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Volatile chemical composition of Octoblepharum albidum Hedw. (Bryophyta) from the Brazilian Amazon
BMC Chemistry volume 16, Article number: 76 (2022)
Bryophytes have a variety of bioactive compounds that can be used in biotechnological processes. The objective of this study was to know the volatile chemical composition of Octoblepharum albidum Hedw. from the Amazon and investigate its association with possible bioactive effects on insects. The volatile concentrate of O. albidum was obtained by micro-scale simultaneous distillation–extraction and analyzed by gas chromatography coupled to mass spectrometry and the identification of the compounds was based on system libraries and specialized literature. Twelve organic compounds (92.44% of the total) were identified. Hexadecanoic acid, oleic acid, E-isoeugenol, 1-octen-3-ol, and stearic acid were the major compounds. Most of the compounds have already been reported from bryophytes, while others have an unprecedented occurrence in the group. All identified compounds have biological activities reported in the literature and may participate in plant defense mechanisms against insects, causing mortality or developmental inhibition. In this study, we describe for the first time the volatile chemical composition of O. albidum from Brazil and provide evidence that this species is a source of bioactive compounds. The identified compounds have been reported in the literature to cause mortality or affect the biological parameters of insects, what suggests the possibility of their usage in the formulation of bioinsecticides.
Bryophytes are avascular plant species taxonomically placed between algae and pteridophytes, divided into three groups: mosses (Bryophyta), liverworts (Marchantiophyta) and hornworts (Anthocerotophyta) . These plants have a high diversity of chemical compounds with biological activities often reported in the literature. These natural bioactive compounds are considered “chemical weapons” that function as a defense mechanism against fungi, bacteria and insects and compensate for the absence of the usual mechanical protection (thick cuticle and bark layers) present in other plant groups [2,3,4].
It is estimated that there are about 12,700 species of mosses worldwide. They are conspicuous floristic components of all terrestrial habitats, from Antarctica to the exuberant tropical forests [5, 6]. The moss family Calymperaceae is composed of pantropical and strictly endemic taxa that occur in disparate environments, such as for example, from the Pacific islands to the Amazon basin . There are 28 genera distributed worldwide and four genera in Brazil, namely, Calymperes Sw. ex Weber, Leucophanes Brid., Syrrhopodon Schwägr., and Octoblepharum Hedw. .
In Brazil, the few phytochemical studies carried out with Calymperes lonchophyllum Schwägr. and Octoblepharum albidum Hedw. have provided satisfactory results with potential application in biotechnological processes, such as the production of antibiotics and pesticides [9, 10].
Octoblepharum albidum occurs in all phytogeographic domains of Brazil (the Amazon, Caatinga, Cerrado, Pantanal, Atlantic Forest and Pampa) [8, 11]. This species is characterized by medium-sized plants that grow in loose tufts or cushions, rarely solitary, presenting an opaque whitish coloration, often vinaceous at the base of the leaves; short, simple, radiculose stems; spreading leaves with ligulate shape, broad oval-obovate, sometimes concave base, with apiculate apex, with entire margins below and serrated margins at the apex, presenting 2–3 layers of leucocysts above and below a medial band of chlorocysts in cross-sectional view; and sporophytes with a short seta and an ovoid-cylindric capsule .
Ten classes of bioactive secondary compounds were found in the ethanolic extract of O. albidum, which showed antibacterial activity alone and in association with antibiotics, mainly against Escherichia coli and Klebsiella pneumoniae . The ethanolic extract of this species had insecticidal activity at different concentrations, killing most third-instar Spodoptera frugiperda (JE Smith, 1797) (Lepidoptera: Noctuidae) caterpillars in cowpea (Vigna unguiculata (L.) Walp.) leaves, especially from 48 h of the experiment onwards (ALVES et al., unpublished data).
Bryophyte species contain primary and secondary compounds that can be easily extracted and used as effective insecticides in agriculture . Therefore, considering the lack of research on the volatile chemical composition of O. albidum and its relationship with various bioactivities, especially on compounds with potential application in bioinsecticide formulations, this study aimed to determine the volatile chemical composition of Amazonian specimens of this species and its association with possible bioactive effects on insects.
