Fractions from Mangifera indica as an Alternative in Meloidogyne incognita Management

Oluwatoyin Adenike Fabiyi

Department of Crop Protection, Faculty of Agriculture, University of Ilorin.

Received | May 30, 2022; Accepted | June 21, 2022; Published | June 24, 2022

*Correspondence | Oluwatoyin Adenike Fabiyi, Department of Crop Protection, Faculty of Agriculture, University of Ilorin; Email: fabiyitoyinike@hotmail.com

Citation | Fabiyi, O.A., 2022. Fractions from Mangifera indica as an alternative in Meloidogyne incognita management. Pakistan Journal of Nematology, 40(1): 65-74.

DOI | https://dx.doi.org/10.17582/journal.pjn/2022/40.1.65.74

Keywords | Mangifera indica, Meloidogyne incognita, Pollution, Nematodes

Copyright: 2022 by the authors. Licensee ResearchersLinks Ltd, England, UK.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

Introduction

Tomato (Lycopersicon esculentum) is a key vegetable crop in Nigeria, after onions and pepper. It is a rich source of vitamins A, B, C, E, iron and phosphorus (Fawusi, 1978; Karen, 2007). Tomato plants are inclined to infestation by plant parasitic nematodes, particularly the Meloidogyne spp. which brings about general reduction in the yield of tomato fruits in western Nigeria (FAO, 2004; Fabiyi, 2018; Fabiyi and Olatunji, 2021a). Meloidogyne incognita invades the root of vegetable plants feed and reproduces, causing galls in the root tissue (Safdar et al., 2012; Fabiyi and Olatunji, 2021b; Fabiyi et al., 2022), which in turn reduces the movement of water and nutrients from the root to the shoot, thus weakening the plant. In severe cases death of the plants and yield loss of about 30% has been recorded (Shakeel et al., 2012). Owing to the devastating effect of M. incognita infestation on vegetables, carbofuran (2,3-dihydro-2,2-dimethyl-benzofuran7-yl-N-methylcarbamate) is customarily employed for control. Carbofuran is relatively mobile in soil and surface runoff because of its high water solubility (351 ppm) and low adsorption coefficient (Lau et al., 2007), consequently contaminating lakes, streams and groundwater (Goad et al., 2004). Environmental issues have increasingly limited the use of carbofuran (Rich et al., 2004), while diverse options of control are put forward (Fabiyi et al., 2020; Atolani and Fabiyi, 2020). This study was initiated as a result of serious concern for the enormous damage resulting from yield reduction associated with M. incognita infestation of tomato plants. The environmental risk sequential upon extensive and indiscriminate use of synthetic nematicides presupposes the pursuit of an alternative plant derived nematicide (Atolani et al., 2014a, b; Fabiyi, 2021a, b, c). Mangifera indica, a member of the family Anacardiaceae is reported to be endowed with medicinal properties. The stem bark of M. indica is known to have antimicrobial and anti-amoebic properties (Das et al., 1989; Tona et al., 2000). The treatment of syphilis and diarrheal is equally associated with extracts from M. indica (Ross, 1999), while Munza et al. (1994) documented the activities of the stem bark extract in the treatment of skin diseases and mouth sores. Accordingly, in this experiment, the toxicity of organic compounds isolated from the stem bark of M. indica on M. incognita infesting tomato plants in screenhouse and field studies is assessed.

Materials and Methods

Collection of plant materials

The stem bark of Mangifera indica was scrapped from the mother tree, the large pieces were meticulously diced into small bits of about 2cm each and were dried at room temperature for four weeks (Fabiyi et al., 2012a). The initial weight was 33.34 Kg and final weight at ambient temperature drying was 29.08 Kg. The materials at equal weight of 9. 69 Kg each was subjected separately to cold extraction in methanol, ethyl acetate and n-hexane for five days. The crude extracts were poured off the jars, it was then filtered and concentrated with rotary evaporator (Buchi Rota Vapour R-300).

