Kong. Res. J. 4(2): 84-88, 2017 ISSN 2349-2694

Kongunadu Arts and Science College, Coimbatore. 8 INSIGHT INTO PHARMACEUTICAL IMPORTANCE OF BRYOPHYTES

Greeshma, G.M.1, G.S. Manoj2 and K. Murugan1* 1Plant Biochemistry and Molecular Biology Lab, Dept. of Botany, University College, Thiruvananthapuram, 2Dept. of Botany, VTM NSS College, Dhanuvachapuram, Thiruvananthapuram, Kerala, India. .26524/krj20 *E.mail: [email protected]

ABSTRACT DOI:10 Historically, Bryophytes were accounted to be a monophyletic group and were placed in an inclusive Bryophyta. Some species are aquatic though some can adapt and live in arid regions. Bryophytes size ranges from microscopic to 12 inches in length, the average size is between 0.5 – 2 inches long and colors vary from green to black and sometimes colorless. Bryophytes plays a vital role in the biosphere even their size is insignificant. As a biotic factor in the environment, they provide food for numerous herbivorous birds and animals. They prevent soil erosion by carpeting the soil. Bryophytes cause the outer portion of rock to slowly crumble as they grow with lichens on rock surfaces. And because of it they contribute and help to soil formation. When mixed with the soil, bryophytes increase the water-holding capacity of the soil and the amount of organic matter in the soil. Some bryophytes like sphagnum or peat has some economic importance. It is used as packing material for breakable or fragile objects such as figurines and dinnerware’s. It is also used as packing materials for transporting plants and plant parts, since sphagnum holds water and hence prevent plants from drying during transport. As a whole, bryophytes are of little economic importance to man. Keywords: Bryophytes, medicinal, non-vascular, microhabitat.

1. INTRODUCTION 40 new carbon skeletal acetogenins, phenolic compounds and terpenoids have been found in this Bryophytes are the ancient terrestrial class, for example, marchantin A, marchantin E and chlorophyllous spore-forming amphibians, although riccardin C and diterpene dialdehyde, sacculatal, by no valid scientific documents for this have been recent development of spectroscopic apparatus, appeared in the literature. This concept was particularly by high resolution NMR techniques. The fundamentally based on the similarity of the present- most interesting chemistry of liverworts is that most day liverworts the first land plant fossils, the spores of sesqui- and diterpenoids are enantiomers of those of which date back almost 400 million years. They found in higher plants and some different species of are taxonomically kept between the algae and the the same genus, like Frullania produce both normal pteridophytes and there are appx. 14000 species of and its enantiomeric sesquiterpenoid. They are the Bryophyta, 6000 of Marchantiophyta (liverworts), source of large variety of secondary metabolites and 300 of Anthoceratophyta, species and 5000 are thus provide a great potential for biotechnological . Generally bryophytes are not infected or and applications. In past few attacked by pathogens, insects, snails, slugs and years more than 400 novel chemical compounds mammals, however, studies on their phytochemistry were isolated from bryophytes and they were have been neglected for centuries. Bryophytes are structurally elucidated. Some of biologically active considered to be useless for human diets, difficulty of compounds isolated from mosses includes identification and collection of a large amount, biflavonoids, terpenes and terpenoids (like di- and although several liverworts have been used as triterpenoids) and flavonoids whereas liverworts medicinal plants in China to cure cut, burns, bruises, reported to contain a large variety of lipophilic pulmonary tuberculosis, neurasthenia, fractures, mono-, di- and sesquiterpenoids as well as aromatic convulsions, ulopathy, neurasthenia etc. compounds like bibenzyls, benzoates, cinnamates 2. REVIEW OF THERAPEUTICAL IMPORTANCE and naphtalenes. Flavonoids such as quercetin (182.5 µg/g), luteolin (464.5 µg/g), and apigenin 2.1. Phytochemicals from bryophytes (297.5 µg/g) were reported to be present in the Bryophytes are known to produce diverse liverwort Marchantia linearis (Remya et al., 2014). secondary metabolites to combat a number of biotic Two moss species Thuidium tamariscellum and and abiotic stress such as predation, UV radiation, Brachythecium buchananii were reported to be rich extreme temperature and microbial decomposition. in secondary metabolites like flavonoids, terpenoids