Plant material and extraction procedure
Specimens of O. albidum were collected from trunks of live trees in domestic backyards in the state of Pará using the techniques proposed by . The identification/confirmation of the species followed the taxonomic classification of . As different bryophyte species often grow intermingled in clusters together with other organisms, such as invertebrates, the specimens of interest were screened and separated with the aid of a magnifying glass and tweezers. Subsequently, a fresh sample (11 g) including gametophytes and sporophytes of O. albidum was subjected to micro-scale simultaneous distillation–extraction (DES) in a Nickerson & Likens extraction apparatus from Chrompack, using n-pentane (3 mL) as solvent, coupled to a refrigeration system to maintain the temperature of the condenser between 5 and 10 °C for 2 h, in duplicate.
The volatile concentrate was analyzed by means of gas chromatography coupled to mass spectrometry (GC/MS) using a Shimadzu QP-2010 Plus system equipped with a Rtx-5MS capillary column (Restek Corporation, Bellefonte, PA USA) (30 m ×0.25 mm; 0.25 μm film thickness) under the following operating conditions: carrier gas: helium, with a linear velocity of 36.5 cm/s; injection type: splitless (2 μL); injector and detector temperature: 250 °C; oven temperature program: 40–60 °C (2 °C/min), 60–250 °C (3 °C/min); MS: electron impact, 70 eV; temperature of ion source and connecting parts: 220 °C. Compound identification was done by comparison of mass spectra and retention indices (RI) with those of substances in the libraries of the system (NIST) and literature data [14, 15]. The RI were obtained using the homologous series of n-alkanes (C8–C40) (Sigma-Aldrich, Milwaukee, WI, USA). The components were quantified by means of GC in a Shimadzu QP-2010 Plus instrument equipped with a flame ionization detector (FID) under the same operating conditions except for the carrier gas, which was hydrogen.
This work has authorization registered with the National System for Management of Genetic Heritage and Associated Traditional Knowledge (SISGEN), with Registration Number: A952E48.
Results and discussion
Twelve organic compounds, corresponding to 92.44% of the total compounds, were identified in the volatile concentrate of O. albidum. The major compounds were hexadecanoic acid (43.97%), oleic acid (17.8%), E-isoeugenol (7.09%), 1-cten-3-ol (5.87%), and stearic acid (4.84%) (Table 1 and Fig. 1). The phytochemical screening of the ethanolic extract of O. albidum conducted by  identified the presence of tannins phlobaphenes, tannins pyrogallates, anthocyanins, flavones, flavonols, flavonones, aurones, proanthocyanidins, alkaloids, and terpenes. Extraction by sample enrichment probe and identification by GC-MS showed aliphatic alcohols and aldehydes (1-hexanol, 7-octen-4-ol, hexanal and nonanal) in O. albidum, in addition to a large amount of fatty acids, especially hexadecanoic acid . Fatty acids produced by bryophytes can be saturated, monounsaturated, polyunsaturated and acetylenic and, together with triglycerides, glycolipids, phospholipids, sterols, wax esters, fatty alcohols and terpenoids, they are classified as lipids because of their hydrophobic nature .
Fatty acids (hexadecanoic, oleic, stearic, and dodecanoic acid) stood out in the volatile concentrate of O. albidum (Table 1 and Fig. 1). Hexadecanoic and stearic acids are some common saturated fatty acids synthesized by bryophytes, and several poly- and monounsaturated fatty acids such as oleic acid are abundant in all bryophyte species . Some of these compounds isolated from bryophytes, such as hexadecanoic and dodecanoic acids, have insecticidal activity against Sitophilus granarius (Coleoptera: Curculionidae) . The viability of S. frugiperda larvae was reduced under the influence of oleic, stearic and hexadecanoic acids, tested separately, and the latter was the most effective . Linoleic acid was more toxic than oleic acid and stearic acid against Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) larvae, and when mixed with commercial insecticidal products, linoleic acid and oleic acid had, respectively, potentiation and additive effects, interfering with the biological parameters of the insects .