Fractionation and spectroscopy

A glass column was filled with silica gel 60 (80-200 mesh), each of the crude extracts was put through fractionation at 500g each for methanol, ethyl acetate and hexane extracts. The mobile phase was hexane, hexane/dichloromethane ratio 2:1, hexane/dichloromethane ratio 1:1, hexane/dichloromethane ratio 1:2, hexane/dichloromethane ratio 2:1, hexane/dichloromethane ratio 3:1, hexane/dichloromethane ratio 3:2. Similar fractions were merged after thin layer chromatography (TLC, silica gel GF254, 0.25mm Merck Germany plates) test of each of the fractions. The infrared of fractions was analysed on Buck 500M spectrophotometer with KBr pellets, while Gas Chromatography-Mass Spectroscopy was carried out with Agilent 7890A GC/MS equipped with a Quadrupole Mass Spectra Detector and an Auto-sampler.

Screenhouse experiment

Pasteurized loamy soil was apportioned into experimental pots at 40 kg each. The experimental design had 4 treatments, 5 replicates and 4 dosages of application. Two weeks’ old tomato (cv Roma) seedlings were transplanted into each experimental pot at a seedling per pot. Fresh, newly hatched eggs of M. incognita was inoculated at the base of each seedling (approximately 1000 each) a week after transplanting (Fabiyi et al., 2019).

Field experiment

A piece of land measuring 55m by 40m was laid out, ploughed and harrowed. It was outlined and segmented into 80 beds of 1m by 7m (7.0m2) in size with a passage way of 0.5m in between (Fabiyi and Olatunji, 2021a). On each bed, a spacing of 50 cm between plants and 75 cm between rows was used (Wageningen, 2005; Gudugi et al., 2012). Plantlets of tomato were later transplanted from the nursery to the field. Each tomato plant on the bed was inoculated with approximately 2500 freshly hatched eggs of M. incognita following the method of Fabiyi et al. (2019).

Treatment application and data collection

Carbofuran 3G, was applied on the field at 1.0, 1.5 and 2.0 kg a.i/ha while the chromatographic fractions were applied at 35 mg/kg soil, 41 mg/kg soil and 47 mg/kg soil. In the screenhouse, carbofuran was used as applied on the field, while fractions were applied at 15 mg/kg soil, 21 mg/kg soil and 27 mg/kg soil. Each quantity was dissolved in 300 mL distilled water. 30 mL of a non-ionic surfactant emulsifier was introduced to attain complete solubility and to come up with a uniform solution of the fractions. Data on vine lenght of tomato plants, number of leaves and days to 50% flowering was taken in the screenhouse and field during the growth period of the tomato plants. Numbers of fruits per plant and fruit weight per plant was recorded progressively at maturity, while nematode population in root and soil in the screenhouse was evaluated by removing 250ml soil from the rhizosphere of each tomato plant. On the field, five core samples (0-25 cm deep) were gathered from the base of tomato plants on each bed, these were pulled together to constitute a single sample for each bed. The soil samples were taken to the laboratory and the nematode population was estimated using Whitehead and Hemming (1965) tray method of nematode extraction. Later, roots were graded for severity of galling using a scale of 0-5, as expressed by Taylor and Sasser (1978). Where 0 = no gall; 1 = 1-20% of the root system galled; 2 = 21-40% of the root system galled; 3 = 41-60% of the root system galled; 4 = 61-80% of the root system galled; and 5 = 81-100% of the root system galled. Subsequently, root samples were neatly washed under running tap water and cut into pieces of 2cm each and macerated in a warring blender for 30 seconds. The contents of the blender were emptied on to a two ply tissue paper in a sieve on a pie-pan. All data taken were then subjected to analysis of variance using GenStat 5.32 and separation of means done with Tukey’s honest significant difference test (Fabiyi, 2020).

