Identification of Nematode Trapping Fungus Monacrosporium eudermatum Based on Genetic Diversity Using RAPD Technique

Ali A. Kasim^1, and Maitham Dragh¹

¹Biological Department, College of Sciences, Misan University, Misan, Iraq

Cite this paper:
Ali A. Kasim and Maitham Dragh. Identification of Nematode Trapping Fungus Monacrosporium eudermatum Based on Genetic Diversity Using RAPD Technique. American Journal of Microbiological Research. 2016; 4(6):178-180. doi: 10.12691/ajmr-4-6-4

Abstract

Nematode-trapping fungus Monacrosporium eudermatum a predacious fungus of nematodes, has been very useful in understanding the most relationship between nematophagous fungi and their nematode hosts. M.eudermatum is by far the common nematode-trapping fungus with the characteristic ability of forming adhesive trapping nets once in contact with nematodes. Total DNA was extracted from M.eudermatum sera and amplified by RAPD-PCR using three random primers (OPA-16, OPB-08, OPF-05). The results show amplification of all markers (OPF-05, OPB-08, OPA-16) with the genomic DNA studied. As evidenced by the results, all the markers covered in the study showed high polymorphism (up to 60%) with the exception of the primer (OPB-08) which showed no polymorphism.

Keywords:
nematophagous Monacrosporium eudermatum RAPD technique PCR

This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

References:

[1]	Williams, J.G.K., Hanafey, M.K., Rafalski, J.A. and Tingey S.V., “Genetic analysis using random amplified polymorphic DNA markers,” Methods Enzymol., 218: 704-741. 1993.
[2]	Mc-Donald, B.A., “The population-genetics of fungi tools and techniques,” Phytopathology, 87: 448-453. April. 1997.
[3]	Williams, J.G., Kubelik, A.R., Livak K.J., Rafalski, J.A. and Tingey, S.V. “DNA polymorphisms amplified by arbitrary primers are useful as genetic markers,” Nucleic Acids Res. 18(22): 6531-6535. Nov.1990.
[4]	Atkins, S.D., Clark, I.M., Sosnowska, D., Hirsch, P.R. and Kerry, B.R. “Detection and quantification of Plectosphaerella cucumerina, a potential biological control agent of potato cyst nematodes, by using conventional PCR, real-time PCR, selective media, and baiting,” Appl. Environ. Microbiol, 69(8):4788-93.Aug.2003.
[5]	Morton, C.O., Mauchline, T.H., Kerry, B.R. and Hirsch, P.R., “PCR-based DNA finger-printing indicates host-related genetic variation in the nematophagous fungus Pochonia chlamydosporia,” Mycol. Res., 107(Pt 2):198-205. Feb.2003.
[6]	Sugimoto, M., Koike, M., Hiyama, N. and Nagao, H., “Genetic, morphological, and virulence characterization of the entomopathogenic fungus Verticillium lecanii.” J Invertebr Pathol, 82(3):176-87. Mar.2003.
[7]	Zhang, Y., Qiao, M., Xu, J., Cao, Y., Zhang, K.-Q.and Yu, Z.F., “Genetic diversity and recombination in natural populations of the nematode-trapping fungus Arthrobotrys oligospora from China,” Ecol Evol., 3(2):312-25.Feb.2013.
[8]	Persson, Y., Erland, S. and Jansson, H. B., “Identification of nematode-trapping fungi using RFLP analysis of the PCR-amplified ITS region of ribosomal DNA.” Mycol. Res, 100: 531-534. May.1996.
[9]	Ahrén, D., Faedo, M., Rajastiekar, B. and Tunlid, A., “Low genetic diversity among isolates of the nematode-trapping fungus Duddingtoniaflagrans: evidence for recent world wide dispersion from a single common ancestor,” Mycol. Res, 108 (10): 1205-1214. Oct.2004.
[10]	Meyer, S. L. F., Carta, L. K. and Rehner, S.A., “Morphological variability and molecular phylogeny of the nematophagous fungus Monacro-sporium drechsleri.” Mycologia, 97(2). 405-415. Mar./Apr. 2005.
[11]	Muhammad, N. U., Saifullah, M. A., Ijaz, A., Aqib, I. and Naqib, U. K., “Genetic characterization of Verticillium chlamydosporium isolated from Pakistan using Random Amplified Polymorphic DNA (RAPD) primers,” Pak. J. Bot., 45(2): 467-472. 2013.
[12]	Herrera-Estrella, A., Casas-Flores, S. and Kubicek, C. P., “Nematophagous Fungi”. Environmental and Microbial Relationships 3rd Edition, The Mycota IV I.S. Druzhinina and C.P. Kubicek, (Eds.© Springer International Publishing Switzerland ), 2016, 247-267.
[13]	Miller, S.A., Dykes, D.D. and Polesky, H.F, “A simple salting out procedure for extracting DNA from human nucleated cells,” Nucleic Acids Res. 11; 16(3): 1215Feb.1988.
[14]	Ahren, D., Ursing B. M., and Tunlid, A., “Phylogeny of nematode-trapping fungi based on 18S rDNA sequences,” FEMS Microbiol. Lett., 158:179-184. Jan.1988.
[15]	Li, Y., Hyde, K. D., Jeewon, R., Cai L., Vijaykrishna, V. and Zhang, K. Q., “Phylogenetics and evolution of nematode-trapping fungi (Orbiliales) estimated from nuclear and protein coding genes,” Mycologia 97(5):1034-46. Sep-Oct. 2005.
[16]	Yang, Y., Yang, E., An, Z. and Liu, X., “Evolution of nematode-trapping cells of predatory fungi of the Orbiliaceae based on evidence from rRNA-encoding DNA and multiprotein sequences,” Proc. Natl Acad. Sci. USA 104: 8379-8384.May. 2007.
[17]	Yang, E., Xu, L., Yang, Y., Zhang, X., Xiang, M., Wang, C. “Origin and evolution of carnivorism in theAscomycota (fungi),” Proc. Natl Acad. Sci. USA 109:10960-10965. Jul. 2012.
[18]	Zhang, W., Cheng, X. , Liu, Y. and Xiang, M., “Genome Studies on Nematophagous and Entomogenous Fungi in China,” J. Fungi, 2, 9 :1-14. Feb.2016.
[19]	Smith, M. E. and Jaffee, B. A.,”PCR Primers with Enhanced Specificity for Nematode-Trapping Fungi (Orbiliales),” Microb Ecol, 58:117-128. Jul.2009.
[20]	Castañeda-Ramírez, G.S., Mendoza-de-Gives, P., Aguilar-Marcelino, L., López-Arellano, M. E. and Hernández-Romano, J., “Phylogenetic Analysis of Nucleotide Sequen-ces from the ITS Region and Biological Characterization of Nematophagous Fungi from Morelos, Mexico,” Journal of Mycology. V. Article ID 8502629, 13 p. 2016.
[21]	Mauchline, T.H., Kerry, B.R. and Hirsch, P.R., “The biocontrol fungus Pochonia chlamy-dosporia shows nematode host preference at the in fraspecific level,” Mycol. Res., 108: 161-169. Feb. 2004.
[22]	Mauchline, T.H., Kerry, B.R. and Hirsch, P.R., “Quantification in Soil and the Rhizosphere of the Nemato-phagous Fungus Verticillium chlamydosporium by Com-petitive PCR,” Appl Environ Microbiol, 68(4): 1846-53. Apr.2002,
[23]	Siebert, P.D., and Larric, J.W. “Competitive PCR,” Nature, 359: 557-558. Oct. 1992.