Meloidogyne arenaria belongs to root-knot nematodes (RKNs) which constitute a group of highly polyphagous nematodes causing serious damages to many crop varieties. Maize (Zea mays) is one of its main hosts. During plant response to RKN infection, many mechanisms are involved. Pathogenesis-related proteins (PRs), which present many functions and en- zymatic activities, such as ribonucleases (RNases), antioxidative enzymes, or proteases are involved in these processes. The aim of this study was to describe changes in peroxidase and RNase activities induced in Z. mays during its response to M. arenaria infection. Moreover, proteins potentially responsible for peroxidase activity were indicated. RNase and peroxi- dase activities were tested on proteins extracted from roots of healthy plants, M. arenaria infected plants, and healthy plants mixed with M. arenaria juveniles, in native polyacryla- mide (PAA) gels. Samples were collected from two varieties of maize at four time points. A selected fraction showing peroxidase activity was excised from the gel and analyzed using mass spectrometry (MS) to determine protein factors responsible for enzymatic activity. As a result, the analyzed varieties showed slight differences in their RNase and peroxidase activities. Higher activity was observed in the Tasty Sweet variety than in the Waza variety. There were no significant differences between healthy and infected plants in RNase activi- ties at all time points. This was in contrast to peroxidase activity, which was the highest in M. arenaria-infected plants 15 days after inoculation. On the basis of protein identification in excised gel fractions using MS it can be assumed that mainly peroxidase 12 is responsible for the observed peroxidase activity. Moreover, peroxidase activity may be presented by glutathione-S-transferase as well.
I wish to thank Prof. A. Obrępalska-Stęplowska from IPP-INR for her valuable help and suggestions during preparation of the manuscript.
This study was supported by the Polish National Scientific Center grant: 2014/13/N/NZ9/00703.
Natasza Borodynko-Filas
The authors have declared that no conflict of interests exist.
Bajaj K., Singh P., Mahajan R. 1985. Changes induced by Meloidogyne incognita in superoxide dismutase, peroxidase and polyphenol oxidase activity in tomato roots. Biochemie und Physiologie der Pflanzen 180: 543−546. DOI: https://doi.org/10.1016/S0015-....
Bariola P.A., Green P.J. 1997. Plant ribonucleases. p. 163−190. In: "Ribonucleases: Structures and Functions” (G. D’Alessio, J.F. Riordan, eds). Academic Press, USA. DOI: https://doi.org/10.1016/B978-0....
Bartling D., Radzio R., Steiner U., Weiler E.W. 1993. A glutathione S-transferase with glutathione-peroxidase activity from Arabidopsis thaliana: Molecular cloning and functional characterization. European Journal of Biochemistry 216: 579−586. DOI: https://doi.org/ 10.1111/j.1432-1033.1993.tb18177.x.
Blank A., Sugiyama R., Dekker C.A. 1982. Activity staining of nucleolytic enzymes after sodium dodecyl sulfate-polyacrylamide gel electrophoresis: use of aqueous isopropanol to remove detergent from gels. Analytical Biochemistry 120: 267−275. DOI: 10.1016/0003-2697(82)90347-5
Bradford M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72: 248−254. DOI: 10.1006/abio.1976.9999
Christensen J.H., Bauw G., Welinder K.G., Van Montagu M., Boerjan W. 1998. Purification and characterization of peroxidases correlated with lignification in poplar xylem. Plant Physiology 118: 125−135. DOI: https://doi.org/10.1104/pp.118....
Edreva A. 2005. Pathogenesis-related proteins: research progress in the last 15 years. General and Applied Plant Physiology 31: 105−124.
Eisenback J.D., Triantaphyllou H.H. 1991. Root-knot nematodes: Meloidogyne species and races. p. 191−274. In: "Manual of Agricultural Nematology" (W.R. Nickle, ed.). CRC Press, USA. DOI: https://doi.org/10.1201/978100....
Elling A.A. 2013. Major emerging problems with minor Melo­ idogyne species. Phytopathology 103: 1092−1102. DOI: https://doi.org/10.1094/PHYTO-....