Results and Discussion

Several functional groups were seen in the result of the infrared analysis of chromatographic fractions. Absorption bands at 3420cm-1, 3211cm-1, 2954cm-1, 2933cm-1, 2855cm-1, 1869cm-1, 1737cm-1, 1707cm-1, 1627cm-1, 1601cm-1, 1450cm-1, 1380cm-1, 1375cm-1, 1274cm-1, 1176cm-1, 1105cm-1, and 1024cm-1 were noted in the spectra of fractions from methanol extract. These depicts the presence of O-H, secondary amines, aliphatic N-H stretch, C-H stretch aldehyde, C-O of carbonyl, C=O acid anhydride, N=O aliphatic nitro, C=C of aromatic ring, C=C alkene, C=C-N stretch, C-H of alkyl group, C-O phenol, C-O acid and a C-H vibration individually. These designates aliphatic chains, aldehydes, amines, amides, acids, esters, ketones and lactones. The ethyl acetate fractions are dominated by vibrations at 3413cm-1, 2928cm-1, 2856cm-1, 1702cm-1, 1646cm-1 and 1456cm-1 which signifies N-H stretch, C-H aliphatic, C-H aldehyde, C=O ketone, C=O amide and alpha CH2 bending singly. From the n-hexane fraction, C-H stretch aliphatic and C-H stretch aldehyde is recurrent. A large number of compounds which ranges from esters, alcohols, terpenes, ketones, phenols, fatty acids, sesquiterpenes, carboxylic acids, terpenes, steroids, aldehydes, hydrocarbons and ethers were detected in the GCMS of the various fractions. Some of these include N-phenyl-1naphtalenamine, 9-beta campesterol, trans-caryophyllene, 3-pentadecylphenol, hexanoic acid, manglupenone, dodecanal, terpinene, 6-methyl-3-heptanol,

 

n-triacontane, 2, 5 dimethyl-4hydroxy-3(2H)-furanone, hexadecane, indicine, 3-octylphenol, mangiferolic acid methyl ester and alpha-thujene (Figure 1).

From the screenhouse, significant differences were not observed between the vine length of tomato plants treated with fractions from methanol extract (MANG/MeOH) and (CBFN) carbofuran (Figures 2, 3). Explains the effect of the quantity of treatments applied on the vine length of tomato plants. Longer vines were recorded in the highest quantity of treatments (27mg/ml), while the lengths of the untreated tomato plants were short. In like manner, observations from the field on vine lengths depicts that vine lengths were also longer in tomato plants

 

 

 

 

 

 

 

 

 

 

treated with CBFN and MANG/MeOH, while there is no notable difference between the two treatments (Figure 10). The highest quantity of treatment materials applied on the field (47mg/ml) equally produced longer vines as opposed to the vine length recorded in untreated tomato plants and those treated with lower quantity of materials (Figure 11). From Figures 4 and 12, numbers of leaves of tomato plants were remarkably more in plants treated with MANG/MeOH and CBFN in the screenhouse and field. Fewer leaves were produced in plants treated with MANG/EtOAc and MANG/Hex (Mangifera indica ethyl acetate fractions and Mangifera indica n-hexane fractions). The effect of quantity of treatment application on numbers of leaves of tomato plants is shown in Figures 5 and 13. More leaves were recorded in treated plants as against untreated tomato plants. The influence of treatment materials on the yield attributes of tomato plants is presented in Figures 6 and 14. In the screenhouse and field trials plants treated with MANG/EtOAc and MANG/Hex flowered lately as contrasted with plants treated with MANG/MeOH and CBFN which flowered earlier. Correspondingly on the field, flowering was late in MANG/EtOAc and MANG/Hex, but early in MANG/MeOH and CBFN. Number of harvested fruits was significantly more in CBFN and MANG/MeOH treated plants in the screenhouse and field trials compared with all other treatments applied (Figures 6 and 14). In the same vein, heavier fruits were recorded in CBFN and MANG/MeOH treated plants in the screenhouse and field rather than the weights obtained in other treatments. The highest dosage of treatment materials in the screenhouse and field had a positive effect on the tomato plants with early flowering. Likewise, significantly more numbers of fruits and heavier fruits were recorded (Figures 7 and 15). Remarkably low soil nematode population was recorded at harvest in treated plants both in the screenhouse and field trials (Figures 8 and 16) in comparison with the population recorded in untreated plants (Figures 9 and 17). Identically, nematode population in 20 g of tomato roots was notably low with a corresponding low gall index (Figures 9 and 17). In general, the influence of M. incognita was more pronounced on the untreated tomato plants. The improvement in vegetative growth of tomato plants administered with fractions from M. indica is accredited to the fragments and integrant component of the various chromatographic fractions that were employed in the treatment of the infected tomato plants in screenhouse and field trials. Chromatographic fractions are acknowledged to be nematicidal (Fabiyi et al., 2012). M. indica is admitted to contain diverse active principles and metabolites which are reported to be nematicidal (Aiyelaagbe and Osamudiamen, 2009; Shah et al., 2010; Zasada, 2010; Joona et al., 2013). Similarly, the bark extract of M. indica is proven to be nematicidal against nematodes parasitizing Ananas comosus (PIP, 2011). The functional groups identified in the fractions through infrared analysis is nearly almost connected with the functional groups of mangiferin a xanthone and isomangiferin which has been described as the vital constituents of M. indica and several other metabolites like polyphenols, flavonoids, triterpenoids, alpha-beta unsaturated ketones, acids, aldehydes, esters and lactones. Stoilova et al. (2005) and Perrucci et al. (2006) stated the anti-parasitic, antifungal and

