Morphological, molecular, and phytochemical characteristics of wild edible fern (Diplazium esculentum) from Jember, East Java, Indonesia
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Abstract. Amalini YR, Muzakhar K, Su’udi M, Setyati D, Dwinianti EF, Ulum FB. 2025. Morphological, molecular, and phytochemical characteristics of wild edible fern (Diplazium esculentum) from Jember, East Java, Indonesia. Biodiversitas 26: 2794-2805. Diplazium esculentum, an edible pteridophyte with medicinal potential, exhibits morphological variation that risks improper medicinal use. This study aimed to comprehensively characterize D. esculentum from Jember, Indonesia, through an integrative approach, including morphological analysis, DNA barcoding, phytochemical profiling, and antioxidant activity assessment. Fifteen specimens, locally referred to as pakis menir, were collected from a single site within a rubber-mixed coffee plantation in Panti, Jember District, East Java, Indonesia. The examination of morphological traits, molecular identification using the rbcL marker, analysis of phytochemical profiles through GC-MS, prediction of biological activities using PASS online, and evaluation of antioxidant activity using the DPPH assay all pointed to the potential of D. esculentum. Morphological analysis confirmed D. esculentum's diagnostic traits, while rbcL DNA barcoding validated a 100% sequence match with D. esculentum (OL536867.1). GC-MS analysis of young fronds identified 25 bioactive metabolites, including 11 phenolics, 9 terpenoids, and 5 alkaloids, with 96% exhibiting high biological activity (Pa>0.7) according to PASS predictions. The dominant compound, 3,6-dimethylquinoline (12.38%), exhibited significant anticancer potential, while the frond extract showed strong antioxidant activity (IC50: 91.25±7.60 ppm), reflecting its phenolic richness. These results highlight the potential of D. esculentum's phytochemical wealth and pharmacological promise, supporting further exploration in pharmacognosy, nutraceuticals, and bioactive compounds.
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References
Abdul-Hammed M, Adedotun IO, Olajide M, Irabor CO, Afolabi TI, Gbadebo IO, Rhyman L, Ramasami P. 2022. Virtual screening, ADMET profiling, PASS prediction, and bioactivity studies of potential inhibitory roles of alkaloids, phytosterols, and flavonoids against COVID-19 main protease (Mpro). Nat Prod Res 36 (12): 3110-3116. DOI: 10.1080/14786419.2021.1935933.
Alamsjah F, Fandini S, Mildawati M. 2024. Evaluation of antibacterial activities and phytochemical composition of ethanolic extract of Diplazium esculentum. Biodiversitas 25 (3): 937-941. DOI: 10.13057/biodiv/d250304.
Amalini YR, Afidah Y. 2023. In-silico PASS prediction for medicinal potency of antioxidant from Ranunculus japonicus Thunb. Life Sci Biotechnol 1 (2): 59-65. DOI: 10.19184/LSB.V1I2.45078.
Amirkia V, Heinrich M. 2014. Alkaloids as drug leads – A predictive structural and biodiversity-based analysis. Phytochem Lett 10: xlviii-liii. DOI: 10.1016/j.phytol.2014.06.015.
Bafeel SO, Arif IA, Bakir M, Khan H, Al Farhan AH, Al Homaidan AA, Ahamed A, Thomas J. 2011. Comparative evaluation of PCR success with universal primers of maturase K (matK) and ribulose-1, 5-bisphosphate carboxylase oxygenase large subunit (rbcL) for barcoding of some arid plants. Plant Omics J 4 (4): 195-198.
Bajaj K, Sakhuja R. 2016. Benzotriazole: Much More Than Just Synthetic Heterocyclic Chemistry. In: Monbaliu JM (eds). The Chemistry of Benzotriazole Derivatives. Topics in Heterocyclic Chemistry. Springer, Cham. DOI: 10.1007/7081_2015_198.
Baliyan S, Mukherjee R, Priyadarshini A, Vibhuti A, Gupta A, Pandey RP, Chang CM. 2022. Determination of antioxidants by DPPH radical scavenging activity and quantitative phytochemical analysis of Ficus religiosa. Molecules 27: 1326. DOI: 10.3390/molecules27041326.