Filipenko E., Kochetov A., Kanayama Y., Malinovsky V., Shumny V. 2013. PR-proteins with ribonuclease activity and plant resistance against pathogenic fungi. Russian Journal of Genetics: Applied Research 3: 474−480. DOI: https://doi.org/10.1134/S20790....
Gheysen G., Fenoll C. 2002. Gene expression in nematode feeding sites. Annual Review of Phytopathology 40: 191−219. DOI: https://doi.org/10.1146/annure....
Hiraga S., Sasaki K., Ito H., Ohashi Y., Matsui H. 2001. A large family of class III plant peroxidases. Plant and Cell Physiology 42: 462−468. DOI: https://doi.org/10.1093/pcp/pc....
Holbein J., Grundler F.M., Siddique S. 2016. Plant basal resistance to nematodes: an update. Journal of Experimental Botany 67: 2049−2061. DOI: https://doi.org/10.1093/jxb/er....
Hussey R. 1973. A comparison of methods of collecting inocula of Meloidogyne spp., including a new technique. Plant Disease Reporter 57: 1025−1028.
Jain D., Khurana J.P. 2018. Role of pathogenesis-related (PR) proteins in plant defense mechanism. p. 265−281. In: "Molecular Aspects of Plant-Pathogen Interaction." (A. Singh, I.K. Singh, eds.). Springer, Singapore. DOI: https://doi.org/10.1007/978-98....
Kyndt T., Nahar K., Haegeman A., De Vleesschauwer D., Höfte M., Gheysen G. 2012. Comparing systemic defencerelated gene expression changes upon migratory and sedentary nematode attack in rice. Plant Biology 14: 73−82. DOI: https://doi.org/10.1111/j.1438....
Mahantheshwara B., Nayak D., Patra M.K. 2019. Protein estimation through biochemical analysis in resistant and susceptible cultivars of cowpea against infection by root-knot nematode, Meloidogyne incognita. Journal of Entomology and Zoology Studies 7 (4): 1191−1193.
MlÝčkovß K., Luhovß L., Lebeda A., Mieslerovß B., Peč P. 2004. Reactive oxygen species generation and peroxidase activity during Oidium neolycopersici infection on Lycopersicon species. Plant Physiology and Biochemistry 42: 753−761. DOI: https://doi.org/10.1016/j.plap....
Mohanty K., Ganguly A., Dasgupta D. 1986. Development of peroxidase (EC 1.11. 1.7) activities in susceptible and resistant cultivars of cowpea inoculated with the root-knot nematode, Meloidogyne incognita. Indian Journal of Nematology 16: 252−256.
Mohsenzadeh S., Esmaeili M., Moosavi F., Shahrtash M., Saffari B., Mohabatkar H. 2011. Plant glutathione S-transferase classification, structure and evolution. African Journal of Biotechnology 10: 8160−8165. DOI: https://doi.org/10.5897/AJB11.....
Przybylska A., Kornobis F., Obrępalska-Stęplowska A. 2018. Analysis of defense gene expression changes in susceptible and tolerant cultivars of maize (Zea mays) upon Meloido­ gyne arenaria infection. Physiological and Molecular Plant Pathology 103: 78−83. DOI: https://doi.org/10.1016/j.pmpp....
Przybylska A., Obrępalska-Stęplowska A. 2020. Plant defense responses in monocotyledonous and dicotyledonous host plants during root-knot nematode infection. Plant and Soil 451: 239–260. DOI: https://doi.org/10.1007/s11104....
Siddiqui Z., Husain S. 1992. Response of twenty chickpea cultivars to Meloidogyne incognita race 3. Nematologia Mediterranea 20: 33−36.
Singh N.K., Paz E., Kutsher Y., Reuveni M., Lers A. 2020. Tomato T2 ribonuclease LE is involved in the response to pathogens. Molecular Plant Pathology 21: 895−906. DOI: https://doi.org/10.1111/mpp.12....
Veronico P., Paciolla C., Pomar F., De Leonardis S., García-Ulloa A., Melillo M.T. 2018. Changes in lignin biosynthesis and monomer composition in response to benzothiadiazole and root-knot nematode Meloidogyne incognita infection in tomato. Journal of Plant Physiology 230: 40−50. DOI: https://doi.org/10.1016/j.jplp....