 

 

 

 

 

 

antibacterial action of mangiferin. Analogously, Guha et al. (1996), Engels et al. (2009, 2010, 2011), Ansari et al. (2000) and Sairam et al. (2003), additionally emphasized the anti-bacterial, anti-diarrhoeal, antimicrobial and anti-HIV of mangiferin and gallotannins from M. indica. The effectiveness of extract from M. indica leaves on malarial parasite was confirmed by Asase et al. (2010). This assertion was corroborated by Bidla et al. (2004), they confirm the action of M. indica extracts on Plasmodium falciparum. In like manner, Malann et al. (2013) gave insight into the effectiveness of the leaf extract on P. berghei, while inhibition of HSV-1 and HSV-2 herpes simplex virus replication was communicated by Zhu et al. (1993) and Zheng and Lu (1990). The stage reliant action of M. indica aqueous extract on animal nematode Trichinella spiralis was accentuated by Garcia et al. (2003) while, Gehad et al. (2013) reported the 100% multiplication inhibition of Strongyloides stercoralis larvae with aqueous extract of unripe M. indica fruit. Several phenolic compounds are documented to be nematicidal. Compounds such as methoxycinnamic acid, protocatechuic acid, juglone, 3-phenylphenol, 7-OH-coumarin, vanillic and syringic acid have been established to have high nematicidal action on M. incognita (Mahanja et al., 1992). Ohri and Kaur (2010) equally reported the potential of polyphenolic compounds in the management of plant parasitic nematodes and this was reiterated by Leontopoulos et al. (2020). The wide range of plant metabolites in M. indica could be harnessed in the management of M. incognita.

Conclusions and Recommendations

Plant bioactive compounds are diverse in the class of phenolic, terpenoid and alkaloids. These could be consolidated in M. incognita management for a safer environment.

Novelty Statement

Metabolites from Mangifera indica could be employed as part of IPM program in the control of Meloidogyne incognita on tomato

Conflict of interest

The authors have declared no conflict of interest.

References

Aiyelaagbe, O.O., and Osamudiamen, P.M., 2009. Phytochemical screening for active compounds in Mangifera. indica leaves from Ibadan, Oyo state. Medwell J., 2(1): 11-13.

Ansari, S.H., Ali, M., Neguruela, V. and Alonaso, M.J.P., 2000. Isolation and characterisation of volatile constituents of M. indica fruits. J. Med. Aromat. Plant Sci., 22(1B): 582-585.

Asasea, A., Akweteya, G.A. and Daniel, G.A., 2010. Ethnopharmacological use of herbal remedies for the treatment of malaria in the Dangme West District of Ghana. J. Ethnopharmacol., 129: 367-376. https://doi.org/10.1016/j.jep.2010.04.001

Atolani, O. and Fabiyi, O.A., 2020. Plant parasitic nematodes management through natural products: Current progress and challenges. In: Management of phytonematodes: Recent advances and future challenges. (eds. Ansari, R.A., Rizvi, R. and Mahmood, I.). pp. 297-315. https://doi.org/10.1007/978-981-15-4087-5_13

Atolani, O., Fabiyi, O.A. and Olatunji, G.A., 2014a. Isovitexin from Kigelia pinnata, a potential eco-friendly nematicidal agent. Trop. Agric., 91(2): 67-74.

Atolani, O., Fabiyi, O.A., and Olatunji, G.A., 2014b. Nematicidal isochromane glycoside from Kigelia pinnata leaves. Acta Agric. Slovenica, 104(1): 25-31. https://doi.org/10.14720/aas.2014.104.1.3

Bidla, G., Titanji, V.P., Jako, B., Bolad, A., and Berzins, K., 2004. Antiplamodial activity of seven plants used in African folk medicine. Indian J. Pharmacol., 36: 245-246.

Das, P.C., Das, A. and Mandal, S., 1989. Anti-inflammatory and anti-microbial activities of the seed kernel of Mangifera indica extract. Phytother. Res. LX. pp. 235-240.