Batoro J, Siswanto D. 2017. Ethnomedicinal survey of plants used by local society in Poncokusumo District, Malang, East Java Province, Indonesia. Asian J Med Biol Res 3 (2): 158-167. DOI: 10.3329/ajmbr.v3i2.33563.
Bellier J, Nokin MJ, Lardé E, Karoyan P, Peulen O, Castronovo V, Bellahcène A. 2019. Methylglyoxal, a potent inducer of AGEs, connects between diabetes and cancer. Diabetes Res Clin Pract 148: 200-211. DOI: 10.1016/J.DIABRES.2019.01.002.
Boiangiu RS, Brinza I, Honceriu I, Mihasan M, Hritcu L. 2023. Insights into pharmacological activities of nicotine and 6-hydroxy-l-nicotine, a bacterial nicotine derivative: A systematic review. Biomolecules 14 (1): 23. DOI: 10.3390/biom14010023.
Bousquet P, Hudson A, García-Sevilla JA, Li JX, France CP. 2020. Imidazoline receptor system: The past, the present, and the future. Pharmacol Rev 72 (1): 50-79. DOI: 10.1124/pr.118.016311.
CBOL Plant Working Group. 2009. A DNA barcode for land plants. Proc Natl Acad Sci USA 106 (31): 12794-12797. DOI: 10.1073/pnas.0905845106.
Choi HS, Kim SL, Kim JH, Ko YC, Lee DS. 2020. Plant volatile, phenylacetaldehyde targets breast cancer stem cell by induction of ROS and regulation of stat3 signal. Antioxidants (Basel) 9 (11): 1119. DOI: 10.3390/ANTIOX9111119.
de Winter W, Amoroso V. 2003. Cryptogams: Ferns and Fern Allies. Backhuys, Leiden.
Del Olmo A, Calzada J, Nuñez M. 2017. Benzoic acid and its derivatives as naturally occurring compounds in foods and as additives: Uses, exposure, and controversy. Crit Rev Food Sci Nutr 57 (14): 3084-3103. DOI: 10.1080/10408398.2015.1087964.
Dey P, Kundu A, Kumar A, Gupta M, Lee BM, Bhakta T, Dash S, Kim HS. 2020. Analysis of Alkaloids (Indole Alkaloids, Isoquinoline Alkaloids, Tropane Alkaloids). In: Nabavi SM, Saeedi M, Nabavi SF, Silva AT (eds). Recent Advances in Natural Products Analysis. Elsevier, Amsterdam.
Doyle J. 1991. DNA Protocols for Plants. In: Hewitt G, Johnson AWB, Young JPW (eds). Molecular Techniques in Taxonomy. NATO ASI Series H Cell Biol 57: 283-293. DOI: 10.1007/978-3-642-83962-7_18.
Durlak P, Jerzykiewicz M, ?wiel?g-Piasecka I. 2019. 1,2,3-propanetriol radicals formed during oxidative stress. Magn Reson Chem 57 (4): S95-S100. DOI: 10.1002/mrc.4822.
Essien EE, Ascrizzi R, Flamini G. 2019. Characterization of volatile compounds of Diplazium esculentum. Chem Nat Compd 55: 958-959. DOI: 10.1007/s10600-019-02860-y.
Fan M, Yuan S, Li L, Zheng J, Zhao D, Wang C, Wang H, Liu X, Liu J. 2023. Application of terpenoid compounds in food and pharmaceutical products. Fermentation 9 (2): P119. DOI: 10.3390/fermentation9020119.
Hadi AA, Ng JY, Shamsuddin M, Matmin J, Malek NANN. 2022. Green synthesis of silver nanoparticles using Diplazium esculentum extract: Catalytic reduction of methylene blue and antibacterial activities. Chem Papers 76: 65-77. DOI: 10.1007/s11696-021-01835-0.
Heinrich M, Mah J, Amirkia V. 2021. Alkaloids used as medicines: Structural phytochemistry meets biodiversity-An update and forward look. Molecules 26 (7): 1836. DOI: 10.3390/molecules26071836.
Ilakiyalakshmi M, Napoleon AA. 2022. Review on recent development of quinoline for anticancer activities. Arab J Chem 15 (11): 104168. DOI: 10.1016/j.arabjc.2022.104168.
