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Parasitism effects on white clover by root-knot and cyst nematodes and molecular separation of Heterodera daverti from H. trifolii

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Abstract

This research carried out an accurate identification of the root-knot and cyst-forming nematode species parasitizing white clover at the Laceno Lake area in Southern Italy. Two species, the root-knot nematode Meloidogyne hapla and the cyst nematode Heterodera daverti were identified by integrative taxonomic approaches (classical, isozyme pattern, and D2-D3 expansion segments of 28S rRNA, ITS rRNA and coxI of mtDNA gene sequences) and found parasitizing white clover roots. These nematodes were detected in stunted plants with a reduced number of rhizobium nodules and the host suitability was confirmed by the high nematode population densities ranging from 53 to 2350 eggs and J2s per g of fresh roots for M. hapla, and 1.36 eggs and J2s/cm3 of soil for H. daverti; and cyst nematode females were also detected on the roots of clover. The studies on the host-parasite relationships of nematode-feeding sites in white clover roots infected by these nematodes showed a high susceptible response. Meloidogyne hapla and H. daverti infections were also observed on nitrogen-fixing root nodules of white clover, where well established feeding sites allowed active nematode reproduction. Histological examination of nitrogen-fixing root nodule tissues revealed that the nematodes established their permanent feeding sites in the vascular bundles of nodules which appeared enlarged deformed and disorganised by the expansion of nematode feeding cells (giant cells and syncytium) and hyperplasia of the nodule cortex. Additionally, coxI of mtDNA gene is an efficient barcoding sequence for discriminating the identification of H. daverti from H. trifolii.

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References

  • Abolafia, J., Liébanas, G., & Peña-Santiago, R. (2002). Nematodes of the order rhabditida from andalucia Oriental, Spain. The subgenus Pseudacrobeles Steiner, 1938, with description of a new species. Journal of Nematode Morphology and Systematics, 4, 137–154.

    Google Scholar 

  • Ambrogioni, L., D’Errico, F. P., & Marinari Palmisano, A. (1986). Observations on a population of cyst forming nematodes from carnation. In F. Lamberti & C. E. Taylor (Eds.), Cyst nematodes (pp. 315–320). New York: Plenum Press.

    Chapter  Google Scholar 

  • Barker, K. (1998). Introduction and synopsis of advancements in nematology. In K. R. Barker, G. A. Pederson, & G. L. Windham (Eds.), Plant nematode interactions (pp. 1–20). Madison: American Society of Agronomy, Crop Science Society of America, Soil Science Society of America.

    Google Scholar 

  • Bird, D. M., Opperman, C. H., & Davies, K. G. (2003). Interactions between bacteria and plant-parasitic nematodes: now and then. International Journal for Parasitology, 33, 1269–1276.

    Article  CAS  PubMed  Google Scholar 

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

  • Castillo, P., Vovlas, N., Subbotin, S., & Troccoli, A. (2003). A new root-knot nematode, Meloidogyne baetica n. sp. (Nematoda: Heteroderidae), parasitizing wild olive in Southern Spain. Phytopathology, 93, 1093–1102.

    Article  PubMed  Google Scholar 

  • Coolen, W. A. (1979). Methods for extraction of Meloidogyne spp. and other nematodes from roots and soil. In F. Lamberti & C. E. Taylor (Eds.), Root-knot nematodes (Meloidogyne species). Systematics, biology and control (pp. 317–329). New York: Academic.

    Google Scholar 

  • Cuany, A., & Dalmasso, A. (1975). Caractères et spécificité de deux especes biologiques d’Heterodera se développant sur Dianthus caryophyllus. Nematologia Mediterranea, 3, 11–21.

    Google Scholar 

  • Derycke, S., Remerie, T., Vierstraete, A., Backeljau, T., Vanfleteren, J., Vincx, M., & Moens, T. (2005). Mitochondrial DNA variation and cryptic speciation within the free living marine nematode Pellioditis marina. Marine Ecology Progress Series, 300, 91–103.

