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Biometrical, biological, biochemical and molecular characteristics of Meloidogyne incognita isolates and related species

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Abstract

Meloidogyne incognita is one of the most polyphagous species of root-knot nematodes occurring in Brazil and worldwide. Eight M. incognita isolates were studied, representing two enzymatic phenotypes (esterase and malate desydrogenase: I1/N1, I2/N1) and four cryptic Meloidogyne sp.1 (S2/N1) isolates, representing one cytological type (3n = 40–46). Three M. hispanica isolates (Hi3/N1, 2n = 32–36) and two of an atypical Meloidogyne sp.2 (S2a/N3, 3n = 40–44) were included in this study for comparison. All isolates were tested with three M. incognita-specific molecular markers. The primer pairs B06F/R, miF/R and incK14F/R amplified three species-specific fragments of 1,200 bp, 955 bp and 399 bp, respectively for M. incognita and Meloidogyne sp.1 isolates. No amplification occurred in the M. hispanica and Meloidogyne sp.2 isolates, except with primers miF/R (1,650 bp). The genetic variability of the Meloidogyne spp. isolates was evaluated, using RAPD and ISSR markers. The phylogenetic analyses revealed two strongly supported monophyletic clades: clade I, consisting of M. hispanica and the atypical Meloidogyne sp.2 isolates, and clade II, clustering together all M. incognita and the Meloidogyne sp.1 isolates. Considering the biometrical, cytological and molecular approaches, it was possible to conclude that the isolates with three enzymatic phenotypes (I1/N1, I2/N1 and S2/N1) presented the characteristics described for M. incognita. Some correlations were detected between the isozymatic phenotypes and the tree topology (S2a/N3, Hi3/N1, I1/N1, S2/N1), but no strict correlation could be observed for the phenotype I2/N1 and one isolate of S2/N1. Morphologically, the Msp.2 isolates differ from M. incognita and M. hispanica by the female stylet features presenting straight cone tip and round pear shaped knobs, posteriorly sloping. The results of this study suggested that the Msp.2 isolates with phenotypes S2aN3 belong to a new or an unidentified species closely related to M. hispanica.

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References

  • Blok, V. C., & Powers, O. (2009). Biochemical and molecular identification. In R. Perry, M. Moens, & J. L. Starr (Eds.), Root-knot nematodes (pp. 98–118). Cambridge: CAB International.

    Chapter  Google Scholar 

  • Blok, V. C., Phillips, M. S., McNicol, J. W., & Fargette, M. (1997). Genetic variation in tropical Meloidogyne spp. as shown by RAPDs. Fundamental and Applied Nematology, 20, 127–133.

    Google Scholar 

  • Carneiro, R. M. D. G., & Almeida, M. R. A. (2001). Técnica de eletroforese usada no estudo de enzimas dos nematóides de galhas para identificação de espécies. Nematologia Brasileira, 25, 555–560.

    Google Scholar 

  • Carneiro, R. M. D. G., & Cofcewicz, E. T. (2008). Taxonomy of coffee-parasitic root-knot nematodes, Meloidogyne spp. In R. M. Souza (Ed.), Plant parasitic nematodes of coffee (pp. 87–122). New York: Springer.

    Chapter  Google Scholar 

  • Carneiro R. M. D. G., Carneiro, R. G., Abrantes, I. M. de O., Santos M. S. N. de A., & Almeida, M. R. A. (1996). Meloidogyne paranaensis n. sp. (Nemata: Meloidogynidae) a root-knot nematode parazitizing coffee from Brazil. Journal of Nematology, 28, 177-189.

  • Carneiro, R. M. D. G., Almeida, M. R. A., & Quénéhervé, P. (2000). Enzyme phenotypes of Meloidogyne spp. populations. Nematology, 2, 645–654.

    Article  CAS  Google Scholar 

  • Carneiro, R. M. D. G., Tigano, M. S., Randig, O., Almeida, M. R. A., & Sarah, J. L. (2004). Identification and genetic diversity of Meloidogyne spp. (Tylenchida: Meloidogynidae) on coffee from Brazil, Central America and Hawaii. Nematology, 6, 37–47.

    Article  CAS  Google Scholar 

  • Carneiro, R. M. D. G., Almeida, M. R. A., Gomes, A. C. M. M., & Hernandez, A. (2005a). Meloidogyne izalcoensis n. sp. (Nematoda: Meloidogynidae), a root-knot nematode parasiting coffee in El Salvador. Nematology, 7, 819–832.

    Article  Google Scholar 

  • Carneiro, R. M. D. G., Randig, O., Almeida, M. R. A., & Gonçalves, W. (2005b). Identificação e caracterização de espécies de Meloidogyne em cafeeiro nos Estados de São Paulo e Minas Gerais através dos fenótipos de esterase e SCAR-Multiplex-PCR. Nematologia Brasileira, 29, 233–241.