Inthachat W, Chantong B, Pitchakarn P, Takoon C, Karinchai J, Suttisansanee U, Temviriyanukul P. 2024. Enhancing therapeutic efficacy of donepezil, an alzheimer's disease drug, by Diplazium esculentum (Retz.) Sw. and its phytochemicals. Pharmaceuticals 17 (3): 341. DOI: 10.3390/ph17030341/s1.
Junejo JA, Gogoi G, Islam J, Rudrapal M, Mondal P, Hazarika H, Zaman K. 2018. Exploration of antioxidant, antidiabetic and hepatoprotective activity of Diplazium esculentum - A wild edible plant from North Eastern India. Futur J Pharm Sci 4 (1): 93-101. DOI: 10.1016/j.fjps.2017.10.005.
Kakhlon O, Ferreira I, Solmesky LJ, Khazanov N, Lossos A, Alvarez R, Yetil D, Pampou S, Weil M, Senderowitz H, Escriba P, Yue WW, Akman HO. 2018. Guaiacol as a drug candidate for treating adult polyglucosan body disease. JCI Insight 3: e99694. DOI: 10.1172/jci.insight.99694.
Kapli P, Yang Z, Telford MJ. 2020. Phylogenetic tree building in the genomic age. Nat Rev Genet 21: 428-444. DOI: 10.1038/s41576-020-0233-0.
Kartikasari SN, Witono Y, Handoyo T, Santoso U. 2024. Hydrolysis with two enzymes in edible fern (Diplazium esculentum) as a source of flavours. Indonesian Food Sci Technol J 7 (2): 148-159. DOI: 10.22437/ifstj.v7I2.30960.
Kongsung S, Inthachat W, Chantong B, Suttisansanee U, On-Nom N, Chupeerach C, Thangsiri S, Pitchakarn P, Temviriyanukul P. 2024. Box–behnken design-based optimization of phytochemical extraction from Diplazium esculentum (Retz.) Sw. associated with its antioxidant and anti-alzheimer's properties. Molecules 29 (10): 2204. DOI: 10.3390/molecules29102204.
Kunkeaw T, Suttisansanee U, Trachootham D, Karinchai J, Chantong B, Potikanond S, Inthachat W, Pitchakarn P, Temviriyanukul P. 2021. Diplazium esculentum (Retz.) Sw. reduces BACE-1 activities and amyloid peptides accumulation in Drosophila models of Alzheimer's disease. Sci Rep 11: 23796. DOI: 10.1038/s41598-021-03142-w.
Kurniawan L, Laili AN, Anggraeni DS, Qurrotu’ain S, Wulandari DR, Ulum FB. 2023. Poiploidy induction of Indonesian black rice Oryza sativa L. var. Cempo Ireng with bio-catharantine. Life Sci Biotechnol 1 (2): 41-47. DOI: 10.19184/lsb.v1i2.43753.
Lima GP, Santos MG, de Paiva SR. 2024. Terpenoid composition of fern and lycophyte essential oils: A chemosystematic approach. NZ J Bot 1-24. DOI: 10.1080/0028825X.2024.2323505.
Manurung H, Susanto D, Kusumawati E, Aryani R, Nugroho RA, Kusuma R, Rahmawati Z, Sari RD. 2022. Phytochemical, GC-MS analysis and antioxidant activities of leaf methanolic extract of lai (Durio kutejensis), the endemic plant of Kalimantan, Indonesia. Biodiversitas 23 (11): 5566-5573. DOI: 10.13057/biodiv/d231104.
Matada BS, Pattanashettar R, Yernale NG. 2021. A comprehensive review on the biological interest of quinoline and its derivatives. Bioorg Med Chem 32: 115973. DOI: 10.1016/j.bmc.2020.115973.
Michaeli DT, Michaeli JC, Albers S, Boch T, Michaeli T. 2023. Established and emerging lipid-lowering drugs for primary and secondary cardiovascular prevention. Am J Cardiovasc Drugs 23 (5): 477-495. DOI: 10.1007/s40256-023-00594-5.
Molyneux P. 2004. The use of the stable free radical Diphenylpicrylhydrazyl (DPPH) for estimating antioxidant activity. Songklanakarin J Sci Technol 26 (2): 211-219.
