EXISTENCE, MOLECULAR DIAGNOSIS AND GENETIC VARIATION OF DIFFERENT ISOLATES OF ROOT – KNOT NEMATODES Meloidogyne spp. PARASITIZING CUCUMBER PLANTS
Keywords:
Meloidogyne spp. Molecular diagnosis, Cucumber
Abstract
The findings indicated that the highest disease incidence (D.I.) was observed in autumn 2021 on cucumber plants grown in the greenhouses of Qustapa, with a rate of 87.5%. This was followed by a D.I. of 70% in the same location during spring 2022 and in the greenhouses of Daraband during autumn 2021.In both growing seasons in Daraband, and in Qustapa during Autumn 2021 more than 100 galls per root system were recorded, indicating a high level of infection and a highest root – knot index (5) . In contrast, no infection (D.I. 0.0%) was found in the greenhouses of Kamosak (Grd Goran) in both growing seasons and in Mastawa location during spring 2022. The results of soil analysis conducted in the surveyed locations revealed that the physical and chemical properties of the soil influence the movement and survival of nematodes. Results of Molecular diagnosis using two universal primers included 10 Picomoles (pmol) of forward primer-TW81 (5’GTTTCCGTAGGTGAACCTGC-3’) and 10 pmol reverse primer-AB28 (5’ATATGCTTAAGTTCAGCGGGT-3’) identified four species of root-knot nematodes Meloidogyne spp., prevalent in cucumber greenhouses across the surveyed locations.
namely. M. incognita, M. javanica, M. arenaria and M. hapla and according to their genetic variation two races for each of the first three species were recorded for the first time by this technique
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References
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AL-Sinjary, S.G.A and Ami, S.N. ( 2022 ). Molecular diagnosis and genetic variability of new isoaltes of root – knot nematodes Meloidogyne spp. on cucumber plants. Journal of University of Duhok, 25, 2 (Agri. and Vet. Sciences) : 238-246
Al-Zubaidy, K.M.D. and AL- Falahy,M.A.H.( 2016). Principles and procedures of statistical and experimental design. Duhok University press, Central library – Duhok – Card number: D- 2039/16, Pp 395.
Ami, S.N.(1985).Studies on ecology and biology of root-knot nematodes (Meloidogyne spp.) and its effect on tomato plant in north of Iraq. M.Sc. thesis, College of Agriculture and Forestry, Mosul University. (In Arabic)
Ami, S.N. and Al-Sabie, R.F.(1989).Study the population density of root-knot nematode Meloidogyne spp. on tomato fields in northern Iraq. Mesopotamia Journal of Agriculture, 21(2):301-321. (In Arabic
Ami, S. N., and Shingaly, S. G. A. (2018). Disease incidence, identification, and monthly fluctuations in the population density of root-knot nematodes on cucumber plants in Semel District, Duhok, Kurdistan Region, Iraq. Acta Universitatis Sapientiae, Agriculture and Environment, 10(1), 52-65
Arnaot, M.(1980). Medical plant and herbs as food and medicine. Lebanese-Egyption AL-Dar. (In Arabic).
Agbenin, O. N.(2004).Potentials of organic amendments in the control of plant parasitic nematodes. Plant Protection Science, 40(1):21-25.
Agu, C. M.(2008).Effects of organic manure types on root-gall nematode disease and African yam bean yield. The Journal of American Science, 4(1):67-77.
Berry, S. D.; Fargette, M.; Morand, S. and Cadet, P.(2007).Reliability of PCR-based techniques for detection and discrimination of plant-parasitic nematodes of sugarcane. Nematology, 9:499-514.
Black G R. (1965). Bulk density. In. CA. Black et al.(Eds). Methods of soil analysis, part 1. Agron. 9: 371-373. Am. soc. Agron. Madison. USA.
Blok, V.C. (2005). Achievements in a future prospect for molecular diagnostics of plant-parAshitic nematodes. Canadian Journal Plant Pathology, 27: 176-185.