Engels, C., Michael, G.G. and Andreas, S., 2010. Fast LC–MS analysis of gallotannins from mango (Mangifera indica L.) kernels and effects of methanolysis on their antibacterial activity and iron binding capacity. Food Res. Int., 45: 422-426. https://doi.org/10.1016/j.foodres.2011.11.008

Engels, C., Knödler, M., Zhao, Y.Y., Carle, R., Gänzle, M.G. and Schieber, A., 2009. Antimicrobial activity of gallotannins isolated from mango (Mangifera indica L.) kernels. J. Agric. Food Chem., 57: 7712-7718. https://doi.org/10.1021/jf901621m

Engels, C., Schieber, A. and Gänzle, M.G., 2011. Studies on the inhibitory spectrum and mode of antimicrobial action of gallotannins from mango kernels (Mangifera indica L.). Appl. Environ. Microbiol., 77: 2215-2223. https://doi.org/10.1128/AEM.02521-10

Fabiyi, O.A., Baker, M.T., Claudius-Cole, A.O., Alabi, R.O and Olatunji, G.A. , 2022. Application of Cyclic Imides in the Management of Root-Knot Nematode (Meloidogyne incognita) on Cabbage. Indian Phytopathol., 75(2):457-466 https://doi.org/10.1007/s42360-022-00480-1

Fabiyi, O.A., 2020. Growth and yield response of groundnut Arachis hypogaea (Linn.) under Meloidogyne incognita infection to furfural synthesized from agro-cellulosic materials. J. Trop. Agric., 58(2): 241-245.

Fabiyi, O.A. and Olatunji, G.A., 2021a. Environmental sustainability: Bioactivity of Leucaena leucocephala leaves and pesticide residue analysis in tomato fruits. Acta Univ. Agric. Silvic. Mendel. Brun., 69(4): 473-480. https://doi.org/10.11118/actaun.2021.042

Fabiyi, O.A. and Olatunji, G.A., 2021b. Toxicity of derivatized citrulline and extracts of water melon rind (Citrullus lanatus) on root-knot nematode (Meloidogyne incognita). Trop. Agric., 98(4): 347-355.

Fabiyi, O.A., 2018. Management of Meloidogyne incognita infected tomato plants with agricultural wastes. Bull. Inst. Trop. Agric., 41: 15-20.

Fabiyi, O.A., 2021a. Evaluation of nematicidal activity of Terminalia glaucescens fractions against Meloidogyne incognita on Capsicum chinense. J. Hortic. Res., 29(1): 67-74. https://doi.org/10.2478/johr-2021-0006

Fabiyi, O.A., 2021b. Sustainable management of Meloidogyne incognita infecting carrot: Green synthesis of silver nanoparticles with Cnidoscolus aconitifolius: (Daucus carota). Vegetos. 34(2): 277-285. https://doi.org/10.1007/s42535-021-00216-y

Fabiyi, O.A., 2021c. Evaluation of plant materials as root-knot nematode (Meloidogyne incognita) suppressant in Okro (Abelmuscous esculentus). Agric. Consp. Sci., 86(1): 51-56.

Fabiyi, O.A., Baker, M.T., Claudius-Cole, A.O., Alabi, R.O. and Olatunji, G.A., 2022. Application of cyclic imides in the management of root-knot nematode (Meloidogyne incognita) on cabbage. Indian Phytopath., https://doi.org/10.1007/s42360-022-00480-1

Fabiyi, O.A., Olatunji, G.A., and Daodu, I.O., 2019. Nematicidal effect of organic extract metal complex on Meloidogyne incognita infecting groundnuts (Arachis hypogea). Sci. Agric. Bohem., 50(3): 191-196. https://doi.org/10.2478/sab-2019-0026

Fabiyi, O.A., Olatunji, G.A. and Atolani, O., 2012. Nematicidal activities of chromatographic fraction from Alstonia boonei and Bridelia ferruginea on Meloidogyne incognita. Pak. J. Nematol., 30(2): 189-198. https://doi.org/10.1016/S2221-1691(12)60347-5

Fabiyi, O.A., Saliu, O.D., Claudius-Cole, A.O., Olaniyi, I.O., Oguntebi, O.V. and Olatunji, G.A., 2020. Porous starch citrate biopolymer for controlled release of carbofuran in the management of root knot nematode Meloidogyne incognita. Biotechnol. Rep., 25(e00428): https://doi.org/10.1016/j.btre.2020.e00428

Fawusi, M.O.A., 1978. Emergence and seedling growth of pepper as influenced by soil compaction nutrient status and moisture regime. Soc. Hortic., 9: 329-335. https://doi.org/10.1016/0304-4238(78)90042-0

Food and Agricultural Organization (FAO), 2004. Food and Agricultural Organization (FAO) 2004. FAO Produc­tion Year Book 2003. FAO, Rome, Italy, pp. 526.