    Article  CAS  Google Scholar 

  • Desaeger, J., Odee, D., Machua, J., & Esitubi, M. (2005). Interactions between Meloidogyne javanica (Treub) chitwood and rhizobia on growth of Sesbania sesban (L.) Merr. Applied Soil Ecology, 29, 252–258.

    Article  Google Scholar 

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

    PubMed Central  CAS  PubMed  Google Scholar 

  • Griffin, G. D., & Rumbaugh, M. D. (1996). Host suitability of twelve leguminosae species to populations of Meloidogyne hapla and M. chitwoodi. Journal of Nematology, 28, 400–405.

    PubMed Central  CAS  PubMed  Google Scholar 

  • Hague, H. G. M. (1980). Nematodes of legume crops. In R. J. Summerfield & A. H. Bunting (Eds.). Advances in legume science (pp. 199–205). Proceedings International Legume Conference, Kew, England, 31 Jul-4 August 1978, Royal Botanic Gardens, Kew, England.

  • Hartman, K. M., & Sasser, J. N. (1985). Identification of Meloidogyne species on the basis of differential host assay and perineal-pattern morphology. In: K. R. Barker, C. C. Carter, & J. J. Sasser, (eds). An Advanced Treatise on Meloidogyne (pp. 67–77). Vol. 2, Methodology. North Carolina State University Graphics, Raleigh.

  • Huelsenbeck, J. P., & Ronquist, F. (2001). MrBAYES: Bayesian inference of phylogenetic trees. Bioinformatics, 17, 754–755.

    Article  CAS  PubMed  Google Scholar 

  • Hussey, R. S., & Williamson, V. M. (1997). Physiological and molecular aspects of nematode parasitism. In K. R. Barker, G. A. Pederson, & G. L. Windham (Eds.), Plant and Nematode Interactions (pp. 87–108). Madison: American Society of Agronomy.

    Google Scholar 

  • Johansen, D. A. (1940). Plant microtechnique. Ney York: McGraw-Hill Book Co. 523 pp.

    Google Scholar 

  • Karssen, G. (2002). The plant parasitic nematode genus Meloidogyne Göldi, 1892 (Tylenchida) in Europe. Leiden: Brill. 157 pp.

    Google Scholar 

  • Ledgard, S., Schils, R., Eriksen, J., & Luo, J. (2009). Environmental impacts of grazed clover/grass pasture. Irish journal of Agricultural and food research, 48, 209–226.

    Google Scholar 

  • Martinuz, A., Schouten, A., & Sikora, R. A. (2013). Post-infection development of Meloidogyne incognita on tomato treated with the endophytes Fusarium oxysporum strain Fo162 and Rhizobium etli strain G12. BioControl, 58, 95–104.

    Article  Google Scholar 

  • Mashela, P. W., & Pofu, K. M. (2012). Interactive effects of Meloidogyne incognita race 2, Bradyrhizobium japonicum and crude extracts of Cucumis myriocarpus fruit on Vigna unguiculata. Crop Protection, 42, 124–127.

    Article  Google Scholar 

  • Massoud, S. I., Rahman, F. H. A., & Ghorab, A. I. (1988). Parasitism of Heterodera daverti on clover root rhizobium nodules in Egypt. Egypt Journal of Phytopathology, 20, 73–78.

    Google Scholar 

  • McLeish, L. J., Berg, G. N., Hinch, J. M., Nambiar, L. V., & Norton, M. R. (1997). Plant parasitic nematodes in white clover and soil from white clover pastures in Australia. Australian Journal of Experimental Agriculture, 37, 75–82.

    Article  Google Scholar 

  • Mercer, C. F. (1994). Plant parasitic nematodes in New Zealand. New Zealand Journal of Zoology, 21, 57–65.

    Article  Google Scholar 

  • Mercer, C. F., Husain, S. W., & Moore, K. K. (2004). Resistance reactions to Meloidogyne trifoliophila in Trifolium repens and T. semipillosum. Journal of Nematology, 36, 499–504.

