Management of root-knot nematode infection by using fly ash and Trichoderma harzianum in Capsicum annum plants by modulating growth, yield, photosynthetic pigments, biochemical substances, and secondary metabolite profiles


  • Gufran AHMAD Aligarh Muslim University, Faculty of Life Sciences, Department of Botany (IN)
  • Amir KHAN Aligarh Muslim University, Faculty of Life Sciences, Department of Botany (IN)
  • Safiuddin ANSARI Aligarh Muslim University, Faculty of Life Sciences, Department of Botany (IN)
  • Abrar A. KHAN Aligarh Muslim University, Faculty of Life Sciences, Department of Botany (IN)
  • Abeer ELHAKEM Prince Sattam Bin Abdulaziz University, Department of (SA)
  • Rokayya SAMI Taif University, Department of Food Science and Nutrition (SA)
  • Heba I. MOHAMED Ain Shams University, Faculty of Education, Biological and Geological Sciences Department (EG)



chili, carbohydrate, flavonoids, Meloidogyne incognita, phenolics, proline


A nematicide is a type of chemical pesticide used to kill plant-parasitic nematodes. Nematicides have tended to be broad-spectrum toxicants, possessing high volatility or other properties that promote migration through the soil. In addition, the nematicides used are more expensive and have adverse effects on health and the environment, so it must use more eco-friendly and less expensive alternative methods to control root-knot nematodes (Meloidogyne incognita). Chili (Capsicum annum) suffers from nematode infestation, which reduces its quality and quantity. Therefore, the goal of this research was to assess the effect of different doses of fly ash (FA) mixed soil (5%, 10%, 15 and 20% FA) with two doses of Trichoderma harzianum (1 g and 2 g) for the management of root-knot nematode infection in chili crop. The results showed that significant enhancement in plant growth, yield, chlorophyll, and carotenoid content, protein, carbohydrate, amino acid, tryptophan, indole acetic acid, phenolics, flavonoids, proline, and nitrate reductase content of chili plants was recorded at 10% fly ash with 2 g of T. harzianum (T6). The inoculated plants registered the greatest damage with galling indexes. The lowest galling index was estimated at the T6 treatment. At higher levels of FA + combined with both doses of T. harzianum, nematode could not survive that’s why eighter galls or egg masses were observed. Nematodes may have ceased to function, lost their activity, and hence been unable to resist the stress of fly ash and T. harzianum set.  The application of T. harzianum with a lower dose (10%) of fly ash to control the nematode favored plant growth in general. In conclusion, 10% fly ash and 2 g of T. harzianum have the ability to operate as growth promoters and biocontrol agents for M. incognita.


Abd El- Rahman SS, Mohamed HI (2014). Application of benzothiadiazole and Trichoderma harzianum to control faba bean chocolate spot disease and their effect on some physiological and biochemical traits. Acta Physiologia Plantarum 36(2):343-354.

Abd-Elgawad MM, Kabeil SS (2012). Biological control of Meloidogyne incognita by Trichoderma harzianum and Serratia marcescens and their related enzymatic changes in tomato roots. African Journal of Biotechnology 11:16247-16252.

Abu-Shahba MS, Mansour MM, Mohamed HI, Sofy MR (2021). Comparative cultivation and biochemical analysis of iceberg lettuce grown in sand soil and hydroponics with or without microbubble and microbubble. Journal of Soil Science and Plant Nutrition 21:389-403.

Ahmad G, Khan AA, Ansari S (2017). Interaction of a fly ash and root-knot nematode pathogens on pumpkin (Cucurbita moschata Duch. ex Lam.). Tropical Plant Research 4(3):449-455.

Ahmad G, Khan AA (2016). Fly ash effect on hatching, mortality and penetration of root-knot nematode (Meloidogyne incognita) in pumpkin roots. International Journal of Environment 5(3):66-73.

Ahmad G, Khan A, Khan AA, Ali A, Mohamed HI (2021a). Biological control: a novel strategy for the control of the plant parasitic nematodes. Antonie van Leeuwenhoek 114:885-912.

Ahmad G, Khan AA, Mohamed HI (2021b). Impact of the low and high concentrations of fly ash amended soil on growth, physiological response, and yield of pumpkin (Cucurbita moschata Duch. Ex Poiret L.). Environmental Science and Pollution Research 28(14):17068-17083.

Akladious SA, Mohamed HI (2018). Ameliorative effects of calcium nitrate and humic acid on the growth, yield component and biochemical attribute of pepper (Capsicum annuum) plants grown under salt stress. Scientia Horticulturae 236:244-250.

