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Anti nematode effect assessment of peganum harmala based-products against meloidogyne javanica on melon

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Anti nematode effect assessment of peganum harmala based-products against meloidogyne javanica on melon

  1. 1. Journal of Biology, Agriculture and HealthcareISSN 2224-3208 (Paper) ISSN 2225Vol.3, No.5, 2013Anti-nematode Effect Assessment ofBased-Products AgainstMayad El Hassan1. Laboratory of Plant Biotechnology, Faculty of Sciences, B.P. 28/S, Agadir, Morocco.2. Department of Plant Protection, Nematology. Hassan II Agriculture and veterinary Institute,* E-mail of the corresponding author:AbstractNatural products from plant origin have been tested for their potential to develop viable components of plantparasitic nematode management strategies especially againstharmala seed evaluated in pot experimentswhen they are used as soil amendmentdamage to melon crops, but also improves significantly growth parameters except for the emulsified oil ofharmala which proved a phytotoxic effect although it gave a complete suppression ofextract and the seeds powder are among the less phytotoxic substances, which are efficient against these pathogens.harmala-seed-based-products have a potential antinematodes effect againstcomponent of an integrated management systeKeywords: nematodes management,1. IntroductionMelon (Cucumis melo L.) is a very sensitive crop to rootof transplants and initial population density of nematodes in the soil, the threshold of tolerance may be locatedbetween 0.48 and 3.53 J2/100g soil (Ploeg 2001). The damage caused byconsiderable (Sasser 1979 Di Vitodepending on the level of infestation (Ploeg and Phillips, 2001). To manage these plant parasite, producers do nothave efficient means though control by chemicaloften effective, they are very expensive and sometimes in reexamination or not registered on melon crop. Foreconomic and environmental reasons, development of new management tools that respectbecome a necessity nowadays. The application of substances from plant origin is a promising option in cropsprotection against plant parasitic nematodes (Chitwood, 2002; KokaliPeganum harmala is a reputed medicinal plant for its high toxicity against several microorganisms and animals(Bellakhdar, 1997; Idrissi Hassani, 2000 and Mahmoudian, 2002). Substances from its different organs have variousbiological activities: antispasmodic, antimitotic,Al-Shamma, 1981; Al Yahya, 1986; Sijilmassi, 1996; Bellakhdar, 1997; Jbilou et al., 2006;et al., 2010). In most cases, the biological activity observed was attributedal. 1978; Duke, 1985 and Ayoub et al., 1994). This study aims to evaluate different products based onseed. Dried powder, emulsified oil, aqueous, methanolic and hexanic extract for their nématotoxicthe most damaging root knot nematodesprotection and its impact on this crop growth.2. Material and Methods2.1 Botanicals products preparationThe hexanic and methanolic extracts ofsoxhlet apparatus and then evaporated on a rotatory evaporator under vacuum at a temperature of 45°C to a smallvolume. The residual syrup was used for the preparation of different concentrations used for bioassays. The residueobtained by hexane has been dissolved in distilled water using Tween 20 was used at 2% while the dry methanolextract was dissolved in distilled water using ultrasound.Journal of Biology, Agriculture and Healthcare(Paper) ISSN 2225-093X (Online)5nematode Effect Assessment of Peganum harmalaProducts Against Meloidogyne javanicaMayad El Hassan1,2*Ferji Zahra2Idrissi Hassani Lalla Minaof Plant Biotechnology, Faculty of Sciences, B.P. 28/S, Agadir, Morocco.Department of Plant Protection, Nematology. Hassan II Agriculture and veterinary Institute,Morocco.mail of the corresponding author: elhassan.mayad@gmail.comNatural products from plant origin have been tested for their potential to develop viable components of plantparasitic nematode management strategies especially against Meloidogyne javanica. Different producin pot experiments against M. javanica associated to Cucumiswhen they are used as soil amendment in pre-planting, not only reduces densities of M. javanicadamage to melon crops, but also improves significantly growth parameters except for the emulsified oil ofwhich proved a phytotoxic effect although it gave a complete suppression ofs powder are among the less phytotoxic substances, which are efficient against these pathogens.products have a potential antinematodes effect against Meloidogyne javanicacomponent of an integrated management system to control