84 and phenols (Greeshma et al., 2016). In spite of the 2.3. Microbicidal potential fact that many plant secondary metabolites are the In recent year extensive studies have been potential therapeutic introduction of novel drugs in conducted for the search of antibacterial properties the market has decreased in past few years. In fact, in different species of plant. Organic extracts of higher plants and bryophytes have similar various medicinal plants containing flavonoids have evolutionary history but search for novel therapeutic been reported to show antimicrobial activity compounds within biodiversity of bryophyte (Praveen Dahiya et al., 2012). The antibacterial remained neglected due to small size and lack of activities of isofavonoids and flavonoids, and awareness among people. These small plants glycosides of luteolin and apigenin have also been remained unexploited so far in drug discovery reported. But most of the investigations were process in spite of few reports from the past depicts centered on angiosperms. Few data are presently some of their ethnomedicinal uses. Studies on available about these smaller groups of plants, secondary metabolites of bryophytes have revealed bryophytes. However, few of the recent study on the presence of few original compounds, some of bryophytes has shown some of the antibacterial which are not synthesized by higher plants (Rashmi activity against gram-positive and gram-negative et al., 2014). bacteria (Meenakshi et al., 2011). 2.2. Antimicrobial aspects of Bryophytes Also, various phenolic compounds isolated Bryophytes have been reported as from Atrichum, Dicranum, Mnium, Polytrichum, and antibiotics (Tedela et al., 2014). Various organic Sphagnum spp. are known to shown antimicrobial solvent extracts of bryophytes have been properties. Apart from this the antimicrobial activity investigated in past. Literature emphasizes that the for three moss species Eurhynchium angustirete, alcoholic and the aqueous extracts or the various Rhytidia delphussquarrosus and Rhodo bryumroseum, compound isolated from different species of and for two liverwort species Frullania dilatata and bryophytes (hepatic and mosses) have shown Lophocolea heterophylla has been reported for the antimicrobial effects against various group of fungi, first time. Similarly, aqueous extract of few as well as Gram negative and Gram positive bacteria. bryophytes have some inhibitory effect on the Recently extracts of few of the selected species of growth of Escherichia coli as tested on plates. bryophytes (seven mosses and three liverworts) viz However, this antibacterial activity seems to be the Radula flacida, Cyatodium africanum, Frullania specific for certain bryophyte species, as the extracts spongiosa, Thuidium gratum, Ectropothecium of Marchantia polymorpha, Porella platyphylla and aeruginosum, Sematophyllum caespitosum, the moss Dicranum scoparium showed antimicrobial Stereophyllum radiculosum, Babulalam berenensis, effects on the gram-positive bacteria namely Bacillus Campilopusa spericuspis and Calympereserosumlam subtilis, Staphylococcus aureus and Sarcina lutea, but berenensis, Campilopusa spericuspis and Calymperes no activity against gram-negative E. coli (Rashmi et erosum have shown interesting antimicrobial activity al., 2014). (Rashmi et al., 2014). Similarly, the methanolic and Many of the bryophyte species are also water fractions of Targionia hypophylla possess known to show antifungal property (Abhijit et al., strong microbicidal activity (Remya et al., 2012). 2011). Different crops growing in greenhouses, like The methanolic and aqueous extracts of Plagiochilla tomatoes, wheat and green pepper were infected beddomei, Leucobryum bowringii and Octoblepharum with the pathogens Phytophora infestans, Erysiphe albidum exhibited different degree of growth graminis and Botrytis cinerea and later treated with inhibition against bacterial species such as alcoholic extracts from different bryophytes. All Salmonella typhimurium, Staphylococcus aureus, bryophyte extracts showed a species-specific Klebsiella pneumonia, E.coli, Bacillus cereus and antifungal activity against the plant pathogenic fungi Pseudomonas aeruginosa and fungal species like depending on the concentration. Ethanolic extract of Candida albicans, Cryptococcus neoformans, Marchantia linearis exerted a potent fungicidal Trichophyton rubrum, Mucor indicus, Apergillus niger activity against Botrytis cinerea and marginally in and A. flavus (Manoj, 2012). Therefore, the potential Rhizoctonia solani i.e., showing varied levels of antimicrobial properties of bryophyte can be growth inhibition with different concentrations. harnessed for the therapeutic purpose against the (Remya et al., 2014). Furthermore, extracts from respective pathogen. The antibacterial and Neckera crispa and Porella obtusata showed antifungal activities of bryophytes have been antifeeding effects against the Portuguese slug discussed in below. Aarion lusitanicus. In view of the above features that bryophyte extracts showed fungicidal and

