Evaluation of Integrated Management Approaches against Citrus Nematode (Tylenchulus semipenetrans) in Pakistan

Waqas A. Khan, Nazir Javed, Muhammad Naveed, Sajid Aleem Khan, Saeed Ahmad


Present study was conducted to explore the nematicidal potential of different synthetic chemicals, biopesticides and antagonists against citrus decline. Effect of eleven chemicals, seven biopesticides and two antagonists on juvenile mortality was evaluated in vitro. Four concentrations (2S, S, S/2 and S/4) of each chemical were prepared on the basis of recommended dose for each chemical. Juvenile mortality of citrus nematodes was calculated after 24, 48 and 72 hour of exposure to chemicals. Rugby expressed maximum percentage of juvenile mortality at 2S, S, S/2 and S/4 concentration. Mortality percentage observed by Rugby, Furadan, Match and Cartap after 72 h at 2S concentration was (100, 100, 95.44, 88.23%) whereas at S/4 concentration mortality percentage was (76.32, 81.18, 62.15, 55.28%) respectively. Among biopesticides maximum percentage mortality observed by Proclaim and Cure after 72 hours at 2S concentration was (83.87, 80.44%) while at S/4 concentration it was (59.87, 57.38%) respectively. Cultural filtrates of two antagonist Trichoderma harzianum and Trichoderma viridi were evaluated at S, S/2 and S/4 concentration under lab conditions. Maximum mortality (88.42%) was observed when both antagonists were applied in combined treatment at S concentration after 72 hours of exposure. Two best performing chemicals, one biopesticide and one antagonist were evaluated under greenhouse against T. semipenetrans on Citrus jambhiri Lush (rough lemon) and their effect on plant growth and nutrient uptake was measured. Nitrogen uptake was measured by following micro Kjeldahl method. For phosphorus and potassium uptake absorbance of samples at 720 nm was measured with the help of spectrophotometer. Phosphorus was calculated by comparing standard curve already prepared while potassium uptake was assessed by flame photometer method after digestion. All the treatments were found significantly effective against citrus nematode but maximum plant height, stem diameter, number of leaves, root length, root weight, shoot weight, and number of feeder roots (90 cm, 2.4 cm, 102, 48.3 g, 12.1 g, 48.8 g and 70) as compared to control when Rugby, Furadan, Proclaim and T. harzianum were applied in combination. Similarly maximum uptake of Nitrogen (3.14) Phosphorous (2.44%) and Potassium (1.95%) was observed when all the treatments were applied in combination. The results of present study will be helpful in selecting the suitable chemicals for growers having problems of citrus nematodes in orchards.


Citrus decline; Antagonists; Biopesticides; Synthetic chemicals; Mortality


Abbasi, M. W., N. Ahmed, M. J. Zaki and S. S. Shaukat. 2008. Effect of Barleria acanthoides Vahl. on root-knot nematode infection and growth of infected okra and brinjal plants. Pakistan Journal of Botany, 40: 2193-98.

Abbott, W. S. 1925. A method of computing the effectiveness of an insecticide. Journal of Economic Entomology, 18: 265-67. https://doi.org/10.1093/jee/18.2.265a

Abd-Elgawad, M., F. Al-Yahya and Z. Stephan. 2010. Nematodes of citrus. In: WA Abu-Gharbieh, AS Al-Hazmi, ZA Stephan and AA Dawabah (eds.), Plant Nematodes in Arab Countries. Arab Society of Plant Protection. Amman, Jordan.

Ali, M. A., M. Shahzadi, A. Zahoor, A. A. Dababat, H. Toktay, A. Bakhsh, M. A. Nawaz and H. Li. 2019. Resistance to cereal cyst nematodes in ingredient with the nematodes and plant root systems: An emphasis on classical and modern approaches. International Journal of Molecular Sciences, 20: 1-18. https://doi.org/10.3390/ijms20020432

Ali, Z., M. Shah, A. Nawaz, M. Shahjahan, H. Butt, M. Shahid and R. Ahmed. 2014. Assessment of induced systemic resistance through antagonistic rhizobacterial potential with salicylic acid against karnal bunt of wheat. Pakistan Journal of Phytopathology, 26: 253-58.

Baines, R., J. Martin, T. DeWolfe, S. Boswell and M. Garber. 1962. Effect of high doses of DD on soil organisms and growth and yield of lemon trees. Journal of Phytopathology, 52: 723-31.

