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Identification of root-knot nematode species occurring on tomatoes in Kenya: use of isozyme phenotypes and PCR-RFLP

Published online by Cambridge University Press:  13 June 2012

Rael Birithia ,
Wanjohi Waceke ,
Peter Lomo  and
Daniel Masiga
Rael Birithia*
Affiliation:
Molecular Biology and Biotechnology Department, International Centre of Insect Physiology and Ecology (icipe –African Insect Science for Food and Health), PO Box 30772-00100, Nairobi, Kenya Department of Biochemistry, Jomo Kenyatta University of Agriculture and Technology, PO Box 62000-00200, Nairobi, Kenya
Wanjohi Waceke
Affiliation:
Department of Agricultural Science and Technology, Kenyatta University, PO Box 43844-00100, Nairobi, Kenya
Peter Lomo
Affiliation:
Department of Biochemistry, Jomo Kenyatta University of Agriculture and Technology, PO Box 62000-00200, Nairobi, Kenya
Daniel Masiga
Affiliation:
Molecular Biology and Biotechnology Department, International Centre of Insect Physiology and Ecology (icipe –African Insect Science for Food and Health), PO Box 30772-00100, Nairobi, Kenya
*
*E-mail: rbirithia@icipe.org
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Abstract

Root-knot nematodes (RKN) are serious pests of tomato production in Kenya. Accurate identification of the plant parasitic nematodes is important for their effective management. A study conducted to assess their prevalence and identify RKN occurring on tomatoes in Central Kenya showed infestation of the crop in all the three districts. Of the total sampled plants (N = 900), the RKN infestation level ranged from 28 to 62%. RKN disease severity ranged from 2.5 to 5.3 in all the locations. Meloidogyne incognita,M. javanica and M. arenaria were the only species found infesting tomatoes in these areas. Female nematodes sampled from the symptomatic root system that had root galls characteristic of RKN were analysed by isozyme phenotypes of esterase (EST) and malate dehydrogenase (MDH). EST phenotype was polymorphic and enabled identification of the three different species, while MDH was monomorphic. Polymerase chain reaction-restriction fragment length polymorphism with region between mitochondrial cytochrome oxidase subunit II and large subunit ribosomal RNA (mtDNA COII-LSUrRNA) using HinfI showed that all isolates of M. incognita could be digested into three restriction fragments of about 1300, 400 and 100 bp, except for one species that showed an additional restriction site, giving four fragments of 900, 420, 380 and 100 bp. The 1800 bp PCR product of M. javanica was not digested by HinfI. Meloidogyne arenaria (EST phenotype A1) PCR product was digested into two restriction fragments of about 1700 and 100 bp, while M. arenaria (EST phenotype A2) had two restriction fragments at 1100 and 100 bp.

Keywords

Meloidogyne spptomatoesisozyme profilesmolecular identification

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Type
Research Paper
Information
International Journal of Tropical Insect Science , Volume 32 , Issue 2 , June 2012 , pp. 78 - 84
Copyright
Copyright © ICIPE 2012