85 antifeedant effects, a commercial product was 2014). According to the some of the recent report developed and is sold as natural pesticide. Apart bryophytes are the source of numerous chemical from this studies conducted in the past have compounds of biotechnological and revealed few of the compounds isolated from biopharmaceutical interest. Several secondary bryophytes extracts have shown reversal of metabolites have been isolated so far from different conventional antibiotic resistance development in species but the mechanisms behind their activity are pathogenic fungi. Therefore, the problem of drug still widely unexplored. resistance development in pathogenic fungi can be 2.5. Antioxidant potential solved easily (Rashmi et al., 2014). Few of the bryophyte species have been Also, in addition to this studies conducted studied in context to antioxidant activity. Recent on one of the model species of moss Physcomitrella study suggests that some of the liverworts and moss patens revealed that this moss under axenic possesses strong antioxidative machinery which condition produces a tetracyclic diterpene, namely helps them to survive in the extreme climate and 16α–hydroxykaurane (16 α-hydroxy-ent-kaurane, stress condition. Heavy metal, desiccation and Kaurenol, C H O) (Anna et al., 2010). Although, the 20 34 ultraviolet radiation have been found to trigger an utility of 16 α–hydroxykaurane is not yet revealed, array of different enzymes in bryophytes (Rashmi et but it is presumed to be bioactive. This, compound is al., 2014). Few of the bryophyte species have been known to be produced by lichen species and fungi found to hyper accumulate metals and few others and it is commonly known from Gibberella fujikuroi, were able to sequester the toxic metals. The study a plant pathogenic fungus that infects rice plants and conducted on antioxidant activity of the Antarctic causes foolish rice seedling disease. As recently mosses Sanionia uncinata and Polytrichastrum shown, 16α–hydroxykaurane is involved in spore alpinum var. alpinum has indicated their potential to germination in and leads to a be used as antioxidants for medicinal and cosmetic complete inhibition of spore germination when purpose. The methanolic extracts of Plagiochilla applied in high concentrations (2-3 µM) (Anna et al., beddomei, Leucobryum bowringii and Octoblepharum 2010). Also, few bryophyte extracts have been found albidum possess potential antioxidant properties to be effective on human pathogenic fungi although (Manoj, 2012). Also, in addition the antioxidant the bioactive compounds may cause allergenic activities of some of the species of bryophyte like effects and dermatitis in few cases. Nevertheless due Atrichum undulatum. Polytrichum formosum, to risk of allergic reactions, bryophyte extracts were Pleurozium schreberi and Thudium tamariscinium not recommended for scientific medicinal use so far. has been screened, and all tested species have 2.4. Phytochemicals vs pharmacological potential showed antioxidant effects lower than the positive control, caffeic acid (Boris et al., 2012). Moreover, Since bryophytes are the reservoir of the screening for the antioxidant property of the complex secondary metabolites, their vast aqueous extract of the three moss namely application in traditional medicine is not astonishing. Brachythecium rutabulum, Calliergonella cuspidate A large number of bryophytes are used as medicines and Hypnum mammillatum in context of their ABTS in homeopathy. About 3.2% of mosses and 8.8% of (2, 2- azino- bis ( 3- ethlybenzthiozoline- 6 sulphonic liverworts taxa have been chemically investigated. acid) cation scavenging activities and phenolic Species like Sphagnum, Marchantia, Riccia, Barbula, content have known to show some positive Bryum, Octeblepharum and Fontinalis are used to response. Out of the three extracts, Brachythecium treat different diseases, including cardiovascular rutabulum have shown the highest of the phenolic diseases, inflammation, fever, lung diseases, content which further suggested potential of this infections, wounds and skin diseases (Rashmi et al., extract in search of many other novel antioxidant 2014). In China more than 30 species can be bought compounds in this moss. Apart from this methanolic at the local pharmacist 66 and around 40 different and and ethyl acetate extract of Marchantia kinds of bryophytes have been used to treat diseases polymorpha have also shown antioxidant property. of cardiovascular system, tonsillitis, tympanitis Summing up, bryophyte could be the source of many cystitis and bronchitis and to cure skin disease and novel antioxidants if screened which could be used burns. Many of the species for example Polytrichum for novel drug discovery. commune which is used as antipyretic and anti- inflammatory agent or boiled as a tea for treating the 2.6. Biopharming for phytochemicals cold. Rhodobryum giganteum is another species Bryophytes have indeed penetrated the fore traditionally used to treat, other diseases like front of modern medicines. Although a vast variety of cardiovascular diseases or angina (Rashmi et al., has been produced in microbial