Mustafa A, El?Kashef DH, Abdelwahab MF, Gomaa AAR, Mustafa M, Abdel?Wahab NM, Ibrahim AH. 2023. Investigation of Antiviral Effects of Essential Oils. In: Inamuddin (ed) Essential Oils: Extraction Methods and Applications. John Wiley & Sons, Inc., New Jersey. DOI: 10.1002/9781119829614.ch5.
Mynssen CM, Sylvestre LdS. 2019. Synopsis of Diplazium (Athyriaceae) from Brazil. American Fern J 109 (4): 283-342. DOI: 10.1640/0002-8444-109.4.283.
National Library of Medicine. 2024. Bethesda (MD): National Library of Medicine (US), National Center for Biotechnology Information www.ncbi.nlm.nih.gov/nuccore/?term=diplazium+esculentum.
Nitta JH, Chambers SM. 2022. Identifying cryptic fern gametophytes using DNA barcoding: A review. Appl Plant Sci 10 (2): e11465. DOI: 10.1002/aps3.11465.
Nurhasnawati H, Sundu R, Sapri, Supriningrum R, Kuspradini H, Arung ET. 2019. Antioxidant activity, total phenolic and flavonoid content of several indigenous species of ferns in East Kalimantan, Indonesia. Biodiversitas 20 (2): 576-580. DOI: 10.13057/biodiv/d200238.
Perwitasari DAG, Rohimah S, Ratnasari T, Sugiharto B, Su’udi M. 2020. DNA barcoding of medicinal orchid Dendrobium discolor Lindl. Tanimbar using rbcL and ITS genes. Buletin Penelitian Tanaman Rempah dan Obat 31 (1): 8-20. DOI: 10.21082/bullittro.v31n1.2020.8-20. [Indonesian]
Poirier D. 2024. Recent advances in the development of 17beta-hydroxysteroid dehydrogenase inhibitors. Steroids 213: 109529. DOI: 10.1016/j.steroids.2024.109529.
POWO [Plants of the World Online Kew Science]. 2025. Diplazium Sw. Plants of the World Online Kew Science. Retrieved April 15, 2025, from https://powo.science.kew.org/taxon/urn:lsid:ipni.org:names: 328225-2.
Praptiwi, Sulistiarini D, Qodrie ENP, Sahroni D. 2021. Antibacterial activity, antioxidant potential, total phenolic and flavonoids of three plant species of Rubiaceae from Banggai Island, Indonesia. Biodiversitas 22 (5): 2773-2778. DOI: 10.13057/biodiv/d220540.
Praptosuwiryo TN. 2008. Biosystematic Study of The Fern Genus Diplazium in West Malesia. [Dissertation]. IPB University, Bogor. [Indonesian]
Purnobasuki H, Rakhashiwi GA, Junairiah, Wahyuni DK, Putra RE, Raffiudin R, Soessilohadi RH. 2022. Morpho-anatomical characterization and DNA barcode of Cosmos caudatus Kunth. Biodiversitas 23 (8): 4097-4108. DOI: 10.13057/biodiv/d230830.
Rahman MM, Rahaman MS, Islam MR, Rahman F, Mithi FM, Alqahtani T, Almikhlafi MA, Alghamdi SQ, Alruwaili AS, Hossain MS, Ahmed M, Das R, Emran TB, Uddin MS. 2021. Role of phenolic compounds in human disease: Current knowledge and future prospects. Molecules 27 (1): 233. DOI: 10.3390/molecules27010233.
Reis J, Massari M, Marchese S, Ceccon M, Aalbers FS, Corana F, Valente S, Mai A, Magnani F, Mattevi A. 2020. A closer look into NADPH oxidase inhibitors: Validation and insight into their mechanism of action. Redox Biol 32: 101466. DOI: 10.1016/j.redox.2020.101466.
Rizqoni MIA, Setyati D, Su’udi M, Dwinianti EF, Ulum FB. 2024. Molecular identification of Cyclosorus parasiticus (L.) Farw. from Gumitir, Jember Indonesia. Bioedukasi 22 (2): 218-223. DOI: 10.19184/bioedu.v22i2.45484.
