Nematicidal activity of gallic acid purified from Terminalia nigrovenulosa bark against the root-knot nematode Meloidogyne incognita

in Nematology
Restricted Access
Get Access to Full Text
Rent on DeepDyve

Have an Access Token?

Enter your access token to activate and access content online.

Please login and go to your personal user account to enter your access token.


Have Institutional Access?

Access content through your institution. Any other coaching guidance?


The nematicidal activity of Terminalia nigrovenulosa bark (TNB) and its purified compound were assayed against Meloidogyne incognita in vitro. The nematicidal compound was isolated from TNB using silica gel column and Sephadex LH-20 chromatography combined with thin-layer chromatography and high performance liquid chromatography. Structural identification of the nematicidal compound was conducted using 1H-nuclear magnetic resonance (NMR), 13C-NMR and liquid chromatography-tandem mass spectrometry. We found that the nematicidal compound purified from TNB was gallic acid (GA) or 3,4,5-trihydroxy benzoic acid. Nematicidal activity bioassays revealed that GA treatment resulted in 20.3, 37.5, 73.3, 88.3 and 95.8% hatch inhibition at 0, 0.25, 0.5, 1.0 and 2.0 mg ml−1 after 3 days, respectively, of incubation. Eggshells appeared to be deformed and destroyed at 2 and 3 days after incubation with a GA concentration of 1.0 mg ml−1, respectively. Additionally, after treatment with a GA concentration of 1.0 mg ml−1, mortality of second-stage juveniles of M. incognita was 65.0, 75.0, 96.7 and 100% at 3, 6, 9 and 12 h incubation, respectively.

Nematicidal activity of gallic acid purified from Terminalia nigrovenulosa bark against the root-knot nematode Meloidogyne incognita

in Nematology



AkhtarM.MahmoodI. (1994). Potentiality of phytochemicals in nematode control: a review. Bioresource Technology 48189-201.

AkhtarY.YeoungR.IsmanM.B. (2008). Comparative bioactivity of selected extracts from Meliaceae and some commercial botanical insecticides against two noctuid caterpillars, Trichoplusia ni and Pseudaletia unipuncta. Phytochemistry Reviews 777-88.

BachayaH.A.IqbalZ.KhanM.N.JabbarA.GilaniA.H.Islam-Ud-Din (2009). In vitro and in vivo anthelmintic activity of Terminalia arjuna bark. International Journal of Agriculture and Biology 11273-278.

BarbosaL.C.A.BarcelosF.F.DemunerA.J.SantosM.A. (1999). Chemical constituents from Mucuna aterrima with activity against Meloidogyne incognita and Heterodera glycines. Nematropica 2981-88.

BarkerK.R. (1985). Nematode extraction and bioassays. In: BarkerK.R.CarterC.C.SasserJ.N. (Eds). An advanced treatise on MeloidogyneVol. 2. Raleigh, NC, USANorth Carolina State University Press pp.  19-35.

BirdA.F.SelfP.G. (1995). Chitin in Meloidogyne javanica. Fundamental and Applied Nematology 18235-239.

BrowningM.WallaceD.B.DawsonC.AlmS.R.AmadorJ.A. (2006). Potential of butyric acid for control of soil-borne fungal pathogens and nematodes affecting strawberries. Soil Biology & Biochemistry 38401-404.

CayrolJ.-C.DjianC.PijarowskiL. (1989). Study on the nematicidal properties of the culture filtrate of the nematophagus fungus Paecilomyces lilacinus. Revue de Nématologie 12331-336.

ChanwitheesukA.TeerawutgulragA.KilburnJ.D.RakariyathamN. (2007). Antimicrobial gallic acid from Caesalpinia mimosoides Lamk. Food Chemistry 1001044-1048.

ChitwoodD.J. (2002). Phytochemical based strategies for nematode control. Annual Review of Phytopathology 40221-249.

ChoH.J.HanS.C. (1986). Survey of plant parasitic nematodes on economic crops. Korean Journal of Plant Protection 25175-182.

EldahshanO.A. (2011). Isolation and structure elucidation of phenolic compounds of carob leaves grown in Egypt. Current Research Journal of Biological Sciences 352-55.

FabryW.OkemoP.O.AnsorgR. (1998). Antibacterial activity of East African medicinal plants. Journal of Ethnopharmacology 6079-84.

FanelliE.Di VitoM.JonesJ.T.De GiorgiC. (2005). Analysis of chitin synthase function in a plant parasitic nematode, Meloidogyne artiellia, using RNAi. Gene 34987-95.

FernándezC.Rodríguez KábanaR.WarriorP.KloepperJ.W. (2001). Induced soil suppressiveness to a root-knot nematode species by a nematicide. Biological Control 22103-114.