this root knot nematode.management, Peganum harmala; extracts, organic amendment, Cucumis meloL.) is a very sensitive crop to root-knot nematodes (Meloidogyne spp.). Depending on the ageof transplants and initial population density of nematodes in the soil, the threshold of tolerance may be locatedbetween 0.48 and 3.53 J2/100g soil (Ploeg 2001). The damage caused by Meloidogyne(Sasser 1979 Di Vito et al., 1983; Netscher and Sikora, 1990). The yield losses can reach 65%depending on the level of infestation (Ploeg and Phillips, 2001). To manage these plant parasite, producers do nothave efficient means though control by chemicals is the most common. Chemical nematicides, although they areoften effective, they are very expensive and sometimes in reexamination or not registered on melon crop. Foreconomic and environmental reasons, development of new management tools that respectbecome a necessity nowadays. The application of substances from plant origin is a promising option in cropsprotection against plant parasitic nematodes (Chitwood, 2002; Kokali-Burelle and Rodrigueza reputed medicinal plant for its high toxicity against several microorganisms and animals(Bellakhdar, 1997; Idrissi Hassani, 2000 and Mahmoudian, 2002). Substances from its different organs have variousbiological activities: antispasmodic, antimitotic, antibacterial, insecticidal and anthelmintic (Ross et al., 1980;Shamma, 1981; Al Yahya, 1986; Sijilmassi, 1996; Bellakhdar, 1997; Jbilou et al., 2006;). In most cases, the biological activity observed was attributed to the alkaloids (Ross et al. 1980; Gupta. 1978; Duke, 1985 and Ayoub et al., 1994). This study aims to evaluate different products based onseed. Dried powder, emulsified oil, aqueous, methanolic and hexanic extract for their nématotoxicthe most damaging root knot nematodes Meloidogyne javanica for melon (Cucumis melo vars.protection and its impact on this crop growth.2.1 Botanicals products preparationThe hexanic and methanolic extracts of Peganum harmala seeds have been prepared using a continous extraction bysoxhlet apparatus and then evaporated on a rotatory evaporator under vacuum at a temperature of 45°C to a smallsed for the preparation of different concentrations used for bioassays. The residueobtained by hexane has been dissolved in distilled water using Tween 20 was used at 2% while the dry methanolextract was dissolved in distilled water using ultrasound. The solids residue have been dissolved and stored at +4°Cwww.iiste.orgPeganum harmalaon MelonIdrissi Hassani Lalla Mina1of Plant Biotechnology, Faculty of Sciences, B.P. 28/S, Agadir, Morocco.Department of Plant Protection, Nematology. Hassan II Agriculture and veterinary Institute, Agadir,elhassan.mayad@gmail.comNatural products from plant origin have been tested for their potential to develop viable components of plantDifferent products based on P.Cucumis melo L. crop showed thatM. javanica in soil and cropdamage to melon crops, but also improves significantly growth parameters except for the emulsified oil of P.which proved a phytotoxic effect although it gave a complete suppression of M. javanica. The aqueouss powder are among the less phytotoxic substances, which are efficient against these pathogens. P.javanica and can be used asCucumis melospp.). Depending on the ageof transplants and initial population density of nematodes in the soil, the threshold of tolerance may be locatedMeloidogyne on melon crop are1983; Netscher and Sikora, 1990). The yield losses can reach 65%depending on the level of infestation (Ploeg and Phillips, 2001). To manage these plant parasite, producers do nots is the most common. Chemical nematicides, although they areoften effective, they are very expensive and sometimes in reexamination or not registered on melon crop. Foreconomic and environmental reasons, development of new management tools that respect the agro-ecosystem hasbecome a necessity nowadays. The application of substances from plant origin is a promising option in cropsBurelle and Rodriguez-Kabana, 2006).a reputed medicinal plant for its high toxicity against several microorganisms and animals(Bellakhdar, 1997; Idrissi Hassani, 2000 and Mahmoudian, 2002). Substances from its different organs have variousantibacterial, insecticidal and anthelmintic (Ross et al., 1980;Shamma, 1981; Al Yahya, 1986; Sijilmassi, 1996; Bellakhdar, 1997; Jbilou et al., 2006; Arshad, 2008 and Edzirito the alkaloids (Ross et al. 1980; Gupta et. 1978; Duke, 1985 and Ayoub et al., 1994). This study aims to evaluate different products based on P. harmalaseed. Dried powder, emulsified oil, aqueous, methanolic and hexanic extract for their nématotoxic potential againstCucumis melo vars. Leonardo) cropseeds have been prepared using a continous extraction bysoxhlet apparatus and then evaporated on a rotatory evaporator under vacuum at a temperature of 45°C to a smallsed for the preparation of different concentrations used for bioassays. The residueobtained by hexane has been dissolved in distilled water using Tween 20 was used at 2% while the dry methanolsolids residue have been dissolved and stored at +4°C