    PubMed Central  PubMed  Google Scholar 

  • Mulvey, R. H. (1958). Parthenogenesis in a cyst-forming nematode, Heterodera trifolii (Nematoda: Heteroderidae). Canadian Journal of Zoology, 36, 91–93.

    Article  Google Scholar 

  • Nordmeyer, D., & Sikora, R. (1983). Studies on the interaction between Heterodera daverti, Fusarium avenaceum and F. oxysporum on Trifolium subterraneum. Revue de Nematologie, 6, 193–198.

    Google Scholar 

  • Nordmeyer, D., Sikora, R. A., & Jaritz, G. (1978). Die Bedeuting von Nematoden und bodenbürtigen Pilzen als Schaderreger in Trifolium subterraneum-Weiden in Nordwest-Tunesien. Mededelingen van de Faculteit Landbouwwetenschappen Rijksuniversiteit Ghent, 43, 747–756.

    Google Scholar 

  • Nunn, G. B. (1992). Nematode molecular evolution. Ph.D. Thesis, University of Nottingham, Nottingham, UK.

  • Pederson, G. A., Windham, G. L., Ellsbury, M. M., McLaughlin, M. R., Pratt, R. G., & Brink, G. E. (1991). White clover yield and persistence as influenced by cypermethrin, bonomyl, and root-knot nematode. Crop Science, 31, 1297–1302.

    Article  CAS  Google Scholar 

  • Ravichandra, N. G. (2014). Horticultural Nematology. New Dehli: Springer. 412 pp.

    Book  Google Scholar 

  • Seinhorst, J. W. (1966). Killing nematodes for taxonomic study with hot f.a. 4:1. Nematologica, 12, 178.

    Article  Google Scholar 

  • Siddiqui, Z. A., Fatima, M., & Alam, S. (2011). Interactions of Meloidogyne incognita, Xanthomonas campestris, and Rhizobium sp. in the disease complex of chickpea. Turkish Journal of Agriculture and Forestry, 37, 173–178.

    Google Scholar 

  • Stelter, H., & Meinl, G. (1972). The effects of the infestation of red and white clover by Heterodera trifolii and Heterodera galeopsidis. Archiv für Pflanzenschutz, 8, 463–470.

    Article  Google Scholar 

  • Subbotin, S. A., Vierstraete, A., De Ley, P., Rower, J., Waeyenberge, L., Moens, M., & Vanfleteren, J. R. (2001). Phylogenetic relationships within the cyst-forming nematodes (Nematoda, Heteroderidae) based on analysis of sequences from the ITS regions of ribosomal DNA. Molecular Phylogenetics and Evolution, 21, 1–16.

    Article  CAS  PubMed  Google Scholar 

  • Subbotin, S. A., Mundo-Ocampo, M., & Baldwin, J. G. (2010). In D. J. Hunt & R. N. Perry (Eds.), Systematics of cyst nematodes (Nematoda: Heteroderinae). Nematology monographs and perspectives 8B. Leiden: Brill. 512 pp.

  • Thompson, J. D., Gibson, T. J., Plewniak, F., Jeanmougin, F., & Higgins, D. G. (1997). The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research, 25, 4876–4882.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Toumi, F., Waeyenberge, L., Viaenne, N., Dababat, A., Nicol, J. M., Ogbonnaya, F., & Moens, M. (2013). Development of a species-specific PCR to detect the cereal cyst nematode, Heterodera latipons. Nematology, 15, 709–717.

    CAS  Google Scholar 

  • Townshend, J. L., & Potter, J. W. (1978). Yield losses among forage legumes infected with Meloidogyne hapla. Canadian Journal of Plant Science, 58, 939–943.

    Article  Google Scholar 

  • Vovlas, N., Castillo, P., & Troccoli, A. (1998). Histology of nodular tissue of three leguminous hosts infected by three root-knot nematode species. International Journal of Nematology, 8, 105–110.

    Google Scholar 

  • Watson, R. N., & Mercer, C. F. (2000). Pasture nematodes: the major scourge of white clover. Proceedings of the New Zealand Association, 62, 195–199.

