Surface Sterilization of Mango’s Mistletoe Leaves (Dendrophtoe pentandra (L.) Miq.) for Endophytic Fungi Isolation

Nur Anizah; Nour Athiroh Abdoes Sjakoer

doi:10.21776/ub.jsmartech.2022.003.01.16

Authors

Nur Anizah Departement of Biology, Faculty of Mathematics and Natural Sciences, Islamic University of Malang, Malang, East Java, Indonesia
Nour Athiroh Abdoes Sjakoer Departement of Biology, Faculty of Mathematics and Natural Sciences, Islamic University of Malang, Malang, East Java, Indonesia

DOI:

https://doi.org/10.21776/ub.jsmartech.2022.003.01.16

Keywords:

Dendrophthoe pentandra, endophytic fungi, Sodium Hypochlorite, surface sterilization

Abstract

Mango mistletoe (Dendrophthoe pentandra L. Miq) is known as a parasitic plant that has been scientifically proven to have the potential as an antihypertensive adjuvant herbal. The content of secondary metabolites in plants is produced by plants andby microorganisms that grow in plant tissues. One of them is endophytic fungi which can induce the host to produce secondary metabolites. The ability of endophytic fungi to synthesize secondary metabolites is an opportunity for the large-scale production of phytopharmaca raw materials in a short time without causing ecological damage. Before isolating endophytic fungi from the plant tissue, one needs to research surface sterilization to remove various kinds of microorganisms that are not desired. This study aims to determine the best method’s surface sterilization of the mango’s mistletoe leaves (Dendrophthoe pentandra (L.) Miq) to isolate endophytic fungi. This study used a two-factor completely randomized design. The concentration of Sodium Hypochlorite (NaOCl) was 5% and 10% as the first factor, and the immersion time of 1, 2, 3, 4, and 5 minutes was the second factor. There is a total of 10 treatments. In each treatment, there were three replications and one control. Observational data were analyzed descriptively. The results showed that application of NaOCl 10% for 3 minutes is the best and recommended method’s surface sterilization of the mango’s mistletoe leaves because the percentage of live explants is 100%, the percentage of contamination is 0%, and the percentage of explants with endophytic fungi is 100%.

References

Anjani, M., Sjakoer, N.A.A., Mubarakati, N.J. 2021. Studi Sub Kronik 28 Hari: Uji Toksisitas Ekstrak Metanolik Kombinasi Scurrulla atropurpurea dan Dendrophthoe pentandra Terhadap Kerusakan Fungsi Ginjal Tikus Wistar Betina. e-JBST. 6(2): 58-63.

Radji, M. 2005. Peranan Bioteknologi Dan Mikroba Endofit Dalam Pengembangan Obat Herbal. J. Pharm. Sci. Res. 2(3): 113-126.

Sugijanto, N.E. 2011. Produksi Bahan Bioaktif Berkhasiat Obat Menggunakan Fungi Endofit. Pidato Pengukuhan Jabatan Guru Besar Pada Fakultas Farmasi Universitas Airlangga: Surabaya.

Oyebanji, O.B., Nweke, O., Odebunmi, O., Galadima, N.B., Idris, M.S., Nnodi, U.N., Afolabi, A.S., Oghadu, G.H.2009. Simple, Effective, and Economical Explant-Surface Sterilization Protocol for Cowpea, Rica and Sorghum Seeds. Afr. J. Biotechnol. 8(20): 5395-5399.

Fitriani, Y., Wijayana, G., Darmawati., I.A.P. 2019. Teknik Sterilisasi dan Efektifitas 2,4 D Terhadap Pembentukan Kalus Eksplan Daun Nilam (Pogostemon cablin Benth.) In Vitro. J. Agr. Sci. Biotech. 8(1): 41-52.

Yildiz, M., Er, C. 2002. The Effect of Sodium Hypochlorite On In Vitro Seedling Growth and Shoor Regeneration of Flax (Linum usitatisimum). J. Naturwissenschaften. 89: 259-261.

Olowe, O.B., Adesoye, A., Ojoba, O., Amusa, O., Liamngee, S. 2014. Effect of Sterilization and Phytohormones on Shoot Tip Culture of Telfairia occidentalis. J. Nat. Sci. Res. 4(2): 53-58.

Wardani, D.P, Solichatun, Setyawan, AD. 2003. Pertumbuhan dan Produksi Saponin Kultur Kalus Talinum paniculatum Gaertn. Pada Variasi Penambahan Asam 2,4 Dikloropenoksi Asetat (2,4 D) dan Kinetin. Biofarm. 2(1): 35-43.

Intan, S. 2013. Isolasi Fungi Endofit Penghasil Senyawa Antimikroba dari Daun Cabai Katokkon (Capsicum annuum (L.) var Chinensis) dan Profil KLT Bioautografi. Skripsi. Fakultas Farmasi. Universitas Hasanuddin: Makassar.