Castagnone-Sereno, P.; Esparrago, G.; Abad, P.; Leroy, F. and Bongiovanni, M.(1995).Satellite DNA as a target for PCR-specific detection of the plant-parasitic nematode Meloidogyne hapla. Current Genetics 28: 566-570.
Coyne, D. L.; Nicol, J. M. and Claudius-Cole, B.(2007).Practical plant nematology: a field and laboratory guide. International Institute of Tropical Agriculture (IITA).
Eisenback, J.D. and Triantaphyllou, H.(1991).Root-knot nematodes: Meloidogyne species and races. In: Nickle, W.R.(ed).Manual of Agricultual nematology. Marces Dekker, New York, pp.191-274.
Esbenshade, P.R. and Triantaphyllou, A.C.(1990).Isozyme phenotypes for the identification of Meloidogyne species. Journal of Nematology, 22: 10–15.
Hartman, K. M. and Sasser, J. N.(1985).Identification of Meloidogyne species on the basis of differential host test and perineal pattern morphology. In: Barker, K. R., Carter, C. C. and Sasser, J.N. (eds) An Advanced Treatise on Meloidogyne. Volume II: Methodology. A cooperative publication of the Department of Plant Pathology and the United States Agency for International Development, North Carolina State University Graphics, Raleigh, North Carolina, pp.69- 77.
https://advancecovercrops.com/resources-advanced-cover-crops/carbon-nitrogen-ratio
Hussain, M. A.; Mukhtar, T. A. R. I. Q.; Kayani, M. Z.; Aslam, M. N. and Haque, M. I.(2012).A survey of okra (Abelmoschus esculentus) in the Punjab province of Pakistan for the determination of prevalence, incidence and severity of root-knot disease caused by Meloidogyne spp. Pakistan Journal of Botany, 44(6): 2071-2075.
Ismail, A. E. (1998). Effect of soil amendments with some hardwood barks on reproduction of Rotylenchulus reiniformis and growth of sunflower, Pak. J. Nematol., 16: 137-144.
Ismail, M.; Anwar, S. A. and Riaz, A.(2012).Incidence of Meloidogyne incognita in cucumber fields. Pakistan Journal of Zoology, 44(5):1383-1387.
Jaiswal P C (2003). Soil Plant and Water analysis. Mrs. Usha Raj Kumar, India, 136Pp.
Jamil A., Riaz S., Ashraf M., Foolad M.R. ( 2011). Gene expression profiling of plants under salt stress. Crit. Rev. Plant Sci. 30 (5) :435 – 458.
Karajeh, M.R. and Al-Nasir, F.M.(2014).Field Utilization of Nitrogen Fertilizers for Controlling Root-knot Nematode and Improving Growth and Yield of Cucumber. International Journal of Agriculture and Forestry. 4(1): 34-40.
Katooli, N., Moghadam, E. M., Taheri, A., and Nasrollahnejad, S. (2010). Management of root-knot nematode (Meloidogyne incognita) on cucumber with the extract and oil of nematicidal plants. International Journal of Agricultural Research, 5(8), 582-586.
Kayani, M. Z.; Mukhtar, T.; Hussain, M. A.; Haque, M. I. and Perveen, R. (2012).Incidence and severity of root-knot nematodes (Meloidogyne spp.) on cucumber in district Rawalpindi. Pakistan Journal of Phytopathology, 24(2): 122-128.
Khalil, M. S.(2013).The potential of five eco-biorational products on the reproduction of root-knot nematode and plant growth. International Journal of Phytopathology, 2(2): 84-91.
Kumar, D.; S. Singh; and N. Singh. (2010). Free radical scavenging and analgesic activities of cucumbers sativa. Fruit extract. J. Young Pharm., 2(4):365-386.
Kumar, N.; Adamu, M.A.; Isah, K.M. and Lawal, A.F.(2014).A Survey of vegetable fields for root-gall disease in Niger State, Nigeria. PAT, 10(1):17-27.