  2. 2. Journal of Biology, Agriculture and HealthcareISSN 2224-3208 (Paper) ISSN 2225Vol.3, No.5, 2013in a refrigerator.The aqueous extract of P. harmala was performed by macerating plant powder distilled water under ultrasonicationfor 10 min and then filtered (Whatman No. 1) after shaking for aboutAqueous solution has been performed by macerating 250 g of seed powder in 2 liters of distilled water. Afterexposing the mixture to ultrasonication for 10 min and agitation for 24 hours, the mixture was centrifugerpm for 20 min and the supernatant was filtered. The resulted solution was stored until use at 4°C.The oil of P. harmala was obtained by cold pressing extraction of seed. This oil was used to formulate an oilemulsion as "oil in water" 2% and st2.2 Preparation of inoculum and inoculationInfested Melon Roots with Meloidogyne javanicaroots were cut and ground in a blenderfrom the gelatinous mass. The mixturemicrons). The refusal of the 40µm sievecellulose tissue and placed in a basincellulose. After 48hours, the suspensionunder a microscope (x40).The inoculation was performed by a suspension of second stage juveniles ofsurface in a circle around the center of jars with a pipette having an initial density of 80 J2/100 g soilafter inoculation, the emulsified oil, hexane, methanolic and aqueousapplied by simply spraying with a corresponding volum to field capacity in three concentration 2%, 0.2% and 0.02%(w/w). The seeds powder was applied by mixing with the soil. Three controls were considered for this study. Apositive control consists in inoculated pot, which received no further treatment. The negative control was representedby only sterilized soil and an adjacentapplied in the same volume as the extracts. The treatments were performed in four replicates and arranged in acompletely randomized design. Observations are made three months after ton M. javanica population and melon plants growth. Watering was performed according to the needs of the crop bykeeping the soil moist.The collected data were statistically analyzed using the statistical software "SPSS 11.5". Treatments means weretested using the Newman and Keuls test (P3. ResultsSeedlings transplanted in soil treated with the seed oil offirst weeks of the experiment. This increased phytotoxicity was expressed by a very slow growth and yellowingleaves. The amendment of soil by different products ofof M. javanica and the gall index (p <0,05). The rate of reduction compared to control increases significantly withincreasing product concentration. The aqueous extract and powder applied at 2% and oil used at 0.2 and 2% are mostactive substance from P. harmala seeds on soil population ofwith the negative control. With the exception of the aqueous extract, the emulsified oil and methanol extract at 0.02%,all other treatments show a significant difference with the positive control (Table 1). The emulsified oil is the mosteffective product against M. javanicaactivity and has not show any phytototoxicity effemulsified oil. The hexane extract remained the least effective3.1 Impacts of treatments on root knot nematodes populationWith the exception of the hexane extract, the majority of the testmelon. Powder, aqueous extracts and oil seeds ofmelo attacks against M. javanica. The number and the index of galls on the roots of pthese extracts is very low compared to the control treatment. The 2% oil completely inhibited the development ofgalls on roots and is the only product that does not reveal any significant difference with the negative contrground material, the aqueous extract and methanolic 2% have a relatively lower inhibition potential than the oil(Table 2)3.2 Impact of treatments on crop growthThe vegetative growth (stem length and dry weight of root) increases very significanJournal of Biology, Agriculture and Healthcare(Paper) ISSN 2225-093X (Online)6was performed by macerating plant powder distilled water under ultrasonicationfor 10 min and then filtered (Whatman No. 1) after shaking for about 24 hours in the dark and stored at 4°C.Aqueous solution has been performed by macerating 250 g of seed powder in 2 liters