    Google Scholar 

  • Williams, K. J. O. (1974). Meloidogyne hapla. Chitwood, 1949. C.I.H. descriptions of plant-parasitic nematodes 1974, set 3, 4 pp.

  • Williams, W. M. (1987). White clover taxonomy and biosystematics. In M. J. Baker & W. M. Williams (Eds.), White clover (pp. 323–342). Wallingford: CAB International.

    Google Scholar 

  • Wouts, W. M. (1978). On the males of Heterodera trifolii Goffart, 1932 (Nematoda: Heteroderidae). Nematologica, 24, 15–120.

    Google Scholar 

  • Wouts, W. M., & Sturhan, D. (1978). The identity of Heterodera trifolii Coffart, 1962 and the description of H. daverti n. sp. (Nematoda, Tylenchida). Nematologica, 24, 121–128.

    Article  Google Scholar 

  • Yeates, G. W., Ross, D. J., Bridger, B. A., & Visser, T. A. (1977). Influence of the nematodes Heterodera trifolii and Meloidogyne hapla on nitrogen fixation by white clover under glasshouse conditions. New Zealand Journal of Agricultural Research, 20, 401–413.

    Article  CAS  Google Scholar 

  • Zahid, M. I., Gurr, G. M., Nikandrow, A., Hodda, M., Fulkerson, W. J., & Nicol, H. I. (2001). Survey of fungi and nematodes associated with root and stolon diseases of white clover in the subtropical dairy region of Australia. Australian Journal of Experimental Agriculture, 41, 1133–1142.

    Article  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). Identication of Meloidogyne incognita, M. javanica and M. arenaria using sequence characterised amplified region (SCAR) based PCR assays. Nematology, 8, 847–853.

    Article  Google Scholar 

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Acknowledgments

The present work has been done in the frame of the Apulia Region Programme: Misura 214/4 sub azione a “Progetti integrati per la Biodiversità” PSR 2007–2013, ASSE II “Miglioramento dell’ ambiente e dello Spazio Rurale” SaVeGraINPuglia. The authors thank Drs W. Wouts and D. Sturhan for assistance in collecting of H. daverti samples in the type locality and morphological identification of this population.

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Authors and Affiliations

  1. Istituto per la Protezione Sostenibile delle Piante (IPSP), Consiglio Nazionale delle Ricerche (CNR), U.O.S. di Bari, Via G. Amendola 122/D, 70126, Bari, Italy

    Nicola Vovlas & Paola Leonetti

  2. A. P. S. Polyxena, Via Donizetti 12, 70014, Conversano, Bari, Italy

    Alessio Vovlas

  3. Departamento de Biología Animal, B. Vegetal y Ecología, Universidad de Jaén, Campus ‘Las Lagunillas’ s/n, Edificio B3, 23071, Jaén, Spain

    Gracia Liébanas

  4. Instituto de Agricultura Sostenible (IAS), Consejo Superior de Investigaciones Científicas (CSIC), Menéndez Pidal s/n, 14004, Córdoba, Spain

    Pablo Castillo & Juan E. Palomares Rius

  5. Plant Pest Diagnostic Center, California Department of Food and Agriculture, 3294 Meadowview Road, Sacramento, CA, 95832-1448, USA

    Sergei A. Subbotin

  6. Center of Parasitology of A.N. Severtsov Institute of Ecology and Evolution of the Russian Academy of Sciences, Leninskii Prospect 33, Moscow, 117071, Russia

    Sergei A. Subbotin

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  1. Nicola Vovlas
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  2. Alessio Vovlas
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  3. Paola Leonetti
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  4. Gracia Liébanas
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  5. Pablo Castillo
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  6. Sergei A. Subbotin
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  7. Juan E. Palomares Rius
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Correspondence to Juan E. Palomares Rius.

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Vovlas, N., Vovlas, A., Leonetti, P. et al. Parasitism effects on white clover by root-knot and cyst nematodes and molecular separation of Heterodera daverti from H. trifolii . Eur J Plant Pathol 143, 833–845 (2015). https://doi.org/10.1007/s10658-015-0735-3

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