Martin, R. R., James, D, and Levesque, C. A. ( 2000 ). Impacts of molecular diagnostic technologies on plant disease management. Annual review of phytopathology 38:207-39.
Michel, L., Sikora, R.A. (2005), Plant parasitic nematodes in subtropical and tropical agriculture.2nd Edition.CABI Bioscience,Egham,UK.. 492 Pp.
Naz, I.; Palomares-Rius, J. E.; Blok, V.; Saifullah; Ali, S. and Ahmed, M. (2012).Prevalence, incidence and molecular identification of root-knot nematodes of tomato in Pakistan. African Journal of Biotechnology, 11: 16546-16556.
Oka Y. (2010 ) Mechanisms of nematode suppression by organic soilamendments – a review Appl. Soil Ecol., 44 (2) : 101-115
Olabiyi, T. I.; Olayiwola, A. O. and Oyediran, G. O. (2009).Influence of soil textures on distribution of phytonematodes in the south western Nigeria. World Journal of Agricultural Sciences, 5(5): 557-560.
Onkendi, E. M. and Moleleki, L. N.(2013).Distribution and genetic diversity of root‐knot nematodes (Meloidogyne spp.) in potatoes from South Africa. Plant Pathology, 62(5): 1184-1192.
Park, S.D.; Khan, Z.; Yeon, K. and Kim, Y.H.(2005).A survey for plant-parasitic nematodes associated with strawberry (Fragaria ananassa Duch.) Crop in Korea. Plant Pathology Journal, 21:387-390.
Powers, T. O.; Mullin, P. G.; Harris, T. S.; Sutton, L. A. and Higgins, R. S. (2005).Incorporating molecular identification of Meloidogyne spp. in to a large-scale regional nematode survey. Journal of Nematology, 37:226-235.
Prot, J. C. and Van Gundy, S. D.(1981).Effect of soil texture and the clay component on migration of Meloidogyne incognita second-stage juveniles. Journal of Nematology, 13(2): 213-217.
Rivera L., Aballay E. ( 2008 ) Nematicide effect of various organic soil amendments on Meloidogyne ethiopica whitehead, 1968, on potted vine plants Chil. J. Agric. Res., 68 (3) : 290-296.
Rowell, .D. L.(1996).Soil science: Methods and applications. Longman Group UK Limited. pp. 50 – 51.
Skantar,A.M; Handoo,Z.A; Zanakis,G.N. and E. A. Tzortzakakis, E.A.(2012). Molecular and Morphological Characterization of the Corn Cyst Nematode, Heterodera zeae, from Greece Journal of Nematology, 44 : 58–66.
Stephan Z.A. (1997). Plant nematode problems and their control in the Near East region. (FAO Plant Production and Protection Paper - 144).
Subbotin, S. and Moens, M.(2006).Molecular Taxonomy and Phylogeny. In: Plant Nematology. Perry, R. and Moens, M. (Eds). Pp: 51-57. CABI Nosworthy Way, Wallingford Oxfordshire OX10 8DE, UK..
Stirling, G. R.(1991). Biological control of plant parasitic nematodes. Wallingford, UK: CAB Int.
Trudgill, D.L.(1995).An assessment of the relevance of thermal time relationships to nematology. Fund. appl. Nematology, 18: 407 –417.
Van Reeuwijk, L. R.(1995).Procedures for soil analysis: 3-8. 5th edition .International Soil Reference and Information Center, technical paper 9.
Waeyenberge, L.; Ryss, A.; Moens, M. ; Pinochet, J..and Vrain, T. ( 2000 ). Molecular characterization of 18 Pratylenchus species using rDNA restriction fragment length polymorphism. Nematology, 2:135–142.