of distilled water. Afterexposing the mixture to ultrasonication for 10 min and agitation for 24 hours, the mixture was centrifugerpm for 20 min and the supernatant was filtered. The resulted solution was stored until use at 4°C.was obtained by cold pressing extraction of seed. This oil was used to formulate an oiloil in water" 2% and stabilized by Tween 20 used at 0,2%.2.2 Preparation of inoculum and inoculationMeloidogyne javanica showing typical galls were washed thoroughlyin a blender in 1% of sodium hypochlorite (NaOCl) solution formixture is filtered through a column of sieves (meshes ranging fromsieve containing eggs of Meloidogyne spp. is depositedbasin filled with water. Neonates (second stage juvenilesuspension containing the J2 is recovered and the larvaes were collectedThe inoculation was performed by a suspension of second stage juveniles of Meloidogynesurface in a circle around the center of jars with a pipette having an initial density of 80 J2/100 g soilafter inoculation, the emulsified oil, hexane, methanolic and aqueous- extract based seed powder ofapplied by simply spraying with a corresponding volum to field capacity in three concentration 2%, 0.2% and 0.02%eds powder was applied by mixing with the soil. Three controls were considered for this study. Apositive control consists in inoculated pot, which received no further treatment. The negative control was representedby only sterilized soil and an adjacent control where the pots were inoculated and treated with Tween 20 used 2%applied in the same volume as the extracts. The treatments were performed in four replicates and arranged in aObservations are made three months after transplantation topopulation and melon plants growth. Watering was performed according to the needs of the crop byThe collected data were statistically analyzed using the statistical software "SPSS 11.5". Treatments means weretested using the Newman and Keuls test (P ≤ 5%).Seedlings transplanted in soil treated with the seed oil of P. harmala at 2% showed phytotoxicity symptoms since thefirst weeks of the experiment. This increased phytotoxicity was expressed by a very slow growth and yellowingleaves. The amendment of soil by different products of Peganum harmala seeds, reduced significantlyand the gall index (p <0,05). The rate of reduction compared to control increases significantly withincreasing product concentration. The aqueous extract and powder applied at 2% and oil used at 0.2 and 2% are mostseeds on soil population of M. javanica and do not show significant differenceswith the negative control. With the exception of the aqueous extract, the emulsified oil and methanol extract at 0.02%,ificant difference with the positive control (Table 1). The emulsified oil is the mostM. javanica. However, the aqueous extract is the product that had an interesting potential ofactivity and has not show any phytototoxicity effect on melon crop. This constitutes an advantage compared to theemulsified oil. The hexane extract remained the least effective.3.1 Impacts of treatments on root knot nematodes populationWith the exception of the hexane extract, the majority of the tested products significantly reduce galling on root ofmelon. Powder, aqueous extracts and oil seeds of P. harmala applied at 2% allow better protection of plant roots of. The number and the index of galls on the roots of plants grown on soil treated withthese extracts is very low compared to the control treatment. The 2% oil completely inhibited the development ofgalls on roots and is the only product that does not reveal any significant difference with the negative contrground material, the aqueous extract and methanolic 2% have a relatively lower inhibition potential than the oil3.2 Impact of treatments on crop growthThe vegetative growth (stem length and dry weight of root) increases very significantly when the soil is amended bywww.iiste.orgwas performed by macerating plant powder distilled water under ultrasonication24 hours in the dark and stored at 4°C.Aqueous solution has been performed by macerating 250 g of seed powder in 2 liters of distilled water. Afterexposing the mixture to ultrasonication for 10 min and agitation for 24 hours, the mixture was centrifuged at 4500rpm for 20 min and the supernatant was filtered. The resulted solution was stored until use at 4°C.was obtained by cold pressing extraction of seed. This oil was used to formulate an oilthoroughly under tap water. Then,for 2 min to release the eggsranging from 250 microns to 40deposited on a screen covered with ajuvenile J2) migrate through theis recovered and the larvaes were collected