Hexadecanoic acid can be converted into unsaturated fatty acid, possibly contributing, at least partially, to the synthesis of longer fatty acids, such as eicosapentaenoic acid, which can deter herbivory . Eicosapentaenoic acid (C20) and arachidonic acid (C20) are produced in high concentrations by bryophytes, but are rarely found in other plant groups . These two very-long-chain acids are precursors for the biosynthesis of some C8 aromatic fatty alcohols, such as 1-octen-3-ol, octan-3-one and octan-3-ol, as observed after mechanical stress in Marchantia polymorpha L. . Neckeropsis undulata (Hedw.) Reichardt has also been reported to produce large amounts of octen-3-ol after injury, which suggests that rapid formation of this compound in wounded sites of mosses and liverworts is necessary for defense against pathogens and herbivores [23, 24]. 1-octen-3-ol and 3-Octanone corresponded to 5.87% and 2.76%, respectively, of the volatile concentrate of O. albidum (Table 1 and Fig. 1).
E-isoeugenol was one of the main compounds found in O. albidum in this study, with a concentration of 7.09%, representing the first record in mosses (Table 1 and Fig. 1). Eugenol and its derivatives have a pungent and spicy aroma and have already been reported in liverwort species and in the essential oils of angiosperms [25,26,27]. Isoeugenol showed contact toxicity against Sitophilus zeamais Motschulsky and Tribolium castaneum (Herbst) (both Coleoptera: Curculionidae) and reduced the growth rate and food consumption of larvae and/or adults of the two species in a concentration-dependent manner . Eugenol and isoeugenol caused high larval mortality, dissuaded oviposition of females, and reduced egg hatchability of Tuta absoluta Meyrick (Lepidoptera: Gelechiidae) . Isoeugenol caused mortality of Drosophila melanogaster Meigen (Diptera; Drosophilidae) and higher volumes of this compound decreased the flying capacity of flies .
2-methoxy-4-methyl-phenol and methyl-p-tert-butyl phenylacetate were also identified in O. albidum, with concentrations of 1.91% and 1.90%, respectively. The occurrence of these compounds is unprecedented in bryophytes (Table 1 and Fig. 1). 2-methoxy-4-methyl-phenol contained in the liquid smoke of coconut fiber is one of the phenolic compounds used in the insecticide industry, whose tested product caused 60–80% mortality of Epilachna sparsa (Hbst.) (Coleoptera: Coccinellidae) . Methyl-p-tert-butyl phenylacetate was the only compound identified in the ethanolic extract of seeds of Annona squamosa Linn. and was observed to cause a mortality rate of more than 50% of brown planthopper Nilaparvata lugens (Stål) (Homoptera: Delphacidae) .
1-octen-3-yl acetate was found in the volatile concentrate of O. albidum at a concentration of 1.33% (Table 1 and Fig. 1). In the literature, this ester has been found only in liverworts, such as Conocephalum, Marchantia, Dumortiera, Pellia, Plagiochila, and Wiesnerella species, being the compound responsible for the mushroom odor of these bryophytes [22, 33,34,35]. 1-octen-3-yl acetate was the predominant C8 volatile present in intact thalli of M. polymorpha, while tissue disruption resulted in the conversion of the acetate to 1-octen-3-ol . C8 volatiles are, in general, known to perform a signaling function when in the vapor phase . 1-octen-3-yl β-primeveroside extracted from soybean (Glycine max L.) has been considered to function as a form of storage of volatile 1-octen-3-ol for immediate response against mechanical damage to leaf tissues, suggesting its participation in plant defense mechanism .