Garcia, D., Escalante, M., Delgado, R., Ubeira, F.M. and Leiro, J., 2003. Antihelmintic and anti-allergic activities of Mangifera indica L. stem bark components vimang and mangiferin. Phytother. Res., 17: 1203-1208. https://doi.org/10.1002/ptr.1343

Gehad, T., El, S. and Samir, M.O., 2013. Anthelmintic activity of unripe Mangifera indica L. (Mango) against Strongyloides stercoralis. Int. J. Curr. Microbiol. App. Sci., 2(5): 401-409.

Goad, R.T., Goad, J.T., Atieh, B.H. and Gupta, R.C., 2004. Carbofuran-induced endocrine disruption in adult male rats. Toxicol. Mech. Methods, 14(4): 233-239. https://doi.org/10.1080/15376520490434476

Gudugi, I.A.S., Odofin, A.J., Adeboye, M.K.A. and Oladiran, J.A., 2012. Agronomic characteristics of tomato as influenced by irrigation and mulching. Adv. Appl. Sci. Res., 3(5): 2539–2543.

Guha, S., Ghosal, S. and Chattopadhyay, U., 1996. Antitumor, immunomodulatory and anti-HIV effect of mangiferin, a naturally occurring glucosylxanthone. Chemotherapy, 42: 443-451. https://doi.org/10.1159/000239478

Joona, K., Sowmia, C., Dhanya, K.P., and Divya, M.J., 2013. Preliminary phytochemical investigation of Mangifera indica leaves and screening of antioxidant and anticancer activity. Res. J. Pharm. Biol. Chem. Sci., 4(1): 1112-1118.

Karen, C., and R.D. 2007. Benefits of eating tomatoes remain bountiful: Lycopene may not reduce cancer risk, but it’s still part of a healthy diet http://www.msnbc.msn.com/id/19979 174/ns/health.

Khan, M.N., Nizami, S.S., Khan, M.A. and Ahmed, Z., 1993. New saponins from Mangifera indica. J. Nat. Prod., 56: 767-770. https://doi.org/10.1021/np50095a016

Lau, T.K., Chu, W. and Graham, N., 2007. Degradation of the endocrine disruptor carbofuran by UV,03 and 03/UV water science and technology: J. Int. Assoc. Water Poll. Res., 55(12): 275-280. https://doi.org/10.2166/wst.2007.416

Leontopoulos, S., Skenderidis, P., and Vagelas, I.K., 2020. Potential use of polyphenolic compounds obtained from olive mill waste waters on plant pathogens and plant parasitic nematodes. In: Mérillon, JM., Ramawat, K.G. (eds) Plant Defence: Biological Control. Progress in Biological Control, vol 22. Springer, Cham., https://doi.org/10.1007/978-3-030-51034-3_6

Mahajan, R., Kaur, D.J. and Bajaj, K.L., 1992. Nematicidal activity of phenolic compounds against Meloidogyne incognita. Nematol. Mediterr., 20: 217-219.

Malann, Y.D., Matur, B.D. and Mailafia, S., 2013. Evaluation of the antimalarial activity of aqueous leaf extracts of Casuarina equistifolia and Mangifera indica against Plasmodium berghei in mice. J. Pharm. Bioresour., 10(1): https://doi.org/10.4314/jpb.v10i1.2

Muanza, D.N., Kim, B.W., Euler, K.L. and Williams, L., 1994. Antibacterial and antifungal activities in nine medicinal plants from Zaire. Int. J. Pharmacogn., 32: 337-345. https://doi.org/10.3109/13880209409083012

Ohri, P. and Kaur, S., 2010. Effect of phenolic compounds on nematodes. A review. J. Appl. Nat. Sci., 2(2): 344-350. https://doi.org/10.31018/jans.v2i2.144

