AGRICULTURE AND BIOLOGY JOURNAL OF ISSN Print: 2151-7517, ISSN Online: 2151-7525, doi:10.5251/abjna.2014.5.5.193.197 © 2014, ScienceHuβ, http://www.scihub.org/ABJNA Evaluation of Nematophagous fungi Dactylaria brochopaga and dactyloides against Meloidogyne incognita infesting peanut plants under field conditions

Noweer E.M.A.1 and Mona E.M. Al-Shalaby2

1- Nematology Laboratory, plant pathology Department, National Research Center, Dokki, Giza, Egypt. 2- Department of Zoology and Agricutlural Nematology, Faculty of Agriculture, Cairo University, Giza, Egypt. ABSTRACT The nematicidal activity of the -trapping fungi Dactylaria brochopaga and Arthrobotrys dactyloides in their compounds named (Dbx-1003) 20% and (Adx-1004) 20% respictevely to control Meloidogyne incognita infesting peanut plant Arachis hypogaea L. CV. Giza 5. were evaluated under field conditions in Egypt. Each compound were added to the soil in two forms once as a liquid using two concentrations 20,40 liter/feddan after germination. And the other treatments were added to the soil as granules around the peanut seed plants at planting time at rates of 100, 200 kg/feddan. A treatment without any addition was used as check treatment. Data indicated two months after application that population densities of M. incognita in soil were significantly reduced in all treatments compared with control, as well as gall formation on peanut roots. The highest reduction in the nematode counts in soil and root galls was recorded with the application of D. brochopaga. Four months after treatments at the end of growing season the percentages reduction were increased espicaialy with using granules of the two fungus. Peanut yield was significantly increased due to the addition of the two fungus to the soil, especially when applied at higher dosage levels.

Key Words: Nematode-trapping fungi - Dactylaria brochopaga - Arthrobotrys dactyloides - Meloidogyne incognita – Peanut - Field conditions.

INTRODUCTION Monacrosporium bembicode were tested by Duponnois et al., (1997) for their trapping ability Hertz (1992) reported that conidia of Arthrobotrys against Meloidogyne mayaguensis. Most of the oligospora germinated directly into adhesive traps when Arthrobotrys strains and one Dactylaria strain applied close to cow faeces on water agar plates. The decreased development of the nematode in tobacco conidial trap is considered a survival structure enabling plants. They found that the growth of the tobacco the fungus to overcome fungistasis. Traps adhere to the plants was consequently improved this refer to that surface of passing , could facilitate spread of certain fungi have proper phytostimulant effects by the fungus, before penetrating the nematode cuticle and acting on the soil structure through commensalism immobilization of the nematode take place. The mechanisms with the plant roots. nematophagous A. oligospora which can live saprophytically as well as predatorily, forms sticky The fungus Dactylaria brochopaga proved to be an reticulate traps in the presence of living nematodes effective nematode antagonist and this may be due to kill and consume them (Scholler and Rubner, to the effect on nematodes population larvae through 1994). Timper and Brodie,(1993) found that production of traps which captured the nematode Arthrobotrys dactyloides, A. oligospora, Monacrosporium larvae, dissolving its outer culticle and then digesting ellipsosporim and M. cionopagum, fungi killed most of the inner contents of the victim (Aboul-Eid et al., the Pratylenchus penetrans adults and juveniles added 2002). to the fungus cultures. Arthrobotrys oligospora, A. Conoides, Dactylaria Shelensis, Dactylaria sp. and Agric. Biol. J. N. Am., 2014, 5(5): 193-197

Recently this fungus has been developed and decanting techniques. Root samples were incubated formulated in a compound named, Dbx-1003 (20%). in tap water for 7 days for egg mass hatching. The Therefore, the objective of this study was to evaluate number of galls on roots was also recorded. The the effects of both nematode-trapping fungi, reduction % of nematode in soil was calculated Arthrobotrys dactyloides and Dactylaria brochopaga in according to Handerson & Tilton formula (Puntener, their compounds named Adx-1004 20% and Dbx -1003 1981). Number of root galls per 5 gm. roots were 20% respectively against the root-knot nematodes M. also determined. At the end of season plants were incognita infecting peanut cv. Giza 5 under field removed and data on plant growth (weights of fruits, conditions in Egypt. shoots,and roots) of peanut were recorded.

