Morphological and molecular identification of cellulolytic fungi associated with local raru species

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DENI ELFIATI
ARIDA SUSILOWATI
CELVIA MODES
HENTI HENDALASTUTI RACHMAT

Abstract

Abstract. Elfiati D, Susilowati A, Modes C, Rachmat HH. 2019. Morphological and molecular identification of cellulolytic fungi associated with local raru species. Biodiversitas 20: 2348-2354. This study is aimed to isolate and identify cellulolytic fungi from different local Raru species in Central Tapanuli, North Sumatra, Indonesia. Cellulolytic fungi are fungi that can degrade natural polymer materials such as cellulose. The fungal species were isolated from soil under three local raru species. The samples inoculating (1 mL) from serial dilutions (10-2 to 10-5) on Carboxy Methyl Cellulose (CMC) plates. The plates were incubated at 28 ?C for one week, then the fungal colonies were observed and pure cultures were maintained. Fungi with cellulolytic activity were characterized by the formation of clear zones around the colonies on CMC media plus congo red (0.1%). The identification of fungi was carried out by molecular identification of the isolated fungi at the species level. The extracted fungal DNA was amplified by PCR using specific internal transcribed spacer primer (ITS1/ITS4). The PCR products were sequenced and compared with the other related sequences in Gen Bank (NCBI). 20 isolates were successfully isolated and nine isolates had cellulolytic activity. Six fungal species from 20 isolates were identified as: Aspergillus sp., Penicillium sp., Aspergillus terreus, Penicillium mariae-crucis, Aspergillus tanneri and Penicillium citrinum. The results showed that Aspergillus terreus was the most abundant fungus obtained from all the raru stand.