Al-Hazmi AS, TariqJaveed M (2016). Effects of different inoculum densities of Trichoderma harzianum and Trichoderma viride against Meloidogyne javanica on tomato. Saudi Journal of Biological Science 23(2):288-292.

Aly AA, Mansour MTM, Mohamed HI, Abd-Elsalam KA (2012). Examination of correlations between several biochemical components and powdery mildew resistance of flax cultivars. Plant Pathology Journal 28(2):149-155.

Aly AA, Mohamed HI, Mansour MTM, Omar MR (2013). Suppression of powdery mildew on flax by foliar application of essential oils. Journal of Phytopathology 161:376-381.

Ashry NA, Ghonaim MM, Mohamed HI, Mogazy AM (2018). Physiological and molecular genetic studies on two elicitors for improving the tolerance of six Egyptian soybean cultivars to cotton leaf worm. Plant Physiology and Biochemistry 130:224-234.

Azher AA Zaghloul AM, Mahmoud SA, Siam HS (2011). Stimulatory effect of kinetin, ascorbic acid and glutamic acid on growth and chemical constituents of Codiaeum variegatum L. plants. American-Eurasian Journal of Agricultural and Environmental Sciences 10:318-323.

Basu A, Sethi U, Mukherjee SP (1989). Regulation of cell proliferation and morphogenesis by amino acids in Brassica tissue cultures and its correlation with threonine deaminase. Plant Cell Report 08:333-335.

Beffa R, Martin HV, Pilet PE (1990). In vitro oxidation of indoleacetic acid by soluble auxin-oxidases and peroxidases from maize roots. Plant Physiology 94:485-491.

Bhattacharjee R, Dey U (2014). An overview of fungal and bacterial biopesticides to control plant pathogens/diseases. African Journal of Microbiological Research 8(17):1749-1762.

Black CA, Evans DD, White JL, Ensmingher LE, Clarke FE, Dinauer RC (1965). Methods of soil analysis. Part I- Physical and mineralogical properties including statistics of measurement and sampling (9 Series Agronomy). Madison: American Society of Agronomy.

Blake GR (2008). Particle density. In: Chesworth W (Ed). Encyclopedia of Soil Science. Encyclopedia of Earth Sciences Series. Springer, Dordrecht.

Bruuinsma J (1963) The quantitative analysis of chlorophylls a and b in plant extracts. Photochemistry and Photobiology 2(2):241-249.

Carter MR, Ball B (1993). Soil porosity. In: Carter MR (Ed). Soil sampling and Methods of Analysis. Canadian Society of Soil Science. Lewis Publishers, CRC Press, Boca Raton, pp 581-588.

Chopra SL, Kanwar JS (1991). Analytical agricultural chemistry. Kalyani Publishers, New Delhi-Ludhiana.

Chopra SL, Kanvar JS (1982). Analytical agricultural chemistry. Kalyani publishers, New Delhi, pp 162.

Costa SR, Ng JLP, Mathesius U (2021). Interaction of symbiotic rhizobia and parasitic root-knot nematodes in legume roots-from molecular regulation to field application. Molecular Plant-Microbe Interactions

Dahiya HS, Budania YK (2018). Prospects of fly ash application in agriculture: a global review. International Journal of Current Microbiology and Applied Sciences 7:397-409.

De SR, Adriana A, Passaglia LMP (2015). Plant growth-promoting bacteria as inoculants in agricultural soils. Genetics and Molecular Biology 38:401-419.

Dickman SR, Bray RH (1940). Colorimetric determination of phosphate. Industrial Engineering Chemistry and Analytical Edition 12:665-668.

Druzhinina IS, Seidl-Seiboth V, Herrera-Estrella A, Horwitz BA, Kenerley CM, Monte E, Kubicek CP (2011). Trichoderma: The genomics of opportunistic success. Nature Reviews Microbiology 9:749-759.

Dubois M, Gilles KA, Hamilton JK, Rebers PT, Smith F (1956). Colorimetric method for determination of sugars and related substances. Analytical Chemistry 28(3):350-356.

El-Beltagi HS, Sofy MR, Aldaej MI, Mohamed HI (2020). Silicon alleviates copper toxicity in flax plants by up-regulating antioxidant defense and secondary metabolites and decreasing oxidative damage. Sustainability 12:4732.

El-Beltagi HS, Mohamed HI, Elmelegy AA, Eldesoky SE, Safwat G (2019). Phytochemical screening, antimicrobial, antioxidant, anticancer activities and nutritional values of cactus (Opuntia ficus Indicia) pulp and peel. Fresenius Environmental Bulletin 28(2A):1534-1551.