Zijlstra, C.; Lever, A.E.M.; Uenk, B.J. and Van Sifhout, C.H.(1995). Differences between ITS regions of isolates of root-knot nematodes Meloidogyne hapla and M. chitwoodi. Phytopathology, 85: 1231–1237
Al-ghamdi, A.A.M. ( 2021). Relationship between Nematodes and some Soil Properties in the Rhizosphere of Banana Plants. International Letters of Natural Sciences 82:1- 12
Al – Rawi, K. M. and Khalafa Allah, A. M. ( 1980 ). Design and analysis of agricultural
experiments.Dar Ibn Al-Atheer for printing and publishing, University of Mosul..(In Arabic).
488Pp.
Al-Sabie, R. F., & Ami, S. N. (1990). Identification of races of RKN Meloidogyne spp. in northern Iraq. Arab Journal of Plant Protection, 8(2), 83-87.
AL–Sinjary, S. G. (2017). Molecular Identification, Bioassay and Integrated Control of Root-Knot Nematode Meloidogyne javanica on Cucumber under Plastic Houses . M.Sc. thesis, College of Agriculture, University of Duhok.
AL-Sinjary, S.G.A and Ami, S.N. ( 2022 ). Molecular diagnosis and genetic variability of new isoaltes of root – knot nematodes Meloidogyne spp. on cucumber plants. Journal of University of Duhok, 25, 2 (Agri. and Vet. Sciences) : 238-246
Al-Zubaidy, K.M.D. and AL- Falahy,M.A.H.( 2016). Principles and procedures of statistical and experimental design. Duhok University press, Central library – Duhok – Card number: D- 2039/16, Pp 395.
Ami, S.N.(1985).Studies on ecology and biology of root-knot nematodes (Meloidogyne spp.) and its effect on tomato plant in north of Iraq. M.Sc. thesis, College of Agriculture and Forestry, Mosul University. (In Arabic)
Ami, S.N. and Al-Sabie, R.F.(1989).Study the population density of root-knot nematode Meloidogyne spp. on tomato fields in northern Iraq. Mesopotamia Journal of Agriculture, 21(2):301-321. (In Arabic
Ami, S. N., and Shingaly, S. G. A. (2018). Disease incidence, identification, and monthly fluctuations in the population density of root-knot nematodes on cucumber plants in Semel District, Duhok, Kurdistan Region, Iraq. Acta Universitatis Sapientiae, Agriculture and Environment, 10(1), 52-65
Arnaot, M.(1980). Medical plant and herbs as food and medicine. Lebanese-Egyption AL-Dar. (In Arabic).
Agbenin, O. N.(2004).Potentials of organic amendments in the control of plant parasitic nematodes. Plant Protection Science, 40(1):21-25.
Agu, C. M.(2008).Effects of organic manure types on root-gall nematode disease and African yam bean yield. The Journal of American Science, 4(1):67-77.
Berry, S. D.; Fargette, M.; Morand, S. and Cadet, P.(2007).Reliability of PCR-based techniques for detection and discrimination of plant-parasitic nematodes of sugarcane. Nematology, 9:499-514.
Black G R. (1965). Bulk density. In. CA. Black et al.(Eds). Methods of soil analysis, part 1. Agron. 9: 371-373. Am. soc. Agron. Madison. USA.
Blok, V.C. (2005). Achievements in a future prospect for molecular diagnostics of plant-parAshitic nematodes. Canadian Journal Plant Pathology, 27: 176-185.
Castagnone-Sereno, P.; Esparrago, G.; Abad, P.; Leroy, F. and Bongiovanni, M.(1995).Satellite DNA as a target for PCR-specific detection of the plant-parasitic nematode Meloidogyne hapla. Current Genetics 28: 566-570.
Coyne, D. L.; Nicol, J. M. and Claudius-Cole, B.(2007).Practical plant nematology: a field and laboratory guide. International Institute of Tropical Agriculture (IITA).
Eisenback, J.D. and Triantaphyllou, H.(1991).Root-knot nematodes: Meloidogyne species and races. In: Nickle, W.R.(ed).Manual of Agricultual nematology. Marces Dekker, New York, pp.191-274.