and countedMeloidogyne spp. by soaking the soilsurface in a circle around the center of jars with a pipette having an initial density of 80 J2/100 g soil. Three daysextract based seed powder of P. harmala wereapplied by simply spraying with a corresponding volum to field capacity in three concentration 2%, 0.2% and 0.02%eds powder was applied by mixing with the soil. Three controls were considered for this study. Apositive control consists in inoculated pot, which received no further treatment. The negative control was representedcontrol where the pots were inoculated and treated with Tween 20 used 2%applied in the same volume as the extracts. The treatments were performed in four replicates and arranged in aransplantation to assess treatments effectpopulation and melon plants growth. Watering was performed according to the needs of the crop byThe collected data were statistically analyzed using the statistical software "SPSS 11.5". Treatments means wereat 2% showed phytotoxicity symptoms since thefirst weeks of the experiment. This increased phytotoxicity was expressed by a very slow growth and yellowingseeds, reduced significantly the populationand the gall index (p <0,05). The rate of reduction compared to control increases significantly withincreasing product concentration. The aqueous extract and powder applied at 2% and oil used at 0.2 and 2% are mostand do not show significant differenceswith the negative control. With the exception of the aqueous extract, the emulsified oil and methanol extract at 0.02%,ificant difference with the positive control (Table 1). The emulsified oil is the most. However, the aqueous extract is the product that had an interesting potential ofect on melon crop. This constitutes an advantage compared to theed products significantly reduce galling on root ofapplied at 2% allow better protection of plant roots of C.lants grown on soil treated withthese extracts is very low compared to the control treatment. The 2% oil completely inhibited the development ofgalls on roots and is the only product that does not reveal any significant difference with the negative control. Theground material, the aqueous extract and methanolic 2% have a relatively lower inhibition potential than the oiltly when the soil is amended by
  3. 3. Journal of Biology, Agriculture and HealthcareISSN 2224-3208 (Paper) ISSN 2225Vol.3, No.5, 2013the homogenate of P. harmala or treated with aqueous and methanolic extract particularly at the dose 2% except wherethe soil is treated with hexane extract and emulsified oil. This treatment causes a strong inhibition of veas result of a pronounced phytotoxicitywas applied at 0.2 and 2% as rate of amendment;methanolic extracts at 2% (Table 3).4. DiscussionMost products made from seeds of P.javanica associated to the melon cropseeds aqueous extract and emulsified oilprovided a complete suppression ofinduced a pronounced phytotoxicityprobably an increase of separating periodthe transplantation date and the moment ofbetween the treated soil with 2% of Tween 20density of Meloidogyne javanica in soilinduced by emulsified oil from P. harmalaaqueous extract, which showed asignificantly reduced and vegetative growthrelease antibiotic components via two major processes: (1) root exudation and (2) release of components during plantdecomposition after incorporation in the soil. Themany studies and was related to the abundanharmine, harmaline, harmalol, harmolcolleagues reported that harmaline andmicroorganisms and these two moleculesto meet the microbial invasion (Baisroot-knot nematodes may also be attributed toare nitrogen-containing molecules andfrom seeds increases the rate of nitrogenand improves crop growth and vigorof nematodes with organic amendmentAmmonium derived from microbialnematodes and fungi for a short period2002). The mode of biocidal actionof the cell membrane (Rush and Lydaammonia against the concentration gradientnitrogen is the necessary amount for1991). Oka (2010) reported that thethe soil environment to reduce the amount5. ConclusionIt is possible to conclude that powder, oil, aqueous and methanolic extract based fromantinematode effect against the population ofimprove its growth. To enhance the performance of these botanicals for nematode control, further investigation isneeded to determine how P. harmalamode of action, identify the active compound and appropriate conditions and methods to use as a component in amultifaceted approach in a sustainable management of agro ecosystem.ReferencesAL Yahya M.A. (1986). Phytochemical studies of the plant used in traditionnel medcine of Saudi Arabia. Fitoterapia.57, (3), 