The diterpene neophytadiene was present at a concentration of 1.11% in the volatile composition of O. albidum. The presence of this compound has already been documented in other moss and liverwort species [37,38,39] (Table 1 and Fig. 1). Neophytadiene is usually found in all green plants, as a product of chlorophyll degradation . This compound was one of the most active obtained from the bio-oil produced from the residues of greenhouse tomato plants, exhibiting toxicity against Leptinotarsa decemlineata Say. (Coleoptera: Chrysomelidae) larvae . In turn, 2E-Hepten-1-ol was the minor component in the volatile concentrate of O. albidum (0.63%) (Table 1 and Fig. 1). Similar aliphatic alcohols, such as 2-hepten-1-ol and 4-hepten-1-ol, have already been reported from moss species, and other alcohols, such as 1-hexanol and 7- octen-4-ol, have been recorded in O. albidum, showing that this class of compounds is part of the oxylipin volatilome of these bryophytes . Oxylipines are oxygenated fatty acids that participate in the defense of bryophytes, being produced in abundance after tissue injury or pathogen attack .
The volatile composition of O. albidum from Brazil is reported for the first time here, using GC/MS. We showed evidence that this specie is a source of fatty acids, phenolic compounds, alcohols, ketones, esters, and terpenes. Further, the occurrence of some compounds in O. albidum is unprecedented in mosses and even in bryophytes, as for example E-isoeugenol, 2-methoxy-4-methyl-phenol, and methyl-p-tert-butyl-phenylacetate. The identified compounds have been reported in the literature to cause mortality or affect the biological parameters of insects, what suggests the possibility of their usage in the formulation of bioinsecticides.
Availability of data and materials
The datasets used and/or analysed during the current study are available from the corresponding author on reasonable request.
Asakawa Y, Nagashima F, Ludwiczuk A. Distribution of bibenzyls, prenyl bibenzyls, bis-bibenzyls, and terpenoids in the liverwort Genus Radula. J Nat Prod. 2020;83:756–69. https://doi.org/10.1021/ACS.JNATPROD.9B01132.
Frahm JP. Recent developments of commercial products from bryophytes. Bryologist. 2004;107:277–83. https://doi.org/10.1639/0007-2745.
Aruna KB, Krishnappa M. Phytochemistry and antimicrobial activities of Pogonatum microstomum (R. Br. ex Schwägr.) Brid. (Bryophyta; Musci: Polytrichaceae). Int J Bot Stud. 2018;3:120–5.
Ludwiczuk A, Asakawa Y. Terpenoids and aromatic compounds from bryophytes and their central nervous system activity. Curr Org Chem. 2020;24:113–28. https://doi.org/10.2174/1385272824666200120143558.
Vanderpoorten A, Goffinet B. Introduction to bryophytes. Cambridge: Cambridge University Press; 2009.
Cox C, Goffinet B, Wickett N, et al. Moss diversity: a molecular phylogenetic analysis of genera. Phytotaxa 2010;9:175–95.
Reese WD. Calymperaceae. Flora Neotrop. 1993;58:1–101.
Costa DP, Peralta D. Lista do Brasil - Briófitas, 2021. http://floradobrasil.jbrj.gov.br/jabot/FichaPublicaTaxonUC/FichaPublicaTaxonUC.do?id=FB128472. Accessed 3 Dec 2021.
de Pinheiro MFS, Lisboa RCL, de Brazão VR. Contribuição ao estudo de briófitas como fonte de antibióticos. Acta Amaz. 1989. https://doi.org/10.1590/1809-43921989191145.
Vidal CAS, Sousa EO, Rodrigues FFG, et al. Phytochemical screening and synergistic interactions between aminoglycosides, selected antibiotics and extracts from the bryophyte Octoblepharum albidum Hedw (Calymperaceae). Arch Biol Sci. 2012;64:465–70. https://doi.org/10.2298/ABS1202465V.
Nascimento GMG do, da Conceição GM, Peralta DF, de Oliveira HC 2020 Bryophytes of sete cidades national park, Piauí Brazil. Check List 16 969 988. https://doi.org/10.15560/16.4.969.
Abay G, Altun M, Karakoc O, et al. Insecticidal activity of fatty acid-rich turkish bryophyte extracts against Sitophilus granarius (Coleoptera: Curculionidae). Comb Chem High Throughput Screen. 2013;16:806–16. https://doi.org/10.2174/13862073113169990049.
Yano O. Briófitas. In: Fidalgo O, Bononi VLR, editors. Técnicas de coleta, preservação e herborização de material botânico. São Paulo: Instituto de Botânica, Estado de São Paulo; 1984. pp. 27–30.