Elhady A, Adss S, Hallmann J, Heuer H (2018). Rhizosphere microbiomes modulated by pre-crops assisted plants in defense against plant-parasitic nematodes. Frontiers in Microbiology 9:1133.

El-Mahdy OM, Mohamed HI, Mogazy AM (2021). Biosorption effect of Aspergillus niger and Penicillium chrysosporium for Cd and Pb contaminated soil and their physiological effects on Vicia faba L. Environmental Science and Pollution Research 28:67608-67631.

Feyisa B, Lencho A, Selvaraj T, Amb PBN, Getaneh G (2016). Evaluation of some botanicals and Trichoderma harzianum for the management of tomato root-knot nematode (Meloidogyne incognita (Kofoid and White) Chit Wood). Advances in Crop Science and Technology 4:201.

Ghonaim MM, Mohamed HI, Omran AAA (2021). Evaluation of wheat salt stress tolerance using physiological parameters and retrotransposon-based markers. Genetic Resources and Crop Evolution 68:227-242.

Halifu S, Deng X, Song X, Song R (2019). Effects of two Trichoderma strains on plant growth, rhizosphere soil nutrients, and fungal community of Pinus sylvestris var. mongolica annual seedlings. Forests 10(9):758.

Haris M, Ahmad G, Shakeel A, Khan AA (2019). Utilization of fly ash to improve the growth and the management of root-knot nematode on carrot. Haya Saudi Journal of Life Science 4(7):221-226.

Heidrich C, Feuerborn HJ, Weir A (2013). Coal combustion products: a global perspective. In: ‘World of Coal Ash’ Conference, pp 22-25.

Hermosa R, Rubio MB, Cardoza RE, Nicolás C, Monte E, Gutiérrez S (2013). The contribution of Trichoderma to balancing the costs of plant growth and defense. International Microbiology 16(2):69-80.

Jackson ML (1973). Soil chemistry analysis. Prentice Hall, New Delhi.

Jang JY, Dang QL, Choi YH, Choi GJ, Jang KS, Cha B, Luu NH, Kim JC (2014). Nematicidal activities of 4-quinolone alkaloids isolated from the aerial part of Triumfetta grandidens against Meloidogyne incognita. Journal of Agricultural and Food Chemistry 63:68-74.

Jaworski EG (1971). Nitrate reductase assay in intact plant tissues. Biochemical and Biophysical Research Communications 43(6):1274-1279.

Ji X, Li J, Meng Z, Dong S, Zhang S, Qiao K (2019). Inhibitory effect of allicin against Meloidogyne incognita and Botrytis cinerea in tomato. Scientia Horticulture 253:203-208.

Kaur DN, Sharma SK, Sultan MS (2011). Effect of different chemicals on root knot nematode in seed beds of tomato. Plant Disease Research 26:170-170.

Katz JJ, Norris JR, Shipman LL, Thurnauer MC, Wasielewski MR (1978). Chlorophyll function in the photosynthetic reaction center. Annual Review of Biophysics and Bioengineering 7:393-434.

Khan MR, Khan MW (1996). Effect of fly ash on plant growth and yield of tomato. Environmental Pollution 92:105-111.

Khan RAA, Najeeb S, Hussain S, Xie BY, Li Y (2020). Bioactive secondary metabolites from Trichoderma spp. against phytopathogenic fungi. Microorganisms 8:817.

Khanna K, Jamwal VL, Kohli SK, Gandhi SG, Ohri P, Bhardwaj R, Wijaya L, Alyemeni MN, Ahmad P (2019). Role of plant growth promoting bacteria (PGPR) as biocontrol agents of Meloidogyne incognita through improved plant defense of Lycopersicon esculentum. Plant Soil 436:325-345.

Kihika R, Murungi LK, Coyne D, Ng’ang’a M, Hassanali A, Teal PEA, Torto1 B (2017). Parasitic nematode Meloidogyne incognita interactions with different Capsicum annum cultivars reveal the chemical constituents modulating root herbivory. Scientific Reports 7: 2903.

Kjeldahl J (1883). New method for the determination of nitrogen. Chemistry News 48:101-102.

Law DM (1987). Gibberellin enhanced indole-3-acetic acid biosynthesis: D-tryptophan as the precursor of indole-3-acetic acid. Plant Physiology 70:626-632.