179-182.Al-Shamma, A., Drake S., Flynn, D.L., Mitscher, L.A., Park, YH, Rao, GSR., Simpson, A., Wayze, J.K., Veysoglu, T.Journal of Biology, Agriculture and Healthcare(Paper) ISSN 2225-093X (Online)7or treated with aqueous and methanolic extract particularly at the dose 2% except wherethe soil is treated with hexane extract and emulsified oil. This treatment causes a strong inhibition of vephytotoxicity when the dose increases. Improved growth was recordedas rate of amendment; as well as in the case when the soil.P. harmala at the high concentration (2%) reduce thecrop and limit galling on roots significantly. At the highest usedemulsified oil of P. harmala are the most effective on M. javanicaroot knot nematodes and total protection of roots. Unfortunatelyphytotoxicity on melon plants expressed by partial inhibitionperiod (needed for the bio-decomposition to release the toxicthe moment of product application would overcome this problemof Tween 20 and the positive control indicates that it has noin soil or growth of melon plants. On the other hand, the nematicidalharmala seeds could be attributed to the oil alone. Concerningsimilar effect, the population of M. javanica in soiland vegetative growth was significantly improved. Various plants are known to produce andrelease antibiotic components via two major processes: (1) root exudation and (2) release of components during plantition after incorporation in the soil. The antimicrobial activity of P. harmalathe abundance of the alkaloids in this organ. These alkaloidsrmol and Harman and quinazolines as vascine andand harmine are endowed with antimicrobial activity againstmolecules are formed and released from the plant OxalisBais et al., 2003). Therefore, the effectiveness of P. harmalaattributed to the direct action of these alkaloids on J2.and very abundant in P. harmala seeds. Probably, soil amendmentnitrogen availability to crop, which could cause a reductioncrop growth and vigor. Mian and Rodriguez-Kabana (1982) reported that the potential fororganic amendment is directly dependent on the nitrogen content andmicrobial decomposition of the amendments play an important roleperiod (days to weeks) after application (Oka et al., 1993of ammonia is not yet clear, but several mechanismsLyda, 1982), depletion of the chemical energy of theconcentration gradient (Britto et al., 2001). The limit of using organic amefor an effective control because it is frequently phytotoxinematicidal activity of this type of amendment can be improvedamount needed for a control practice.It is possible to conclude that powder, oil, aqueous and methanolic extract based from P. harmalaantinematode effect against the population of Meloidogyne javanica on melon crop, can limit their damage andimprove its growth. To enhance the performance of these botanicals for nematode control, further investigation isP. harmala seed products act on nematodes (lethal, nematostatic, repellent or attractant) theirmode of action, identify the active compound and appropriate conditions and methods to use as a component in amultifaceted approach in a sustainable management of agro ecosystem.hytochemical studies of the plant used in traditionnel medcine of Saudi Arabia. Fitoterapia.Shamma, A., Drake S., Flynn, D.L., Mitscher, L.A., Park, YH, Rao, GSR., Simpson, A., Wayze, J.K., Veysoglu, T.www.iiste.orgor treated with aqueous and methanolic extract particularly at the dose 2% except wherethe soil is treated with hexane extract and emulsified oil. This treatment causes a strong inhibition of vegetative growthrecorded when seed powderis treated with aqueous orpopulation of Meloidogynehighest used concentration,javanica. The emulsified oilUnfortunately this product hasof vegetative growth. Butto release the toxic products) betweenproblem. The lack of differencehas no effect on the populationhand, the nematicidal effectsConcerning the powder and thesoil and galling have beenVarious plants are known to produce andrelease antibiotic components via two major processes: (1) root exudation and (2) release of components during plantseeds was demonstrated inalkaloids include β-carboline as:and vasicinone. Bais and hisagainst terrestrial pathogenictuberosa in the rhizosphereP. harmala on the population of. In addition, these alkaloidssoil amendment by powdersreduction in nematode populationthat the potential for managementand inversely related to C/N.an important role in control of1993; Tenuta and Lazarovits,may be involved: alterationcells carrying the cytosolicusing organic amendment that releasephytotoxic or not economic (Stirling,be improved by manipulatingP. harmala seeds have a potentialon melon crop, can limit their damage andimprove its growth. To enhance the performance of these botanicals for nematode control, further investigation istostatic, repellent or attractant) theirmode of action, identify the active compound and appropriate conditions and methods to use as a component in ahytochemical studies of the plant used in traditionnel medcine of Saudi Arabia. Fitoterapia.Shamma, A., Drake S., Flynn, D.L., Mitscher, L.A., Park, YH, Rao, GSR., Simpson, A., Wayze, J.K., Veysoglu, T.