Adams RP. Identification of essential oil components by gas chromatography/mass spectrometry. Ilinois: Allured Publishing Corporation; 2007.
Mondello L. FFNSC 2: flavors and fragrances of natural and synthetic compounds mass spectral database. New York: John Wiley & Sons; 2011.
Mitra S, Burger BV, Poddar-Sarkar M. Comparison of headspace-oxylipin-volatilomes of some Eastern Himalayan mosses extracted by sample enrichment probe and analysed by gas chromatography-mass spectrometry. Protoplasma. 2017;254:1115–26. https://doi.org/10.1007/s00709-016-1018-3.
Lu Y, Eiriksson FF, Thorsteinsdóttir M, Simonsen HT. Valuable fatty acids in bryophytes—production, biosynthesis, analysis and applications. Plants. 2019;8:1–18. https://doi.org/10.3390/plants8110524.
Pérez-Gutiérrez S, Zavala-Sánchez MA, González-Chávez MM, et al. Bioactivity of Carica papaya (Caricaceae) against Spodoptera frugiperda (Lepidoptera: Noctuidae). Molecules. 2011;16:7502–9. https://doi.org/10.3390/molecules16097502.
Eldesouky SE, Khamis WM, Hassan SM. Joint action of certain fatty acids with selected insecticides against cotton leafworm, Spodoptera littoralis and their effects on biological aspects. J Basic Environ Sci. 2019;6:23–32.
Soriano G, Kneeshaw S, Jimenez-Aleman G, et al. An evolutionarily ancient fatty acid desaturase is required for the synthesis of hexadecatrienoic acid, which is the main source of the bioactive jasmonate in Marchantia polymorpha. New Phytol. 2022;233:1401–13. https://doi.org/10.1111/nph.17850.
Shanab SMM, Hafez RM, Fouad AS. A review on algae and plants as potential source of arachidonic acid. J Adv Res. 2018;11:3–13. https://doi.org/10.1016/j.jare.2018.03.004.
Kihara H, Tanaka M, Yamato KT, et al. Arachidonic acid-dependent carbon-eight volatile synthesis from wounded liverwort (Marchantia polymorpha). Phytochemistry. 2014;107:42–9.
Wichard T, Göbel C, Feussner I, Pohnert G. Unprecedented lipoxygenase/hydroperoxide lyase pathways in the moss Physcomitrella patens. Angew Chemie—Int Ed. 2004;44:158–61. https://doi.org/10.1002/anie.200460686.
Miranda TG, Alves RJM, de Souza RF, et al. Volatile concentrate from the neotropical moss Neckeropsis undulata (Hedw.) Reichardt, existing in the brazilian Amazon. BMC Chem. 2021;15:3–7. https://doi.org/10.1186/s13065-021-00736-3.
Asakawa Y, Yoyota M, Takemoto T, et al. Insect antifeedant secoaromadendrane-type sesquiterpenes from Plagiochila species. Phytochemistry. 1980;19:2147–54. https://doi.org/10.1016/S0031-9422(00)82212-8.
Figueiredo AC, Sim-Sim M, Barroso JG, et al. Composition of the essential oil from the liverwort Marchesinia mackaii (Hook.) S. F. Gray grown in Portugal. J Essent Oil Res. 2002;14:439–42. https://doi.org/10.1080/10412905.2002.9699915.
Chowdhry BZ, Ryall JP, Dines TJ, Mendham AP. Infrared and Raman spectroscopy of eugenol, isoeugenol, and methyl eugenol: conformational analysis and vibrational assignments from density functional theory calculations of the anharmonic fundamentals. J Phys Chem A. 2015;119:11280–92. https://doi.org/10.1021/acs.jpca.5b07607.
Huang Y, Ho SH, Lee HC, Yap YL. Insecticidal properties of eugenol, isoeugenol and methyleugenol and their effects on nutrition of Sitophilus zeamais Motsch. (Coleoptera: Curculionidae) and Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae). J Stored Prod Res. 2002;38:403–12. https://doi.org/10.1016/S0022-474X(01)00042-X.