    Google Scholar 

  • Carneiro, R. M. D. G., Santos, M. F. A., Almeida, M. R. A., Mota, F. C., & Tigano, M. S. (2008a). Diversity of Meloidogyne arenaria using morphological, cytological and molecular approaches. Nematology, 10, 819–834.

    Article  CAS  Google Scholar 

  • Carneiro, R. M. D. G., Mendes, M. L., Almeida, M. R. A., Santos, M. F. A., Gomes, A. C. M. M., & Karssen, G. (2008b). Additional information on Meloidogyne inornata Lordello, 1956 (Tylenchida: Meloidogynidae) and its characterization as a valid species. Nematology, 10, 123–136.

    Article  Google Scholar 

  • Carneiro, R. M. D. G., Santos, D. F., Costa, S. B., Sousa, F. R., Almeida, M. R. A., Santos, M. F. A., Siqueira, K. M. S., Tigano, M. S., & Fonseca, A. F. A. (2009). Reaction of ‘Conilon’ coffees to different populations of Meloidogyne spp. Nematologia Brasileira, 33, 311.

    Google Scholar 

  • Castagnone-Sereno, P. (2006). Genetic variability and adaptative evolution in parthenogenetic root-knot nematodes. Heredity, 96, 282–289.

    Article  PubMed  CAS  Google Scholar 

  • Castagnone-Sereno, P., Vanlerberghe-Masutti, F., & Leroy, F. (1994). Genetic polymorphism between and within Meloidogyne species detected with RAPD markers. Genome, 37, 904–909.

    Article  PubMed  CAS  Google Scholar 

  • Castro, J. M. C., Lima, R. D., & Carneiro, R. M. D. G. (2003). Variabilidade isoenzimática de populações de Meloidogyne spp. de regiões brasileiras produtoras de soja. Nematologia Brasileira, 27, 1–12.

    Google Scholar 

  • Cenis, J. L. (1993). Identification of four major Meloidogyne spp. by random amplified polymorphic DNA (RAPD-PCR). Phytopathology, 83, 76–80.

    Article  CAS  Google Scholar 

  • Charchar, J. M., Eisenback, J. D., & Hirschmann, H. (1999). Meloidogyne petuniae n. sp. (Nemata: Meloidogynidae), a root-knot nematode parasitic on petunia in Brazil. Journal of Nematology, 31, 81–91.

    PubMed  CAS  Google Scholar 

  • Cofcewicz, E. T., Carneiro, R. M. D. G., Randig, O., Chabrier, C., & Quénéhervé, P. (2005). Diversity of Meloidogyne spp. on Musa in Martinique, Guadeloupe and French Guiana. Journal of Nematology, 37, 313–322.

    PubMed  CAS  Google Scholar 

  • Cofcewicz, E. T., Carneiro, R. M. D. G., Castagnone-Sereno, P., & Quénéhervé, P. (2004). Enzyme phenotypes and genetic diversity of root-knot nematodes parasitising Musa in Brazil. Nematology, 6, 85–95.

    Article  CAS  Google Scholar 

  • Eisenback, J. D., & Hunt, D. J. (2009). General morphology. In R. Perry, M. Moens, & J. L. Starr (Eds.), Root-knot Nematodes (pp. 18–54). Cambridge: CABI North America Ofice.

    Chapter  Google Scholar 

  • Eisenback, J. D., & Triantaphyllou, H. H. (1991). Root-knot nematode: Meloidogyne spp. and races. In W. R. Nickle (Ed.), Manual of agricultural nematology (pp. 191–274). New York: Marcel Dekker.

    Google Scholar 

  • Esbenshade, P. R., & Triantaphyllou, A. C. (1990). Isozyme phenotypes for the identification of Meloidogyne species. Journal of Nematology, 22, 10–15.

    PubMed  CAS  Google Scholar 

  • Esbenshade, P. R., & Triantaphyllou, A. C. (1985). Use of enzyme phenotypes for identification of Meloidogyne species (Nematoda: Tylenchida). Journal of Nematology, 17, 6–20.

    PubMed  CAS  Google Scholar 

  • Fargette, M. (1987). Use of esterase phenotype in the taxonomy of the genus Meloidogyne. 2. Esterase phenotypes observed in West African populations and their characterization. Révue de Nématologie, 10, 45–56.

    Google Scholar 

  • Fassuliotis, G. (1985). The role of the nematologist on the development of resistant cultivars. In J. N. Sasser & C. C. Carter (Eds.), An advanced treatise on Meloidogyne. Biology and control Vol. 1, (pp. 233–240). Raleigh: North Carolina State University Graphics.

  • Felsenstein, J. (1985). Confidence limits on phylogenies: an approach using the bootstrap. Evolution, 39, 783–791.