  4. 4. Journal of Biology, Agriculture and HealthcareISSN 2224-3208 (Paper) ISSN 2225Vol.3, No.5, 2013& Wu, STS. (1981). Antimicrobial agentd from higher plants. Antimicrobial agents fromNat. prod. 44 (6) p:745-747.Arshad, N., Zitterl-Eglseer, K., Hasnain, S. & Hess, M. (2008). Effect ofalkaloids on certain antibiotic resistant strains of bacteria and protozoa from poultry. Phytother. Res. 22, 1533Ayoub, MT., AL-Allaf, TAK. & Rshan, LJ. (1994). Antiprofilative activity of harmalol. Fitotérapia, 65, (1), 14Bais, HP., Vepachedu, R. & Vivanco, JM. (2003). Root spin transformed root cultures of OxalisBellakhdar, J. (1997). La pharmacopée marocaine traditionnelle.Britto, DT., Siddiqi, MY., Glass, ADM. & Kronzucker, HJ. (2001). Futile transmembrane NH4+ cycling: a cellularhypothesis to explain ammonium toxicity in plants. Proc. Natl. Acad. Sci. U.S.A. 98, 4255Chitwood, DJ. (2002). Phytochemical based strateDi Vito, M., Greco, N. & Carelu, A. (1983). The effect of population densities ofof cantaloupe and tobacco. Nematol. medit.,Duke, AJ. (1985). Handbook of medicinale herbs. Editions CRC Press Inc., Florida, 676p.Edziri, H., Mastouri, M., Matieu, M., Zine, M., Gutman, L. & Aouni M, (2010). Biological activities ofharmala leaves. African J. of Biotechnology Vol. 9(48), pp. 8199Gupta OP., Anand KK., Ghatak, BJR. & Atal, CK. (1978). Vasicine, alkloid ofuteroteonic abortifacient. Indian J. Of Experimental Biology, 16, 1075Idrissi Hassani LM. (2000). Analyse phytochimique de lharmeleffets sur le criquet pèlerin Schistocerca gregariaAgadir, 214pJbilou, R., Ennabili, A. & Sayah, F. (2006). Insecticidal activity of four medicinal plant extrcastaneum (Herbst) (Coleoptera: Tenebrionidae). African J. of Biotechnology Vol. 5 (10), pp. 936Kokalis-Burelle, N., & Rodriguezplant-parasitic nematodes. Allelochemicals: Biological Control of Plant Pathogens and Diseases. Eds. Springer, pp.15-29Mahmoudian, M., Jalilpour, H., Salehian, P. 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Root specific elicitation and exudation of fluorescent bOxalis tuberosa. Plant Physiol Biochem 41:345–353La pharmacopée marocaine traditionnelle. Ibis Press. 770p.Britto, DT., Siddiqi, MY., Glass, ADM. & Kronzucker, HJ. (2001). Futile transmembrane NH4+ cycling: a cellularhypothesis to explain ammonium toxicity in plants. Proc. Natl. Acad. Sci. U.S.A. 98, 4255Chitwood, DJ. (2002). Phytochemical based strategies for nematode control. Annu. Rev. Phytopathol. 40: 221Di Vito, M., Greco, N. & Carelu, A. (1983). The effect of population densities of Meloidogyne incognitaNematol. medit., II: 169-174.Handbook of medicinale herbs. Editions CRC Press Inc., Florida, 676p.Edziri, H., Mastouri, M., Matieu, M., Zine, M., Gutman, L. & Aouni M, (2010). Biological activities ofleaves. African J. of Biotechnology Vol. 9(48), pp. 8199-8205.a OP., Anand KK., Ghatak, BJR. & Atal, CK. (1978). Vasicine, alkloid of Adhatoda vasica,Indian J. Of Experimental Biology, 16, 1075-1077.Idrissi Hassani LM. (2000). 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Mechanisms of nematode suppression by organic soil amendments- A review. Appl. Soil Ecol. 44:(1993). Control of the root-knot nematode Meloidogyne javanica1262001). Damage to melon (Cucumis melo L.) cv. Durango bySouthern California. J. Nematology, Vol. 3, N. 2 , : 151- 157.Ratnadass, A., Fernandes, P., Avelino, J., Habib, R. (2012). Plant species diversity for sustainable management ofand diseases in agroecosystems: a review. Agron. Sustain. Dev. 32:273–303Ross, SA., Megalla, SE., Bishay, DW., Awad, AH. (1980). Studies for determining antibiotic substances in someegyptian plants. Part II. Antimicrobial alkaloids from the seed of Peganum harmala L. Fitoterapia, 51, 309Rush, CM. & Lyda, SD. (1982). The effects of anhydrous ammonia on mycelium and sclerotia of. Phytopathology. 72: 1085-1089Sasser, JN. & Kirby, MF. (1979). Crop cultivars resistant to root-knot nematodes, Meloidogyneof Plant Pathology, North Carolina State University, Raleigh, NCSijilmassi, A. (1996). 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  5. 