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References

Anwar, S. A., Zia, A., Hussain, M. and Kamran, M. (2007) Host suitability of selected plants to Meloidogyne incognita in the Punjab, Pakistan. International Journal of Nematology 17, 144–150.Google Scholar
Atessahin, A., Türk, G., Karahan, I., Yilmaz, S., Ceribasi, A. O. and Bulmus, O. (2006) Lycopene prevents adriamycin-induced testicular toxicity in rats. Fertility and Sterility 85, 1216–1222.CrossRefGoogle ScholarPubMed
Baum, T., Gresshoff, P. M., Lewis, S. A. and Dean, R. A. (1994) Characterization and phylogenetic analysis of four root-knot nematode species using DNA amplification fingerprinting and automated polyacrylamide gel electrophoresis. Molecular Plant–Microbe Interaction 7, 39–47.CrossRefGoogle Scholar
Blok, V. C., Wishart, J., Fargette, M., Berthier, K. and Phillips, M. S. (2002) Mitochondrial DNA differences distinguishing Meloidogyne mayaguensis from the major species of tropical root-knot nematodes. Nematology 4, 773–781.CrossRefGoogle Scholar
Carneiro, R. M. D. G., Almeida, M. R. A. and Quénéhervé, P. (2000) Enzyme phenotypes of Meloidogyne spp. populations. Nematology 2, 645–654.CrossRefGoogle Scholar
Castagnone-Sereno, P. (2006) Genetic variability and adaptive evolution in parthenogenetic root-knot nematodes. Heredity 96, 282–289.CrossRefGoogle ScholarPubMed
Castagnone-Sereno, P., Vanlerberghe-Masutti, F. and Leroy, F. (1994) Genetic polymorphism between and within Meloidogyne species detected with RAPD markers. Genome 37, 904–909.CrossRefGoogle ScholarPubMed
Curran, J. (1991) Application of DNA analysis to nematode taxonomy, pp. 125–143. In Manual of Agricultural Nematology (edited by Nickle, W. R.). Marcel Dekker, New York.Google Scholar
Desaeger, J. and Rao, M. R. (2000) Infection and damage potential of Meloidogyne javanica on Sesbania sesban in different soil types. Nematology 2, 169–178.CrossRefGoogle Scholar
Esbenshade, P. R. and Triantaphyllou, A. C. (1985) Use of enzyme phenotypes for identification of Meloidogyne species. Journal of Nematology 17, 6–20.Google ScholarPubMed
Esbenshade, P. R. and Triantaphyllou, A. C. (1990) Isozyme phenotypes for the identification of Meloidogyne species. Journal of Nematology 22, 10–15.Google ScholarPubMed
Floyd, R., Abebe, E., Papert, A. and Blaxter, M. (2002) Molecular barcodes for soil nematode identification. Molecular Ecology 11, 839–850.CrossRefGoogle ScholarPubMed
Hartman, K. M. and Sasser, J. N. (1985) Identification of Meloidogyne species on the basis of differential host test and perineal-pattern morphology, pp. 69–77. In An Advanced Treatise on Meloidogyne: Volume 2. Methodology (edited by Barker, C. R., Carter, C. C. and Sasser, J. N.). North Carolina State University/USAID, Raleigh, North Carolina.Google Scholar
Hugall, A., Moritz, C., Stanton, J. and Wolstenholme, D. R. (1994) Low, but strongly structured mitochondrial DNA diversity in root knot nematodes (Meloidogyne). Genetics 136, 903–912.CrossRefGoogle ScholarPubMed
Hunt, D. J., Luc, M. and Manzanilla-Lopez, R. H. (2005) Identification, morphology and biology of plant parasitic nematodes, pp. 11–52. In Plant Parasitic Nematodes in Subtropical and Tropical Agriculture (edited by Luc, M., Sikora, R. A. and Bridge, J.), 2nd edn. CABI Publishing, Wallingford.CrossRefGoogle Scholar
Ibrahim, S. K., Perry, R. N. and Webb, R. M. (1995) Use of isoenzyme and protein phenotypes to discriminate between six Pratylenchus species from Great Britain. Annals of Applied Biology 126, 317–327.CrossRefGoogle Scholar
Jacquet, M., Bongiovanni, M., Martinez, M., Verschave, P., Wajnberg, E. and Castagnone-Sereno, P. (2005) Variation in resistance to the root-knot nematode Meloidogyne incognita in tomato genotypes bearing the Mi gene. Plant Pathology 54, 93–99.CrossRefGoogle Scholar
Jones, J. T., Phillips, M. S. and Armstong, M. A. (1997) Molecular approaches in plant nematology. Fundamental and Applied Nematology 20, 1–14.Google Scholar
KARI (2005) Research priorities at KARI-Thika, Report of the Prioritization Workshop held at KARI-Thika, 6–8 December, 2005. KARI, Thika.Google Scholar
Karssen, G. (2002) The Plant-Parasitic Nematode Genus Meloidogyne Goeldi, 1892 (Tylenchida) in Europe. Brill Academic Publishers, Leiden. 160 pp.CrossRefGoogle Scholar
Mehdi, N. E. (2009) Distribution and identification of root-knot nematode species in tomato fields. Mycopathology 7, 45–49.Google Scholar
Milligan, S. B., Bodeau, J., Yaghoobi, J., Kaloshian, I., Zabel, P. and Willamson, V. M. (1998) The root knot nematode resistance gene Mi from tomato is a member of the leucine zipper, nucleotide binding, leucine-rich repeat family of plant genes. The Plant Cell 10, 1307–1319.CrossRefGoogle ScholarPubMed
Nono-Womdim, R., Swai, I. S., Mrosso, L. K., Chadha, M. L. and Opeña, R. T. (2002) Identification of root-knot nematode species occurring on tomatoes in Tanzania and resistant lines for their control. Plant Disease 86, 127–130.CrossRefGoogle ScholarPubMed
Powers, T. O. and Harris, T. S. (1993) A polymerase chain reaction method for identification of five major Meloidogyne species. Journal of Nematology 25, 1–6.Google ScholarPubMed
Shahid, M., Rehman, A. U., Khan, A. U. and Mahmood, A. (2007) Geographical distribution and infestation of plant parasitic nematodes on vegetables and fruits in the Punjab province of Pakistan. Pakistan Journal of Nematology 25, 59–67.Google Scholar
Siddiqui, Z. A. and Mahmood, I. (1998) Effect of a plant growth promoting bacterium, an AM fungus and soil types on the morphometrics and reproduction of Meloidogyne javanica on tomato. Applied Soil Ecology 8, 77–84.CrossRefGoogle Scholar
Sikora, R. A. and Fernandez, E. (2005) Nematode parasites of vegetables, pp. 319–392. In Plant-Parasitic Nematodes in Subtropical and Tropical Agriculture (edited by Luc, M., Sikora, R. A. and Bridge, J.), 2nd edn.CABI Publishing, Wallingford.CrossRefGoogle Scholar
Stanton, J., Hugall, A. and Moritz, C. (1997) Nucleotide polymorphisms and an improved PCR-based mtDNA diagnostic for parthenogenetic root-knot nematodes (Meloidogyne spp). Fundamental and Applied Nematology 20, 261–268.Google Scholar
Taylor, A. L. and Sasser, J. N. (1978) Biology, Identification and Control of Root-knot Nematodes (Meloidogyne species). Department of Plant Pathology North Carolina State University and U.S. Agency for International Development, Raleigh, North Carolina. 111 pp.Google Scholar
Thomas, D. B. (1997) Soil and Water Conservation Manual for Kenya. NairobiSoil and Water Conservation Branch, Ministry of Agriculture, Livestock Development and Marketing. 296 pp..Google Scholar
Varela, A. M., Seif, A. and Löhr, B. (2003) A Guide to IPM in Tomato Production in Eastern and Southern Africa (edited by Ng'eny-Mengech, A.). 128 pp.+full colour insert. icipe Science Press, Nairobi.Google Scholar
Wang, L. P., Liu, S. R. and Mahson, J. E. (2006) The consumption of lycopene and tomato-based food is not associated with the risk of type 2 diabetes in women. Journal of Nutrition 136, 620–625.CrossRefGoogle Scholar