Study of the activity of β -1,4 endoglucanase enzyme in three plant parasites nematodes*

Document Type : Research Article

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Abstract

Host plant cell wall is the main mechanical barrier to penetration and migration by endoparasitic nematodes. In order to degradation of plant cell walls, the nematodes secrete various types of cell wall degrading enzymes. β‐1, 4‐endoglucanase was the first known cell wall degrading enzyme. In this research, the enzyme activites were studied in three species; Meloidogyne incognita (Kofoid & White1919)Chitwood, 1949, Pratylenchus loosi Loof, 1960 and Aphelenchoides besseyi Christie, 1942. Initially, sampling, identification, purification and multiplication of the nematodes were performed and after extraction of their DNA, the encoding β - 1, 4 endoglucanase gene was detected in the nematodes  by using pairs of specific degenerate primers in polymerase chain reaction. After extraction of total RNA from three species of nematodes, cDNA was synthesized and determination of cDNA goodness was carried out by a reference gene. Gene expression of the β -1,4 endoglucaase was done for all samples. The results showed that the gene encoding β -1 ,4 endoglucanase had the highest activity and expression in the root- knot nematode (M. incognita). Tea root lesion nematode (P. loosi) and Rice white tip nematode (A. besseyi) were classified in  next  step, respectively.