Lichtenthaler HK, Wellburn AR (1983). Determinations of total carotenoids and chlorophylls a and b of leaf extracts in different solvents. Portland Press Limited, London.

Menon MP, Ghuman GS, James J, Chandra K (1990). Physicochemical characterization of water extracts of different coal fly ashes and fly ash amended composts. Water, Air Soil Pullulation 50:343-353.

Meyer SL, Roberts DP, Chitwood DJ, Carta LK, Lumsden RD, Mao W (2001). Application of Burkholderia cepacia and Trichoderma virens, alone and in combinations, against Meloidogyne incognita on bell pepper. Nematropica 75-86.

Mohamed HI, El-Beltagi HS, Aly AA, Latif HH (2018). The role of systemic and non systemic fungicides on the physiological and biochemical parameters in Gossypium hirsutum plant, implications for defense responses. Fresenius Environmental Bulletin 27(12):8585-8593.

Mohamed HI, Abd–El Hameed AG (2014). Molecular and biochemical markers of some Vicia faba L. genotype in response to storage insect pests infestation. Journal of Plant Interactions 9(1):618-626.

Mohamed HI (2011). Molecular and biochemical studies on the effect of gamma rays on lead toxicity in cowpea (Vigna sinensis) plants. Biological Trace Element Research 144:1205-1218.

Moustafa-Farag M, Mohamed HI, Mahmoud A, Elkelish A, Misra AN, Guy KM, … Zhang M (2020). Salicylic acid stimulates antioxidant defense and osmolyte metabolism to alleviate oxidative stress in watermelons under excess boron. Plants 9:724.

Mohamed HI, Elsherbiny EA, Abdelhamid MT (2016). Physiological and biochemical responses of Vicia faba plants to foliar application with zinc and iron. Gesunde Pflanzen 68:201-212.

Moosavi MR (2015). Damage of the root-knot nematode Meloidogyne javanica to bell pepper, Capsicum annuum. Journal of Plant Diseases and Protection 122(5-6):244-249.

Nakagawa H, Poulle M, Oaks A (1984). Characterization of nitrate reductase from corn leaves (Zea mays cv W64A* W182E): two molecular forms of the enzyme. Plant Physiology 75:285-289

Neals TF (1956). Components of total magnesium content within the leaves of white clover and perennial rye grass. Nature 177:388-389.

Nieto-Jacobo MF, Steyaert JM, Salazar-Badillo FB, Nguyen DV, Rostás M, Braithwaite M, … Mendoza-Mendoza A (2017). Environmental growth conditions of Trichoderma spp. affects indole acetic acid derivatives, volatile organic compounds, and plant growth promotion. Frontiers in Plant Science 8:102.

Niu J, Su X, Tang Z (2021). Influence of fly ash and polyacrylamide mixtures on growth properties of Artemisia ordosica in the desert region of north China. Agronomy 11:590.

Nyaku ST, Affokpon A, Danquah A, Brentu FC (2017). Harnessing useful rhizosphere microorganisms for nematode control. Nematology-Concepts, Diagnosis and Control 153-182.

Panda RB, Biswal T (2018). Impact of fly ash on soil properties and productivity. International Journal of Agriculture Environment and Biotechnology 11:275-283.

Patel VS, Shukla YM, Dhruve JJ (2017). Influence of root knot nematode (Meloidogyne spp.) on phenolic acid profile in root of tomato (Solanum lycopersicum L.). International Journal of Current Microbiology and Applied Sciences 6:840-848.

Paradikovic N, Vinkovic T, Vrcek IV, Zuntar I, Bojic M, Medic- Saric M (2011). Effect of natural biostimulants on yield and nutritional quality: as example of sweet yellow pepper (Capsicum annuum L.) plants. Journal of the Science of Food and Agriculture 91:2146-2152.

Penella C, González Nebauer S, López Galarza SV, San Bautista Primo A, Gorbe E, Calatayud A (2013). Evaluation for salt stress tolerance of pepper genotypes to be used as rootstocks. Journal of Food, Agriculture and Environment 11(3):1101-1107.

Penella C, Nebauer SG, López-Galarza S, San Bautista A, Rodríguez-Burruezo A, Calatayud Á (2014). Evaluation of some pepper genotypes as rootstocks in water stress conditions. Horticulture Research 41(4):192-200.

Richard LA (1954). Diagnosis and improvement of saline and alkali soils. Agric. Handbook, 60, U.S. Dept, Agric. Washington DC, pp 160.

Ruanma K, Shank L, Chairote G (2010). Phenolic content and antioxidant properties of green Chili paste and its ingredients. Maejo International Journal of Science and Technology 4(2):193-200.