    Article  Google Scholar 

  • Fourie, H., Zijlstra, C., & McDonald, A. H. (2001). Identification of root-knot nematode species occurring in South Africa, using the SCAR-PCR technique. Nematology, 3, 675–80.

    Article  CAS  Google Scholar 

  • Hartman, R. M., & Sasser, J. N. (1985). Identification of Meloidogyne species on the basis of differential host test and perineal pattern morphology. In K. R. Barker, C. C. Carter, & J. N. Sasser (Eds.), An advanced treatise on Meloidogyne. Vol. 2, Methodology (pp. 69–77). Raleigh: North Carolina State University Graphics.

    Google Scholar 

  • Hirschmann, H. (1986). Meloidogyne hispanica n. sp. (Nematoda: Meloidogynidae) the ‘Seville Root-Knot Nematode’. Journal of Nematology, 18, 520–532.

    PubMed  CAS  Google Scholar 

  • Hirschmann, H. (1984). Morphological variability of Meloidogyne incognita revealed by light and scanning microscopy. Proceedings of the 1st International Congress of Nematology. (pp.35). Guelph, Ontario, Canada.

  • Janati, A. A., Bergé, A., Triantaphyllou, A. C., & Dalmasso, A. (1982). Nouvelles données sur l’ utilisation des isoestérases pour l’ identification des Meloiodogyne. Révue de Nématologie, 5, 147–154.

    CAS  Google Scholar 

  • Karssen, G., & Moens, M. (2006). Root-knot nematodes. In R. L. Perry & M. Moens (Eds.), Plant nematology (pp. 59–90). Wallingford: CABI Publishing.

    Chapter  Google Scholar 

  • Landa, B. B., Palomares Rius, J. E., Vovlas, N., Carneiro, R. M. D. G., Maleita, C. M. N., Abrantes, I. M. de O., & Castillo, P. (2008). Molecular characterization of Meloidogyne hispanica (Nematoda, Meloidogynidae) by phylogenetic analysis of genes within the rDNA in Meloidogyne spp. Plant Disease, 92, 1104–1110.

  • Lax, P., Dueñas, J. C. R., Gardenal, C. N., & Doucet, M. E. (2007). Assessment of genetic variability in populations of Nacobbus aberrans (Thorne, 1935) Thorne & Allen, 1944) Nematoda: Pratylenchidae) from Argentina. Nematology, 9, 261–270.

    Article  Google Scholar 

  • Medina, I. L., Gomes, C. B., Rossi, C., & Carneiro, R. M. D. G. (2006). Characterization and identification of root-knot nematodes (Meloidogyne spp.) from fig trees in Rio Grande do Sul and São Paulo States of Brazil. Nematologia Brasileira, 30, 179–187.

    Google Scholar 

  • McClure, M., Kruk, T. H., & Migashi, I. (1973). A method for obtaining quantities of clear Meloidogyne eggs. Journal of Nematology, 5, 230.

    PubMed  CAS  Google Scholar 

  • Meng, Q. P., Long, H., & Xu, J. H. (2004). PCR assays for rapid and sensitive identification of three major root-knot nematodes, Meloidogyne incognita, M. javanica and M. arenaria. Acta Phytopathologica Sinica, 34, 2004–2010.

    Google Scholar 

  • Moens, M., Perry, R. N., & Starr, J. L. (2009). Meloidogyne Species—a diverse group of novel and important plant parasites. In R. N. Perry, M. Moens, & J. L. Starr, (Eds.), Root-knot Nematodes, (pp. 1–17). Cambridge: CABI North America Office.

  • Muniz, M. F. S., Campos, V. P., Moita, A. W., Gonçalves, W., Almeida, M. R. A., Sousa, F. R., & Carneiro, R. M. D. G. (2009). Reaction of coffee genotypes to different populations of Meloidogyne spp.: detection of a naturally virulent M. exigua population. Tropical Plant Pathology, 34, 370–378.

    Article  Google Scholar 

  • Oliveira, D. S., Oliveira, R. D. L., & Gonçalves, W. (2006). Fenótipo S1 de esterase em Meloidogyne incognita no Brasil. Fitopatologia Brasileira, 31, 207.

    Article  Google Scholar 

  • Randig, O., Bongiovanni, M., Carneiro, R. M. D. G., & Castagnone-Sereno, P. (2002). Genetic diversity of root-knot nematodes from Brazil and development of SCAR markers specific for the coffee-damaging species. Genome, 45, 862–870.

    Article  PubMed  CAS  Google Scholar 

  • Ronquist, F., & Huelsenbeck, J. P. (2003). MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics, 19, 1572–1574.