Esbenshade, P.R. and Triantaphyllou, A.C.(1990).Isozyme phenotypes for the identification of Meloidogyne species. Journal of Nematology, 22: 10–15.
Hartman, K. M. and Sasser, J. N.(1985).Identification of Meloidogyne species on the basis of differential host test and perineal pattern morphology. In: Barker, K. R., Carter, C. C. and Sasser, J.N. (eds) An Advanced Treatise on Meloidogyne. Volume II: Methodology. A cooperative publication of the Department of Plant Pathology and the United States Agency for International Development, North Carolina State University Graphics, Raleigh, North Carolina, pp.69- 77.
https://advancecovercrops.com/resources-advanced-cover-crops/carbon-nitrogen-ratio
Hussain, M. A.; Mukhtar, T. A. R. I. Q.; Kayani, M. Z.; Aslam, M. N. and Haque, M. I.(2012).A survey of okra (Abelmoschus esculentus) in the Punjab province of Pakistan for the determination of prevalence, incidence and severity of root-knot disease caused by Meloidogyne spp. Pakistan Journal of Botany, 44(6): 2071-2075.
Ismail, A. E. (1998). Effect of soil amendments with some hardwood barks on reproduction of Rotylenchulus reiniformis and growth of sunflower, Pak. J. Nematol., 16: 137-144.
Ismail, M.; Anwar, S. A. and Riaz, A.(2012).Incidence of Meloidogyne incognita in cucumber fields. Pakistan Journal of Zoology, 44(5):1383-1387.
Jaiswal P C (2003). Soil Plant and Water analysis. Mrs. Usha Raj Kumar, India, 136Pp.
Jamil A., Riaz S., Ashraf M., Foolad M.R. ( 2011). Gene expression profiling of plants under salt stress. Crit. Rev. Plant Sci. 30 (5) :435 – 458.
Karajeh, M.R. and Al-Nasir, F.M.(2014).Field Utilization of Nitrogen Fertilizers for Controlling Root-knot Nematode and Improving Growth and Yield of Cucumber. International Journal of Agriculture and Forestry. 4(1): 34-40.
Katooli, N., Moghadam, E. M., Taheri, A., and Nasrollahnejad, S. (2010). Management of root-knot nematode (Meloidogyne incognita) on cucumber with the extract and oil of nematicidal plants. International Journal of Agricultural Research, 5(8), 582-586.
Kayani, M. Z.; Mukhtar, T.; Hussain, M. A.; Haque, M. I. and Perveen, R. (2012).Incidence and severity of root-knot nematodes (Meloidogyne spp.) on cucumber in district Rawalpindi. Pakistan Journal of Phytopathology, 24(2): 122-128.
Khalil, M. S.(2013).The potential of five eco-biorational products on the reproduction of root-knot nematode and plant growth. International Journal of Phytopathology, 2(2): 84-91.
Kumar, D.; S. Singh; and N. Singh. (2010). Free radical scavenging and analgesic activities of cucumbers sativa. Fruit extract. J. Young Pharm., 2(4):365-386.
Kumar, N.; Adamu, M.A.; Isah, K.M. and Lawal, A.F.(2014).A Survey of vegetable fields for root-gall disease in Niger State, Nigeria. PAT, 10(1):17-27.
Martin, R. R., James, D, and Levesque, C. A. ( 2000 ). Impacts of molecular diagnostic technologies on plant disease management. Annual review of phytopathology 38:207-39.
Michel, L., Sikora, R.A. (2005), Plant parasitic nematodes in subtropical and tropical agriculture.2nd Edition.CABI Bioscience,Egham,UK.. 492 Pp.
Naz, I.; Palomares-Rius, J. E.; Blok, V.; Saifullah; Ali, S. and Ahmed, M. (2012).Prevalence, incidence and molecular identification of root-knot nematodes of tomato in Pakistan. African Journal of Biotechnology, 11: 16546-16556.