Bashir, A., M. T. Khan, R. Ahmed, B. Mehmood, M. T. Younas, H. M. Rehman and S. Hussain. 2020. Efficiency of selected botanicals against (Alternaria solani) causing early blight disease on tomato in Azad Jammu and Kashmir. Pakistan Journal of Phytopathology, 32: 179-86. https://doi.org/10.33866/phytopathol.030.02.0587

Bhaduri, D., R. Rakshit and K. Chakraborty. 2014. Primary and secondary nutrients: A boon to defense system against plant diseases. International Journal of Bio-resource and Stress Management, 5: 461-66. https://doi.org/10.5958/0976-4038.2014.00597.1

Bhargava, B. and H. Raghupathi. 1995. Current status and new norms of nitrogen nutrition for grapevine (Vitis vinifera). Indian Journal of Agricultural Sciences, 65: 165-69.

Bibi, S., M. Inam-Ul-Haq, A. Riaz, S. I. Malik, M. I. Tahir and R. Ahmed. 2017. Screening and characterization of Rhizobacteria antagonistic to Pseudomonas syringae causing bacterial canker of stone fruits in Punjab and KPK. International Journal of Biosciences, 10: 405-12.

Casas-Flores, S. and A. Herrera-Estrella. 2007. Antagonism of plant parasitic nematodes by fungi. The Mycota, 6: 147-57.

Cayrol, J.-C., C. Djian and J. Frankowski. 1993. Efficacy of abamectin B1 for the control of Meloidogyne arenaria. Fundamental and Applied Nematology, 16: 239-46.

Cigdem, K. and M. Kivanc. 2005. Effect of formulation on the viabilıty of biocontrol agent, Trıchoderma harzıanum conidia. African Journal of Biotechnology, 4: 483-86.

Desaeger, J., M. R. Rao and J. Bridge. 2004. Nematodes and other soilborne pathogens in agroforestry. Below-ground interactions in tropical agroecosystems: concepts and models with multiple plant components: 263-83. https://doi.org/10.1079/9780851996738.0263

Dordas, C. 2008. Role of nutrients in controlling plant diseases in sustainable agriculture. A review. Agronomy for sustainable development, 28: 33-46. https://doi.org/10.1051/agro:2007051

Dubey, W. and P. Trivedi. 2011. Evaluation of some nematicides for the control of Meloidogyne incognita on okra. Indian Journal of Fundamental and Applied Life Science, 1: 264-70.

El-Rokiek, K. G. and W. M. El-Nagdi. 2011. Dual effects of leaf extracts of Eucalyptus citriodora on controlling purslane and root-knot nematode in sunflower. Journal of plant protection research, 51: 121-29. https://doi.org/10.2478/v10045-011-0021-0

Fisk, C. H. and Y. J. Subbarow. 1925. The colorimetric determination of phosphorus. Journal of Biological Chemistry, 66: 375-400. https://doi.org/10.1016/S0021-9258(18)84756-1

Gade, R. and S. Armarkar. 2011. Growth promotion and disease suppression ability of Pseudomonas fluorescens in Acid lime. Archives of Phytopathology and Plant Protection, 44: 943-50. https://doi.org/10.1080/03235400903458761

Gašić, S. and B. Tanović. 2013. Biopesticide formulations, possibility of application and future trends. Pesticidi i fitomedicina, 28: 97-102. https://doi.org/10.2298/PIF1302097G

Goswami, J., R. K. Pandey, J. Tewari and B. Goswami. 2008. Management of root knot nematode on tomato through application of fungal antagonists, Acremonium strictum and Trichoderma harzianum. Journal of Environmental Science and Health, 43: 237-40. https://doi.org/10.1080/03601230701771164

Graham, J. H. 2004. Biological control of soil borne plant pathogens and nematodes. In: D. M. Sylvia, J. J. Fuhrmann, P. G. Hartel and D. Zuberer (eds.), Principals and Applications of Soil Microbiology. Pearson Prentice Hall. Upper Saddle River, NJ.