Rosas-Rodríguez JA, Soñanez-Organis JG, Godoy-Lugo JA, Espinoza-Salazar JA, López-Jacobo CJ, Stephens-Camacho NA, González-Ochoa G. 2017. Betaine aldehyde dehydrogenase expression during physiological cardiac hypertrophy induced by pregnancy. Biochem Biophys Res Commun 490: 623-628. DOI: 10.1016/j.bbrc.2017.06.087.
Russo CAM, Selvatti AP. 2018. Bootstrap and rogue identification tests for phylogenetic analyses. Mol Biol Evol 35 (9): 2327-2333. DOI: 10.1093/molbev/msy118.
Saha J, Kumari T, Nickhil C, Deka SC. 2024. Studies on the cytotoxicity of leaf protein concentrate from Diplazium esculentum, pasta preparation and assessment of its quality during storage. Food Chem Adv 4: 100667. DOI: 10.1016/j.focha.2024.100667.
Semwal P, Painuli S, Painuli KM, Antika G, Tumer TB, Thapliyal A, Setzer WN, Martorell M, Alshehri MM, Taheri Y, Da?tan SD, Ayatollahi SA, Petkoska AT, Sharifi-Rad J, Cho WC. 2021. Diplazium esculentum (Retz.) Sw.: Ethnomedicinal, phytochemical, and pharmacological overview of the Himalayan Ferns. Oxid Med Cell Longev 2021: 1917890. DOI: 10.1155/2021/1917890.
Setyati D, Adawiyah RA, Ratnasari T, Su'udi M, Ulum FB. 2023. Phenolic profile and antimicrobe of the Asplenium Nidus L. from Mount Gumitir, Jember, East Java, Indonesia. Bioedukasi 21 (3): 189-193. DOI: 10.19184/bioedu.v21i3.43368.
Setyati D, Su’udi M, Ravitamala ES, Miladina FF, Babudin B, Utarti E, Arimurti S, Nugraha AS, Putri YA, Farhan AM, Ulum FB. 2024. Antimicrobial and phytochemistry study of Dendrobium linearifolium Teijsm. & Binn. from Gumitir, Jember, Indonesia. BIO Web Conf 01001: 12. DOI: 10.1051/bioconf/202410101001.
Shan KS, Rehman TU, Ivanov S, Domingo G, Raez LE. 2024. Molecular targeting of the BRAF proto-oncogene/mitogen-activated protein kinase (MAPK) pathway across cancers. Intl J Mol Sci 25 (1): 624. DOI: 10.3390/ijms25010624.
Sofiyanti N, Iriani D, Fitmawati F, Marpaung AA. 2019. Morphology, palynology, and stipe anatomy of four common ferns from Pekanbaru, Riau Province, Indonesia. Biodiversitas 20 (1): 327-336. DOI: 10.13057/biodiv/d200138.
Sridhar KR. 2023. On the Bioactive Potential of Ferns: An Overview. In: Murthy HN (eds). Bioactive Compounds in Bryophytes and Pteridophytes. Reference Series in Phytochemistry. Springer, Cham. DOI: 10.1007/978-3-031-23243-5_11.
Stover NA, Cavalcanti ARO. 2017. Using NCBI BLAST. Curr Protoc Essent Lab Tech 14 (1): 1-11. DOI: 10.1002/cpet.8.
Su’udi M, Budyartini DW, Ramadany Z. 2022. DNA Barcoding of Dendrobium linearifolium Teijsm. & Binn. Orchid based on ITS2 molecular marker. Al-Kauniyah 15 (1): 53-61. DOI: 10.15408/kauniyah.v15I1.16710. [Indonesian]
Sun W, Shahrajabian MH. 2023. Therapeutic potential of phenolic compounds in medicinal plants-Natural health products for human health. Molecules 28 (4): 1845. DOI: 10.3390/molecules28041845.
Sururin FW, Khafiyya NA. 2024. Prediction prediction of the insecticidal potency of biduri plants (Calotropis gigantea) using the PASS online web resource. Life Sci Biotechnol 2 (1): 13-19. DOI: 10.19184/LSB.V2I1.47444.