Sadasivam S, Manickam A (1996). Biochemical Methods for Agricultural Science. Wiley Eastern Limited, New Delhi, pp 1.

Samolski I, Rincon AM, Pinzon LM, Viterbo A, Monte E (2012). The qid74 gene from Trichoderma harzianum has a role in root architecture and plant biofertilization. Microbiology 158(1):129-138.

Seid A, Fininsa C, Mekete T, Decraemer W, Wesemael WM (2015). Tomato (Solanum lycopersicum) and root-knot nematodes (Meloidogyne spp.)–a century-old battle. Nematology 17(9):995-1009.

Shakeel A, Khan AA, Ahmad G (2019). The potential of thermal power plant fly ash to promote the growth of Indian mustard (Brassica juncea) in agricultural soils. SN Applied Science 1(4):1-5.

Slyke V, Dillon DD, MacFadyen DA, Hamilton P (1941). Determination of carboxyl groups in free amino acids. Journal of Biological Chemistry 142:627-668.

Sofy MR, Mohamed HI, Dawood MFA, Abu-Elsaoud AM, Soliman MH (2021a). Integrated usage of arbuscular mycorrhizal fungi and chicken waste biochar as economic potential tools to ameliorate antioxidant activity, osmolyte accumulation and salt endogenous hormone-stressed spinach plants. Archives of Agronomy and Soil Science.

Sofy AR, Sofy MR, Hmed AA, Dawoud RA, Refaey EE, Mohamed HI, El-Dougdoug NK (2021b). Molecular characterization of the Alfalfa mosaic virus infecting Solanum melongena in Egypt and control of its deleterious effects with melatonin and salicylic acid. Plants 10(3):459.

Sokhandani Z, Moosavi MR, Basirnia T (2016) Optimum concentrations of Trichoderma longibrachiatum and cadusafos for controlling Meloidogyne javanica on zucchini plants. Journal of Nematology 48(1):54.

Taylor AL, Sasser JN (1978). Biology, identification and control of root-knot nematodes. North Carolina State University Graphics, pp 111.

Van Acker SA, Tromp MN, Griffioen DH, Van Bennekom WP, Van Der Vijgh WJ, Bast A (1996). Structural aspects of antioxidant activity of flavonoids. Free Radical Biology and Medicine 20(3):331-342.

Vasanth K, Lakshmiprabha A, Jayabalan N (2006). Amino acids enhancing plant regeneration from cotyledon and embryonal axis of peanut (Arachis hypogaeol). Indian Journal of Crop Science 01:79-83.

Vierstra RD (1993). Protein degradation in plants. Annual Review in Plant Physiology and Plant Molecular Biology 44:385-410.

Wurtman RJ, Anton-Tay F (1969). The mammalian pineal as a neuroendocrine transducer. Recent Progress in Hormone Research 25:493-522.

Yan Y, Mao Q, Wang Y, Zhao J, Fu Y, Yang Z, … Ahammed GJ (2021). Trichoderma harzianum induces resistance to root-knot nematodes by increasing secondary metabolite synthesis and defense-related enzyme activity in Solanum lycopersicum L. Biological Control 158:104609.

Yao ZT, Ji XS, Sarker PK, Tang JH, Ge LQ, Xia MS, Xi YQ (2015). A comprehensive review on the applications of coal fly ash. Earth-Science Reviews 141:105-121.

Zhang S, Gan Y, Xu B (2015). Biocontrol potential of a native species of Trichoderma longibrachiatum against Meloidogyne incognita. Applied Soil Ecology 94:21-29.

Zhang S, Gan Y, Ji W, Xu B, Hou B, Liu J (2017). Mechanisms and characterization of Trichoderma longibrachiatum T6 in suppressing nematodes (Heterodera avenae) in wheat. Frontiers in Plant Science 8:1491.

Zhang S, Gan Y, Xu B, Xue Y (2014). The parasitic and lethal effects of Trichoderma longibrachiatum against Heterodera avenae. Biological Control 72:1-8.



How to Cite

AHMAD, G., KHAN, A., ANSARI, S., KHAN, A. A., ELHAKEM, A., SAMI, R., & MOHAMED, H. I. (2022). Management of root-knot nematode infection by using fly ash and Trichoderma harzianum in Capsicum annum plants by modulating growth, yield, photosynthetic pigments, biochemical substances, and secondary metabolite profiles. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 50(1), 12591.



Research Articles
DOI: 10.15835/nbha50112591

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