Huang, X., N. Zhao and K. Zhang. 2004. Extracellular enzymes serving as virulence factors in nematophagous fungi involved in infection of the host. Research in Microbiology, 155: 811-16. https://doi.org/10.1016/j.resmic.2004.07.003

Hyder, S., A. S. Gondal, Z. F. Rizvi, R. Ahmad, M. M. Alam, A. Hannan, W. Ahmed, N. Fatima and M. Inam-ul-Haq. 2020. Characterization of native plant growth promoting rhizobacteria and their anti-oomycete potential against Phytophthora capsici affecting chilli pepper (Capsicum annum L.). Scientific Reports, 10: 1-15. https://doi.org/10.1038/s41598-020-69410-3

Institute, S. 1990. SAS/STAT User's Guide: Version 6. SAS Institute Incorporated.

Johnson, C. D. and A. Stretton. 1987. GABA-immunoreactivity in inhibitory motor neurons of the nematode Ascaris. Journal of Neuroscience, 7: 223-35. https://doi.org/10.1523/JNEUROSCI.07-01-00223.1987

Khan, A. R., W. Ashraf, W. A. Khan, H. ur Rehman, A. A. Khan and M. Mehdi. 2017. Role of Glomus mosseae and neemex in the management of Meloidogyne incognita on tomato. Plant Protection, 1: 69-73.

Khan, F., M. Mazid, T. A. Khan, H. K. Patel and R. Roychowdhury. 2014. Plant derived pesticides in control of lepidopteran insects: Dictum and directions. Research Journal of Biology, 2: 1-10.

Kjeldahl, C. 1883. A new method for the determination of nitrogen in organic matter. Analytical Chemistry, 22: 366-82.

Koche, M. 2011. Integrated disease management for root rot and gummosis in Nagpur mandarin.

Lamberti, F., T. D'addabbo, P. Greco, A. Carella and P. De Cosmis. 2000. Management of root-knot nematodes by combination of soil solarization and fenamiphos in southern Italy. Nematologia Mediterranea, 28: 31-45.

Li, B., G.-l. Xie, A. Soad and J. Coosemans. 2005. Suppression of Meloidogyne javanica by antagonistic and plant growth-promoting rhizobacteria. Journal of Zhejiang University Science, 6: 496-501. https://doi.org/10.1631/jzus.2005.B0496

Mahmood, I., A. Masood, S. Saxena and S. Husain. 1979. Effect of some plant extracts on the mortality of Meloidgyne incognita and Rotylenchulus reniformis. Acta Botanica Indica, 7: 129-31.

McKenry, M. V. and P. Roberts. 1985. Phytopathology: Study Guide. Agriculture and Natural Resources, California Public University. Oakland.

Meola, R. W., S. R. Dean, S. M. Meola, H. Sittertz-Bhatkar and R. Schenker. 1999. Effect of lufenuron on chorionic and cuticular structure of unhatched larval Ctenocephalides felis (Siphonaptera: Pulicidae). Journal of medical entomology, 36: 92-100. https://doi.org/10.1093/jmedent/36.1.92

Mishra, R. and R. Gupta. 2012. In vitro evaluation of plant extracts, bio-agents and fungicides against purple blotch and Stemphylium blight of onion. Journal of Medicinal Plants Research, 6: 5658-61.

Moosavi, M. R. and R. Zare. 2020. Fungi as biological control agents of plant-parasitic nematodes. In, Plant Defence: Biological Control. Springer Science Business Media. Dordrecht. https://doi.org/10.1007/978-3-030-51034-3_14

Nefzi, A., B. A. R. Abdullah, H. Jabnoun-Khaireddine, S. Saidiana-Medimagh, R. Haouala and M. Danmi-Remadi. 2016. Antifungal activity of aqueous and organic extracts from Withania somnifera L. against Fusarium oxysporum f. sp. radices-lycopersici. Journal of Microbial and Biochemical Technology, 8: 144-50. https://doi.org/10.4172/1948-5948.1000277

Osei, K., R. Addico, A. Nafeo, A. Edu-Kwarteng, A. Agyemang, Y. Danso and J. Sackey-Asante. 2011. Effect of some organic waste extracts on hatching of Meloidogyne incognita eggs. African Journal of Agricultural Research, 6: 2255-59.

Ploetz, R. C. 2006. Fusarium-induced diseases of tropical, perennial crops. Phytopathology, 96: 648-52. https://doi.org/10.1094/PHYTO-96-0648

Qifei, Y., X. Fuwu and Y. Xiulin. 2003. Effects of increasing phosphate and potassium fertilizers on the control of Cymbidium anthracnose. Journal of Tropical and Subtropical Botany, 11: 157-60.