86 or mammalian cells but plants based production of herbal compounds have been discovered with system possesses several advantages over the immense therapeutic potential. Therefore, to meet mammalian and microbial system thus, making them the potential future demand for various bioactive interesting alternatives. Microbial systems are compounds used as drugs, a new production system favored because of easy cultivation and high is required significantly. Bryophyte, a small and productivity, whereas mammalian cell lines apparently insignificant group of plants may serve as (preferentially Chinese Hamster Ovary cells) are a source of some unique biologically active favored for complex multimeric proteins or those molecules. Many of the bryophytes are important requiring posttranslational modifications (Rashmi et source of medicine, antibacterial and antifungal al., 2014). In contrast to these currently used agents. Antifungal efficacy of certain liverworts and systems, plants based production system possesses mosses can substitute the conventional synthetic several advantages over these system thus making fungicides used in crop protection especially in the them interesting alternatives. As higher eukaryotes, countries where fugal invasion in the crop fields is a they perform posttranslational modifications closely common phenomenon. The problem of development resembling those of humans, thus minimizing the of drug resistance in common human pathogenic risk of product contamination by infectious agents fungi can be solved by using antifungal compounds derive from the used cells or media (Rashmi et al., harvested from uncommon sources like bryophytes. 2014). Also bryophytes offer the researchers and the Several bryophytes are able to produce antifungal company a high production system which can be compounds. Furthermore, the use of moos grown without antibiotics, hence avoiding the has opened new possibilities for the danger of contamination of the final product. Apart production of many plant and animal metabolites. from these advantages, mosses are the only plants Future scope: In the past few years, rapid known to show a high frequency of homologous progress has been made to isolate various plant recombination. They allow stable integration of based therapeutic compounds. Bryophytes being inserted genes into the host cell. Furthermore, the rich source of a variety of secondary metabolites highly complex moss system, compared to bacteria could be a promising source of the bioactive and fungi, permits a much wider array of expression compounds with immense therapeutic potential. than is possible in other systems. In view of the Being present in varied niche and occupying the above advantages of mosses over other production most diverse group of plant kingdom, they could be system, today, many complex biopharmaceuticals the source of various evolved metabolic pathways are being produced by moss . The Chair that could be wisely manipulated for the of plant Biotechnology from the University of development of various novel therapeutic Freiburg, Germany, and the biopharmaceutical compounds. Therefore, bioprospecting of company Greenovation Biotech Gmbh in Heilbronn, bryophytes is required to discover the natural Germany; have started a cooperation to enhance the wealth of bryophytes. Creation and development of yield of recombinant proteins from moss. The moss production system by using bryophyte cells could Physcomitrella patens has been successfully grown in solve the future demand of novel plant based a bioreactor which require only water and minerals production system. Hence, engineering of metabolic to nourish the moss, in the presence of light and CO 2 pathway for production of novel metabolites, and (Greenovation). Consequently many complex strategies for the development of the bioprocess for proteins can be produced in moss bioreactor. Other bryophyte cell system is the need of time to dig out products are human growth factor that is required some more information to satisfy the thirst of novel by the researcher for tissue culture. This plant has drug discovery. successfully been able to produce human proteins and is the only plant being used to produce the REFERENCES blood-clotting factor IX for pharmaceutical use. Abhijit, Dey and Jithendra Nath De, (2011). 3. CONCLUSION Antifungal bryophytes. Possible role against human pathogens and in plant protection. Res. J. Use of medicinal plants has been Bot. 6(4):129-140. appreciated due to low cost and lesser side effects. Herbal drugs have been used successfully in the Anna, K.B., L.D. Eva, F.Wolfgang, L.Daniel, V.S. Marco, treatment of various ailments over the last few D.Z. Andreas and , (2010). Applied decades. Development of drug resistance in Bryology – Bryotechnology. Trop. Bryol. 31: 22- pathogens is one of the major problems in medicine. 32. Natural products derived from the botanicals can be used as a substitute to solve the problem. A number

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