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References
Ahmed MM, El-Zayat SA, El-Sayed MA. 2018. Celluloytiv activity of cellulose-decomposing fungi isolated from Aswan hot desert soil, Egypt. Journal of Biological Studies 1(2):35-48
Anderson IC, Parkin PI. 2007. Detection of active soil fungi by RT-PCR amplification of precursor rRNA molecules. Journal of Microbiological Methods, 68:248-253
Ashton, PS. 1998. Cotylelobium melanoxylon. The IUCN Red List of threatened species 1998:e. T33070A9746855
Bennett JW .1998. Mycotechnology: the role of fungi in biotechnology. J Biotechnol 66: 101–107. doi:10.1016/S0168-1656(98)00133-3.
Bao Z, Ikunaga Y, Matsushita Y, Morimoto S, Takada-Hoshino Y, et al. 2012. Combined analyses of bacterial, fungal and nematode communities in Andosolic agricultural soils in Japan. Microbes Environ., 27:72–79.
Barstow M. 2019. Cotylelobium melanoxylon. The IUCN Red List of Threatened Species 2019: e.T33070A68069829. DOI: 10.2305/IUCN.UK.2019-1.RLTS.T33070A68069829.en.
Brigg JC. 2015. Species extinction: Frequency and biogeography . Environmental Skeptics and Critics : 4(4): 96-105
David H, Hofmann G, Oliveira AP, Jarmer H. 2006. Metabolic net- work driven analysis of genome-wide transcription data from Asper- gillus nidulans. Genome Biol 7. doi:10.1186/gb-2006-7-11-r108.
Gilman JC. 1971. A Manual of Soil Fungi. The Low a State University Press. USA
Gaddeyya G, Niharika PS, Bharathi P and Kumar PKR. 2012. Isolation and identification of soil mycoflora in different crop fields at Salur Mandal. AdvAppl Sci Res., 3:2020-2026
Gandjar I, Samson RA, van den Tweel-Vermeulen K, Oetari A, Santoso I. 1999. Introduction of General Tropical Fungi. University of Indonesia. Depok. Indonesia
Hawksworth DL (2011) Naming Aspergillus species: progress to- wards one name for each species. Med Mycol 49: 70–76. doi: 10.3109/13693786.2010.504753.
Henselt M and Holden DW. 1996. Molecular genetic approaches for the study of virulence in both pathogenic bacteria and fungi. Microbiol., 142:1049-1058.
Herculano PN, Lima DMM, Fernandes MJS, Neves RP, Souza-Motta CM, Porto ALF. 2011. Isolation of cellulolytic fungi from waste of castor (Ricinus communis L.) Curr. Microbiol 62:1416-1422
Hossain, M. M.; Sultana, F.; Kubota, M.; Koyama, H. and M. Hyakumachi 2007. The plant growth-promoting fungus Penicillium simplicissimum GP17-2 induces resistance in Arabidopsis thaliana by activation of multiple defense signals. Plant Cell Physiol. 48: 1724–1736.
Hyakumachi, M. 1994. Plant-growth-promoting fungi from turfgrass rhizosphere with potentials for disease suppression. Soil Microorg. 44: 53–68.
Khan SA, Hamayun M, Yoon H, Kim HY, Suh SJ, Hwang SK. 2008. Plant growth promotion and Penicillium citrinum. BMC Microbiol. 8:231. doi: 10.1186/1471-2180-8-231
Kim MJ, Lee H, Choi YS, Kim, GH, Huh NY., Lee, S, Lim, Y.W., Kim, J.-J. 2011. Diversity of fungi in creosote-treated crosstie wastes and their resistance to polycyclic aromatic. hydrocarbons. A. Van Leeuw. J. Microb. 97:377–387
Krijgsheld P, Bleichrodt R, van Veluw GJ, Wang F, Mu?ller WH, Di- jksterhuis J, Wo?sten HAB. 2013. Development in Aspergillus. Stud Mycol 74: 1–29. DOI:10.3114/sim0006.
Kuhad RC, Gupta R, Singh A. 2011. Microbial Cellulases and their Industrial Applications. Enzyme Research. 1-10. DOI: 10.4601/2011/280696
Land Research Center. 2005. Chemical Analysis of Soil, Plants, Water and Fertilizer. Agricultural Research and Development Center. Agricultural Department.
Landeweert R, Veenman C, Kuyper TW, Fritze H, Werners K & Smit E (2003b) Quantification of ectomycorrhizal mycelium in soil by real time PCR compared to conventional quantification techniques. FEMS Microbiol Ecol 45: 283–292
Lima RF and Borba 2001. Viability, morphological characteristics and dimorphic ability of fungi preserved by different methods. Rev Iberoam Mycol., 18: 191-196.
Madigan, MT; Martinko, JM and Parker, J. 2012. Biology of Microorganism Eight Edition Prentice Hall International, Inc.
Maheswari NU and Komalavalli R. 2013. Diversity of soil fungi from Thiruvarur District, Tamil Nadu, India. Int J Curr Microbiol App Sci., 2:135-141
Meena VS, Meena SK, Verma JP, Kumar A, Aeron A, Mishra PK, Bisht JK, Pattanayak A, Naveed M, Dotaniya MI. 2017. Plant beneficial rhizospheric microorganism (PBRM) strategies to improve nutrient use efficiency: A review. Ecological Engineering 107:8-32