    Article  PubMed  CAS  Google Scholar 

  • Semblat, J. P., Wajnberg, E., Dalmasso, A., Abad, P., & Castagnone-Sereno, P. (1998). High-resolution DNA fingerprinting of parthenogenetic root-knot nematodes using AFLP analysis. Molecular Ecology, 7, 119–125.

    Article  PubMed  CAS  Google Scholar 

  • Swofford, D. L. (2002). PAUP*. Phylogenetic analysis using UPGMA (*and other methods). Version 4 b10. Sinauer Associates, Sunderland, Massachusetts.

  • Taylor, D. P., & Netscher, C. (1974). An improved technique for preparing perineal patterns of Meloidogyne spp. Nematologica, 20, 268–269.

    Article  Google Scholar 

  • Tigano, M., Siqueira, K., Castagnone-Sereno, P., Mulet, K., Queiroz, P., Santos, M., Teixeira, C., Almeida, M., Silva, J., & Carneiro, R. (2010). Genetic diversity of the root-knot nematode Meloidogyne enterolobii and development for SCAR marker for this guava-damaging species. Plant Pathology, 59, 1054–1061.

    Article  CAS  Google Scholar 

  • Tigano, M. S., Carneiro, R. M. D. G., Jeyaprakash, A., Dickson, D., & Adams, B. J. (2005). Phylogeny of Meloidogyne spp. based on 18S rDNA and the intergenic region of mitochondrial DNA sequences. Nematology, 7, 851–862.

    Article  CAS  Google Scholar 

  • Triantaphyllou, A. C. (1985a). Cytogenetics, cytotaxonomy and phylogeny of root-knot nematodes. In K. R. Barker, C. C. Carter, & J. N. Sasser (Eds.), An advanced treatise on Meloidogyne, vol. 1, biology and control (pp. 113–126). Raleigh: North Carolina State University Graphics.

    Google Scholar 

  • Triantaphyllou, A. C. (1985b). Cytological methods for the study of oogenesis and reproduction of root-knot nematode. In K. R. Barker, C. C. Carter, & J. N. Sasser (Eds.), An advanced treatise on meloidogyne, vol. 2, methodology (pp. 107–114). Raleigh: North Carolina State University Graphics.

    Google Scholar 

  • Triantaphyllou, A. C. (1981). Oogenesis and the chromosomes of the parthenogenetic root-knot nematode Meloidogyne incognita. Journal of Nematology, 13, 95–104.

    PubMed  CAS  Google Scholar 

  • Zijlstra, C. (2000). Identification of Meloidogyne chitwoodi, M. fallax and M. hapla based on SCAR-PCR: a powerful way of enabling reliable identification of populations or individuals that share common traits. European Journal of Plant Pathology, 106, 283–290.

    Article  CAS  Google Scholar 

  • Zijlstra, C., Donkers-Venne, D. T. H. M., & Fargette, M. (2000). Identification of Meloidogyne incognita, M. javanica and M. arenaria using sequence characterised amplified regions (SCAR) based PCR assays. Nematology, 2, 847–53.

    Article  CAS  Google Scholar 

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Acknowledgements

This work was funded by Consórcio Brasileiro de Pesquisa e Desenvolvimento do Café, Embrapa and Conselho Nacional de Pesquisa (CNPq).

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Authors and Affiliations

  1. EMBRAPA-Recursos Genéticos e Biotecnologia, Brasília, DF 70849-970, Brazil

    Marcilene Fernandes Almeida dos Santos, Maria Rita Alves Almeida, Marina Dechechi Gomes Carneiro, Fabiane Castro Mota, Ana Cristina Meneses Mendes Gomes, Natália Orrú Reis Silveira, Joelma Gardênia Pereira Silva, Myrian Silvana Tigano & Regina Maria Dechechi Gomes Carneiro

  2. Departamento de Fitopatologia, Universidade de Brasília, Brasília, DF 70910-900, Brazil

    Marcilene Fernandes Almeida dos Santos, Cleber Furlanetto & Joelma Gardênia Pereira Silva

  3. INRA-UMR1355, UNSA, CNRS UMR7354, Institut Sophia Agrobiotech, 400 route des Chappes, BP167, 06903, Sophia Antipolis, Frances

    Philippe Castagnone-Sereno

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  1. Marcilene Fernandes Almeida dos Santos
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  2. Cleber Furlanetto
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  3. Maria Rita Alves Almeida
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  4. Marina Dechechi Gomes Carneiro
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  6. Ana Cristina Meneses Mendes Gomes
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  11. Regina Maria Dechechi Gomes Carneiro
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Correspondence to Regina Maria Dechechi Gomes Carneiro.

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dos Santos, M.F.A., Furlanetto, C., Almeida, M.R.A. et al. Biometrical, biological, biochemical and molecular characteristics of Meloidogyne incognita isolates and related species. Eur J Plant Pathol 134, 671–684 (2012). https://doi.org/10.1007/s10658-012-0018-1

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