GommersF.J. (1981). Biochemical interactions between nematodes and plants and the relevance to control: a review. Helminthological Abstracts 509-24.

HarishS.SaravanakumarD.RadjacommareR.EbenezarE.G.SeetharamanK. (2008). Use of plant extracts and biocontrol agents for the management of brown spot disease in rice. BioControl 53555-567.

HarrisM.T.FuhrmanJ.A. (2002). Structure and expression of chitin synthase in the parasitic nematode Dirofilaria immitis. Molecular and Biochemical Parasitology 122231-234.

IsmanM.B. (2000). Plant essential oils for pests and diseases management. Crop Protection 19603-608.

IsmanM.B. (2006). Botanical insecticides, deterrents, and repellents in modern agriculture and an increasingly regulated world. Annual Review of Entomology 5145-66.

JavedN.GowenS.R.Inam Ul HaqM.AbdullahK.ShahinaF. (2006). Systemic and persistent effect of neem (Azadirachta indica) formulations against root-knot nematodes, Meloidogyne javanica and their storage life. Crop Protection 26911-916.

KhanZ.KimY.H. (2007). A review on the role of predatory soil nematodes in the biological control of plant parasitic nematodes. Applied Soil Ecology 35370-379.

KhanZ.KimY.H.KimS.G.KimH.W. (2007). Observations on the suppression of root-knot nematode (Meloidogyne arenaria) on tomato by incorporation of cyanobacterial powder (Oscillatoria chlorina) into potting field soil. Bioresource Technology 9869-73.

KroesB.H.van den BergA.J.J.Quarles van UffordH.C.van DijkH.LabadieR.P. (1992). Anti-inflammatory activity of gallic acid. Planta Medica 58499-504.

KwonT.Y.JungK.C.ParkS.D.SimY.G.ChoiB.S. (1998). Cultural and chemical control of root-knot nematodes, Meloidogyne sp. on oriental melon in plastic film house. RDA Journal of Crop Protection 4096-101.

LuZ.NieG.BeltonP.S.TangH.ZhaoB. (2006). Structure-activity relationship analysis of antioxidant ability and neuroprotective effect of gallic acid derivatives. Neurochemistry International 48263-274.

MeyerS.L.F.ZasadaI.A.RobertsD.P.VinyardB.T.LakshmanD.K.LeeJ.K.ChitwoodD.J.CartaL. (2006). Plantago lanceolata and Plantago rugelii extracts are toxic to Meloidogyne incognita but not to certain microbes. Journal of Nematology 38333-338.

OkaY. (2000). Nematicidal activity of essential oils and their components against the root-knot nematode. Nematology 90710-715.

OkaY.KoltaiH.Bar EyalM.MorM.SharonE.ChetI.SpiegelY. (2000). New strategies for the control of plant-parasitic nematodes. Pest Management Science 56983-988.

PandeyR.KalraA.TandonS.MehrotraN.SinghH.N.KumarS. (2000). Essential oils as potent sources of nematicidal compounds. Journal of Phytopathology 148501-502.

RangsriwongP.RangkadilokN.SatayavivadJ.GotoM.ShotiprukA. (2009). Subcritical water extraction of polyphenolic compounds from Terminalia chebula Retz. fruits. Separation and Purification Technology 6651-56.

ShahrzadS.AoyagiK.WinterA.KoyamaA.BitschI. (2001). Pharmacokinetics of gallic acid and its relative bioavailability from tea in healthy humans. Journal of Nutrition 1311207-1210.

SharmaR.N.SaxenaK.N. (1974). Orientation and developmental inhibition in the housefly by certain terpenoids. Journal of Medical Entomology 11617-621.

SiddiquiZ.A.IqbalA.MahmoodI. (2001). Effects of Pseudomonas fluorescens and fertilizers on the reproduction of Meloidogyne incognita and growth of tomato. Applied Soil Ecology 16179-185.

SoutheyJ.F. (1986). Laboratory methods for work with plant and soil nematodes6th edition. Reference Book 402London, UKMinistry of Agriculture, Fisheries and Food.

SultanaN.AkhterM.KhatoonZ. (2010). Nematicidal natural products from the aerial parts of Rubus niveus. Natural Product Research 24407-415.

UjvaryI. (2001). Pest control agents from natural products handbook of pesticide toxicology2nd edition. San Diego, USAAcademic Press.

VeronicoP.GrayL.J.JonesJ.T.BazzicalupoP.ArbucciS.CorteseM.R.Di VitoM.De GiorgiC. (2001). Nematode chitin synthases: gene structure, expression and function in Caenorhabditis elegans and the plant parasitic nematode Meloidogyne artiellia. Molecular Genetics and Genomics 26628-34.

WalkerJ.T.MelinJ.B. (1996). Mentha × piperita, Mentha spicata and effects of their essential oils on Meloidogyne in soil. Journal of Nematology 28629-635.