Monod M, Bontems O, Zaugg C, Chenne BL, Fratti M and Panizzon R. 2006. Fast and reliable PCR/sequencing/RFLP assay for identification of fungi in onychomycoses. J Medical Microbiol., 55: 1211–1216.
Mulani RM and Turkmane KL. 2014. Diversity of hizospheric fungi of Ceropegia bulbosa Var. bulbosa Roxb. J Global Biosci., 3(4): 1089– 1093.
Nath D, Mauriya BR, Meena VS. 2017. Documentation of fire potassium and phosphosphorus solubilizing bacteria for their K and P solubilization ability from various mineral. Biocatal.Agric.Biotechnol.10:174-181
Punt PJ, van Biezen N, Conesa A, Albers A, Mangnus J, van den Hondel C .2002. Filamentous fungi as cell factories for heterolo- gous protein production. Trends Biotechnol 20: 200–206. doi:10.1016/ S0167-7799(02)01933-9.
Radhakrishnan R, Shim KB, Lee BW, Hwang CD, Pae SB, Park CH, Kim SU, Lee CK, Baek IY. 2013. IAA- producing Penicillium sp. NICS01 triggers plant growth and suppresses Fusarium sp.-induced oxidative stress in sesame (Sesamum indicum L.). J Microbiol Biotechnol. 23:856–863.
Rebecca LJ, Dhanalakshmi V, Sharmila S, Susithra G, Kumar S and Bala S. 2012. Isolation, identification and characterization of fungi from rhizosphere soil of Barleria Cristata. Inter J Hort Crop Sci Res., 2: 1-6.
Reddy PLN, Babu BS, Radhaiah A, Sreeramulu A. 2014. Screening, identification, and isolation of cellulolytic fungi from soils of Chittoor District, India. Int. J. Curr. Microbial. App.Sci. 3(7):761-771
Rodrigues, K. F., Hesse, M. & Werner, C. 2008. Antimicrobial activities of secondary metabolites produced by endophytic fungi from Spondias mombin. Journal of basic microbiology, 40(4), 261- 267
Saraswati R, Husen E, Simanungkalit RDM. 2007. Soil Biological Ananlysis Method. Agricultural Research and Development Center. Agricultural Department.
Sari SLA, Setyaningsih R, Wibowo NFA. 2017. Isolation and screening of cellulolytic fungi from Salacca zalacca leaf litter. Biodiversitas. 18(3):1282-1288
Sartori FG, Leandro LF, Montanari LB, de Souza et al., .2013. Isolation and identification of environmental mycobacteria in the waters of a hemodialysis center. Curr Microbiol., 67: 107-111
Schoch CL, Seifert KA, Huhndorf S, Robert V, Spouge JL, Levesque CA. 2012.Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi. Proceedings of the National academy of Sciences of the United States of America, 109: 6241-6246
Shivanna, M B, Meera M S, Hyakumachi M. 1996. Role of root colonization ability of plant growth promoting fungi in suppression of take-all and common root rot of wheat. Crop Protect 15: 497–504.
Susilowati A, Rachmat HH, Elfiati D, Kholibrina CR, Kusuma YS, Siregar H. 2019. Population structure of Cotylelobium melanoxylon within vegetation community in Bona Lumban Forest, Central Tapanuli, North Sumatra, Indonesia. Biodiversitas 20: 1681-1687
Torres LMA, Pulido CPG, Rojas ET. 2014. Assesssment of cellulolytic microorganisms in soils of Nevados Park, Colombia. Brazilian Journal of Microbiology 45(4):1211-1220
Torres J. L., Wilbers R. H. P., Gawronski P., Boshoven J. C., Finkers-Tomczak A., Cordewener J. H. G., 2012. Dual disease resistance mediated by the immune receptor Cf-2 in tomato requires a common virulence target of a fungus and a nematode. Proc. Natl. Acad. Sci. U.S.A. 109, 10119–10124. 10.1073/pnas.1202867109
Tuli MS, Gurumayum S, Kaur S, Nagal S, Attri I. 2015. Isolation and screening of cellulolytic fungi by Baiting Method from Soil of Jalandar. Research Journal of Pharmaceutical, Biological and Chemical Sciences 6(2):375-380
Verma JP, Yadav J, Tiwari KN, Lavakush SV. 2010. Impact of plant growth promoting rhizobacteria on crop production. Int. J. Agric. Res.5:954-983
Verma R, Maurya BR, Meena VS, Dotaniya MI, Deewan P. 2017. Enhancing production potential of cabbage and improves soil fertility status of Indo-Gangetic Plant through application of bio-organics and mineral fertilizer. Int. J. Curr. Microbiol. App. Sci. 6(3):301-309
Yadav K, Singh N, Aggarwal A. 2011. Influence of arbuscular mycorrhizal (AM) fungi on survival and development of micropropagated acorus calamus L. during acclimatization. Journal of Agricultural Technology, 7: 775–781.

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