Moawad SS, Ebadah IMA, Mahmoud YA. Biological and histological studies on the efficacy of some botanical and commercial oils on Tuta absoluta Meyrick (Lepidoptera: Gelechiidae). Egypt J Biol Pest Control. 2013;23:301–8.
de SousaJúnior DL, Cordeiro PPM, dos SantosBarbosa CR, et al. Evaluation of isoeugenol in inhibition of Staphylococcus aureus efflux pumps and their toxicity using Drosophila melanogaster model. Life Sci. 2021. https://doi.org/10.1016/j.lfs.2021.119940.
Anom DK, Mamangkey JJ. Utilization of coconut fiber waste as insecticides against Epilachna sparsa. Chem Mater Res. 2016;8:70–6.
Deewatthanawong R, Kongchinda P, Deewatthanawong P, et al. GC-MS analysis and biopesticide properties of different crude extracts of Annona squamosa and Annona muricata. Int J Agric Technol. 2019;15:859–68.
Asakawa Y, Toyota M, Nagashima F, Hashimoto T. Chemical constituents of selected Japanese and New Zealand liverworts. Nat Prod Commun. 2008;3:289–300. https://doi.org/10.1177/1934578X0800300238.
Ludwiczuk A, Nagashima F, Gradstein RS, Asakawa Y. Volatile components from selected Mexican, Ecuadorian, Greek, German and Japanese liverworts. Nat Prod Commun. 2008;3:133–40. https://doi.org/10.1177/1934578x0800300205.
Ghani NA, Ludwiczuk A, Ismail NH, Asakawa Y. Volatile components of the stressed liverwort Conocephalum conicum. Nat Prod Commun. 2016;11:103–4. https://doi.org/10.1177/1934578x1601100130.
Ntoruru JM, Ohnishi T, Katsumata F, et al. 1-Octen-3-ol is formed from its primeveroside after mechanical wounding of soybean leaves. Plant Mol Biol. 2021. https://doi.org/10.1007/s11103-021-01226-9.
Komala I, Ito T, Nagashima F, et al. Zierane sesquiterpene lactone, cembrane and fusicoccane diterpenoids, from the Tahitian liverwort Chandonanthus hirtellus. Phytochemistry. 2010;71:1387–94. https://doi.org/10.1016/j.phytochem.2010.04.023.
Üçüncü O, Cansu TB, Özdemlr T, et al. Chemical composition and antimicrobial activity of the essential oils of mosses (Tortula muralis Hedw., Homalothecium lutescens (Hedw.) H. Rob., Hypnum cupressiforme Hedw., and Pohlia nutans (Hedw.) Lindb.) from Turkey. Turkish J Chem. 2010;34:825–34. https://doi.org/10.3906/kim-1002-62.
Klavina L, Springe G, Steinberga I, et al. Seasonal changes of chemical composition in boreonemoral moss species. Environ Exp Biol. 2018;16:9–19. https://doi.org/10.22364/eeb.16.02.
Stelmasiewicz M, Światek Ł, Ludwiczuk A. Phytochemical profile and anticancer potential of endophytic microorganisms from liverwort species, Marchantia polymorpha L. Molecules. 2022. https://doi.org/10.3390/molecules27010153.
Cáceres LA, McGarvey BD, Briens C, et al. Insecticidal properties of pyrolysis bio-oil from greenhouse tomato residue biomass. J Anal Appl Pyrolysis. 2015;112:333–40. https://doi.org/10.1016/j.jaap.2015.01.003.
de León IP, Hamberg M, Castresana C. Oxylipins in moss development and defense. Front Plant Sci. 2015;6:1–12. https://doi.org/10.3389/fpls.2015.00483.
To the Universidade do Estado do Pará and Fundação Amazônia de Amparo a Estudos e Pesquisas.
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Alves, R.J.M., Miranda, T.G., Pinheiro, R.O. et al. Volatile chemical composition of Octoblepharum albidum Hedw. (Bryophyta) from the Brazilian Amazon. BMC Chemistry 16, 76 (2022). https://doi.org/10.1186/s13065-022-00872-4
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