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References

Bellafiore S., Shen Z.X., Rosso M.N., Abad P., Shih P. and Briggs S.P. 2008.Direct Identification of the Meloidogyne incognita Secretome Reveals Proteins with Host Cell Reprogramming Potential. Plos Pathogen. 4(10): 1-12.
Bridge J., Plowright R. A. and Peng D. 2005. Nematode parasites of rice. In: Luc. M, R.A. Sikora and J. Bridge. (Eds.), Plant Parasitic Nematodes in Subtropical and Tropical Agriculture. 2nd ed., CABI Publishing,Wallingford, UK. pp 87-130.
Coolen W.A. and  D'Herde, C. J. 1972. A method for the quantitative extraction of nematodes from plant tissue. Ghent Agri. Res. Cent., Belgium, 77p.
Curtis R.H.C. 2007.Plant parasitic nematode proteins and the host-parasite interaction. Briefings in Functional Genomics and Proteomics 6: 50-58.
Davis E. L., Hussey R. S., Baum T. J., Bakker J., Schots A., Rosso M.N. and Abad P. 2000. Nematode parasitism genes. Annual Review of Phytopathology 38:365-396.
Davis E., Haegeman A. and Kikuchi T. 2011. Degradation of the plant cell wall by nematodes. In Genomics and Molecular Genetics of Plant-Nematode Interactions. 225-272.
Doyle E.A., and Lambert K.N. 2002. Cloning and characterization of an esophageal-gland-specific pectate lyase from the root-knot nematode Meloidogyne javanica. Molecular Plant-Microbe Interaction 15:549-556.
Fanelli E, Troccoli A., Picardi E., Pousis C. and De Luca F. 2014. Molecular characterization and functional analysis of four b-1,4-endoglucanases from the root-lesion nematode Pratylenchus vulnus. Plant Pathology 63: 1436-1445.
Fu Z, Agudelo P. and Wells CE. 2012. Differential expression of a β-1, 4-endoglucanase induced by diet change in the foliar nematode Aphelenchoides fragariae. Phytopathology 102 (8): 804-811.
Gao B.L., Allen R., Maier T., Davis E.L., Baum T.J. and Hussey R.S. 2003. The parasitome of the phytonematode Heterodera glycines. Molecular Plant-Microbe Interactions 16: 720-726.
Goellner M, Smant G., De Boer J.M., Baum T.J. and Davis E.L. 2000. Isolation of β-1,4 endoglucanase genes from Globodera tabacum and their expression during parasitism. Nematology 32: 154-165.
Haegeman A., Jacob J., Van Holme B., Kyndt T. and Gheysen G. 2008. A family of GHF5 endo-1,4-β-glucanases in the migratory plantparasitic nematode Radopholus similis. Plant Pathology 57: 581-590.
Hussey R. S. and Barker K. R. 1973. Comparison of methods for collecting inocula of Meloidogyne spp., including a new technique. Plant Disease Reporter 57:1025-1028.
Jabbari H., Niknam G., Tanha Maafi Z., Elashry A. and Grundler F.M.W.  2018. Identification of some putative parasitism effectors of Heterodera cruciferae, Franklin, 1945 on cabbage. Iranian Journal of Plant Pathology 53(3):287-302.
Jamali S., Pourjam E., Alizadeh A. and Alinia F. 2008. Reproduction of white tip nematode (Aphelenchoides besseyi Christie, 1942) in different monoxenic cultures. Agricultural Science and Technology 10: 165-171.
Kikuchi T., Jones J.T., Aikawa T., Kosaka H. and Ogura N. 2004. A family of glycosyl hydrolase family 45 cellulases from the pine wood nematode Bursaphelenchus xylophilus. FEBS Letters 572: 201-205.
Ledger T.N., Jaubert S., Bosselut N., Abad P. and Rosso M.N. 2006. Characterization of a new β-1,4-endoglucanase gene from the root-knot nematode Meloidogyne incognita and evolutionary scheme for phytonematode family 5 glycosyl hydrolases. Gene 382: 121-128.
Livak K.J. and Schmittgen T.D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2−11CT method. Methods 25: 402-408.
Long H., Peng D., Huang W., Peng H. and Wang G. 2013. Molecular characterization and functional analysis of two new b- 1,4-endoglucanase genes (Ha-eng-2, Ha-eng-3) from the cereal cyst nematode Heterodera avenae, Plant Pathology 62: 953-960.
Lowry O.H., Rosebrough N.J., Farr A.L. and Randall R.J. 1951. Protein measurement with the Folin phenol reagent. The Journal of Biological Chemistry 193: 265-275.
Moody E.H., Lownsbery B.F. and Ahmad J.M. 1973. Culture of the root- lesion nematode Pratylenchus vulnus on carrot discs. Nematology 3:225-226.
Muturi J., Gichuki C., Waceke J.W. and Runo S.M. 2010. Use of isoenzymes phenotypes to 22 characterize the major root knot nematodes (Meloidogyne spp.) parasitizing indigenous leafy vegetables in Kisii, 12th 23 KARI Biennial Scientific Conference, Kenya, Kenya 24 Agricultural Research Institute, Kenya, pp. 605-612.
Puissant C. and Houdebine L. 1991. An improvement of the single step method of RNA isolation by acid Guanidinium Thiocyanate Phenol chloroform Extraction. Bio-Techniques 8: 148-149.
Rosso M. N., Favery B., Piotte C., Arthaud L., De Boer J. M., Hussey R. S., Bakker J., Baum T. J. and Abad P. 1999. Isolation of a cDNA encoding a β-1,4-endoglucanase in the root-knot nematode Meloidogyne incognita and expression analysis during plant parasitism. Molecular Plant-Microbe Interactions 12: 585-591.
Sambrooke J., Frritsch EF.and Manitis T. 2000. Molecular cloning: a laboratory manual, 3rd ed. Cold Spring Harbor laboratory press. Cold Spring Harbor, N.Y. Chapter 7 pp: 1-77.
Seraji A., Pourjam E., Safaie N. and Maafi Z.T. 2010. Effect of tea root lesion nematode (Pratylenchus loosi) on tea quality in Iran. The 4th International Conference on O – CHA (Tea) Culture and Science, 26 -28 October, Shizuoka, Japan.
Smant G., Stokkermans J.P.W.G., Yan Y.T., De Boer J.M., Baum T.J., Wang X.H., Hussey R.S., Gommers F.J., Henrissat B., Davis E.L., Helder J., Schots A. and Bakker J. 1998. Endogenous cellulases in animals: Isolation of β-1,4-endoglucanase genes from two species of plant-parasitic cyst nematodes. Procceding of National Academy Science of the USA 95: 4906-4911.
Taylor D.P. and Netscher C. 1974. An improved technique for preparing perineal patterns of Meloidogyne spp. Nematologica 20: 268-269.
Tondje P.R., Roberts D.P., Bon M.C., Widmer T., Samuels G.J., Ismaiel A., Begoude A.D., Tchana T., Nyemb-Tshomb E., Ndoumbe-Nkeng M., Batman R., Fontem D. and Hebbar K.P. 2007. Isolation and identification of mycoparasitic isolates of Trichoderma asperellum with potential for suppression of black pod disease of cacao in Cameron. Biological Control 43: 202-212.
Uehara T., Kushida A. and Momota Y. 2001. PCR-based cloning of two β-1,4-endoglucanases from the root-lesion nematode Pratylenchus penetrans. Nematology 3: 335-341.
Wubben M.J., Callahan F.E. and Scheffler B.S. 2010. Transcript analysis of parasitic females of the sedentary semi-endoparasitic nematode Rotylenchulus reniformis. Molecular and Biochemical Parasitology 172: 31-40.
Wu G.L., Kuo T.H., Tsay T.T., Tsai I.J., Chen P. J. 2016. Glycoside Hydrolase (GH) 45 and 5 Candidate Cellulases in Aphelenchoides besseyi Isolated from Bird’s-Nest Fern . PLoS ONE 11(7): DOI:10.1371.
Iranian Journal of Plant Pathology
Volume 55, Issue 1
October 2019
Pages 1-12

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