Rehman, A., R. Bibi and M. H. Ullah. 2006. Evaluation of different chemicals against root knot nematode (Meloidogyne incognita) on sunflower. Journal of Agriculture and Social Sciences, 2: 185-88.

Rich, J. R., R. Dunn and J. Noling. 2004. Nematicides: Pest and present uses. In: Z.X. Chen, S. Y. Chen and D. W. Dickson (eds.), Nematology: Advances and Perspective, Nematode Management and Utilization. CABI Publishing. Wallingford, UK. https://doi.org/10.1079/9780851996462.1179

Safdar, H., N. Javed, S. A. Khan, I. ul Haq, A. Safdar and N. Khan. 2012. Control of Meloidogyne incognita (Kofoid and White) chitwood by cadusafos (Rugby®) on tomato. Pakistan journal of Zoology, 44: 1703-10.

Samson, R. A., E. S. Hoekstra and C. A. Van Oorschot. 1996. Introduction to Food-Borne Fungi. Centraal Bureau voor de Statistiek: Netherlands.

Samuels, G. J. 1996. Trichoderma: A review of biology and systematics of the genus. Mycological research, 100: 923-35. https://doi.org/10.1016/S0953-7562(96)80043-8

Semeniuc, C. A., C. R. Pop and A. M. Rotar. 2017. Antibacterial activity and interactions of plant essential oil combinations against Gram-positive and Gram-negative bacteria. journal of food and drug analysis, 25: 403-08. https://doi.org/10.1016/j.jfda.2016.06.002

Shishir, A., A. Bhowmik, N. Akanda, A. Al Mamun, S. Khan and M. Hoq. 2015. Efficacy of indigenous Bacillus thuringiensis strains for controlling major vegetable pests in Bangladesh. Egyptian Journal of Biological Pest Control, 25: 729-34s.

Singh, A. K., B. Bhatt, D. Singh, R. Gade, A. K. Singh and U. Sangle. 2012. Good Agronomic Practices (GAP)-An efficient and eco-friendly tool for sustainable management of plant diseases under changing climate scenario. Journal of Plant Disease Sciences, 7: 1-8.

Singh, B. 2004. Control of citrus nematode, Tylenchulus semipenetrans in Nagpur mandarin orchard. Indian Journal of Nematology, 34: 70-74.

Sorribas, F. J., S. Verdejo-Lucas, J. Pastor, C. Ornat, J. Pons and J. Valero. 2008. Population densities of Tylenchulus semipenetrans related to physicochemical properties of soil and yield of clementine mandarin in Spain. Plant Disease, 92: 445-50. https://doi.org/10.1094/PDIS-92-3-0445

Spann, T. M. and A. W. Schumann. 2010. Mineral nutrition contributes to plant disease and pest resistance (2576-0009). University of Florida, IFAS Extention. Florida, USA, pp. 1-5. https://doi.org/10.32473/edis-hs1181-2010

Spiegel, Y., I. Chet and E. Sharon. 2004. Mechanisms and improved biocontrol of the root-knot nematodes by Trichoderma spp. Acta Horticulturae, 698: 225-28. https://doi.org/10.17660/ActaHortic.2005.698.30

Ullah, Z., N. Javed, S. A. Khan, I. U. Haq and Q. Shakeel. 2015. Efficacy of biopesticides for management of root knot nematode on tomato. International Journal of Vegetable Science, 21: 215-22. https://doi.org/10.1080/19315260.2013.829897

Vandermeer, J., I. Perfecto and S. Philpott. 2010. Ecological complexity and pest control in organic coffee production: Uncovering an autonomous ecosystem service. BioScience, 60: 527-37. https://doi.org/10.1525/bio.2010.60.7.8

Verdejo-Lucas, S. and M. McKenry. 2004. Management of the citrus nematode, Tylenchulus semipenetrans. Journal of Nematology, 36: 424.

Whitehead, A. and J. Hemming. 1965. A comparison of some quantitative methods of extracting small vermiform nematodes from soil. Annals of applied Biology, 55: 25-38. https://doi.org/10.1111/j.1744-7348.1965.tb07864.x

Zaitoun, F., M. M. Fahmy and S. M. Ziyada. 2015. Role of certain bioagents against guava decline disease and in enhancement of the growth of guava trees. Journal of Phytopathology and Pest Management, 2: 43-54.

Full Text: PDF

DOI: 10.33687/phytopath.011.01.4092


  • There are currently no refbacks.

Copyright (c) 2022 Sajid Aleem Khan

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.