MATERIAL AND METHODS Statistical analysis: Data were analyzed following standard procedures for analysis of variance. This work was carried out in sandy soil with natural Differences between means were evaluated for infestation of root-knot nematode Meloidogyne significance according to Duncan's New Multiple incognita through the period of May to September Range Test at the 5% and 1% level of probability 2009 in Mansouria village, Giza Governorate, Egypt. using MSTAT Version 4 (1987).

The area of the experiment was 1. kirate(175m)for each RESULTS: treatment, the seed of peanut Arachis hypogaea L.CV. Giza 5. were planted in the soil in May 2009. The Effects of the nematode trapping fungi on following treatments were applied to the soil as follows: M.incognita larvae in soil: Numbers of root-knot Dbx-1003 (20%) as a liquid containing Dactylaria nematodes M.incognita larvae before treatment in brochopaga fungi and ADX-1004 (20%) containing May 2009, after tow month in july 2009 at Mid Arthrobotrys dactyloides fungi were applied once to the Season and at end of the growing season in soil at the rate of 1 litre/kirate (20 litre/feddan) and at the September 2009 were recorded in Table 1. Data rate of 2 litre/kirate (40 litre/feddan) these treatments showed that the expression of nematicidal activity were added to the soil around the peanut plants at varied with type of application, concentration and previous two concentrations. The first concentration time of exposure. Two months after treatments in prepared by diluted 1 litre from two liquid component in July 2009 (Mid season) population densities of M. 49 litre of water and the second concentration was incognita in soil were significantly reduced by all prepared by diluted 2 litre from two component in 48 treatments compared with untreated control. As the litre of water, then two concentrations added to the soil reduction percents recorded were 63% and 83% at the rate 25 ml/plant after completely germination with the application of D. brochopaga (Dbx- compontent Dbx-1003 (20%) and ADX-1004 (20%) as 1003)liquid and were 76% and 74% with Dbx-1003 agranules. The spores of two fungus was carried on granules. These percents increased by the end of the vermoculite substrate. Treatments were applied once to growing season where they reached 85% , 93% for the soil at the rate 5kg/kirate (100 kg/feddan) and at 10 liquid form and 92%, 93% for granules form. As for the kg/kirate (200 kg/feddan) and mixed to the soil around fungus A. dactyloides (ADX-1004) the reduction the plants after 15 days from planting. percents recorded in Mid season were 65% and 74% by 1&2 liter concentration respectively for liquid form Untreated plants as check treatments. Each and 79% and 77% by the 5&10 Kg concentration treatment was replicated ten times. All treatments respectively for granules form. On the other hand in the were completely randomized. Also chemical end of the growing season the reduction percents were fertilizers and all other culture ractices were made 77% and 89% by the liquid form 1&2 liter concentration according to the technical recommendations of respectively and 90% and 94% by the granules form peanut growing. Irrigation was executed. Soil 5&10 Kg concentration respectively . samples were taken at three intervals; before adding the tested materials May 2009 (the initial population),Mid season (after two months, July 2009) and the end of growing season (after four months, september 2009). Also root samples were taken in mid season and the end of the growing season. Soil samples were extracted by using sieving and