Oka Y. (2010 ) Mechanisms of nematode suppression by organic soilamendments – a review Appl. Soil Ecol., 44 (2) : 101-115
Olabiyi, T. I.; Olayiwola, A. O. and Oyediran, G. O. (2009).Influence of soil textures on distribution of phytonematodes in the south western Nigeria. World Journal of Agricultural Sciences, 5(5): 557-560.
Onkendi, E. M. and Moleleki, L. N.(2013).Distribution and genetic diversity of root‐knot nematodes (Meloidogyne spp.) in potatoes from South Africa. Plant Pathology, 62(5): 1184-1192.
Park, S.D.; Khan, Z.; Yeon, K. and Kim, Y.H.(2005).A survey for plant-parasitic nematodes associated with strawberry (Fragaria ananassa Duch.) Crop in Korea. Plant Pathology Journal, 21:387-390.
Powers, T. O.; Mullin, P. G.; Harris, T. S.; Sutton, L. A. and Higgins, R. S. (2005).Incorporating molecular identification of Meloidogyne spp. in to a large-scale regional nematode survey. Journal of Nematology, 37:226-235.
Prot, J. C. and Van Gundy, S. D.(1981).Effect of soil texture and the clay component on migration of Meloidogyne incognita second-stage juveniles. Journal of Nematology, 13(2): 213-217.
Rivera L., Aballay E. ( 2008 ) Nematicide effect of various organic soil amendments on Meloidogyne ethiopica whitehead, 1968, on potted vine plants Chil. J. Agric. Res., 68 (3) : 290-296.
Rowell, .D. L.(1996).Soil science: Methods and applications. Longman Group UK Limited. pp. 50 – 51.
Skantar,A.M; Handoo,Z.A; Zanakis,G.N. and E. A. Tzortzakakis, E.A.(2012). Molecular and Morphological Characterization of the Corn Cyst Nematode, Heterodera zeae, from Greece Journal of Nematology, 44 : 58–66.
Stephan Z.A. (1997). Plant nematode problems and their control in the Near East region. (FAO Plant Production and Protection Paper - 144).
Subbotin, S. and Moens, M.(2006).Molecular Taxonomy and Phylogeny. In: Plant Nematology. Perry, R. and Moens, M. (Eds). Pp: 51-57. CABI Nosworthy Way, Wallingford Oxfordshire OX10 8DE, UK..
Stirling, G. R.(1991). Biological control of plant parasitic nematodes. Wallingford, UK: CAB Int.
Trudgill, D.L.(1995).An assessment of the relevance of thermal time relationships to nematology. Fund. appl. Nematology, 18: 407 –417.
Van Reeuwijk, L. R.(1995).Procedures for soil analysis: 3-8. 5th edition .International Soil Reference and Information Center, technical paper 9.
Waeyenberge, L.; Ryss, A.; Moens, M. ; Pinochet, J..and Vrain, T. ( 2000 ). Molecular characterization of 18 Pratylenchus species using rDNA restriction fragment length polymorphism. Nematology, 2:135–142.
Zijlstra, C.; Lever, A.E.M.; Uenk, B.J. and Van Sifhout, C.H.(1995). Differences between ITS regions of isolates of root-knot nematodes Meloidogyne hapla and M. chitwoodi. Phytopathology, 85: 1231–1237
Published
2023-09-25
How to Cite
AMI, S. N., & ABDULQAHAR, F. I. (2023). EXISTENCE, MOLECULAR DIAGNOSIS AND GENETIC VARIATION OF DIFFERENT ISOLATES OF ROOT – KNOT NEMATODES Meloidogyne spp. PARASITIZING CUCUMBER PLANTS. Journal of Duhok University, 26(2), 20-33. https://doi.org/10.26682/ajuod.2023.26.2.3
Section
Agriculture and Veterinary Science
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