Takamiya M, Takaoka C, Ohta N. 1999. Cytological and reproductive studies on Japanese Diplazium (Woodsiaceae; Pteridophyta): Apomictic reproduction in Diplazium with evergreen bi- to tripinnate leaves. J Plant Res 112: 419-436. DOI: 10.1007/PL00013897.
Tamura K, Stecher G, Kumar S. 2021. MEGA11: Molecular evolutionary genetics analysis version 11. Mol Biol Evol 38 (7): 3022-3027. DOI: 10.1093/molbev/msab120.
Tongco JV, Villaber RA, Aguda RM, Razal RA. 2014. Nutritional and phytochemical screening, and total phenolic and flavonoid content of Diplazium esculentum (Retz.) Sw. from Philippines. J Chem Pharm Res 6 (8): 238-242.
Trujillo-Argueta S, del Castillo RF, Tejero-Diez D, Matias-Cervantes CA, Velasco-Murguía A. 2021. DNA barcoding ferns in an unexplored tropical montane cloud forest area of southeast Oaxaca, Mexico. Sci Rep 11: 22837. DOI: 10.1038/s41598-021-02237-8.
Ulum FB, Setyati D, Rizqoni A, Su’udi M. 2023. Antimicrobial and phytochemistry study of Liparis resupinata Ridl. from Mount Gumitir, East Java, Indonesia. J Pharm Sci Clinic Res 8 (3): 373-384. DOI: 10.20961/jpscr.v8I3.70856.
Uraku AJ. 2015. Leaves by Gas Chromatography-Mass Spectrometry (GC-MS) method. Res J Phytochem 9 (4): 175-187. DOI: 10.3923/rjphyto.2015.175.187.
Wang FH, Lu JM, Wen J, Ebihara A, Li DZ. 2016. Applying DNA barcodes to identify closely related species of ferns: A case study of the Chinese Adiantum (Pteridaceae). PLoS One 11 (9): e0160611. DOI: 10.1371/journal.pone.0160611.
Watanabe M, Miyashita T, Devkota HP. 2021. Phenolic compounds and ecdysteroids of Diplazium esculentum (Retz.) Sw. (Athyriaceae) from Japan and their chemotaxonomic significance. Biochem Syst Ecol 94: 104211. DOI: 10.1016/j.bse.2020.104211.
Wei Z, Wei M, Yang X, Xu Y, Gao S, Ren K. 2022. Synaptic secretion and beyond: Targeting synapse and neurotransmitters to treat neurodegenerative diseases. Oxid Med Cell Longev 2022: 9176923. DOI: 10.1155/2022/9176923.
Yao H, Song J, Liu C, Luo K, Han J, Li Y, Pang X, Xu H, Zhu Y, Xiao P, Chen S. 2010. Use of ITS2 region as the universal DNA barcode for plants and animals. PLoS One 5 (10): e13102. DOI: 10.1371/journal.pone.0013102.
Yildiz I, Yildiz BS. 2021. Mechanistic study of L-6-hydroxynicotine oxidase by DFT and ONIOM methods. J Mol Model 27 (2): 53. DOI: 10.1007/s00894-020-04646-4.
Yu J, Wu X, Liu C, Newmaster S, Ragupathy S, Kress WJ. 2021. Progress in the use of DNA barcodes in the identification and classification of medicinal plants. Ecotoxicol Environ Saf 208: 111691. DOI: 10.1016/j.ecoenv.2020.111691.
Zeb MA, Rahman TU, Sajid M, Xiao W, Musharraf SG, Bibi S, Akitsu T, Liaqat W. 2021. GC-MS analysis and in silico approaches of Indigofera heterantha root oil chemical constituents. Compounds 1 (3): 116-124. DOI: 10.3390/compounds1030010.
Zhi X, Fang C, Gu Y, Chen H, Chen X, Cui J, Hu Y, Weng W, Zhou Q, Wang Y, Wang Y, Jiang H, Li X, Cao L, Chen X, Su J. 2020. Guaiacol suppresses osteoclastogenesis by blocking interactions of RANK with TRAF6 and C-Src and inhibiting NF-?B, MAPK and AKT pathways. J Cell Mol Med 24 (9): 5122-5134. DOI: 10.1111/jcmm.15153.