5. Journal of Biology, Agriculture and HealthcareISSN 2224-3208 (Paper) ISSN 2225Vol.3, No.5, 2013Table 1. Effect of different products fromTreatmentsDryed powderAquous extractHexane extractEmulsified oilMethanolic extractPositif controlTween 20 (0,2%)* Numbers followed by the same letter in the same column are not significantly different according to Newman andKeuls test (P ≤ 5%).Table 2. Effect of different products fromCucumus melo vars. LeonardoTreatmentsDryed powderAquous extractHexane extractEmulsified oilMethanolic extractPositif controlTween 20 (0,2%)* Numbers followed by the same letter inKeuls test (P ≤ 5%).Journal of Biology, Agriculture and Healthcare(Paper) ISSN 2225-093X (Online)9products from P. harmala seeds on the Meloidogyne javanica populationConcentration (%) final Densit0,02 29,25b,c,d*0,20 34,00b,c,d2,00 17,00a,b,c0,02 38,25d,e0,20 23,25b,c,d2,00 17,00a,b,c0,02 32,75 b,c,d0,20 32,25 b,c,d2,00 21,25b,c,d0,02 36,67c,d,e0,20 15,00a,b2,00 0,00a0,02 40,50d,e0,20 24,75b,c,d2,00 29,50b,c,d52,25e63,50e* Numbers followed by the same letter in the same column are not significantly different according to Newman andTable 2. Effect of different products from P. harmala seeds on root galling induced byConcentration (%) Galls number0,02 14,75c,d*0,20 15,67c,d2,00 01,50a0,02 29,25e,f0,20 11,50b,c2,00 04,75a,b0,02 29,25e,f0,20 19,75c,d,e2,00 18,00c,d,e0,02 22,33c,d,e0,20 23,00e2,00 0,00a0,02 26,25e0,20 24,75e2,00 13,00b,c37,00f37,25fletter in the same column are not significantly differentwww.iiste.orgpopulation density in soilDensity J2/100 cm329,25b,c,d*34,00b,c,d23,25b,c,db,c,d32,25 b,c,d21,25b,c,d24,75b,c,d29,50b,c,d* Numbers followed by the same letter in the same column are not significantly different according to Newman andseeds on root galling induced by Meloidogyne javanica onGall index02,75c,d03,00c,d,e01,00b03,50c,d,e02,50c01,75b03,75d,e03,00c,d,e03,00c,d,e03,00c,d,e03,33c,d,e0,00a03,25c,d,e03,25c,d,e02,33b04,00e04,00edifferent according to Newman and
  6. 6. Journal of Biology, Agriculture and HealthcareISSN 2224-3208 (Paper) ISSN 2225Vol.3, No.5, 2013Table 3: Effect of different products fromTreatments ConcentrationDryed powder0,020,202,00Aquous extract0,020,202,00Hexane extract0,020,202,00Emulsified oil0,020,202,00Methanolic extract0,020,202,00Negatif controlPositif controlTween 20 (0,2%)* Numbers followed by the same letter inKeuls test (P ≤ 5%).Journal of Biology, Agriculture and Healthcare(Paper) ISSN 2225-093X (Online)10products from P. harmala seeds on growth parameters of Cucumus meloConcentration (%)Stem dry weight(g)Stem length (cm)4,971b,c,d* 41,50b,c,d6,429a,b 53,67a,b7,936a 62,00a5,599b,c,d 51,00a,b,c5,031b,c,d 44,00a,b,c,d7,637a 61,25a5,352b,c,d 48,75a,b,c,d5,343b,c,d 48,67a,b,c,d4,941b,c,d 45,00a,b,c,d,3,586d 32,66d3,769c,d 34,33c,d1,949e 17,75e5,243b,c,d 47,75a,b,c,d5,737b,c,d 52,25a,b6,631a,b 58,25a,b5,929b,c 54,00a,b4,314c,d 52,25b4,045c,d 53,01bletter in the same column are not significantly differentwww.iiste.orgCucumus melo vars. Leonardo.Stem length (cm)Root dryweight (g)00,26e,f00,33d,e00,71a00,50b,c00,48b,c,d00,69a00,49b,c,d00,49b,c,d00,45b,c,d00,33d,e00,34d,e00,18f00,48b,c,d00,52b,c,00,58b00,38c,d,e00,41c,d,e00,40c,d,edifferent according to Newman and
  7. 7. This academic article was published by The International Institute for Science,Technology and Education (IISTE). The IISTE is a pioneer in the Open AccessPublishing service based in the U.S. and Europe. The aim of the institute isAccelerating Global Knowledge Sharing.More information about the publisher can be found in the IISTE’s homepage:http://www.iiste.orgCALL FOR PAPERSThe IISTE is currently hosting more than 30 peer-reviewed academic journals andcollaborating with academic institutions around the world. There’s no deadline forsubmission. Prospective authors of IISTE journals can find the submissioninstruction on the following page: http://www.iiste.org/Journals/The IISTE editorial team promises to the review and publish all the qualifiedsubmissions in a fast manner. All the journals articles are available online to thereaders all over the world without financial, legal, or technical barriers other thanthose inseparable from gaining access to the internet itself. Printed version of thejournals is also available upon request of readers and authors.IISTE Knowledge Sharing PartnersEBSCO, Index Copernicus, Ulrichs Periodicals Directory, JournalTOCS, PKP OpenArchives Harvester, Bielefeld Academic Search Engine, ElektronischeZeitschriftenbibliothek EZB, Open J-Gate, OCLC WorldCat, Universe DigtialLibrary , NewJour, Google Scholar

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