Maulana, M.R., Restanto, D.P., Slameto. 2019. Pengaruh Konsentrasi 2,4-Dicholorophenoxyaceticacid (2,4-D) Terhadap Induksi Kalus Tumbuhan Sorgum (Sorghum bicolor (L.) Moench). J. Bioindustri. 1(2): 138-148.

Utami, U., Harianie, L., Kusmiyati, N., Fitriasari, P.D. 1988. Buku Panduan Praktikum Mikrobiologi Umum. Malang: Universitas Negeri Maulana Malik Ibrahim. 2018.

Fardiaz, S. Mikrobiologi Pengolahan Pangan Lanjut. IPB Press: Bogor.

Asep, A.P., Timur, H.D.L., Jaziri, A.A., Nurdiani, R., Pradarameswari, K.A. 2018. Isolasi dan Identifikasi Bakteri Endofit Mangrove Sonneratia alba Penghasil Enzim Gelatinase Dari Pantai Sendang Biru, Malang, Jawa Timur. Ind. J. Halal. 1(1): 31-42.

Suhartina, Kandou, F.E.F., Singkoh, M.F.O. 2018. Isolasi dan Identifikasi Fungi Endofit Pada Tumbuhan Paku Asplenium nidus. J. MIPA UNSRAT. 7 (2): 24-28.

Ribeiro, S.F.L., Garcia, A.D.C., Santos, H.E.D.D., Montoya, Q.V., Rodrigues, A., Oliveira, J.M.D., Oliviera, C.M.D. 2018. Antimicrobial Activity OF Crude Extract OF Endophytic Fungi From Oryctanthus alveolatus (Kunth) Kuijt (Mistletoe). Afr. J. Microbiol. Res. 12 (1): 263-268.

Slama, H.B., Bouket, A.C., Alenezi, F.N., Pourhassan, Z., Golinska, P., Oszako, T., Belbahri, L. 2021. Potentials of Endophytic Fungi in the Biosynthesis of Versatile Secondary Metabolites and Enzymes. Forest. 12(1784): 1-13.

Shofiyani, A., Purnawanto, A.M., Aziz, R.Z.A. 2020. Pengaruh Berbagai Jenis Sterilan dan Waktu Perendaman Terhadap Keberhasilan Sterilisasi Eksplan Daun Kencur (Kaempferia galanga L.) Pada Teknik Kultur Jaringan. J. Agritech. 22 (1): 29-39.

Zhang, H.W., Song, Y.C., Tan, R.X. 2006. Biology and Chemistry of Endophytes. Nat. Prod. Rep. 23(5): 753-771.

Tejasvi, M.V., Nalini, M.S., Mahesh, B., Prakash, H.S., Kini, K.R., Shetty, H.S., Sabbiah, V. 2007. New Hopes from Endophytic Fungal Secondary Metabolites. Bol. Soc. Quim. Mex. 1(1): 19-26.

Kuncoro, H., Sugijanto, N.E.2011. Fungi Endofit, Biodiversitas, Potensi dan Prospek Penggunaannya Sebagai Sumber Bahan Obat Baru. J. Trop. Pharm. Chem. 1(3): 247-262.

Bhojwana, S.S, Razdan M.K. 1996. Plant Tissue Culture Theory and Practice, a Revised Edition. Elsevier: Amsterdam.

Hendaryo, D.P., Wijayani. 1994.Teknik Kultur Jaringan dan Petunjuk Perbanyakan Tumbuhan Secara Vegetatif Modern. Kanisius: Yogyakarta.

Admojo, L., Indrianto, A. 2016. Pencegahan Browning Fase Inisiasi Kalus Pada Kultur Midrib Daun Klon Karet (Hevea brasiliensis Muell. Arg) PB 330. J. Pen. Kar. 34(1): 25-34.

Dian, R., Fatimawali, Fona, B. 2015. Uji Resistensi Bakteri Escherichia coli Yang Diisolasi Dari Plak Gigi Terhadap Merkuri dan Antibiotik Kloramfenikol. J. e-Biomed. 3(1): 59-63.

Gunawan, L.W. 1992. Teknik Kultur Jaringan Tumbuhan. PAU Bioteknologi. Departemen Pendidikan dan Kebudayaan. Direktorat Jenderal Pendidikan Tinggi. Institute Pertanian Bogor: Bogor.

Setiani, N.A., Nurwinda, F., Astriany. D. 2018. Pengaruh Desinfektan Dan Lama Perendaman Pada Sterilisasi Eksplan Daun Sukun (Artocarpus altilis (Parkinson ex. F.A Zorn) Fosberg). J. Biotrop. 6(3): 78-82.

Nurhelmi, Putri, D.H. 2021. Optimasi Sterilisasi Permukaan Jaringan Daun Andalas (Morus macroura Miq.) Dengan NaOCl Untuk Isolasi Mikroba Endofit. J. Ser. Biol. 6(1): 13-18.