). cytochalasins have shown inhibitory activities Phoma infections: classification, potential food towards lung cancer A-549 (Bladt et al., 2013). sources, and their clinical impact. Microorganisms, 6(3): 58.‏ CONCLUSION [4] Bhimba BV, Pushpam AC, Arumugam P, Searching for sources of bioactive metabolites is Prakash S. (2012). Phthalate derivatives from of current serious importance in order to the marine fungi Phoma herbarum

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Citation: Waill A. Elkhateeb, Ghoson M. Daba, "The Exceptional Endophytic Fungi, Emericella (Berk.) and Phoma (Sacc.) Genera ", International Journal of Research in Pharmacy and Biosciences, 2020, 7(1), pp. 10-15. Copyright: © 2019 Waill A. Elkhateeb. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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