WangW.Ben DanielB.H.CohenY. (2004). Control of plant diseases by extracts of Inula viscosa. Phytopathology 941042-1047.

WhartonD. (1980). Nematode eggshells. Parasitology 81447-463.

WhartonD.A. (2002). Nematode survival strategies. In: LeeD.L. (Ed.). The biology of nematodes. New York, NY, USATaylor & Francis pp.  389-411.

WHO/WPRO (1990). Medicinal plants in Vietnam. Hanoi, VietnamInstitute of Materia444 pp.

WilliamsonV.M.GleasonC.A. (2003). Plant-nematode interactions. Current Opinion in Plant Biology 6327-333.

WilliamsonV.M.KumarA. (2006). Nematode resistance in plants: the battle underground. Trends in Genetics 22396-403.

WinkM. (1993). Production and application of phytochemicals from an agricultural perspective. In: van BeekT.A.BretelerH. (Eds). Phytochemistry and agricultureVol. 34. Oxford, UKClarendon Press pp.  171-213.

WiratnoTaniwiryonocD.van den BergH.RiksenJ.A.G.RietjensI.M.C.M.DjiwantiS.R.KammengaJ.E.MurkA.J. (2009). Nematicidal activity of plant extracts against the root-knot nematode, Meloidogyne incognita. The Open Natural Products Journal 277-85.

ZasadaI.A.KlassenW.MeyerS.L.F.CodalloM.Abdul BakiA.A. (2006). Velvetbean (Mucuna pruriens) extracts: impact on Meloidogyne incognita survival and on Lycopersicon esculentum and Lactuca sativa germination and growth. Pest Management Science 621122-1127.

Zia-Ul-HaqM.AhmadM.AkhterM. (2010). Nematicidal activity of selected flora of Pakistan (2010). Pakistan Journal of Botany 422119-2123.


  • View in gallery

    Schematic representation of the extraction and isolation of gallic acid from Terminalia nigrovenulosa bark.

  • View in gallery

    High performance liquid chromatography (HPLC) of gallic acid purified from Terminalia nigrovenulosa bark. Retention time was 14.6 min.

  • View in gallery

    A: 1H-NMR; B: 13C-NMR spectra of gallic acid purified from Terminalia nigrovenulosa bark. 1H- and 13C-spectra were measured in CD3OD at 500 and 125 MHz, respectively.

  • View in gallery

    Liquid chromatography-tandem mass spectroscopy (LC-MS/MS) spectrum of gallic acid (GA) purified from Terminalia nigrovenulosa bark.

  • View in gallery

    Juvenile mortality after treatment with A: the crude extract; B: ethyl acetate fraction; or C: gallic acid purified from Terminalia nigrovenulosa bark at various concentrations (0, 0.25, 0.5, 1.0 and 2.0 mg ml−1) after 0, 3, 6, 9 and 12 h incubations. Values are mean ± standard deviation.

  • View in gallery

    Effect of purified gallic acid (GA) (1.0 mg ml−1) on the morphology of Meloidogyne incognita eggs. Meloidogyne incognita egg shapes in the water control under light microscopy (A0, 1, 2, 3) and fluorescence microscopy (B0, 1, 2, 3) at days 0, 1, 2 and 3. On the second day in the water control, M. incognita egg developed to the first-stage juvenile (A2). On day 3 in the water control, the second-stage juvenile appeared (A3). M. incognita egg shapes in the GA incubation under light microscopy (C0, 1, 2, 3) and fluorescence microscopy (D0, 1, 2, 3) at days 0, 1, 2 and 3. On day 1 after GA incubation, development ceased soon after deposition of GA (C1). On day 2 after GA incubation, a deformed eggshell was observed (arrow) (C2) and on day 3 after GA incubation, some eggshells were totally destroyed (circle) (C3) under light microscopy. The GA-treated egg at days 0, 1, 2 and 3 (D0, 1, 2, 3) under fluorescence microscopy after staining with 0.01% Fluorescence Brightener 28. This figure is published in colour in the online edition of this journal, which can be accessed via

  • View in gallery

    Light microscopy images of second-stage Meloidogyne incognita juveniles after treatment with gallic acid purified from Terminalia nigrovenulosa bark. A: Untreated; B-D: Treated with 1 mg ml−1 gallic acid for 12 h. Characteristic shapes of dead nematodes: straight (I-shape) (B); bent (banana-shape) (C); sigmoid (Σ-shape) (D). Scale bar is 50 μm. This figure is published in colour in the online edition of this journal, which can be accessed via


Content Metrics

Content Metrics

All Time Past Year Past 30 Days
Abstract Views 33 33 6
Full Text Views 48 48 36
PDF Downloads 3 3 1
EPUB Downloads 0 0 0