Perrucci, S., Fichi, G., Buggiani, C., Rossi, G. and Flamini, G., 2006. Efficacy of mangiferin against Cryptosporidium parvum in a neonatal mouse model. Parasitol. Res., 99: 184-188. https://doi.org/10.1007/s00436-006-0165-4

Pino, J.A., Mesa, J., Yamilie, M.M., Pilar, M., and Rolando, M., 2005. Volatile components from mango (Mangifera indica L.) cultivars. J. Agric. Food Chem., pp. 53. https://doi.org/10.1021/jf0402633

PIP-Guide to good crop protection practices 2011.www.coleacp.org/pip.15.

Raymond, G.R. and Sarker, S.D., 2006. Isolation of natural products by low pressure column chromatography. In natural products isolation, 2nd Ed. Humana Press, New Jersey. pp. 77-116.

Rich, J.R., Dunn, R.A. and Noling, J.W., 2004. Nematicides: Past and present use. In: Chen, Z.X., Chen, S.Y. and Dickson, D.W. (Eds). Nematology, advances and perspectives. Wallingford, UK, CAB International, pp. 1179-1200. https://doi.org/10.1079/9780851996462.1179

Ross, I.A., 1999. Medicinal plants of the world, chemical constituents, traditional and modern medicinal uses. Humana Press, Totowa, pp. 197-205.

Safdar, A. Anwar, and McKenry, M.V., 2012. Incidence and population density of plant parasitic nematodes infecting vegetable crops and associated yield losses in Punjab, Pakistan. Pak. J. Zool., 44(2): 327-333.

Sairam, K., Hemalatha, S., Ashok, K., Srinivasan, T., Jai, G., Shankar, M. and Venkataraman, S., 2003. Evaluation of anti-diarrhoeal activity in seed extracts of Mangifera indica. J. Ethnopharmacol., 84: 11-15. https://doi.org/10.1016/S0378-8741(02)00250-7

Shah, K.A., Patel, M.B., Patel, R.J., and Parmar, P.K., 2010. Mangifera indica (Mango). Pharmacogn. Rev., 4(7): 42-48. https://doi.org/10.4103/0973-7847.65325

Shakeel, Q., Javed, N., Iftikhar, Y., Haq, I.U., Khan, S.A. and Ullah, Z., 2012. Association of plant parasitic nematodes with four vegetable crops. Pak. J. Phytopathol., 24(2): 143-148.

Stoilova, I., Gargova, S., Stoyanova, A. and Ho, L., 2005. Antimicrobial and antioxidant activity of the polyphenol mangiferin. Herbal Polonica, 51: 37-44.

Taylor, A.L. and Sasser, J.N., 1978. Biology, identification and control of root-knot nematodes Meloidogyne spp. North Carolina University USAID. pp. 111.

Tona, L., Kambu, K., and Ngimbi, N., 2000. Anti-amoebic and spasmolytic activities of extract from some antidiarrhoeal traditional preparations used in Kinshasa Congo. Phytomedicina, 7: 31-38. https://doi.org/10.1016/S0944-7113(00)80019-7

Wageningen, 2005. Cultivation of tomato. Agromisa foundation and CTA, ISBN Agromisa: 90-8573-039-2, ISBN CTA: 92-9081-299-0

Whitehead, A.G. and Hemning, J.R. 1965. A Comparison of Some Quantitative Methods of Extracting Small Vermiform Nematodes from Soil. Ann. Appl. Biol., 55(1): 2538.

Yudelman, M.A., Ratta, and Nygaard, D., 1998. Pest management and food production: Looking to the future. Food, agriculture and the environment. Discussion paper 25, International Food Policy Research Institute, USA. 50.

Zasada, I.A., Walters, T.W. and Pinkerton, J.N., 2010. Post plant nematicides for the control of root lesion nematode in raspberry. Hortic. Technol., 20(5): 856-862

Zheng, M.S. and Lu, Z.Y., 1990. Antiviral effect of mangiferin and isomangiferin on herpes simplex virus. Chinese Med. J., 103: 160-165.

Zhu, X.M., Song, J.X. and Huang, Z.Z., 1993. Antiviral activity of mangiferin against herpes simplex virus type 2 in vitro. Zhongguo Yaoli Xuebao 14: 452-454