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Table (1): Effects of D. brochopaga (Dbx-1003) 1 Liter 114 49.2 98 63.2 and A. dactyloides (ADX-1004), a nematode 2 Liter 165 26.5 87 67.3 5 k.g. 88 60.8 62 76.7 trapping fungus on M. incognita Larvae in soil. 10 k.g. 24 80.3 45 83.1 Check 224.5 - 266 - No. of Juveniles/1 kg soil L.S.D. End of the 0.05 45 - 42 - Mid season Treatments Initial growing season 0.01 65 - 67 - population No. of No. of Red. % Red. % No. of galls in treatment J2 J2 Red % = 1- X 100 DBx-1003 No. of galls in check 1 liter 1256 810 63 422 85 2 liter 2120 625 83 316 93 5 k.g. 1930 816 76 344 92 Effects of nematode trapping fungi on peanut 10 k.g. 1410 624 74 218 93 ADX-1004 yield: Besides reducing nematode infestation, the 1 Liter. 2105 1200 65 890 77 biocontrol agents also enhanced the plant growth 2 Liter. 3120 1315 74 624 89 parameters (Table 3). It is evident from the table that 5 k.g. 2875 965 79 535 90 all treatments improved the Shoot and Root fresh 10 k.g. 1870 670 77 214 94 Check 1659 2725 - 3065 - weight of the plant, weight of peanut pods per plant L.S.D. and peanut yield per feddan. 0.05 - 412 - 514 - 0.01 - 624 - 710 - As shwon in Table 3 application of granules form fa two fungi more effective than that of liquid form. No. in treatment after No. in check Reduction % = application before X X 1- No. in treatment No- in 100 Table (3): Effects of D. brochopaga (Dbx-1003) and A. before application check after dactyloides (ADX-1004) on peanut fruits production infested with root-knot nematode. Mean of plant weight Yield per Effects of nematode trapping fungi on (Shoot- Treatments Pods/plant feddan M.incognita the root galls number: All treatments Root)/plant (gm) (kg) significantly reduced M. incognita gall formation (gm) compared with the untreated control. Numbers of DBx-1003 1 Liter 128 43 1290 root galls in 5 gm root were differed according to the 2 Liter 162 52 1560 type of treatments (Table 2). At the mid season, the 5 k.g. 158 52 1560 percentages reductions recorded were 67.9, 80.4% 10 k.g. 206 54 1620 with the application of the two concentrations of Dbx- ADX-1004 1 Liter 134 42 1260 1003 granules while the two concentrations of the 2 Liter 178 48 1440 liquid form which induceed only 56.3 and 66.1% 5 k.g. 182 52 1560 reduction. these percentages increased at the end of 10 k.g. 214 56 1680 the growing season to reache 79.7, 88% with Check 96 32 960 L.S.D. granules form and 72.9, 76.7% with liquid form. Also, 0.05 32 8 465 obtained with the fungus application of A. dactyloides 0.01 46 12 630 (Adx-1004) form. The percentages reductions were 76.7% and 83.1% granules while that of the liquid DISCUSSION form were 63.2 and 67.3% at the end of growing season. Nematophagous fungi are an interesting ecological group of fungi because they can be used to control

Table (2): Effects of D. brochopaga (Dbx-1003) and A. phytoparasitic nematodes. The species from the dactyloides (ADX-1004) on M. incognita root galls. Arthrobotrys, Dactylaria genera are predators and No. of galls/5gm roots create organs of capture (Adhesive Knobs or nets, Mid season Final season Treatments constricting and non-constricting rings). No. of No. of Red % Red % galls galls DBx-1003 These fungi release chemo-attractants which bring 1 Liter 98 56.3 72 72.9 nematodes close to the fungal mycelia where they 2 Liter 76 66.1 62 76.7 are immobilized in special trapping organs such as 5 k.g. 72 67.9 54 79.7 10 k.g. 44 89.4 32 88 sticky pads or constricting/non constricting rings. One ADX-1004 of such fungi is Arthrobotrys, in which researchers

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have identified two pathogenicity factors. a granules containing the fungus are applied around carbohydrate binding protein (Lectin) and an the roots, they may be active enough to substantially extracellular serine protease (Ahman et al., 2002). reduce the number of root-knot nematodes entering Proteases have been found to be involved in the roots during the first few weeks of growth immobilization of nematodes captured by A. (Stirling, et al., 1998). oligosora. The adhesion of fungal structure, penetration and immobilization of nematodes take REFERENCES not more than one hour as suggestes by Tunlid and Jansson (1991). Aboul-Eid, H.Z., M. Fatma Hammad and E.M.A. Noweer, (2002). Effects of the nematode biocide Dbx-1003 in In the present study, the effectiveness of the controlling citrus nematode infecting lemon, and compounds Dbx-1003 20% containing the nematode interrelationship with the co-inhabitant fungi. Egyptian Journal of Agronematology 6, 1-13. trapping fungus, Dactylaria brochopaga and Adx- 1004 20% containing the nematode trapping fungus Aboul-Eid H.Z., Susan A. Hasabo and E.M.A. Noweer Arthrobotrys dactyloides were evaluated against root- (2006). Effect of a nematode trapping fungus knot nematode Meloidogyne incognita infesting Dactylaria brochopaga on Meloidogyne incognita peanut plant under field conditions. Data clearly infesting olives and coconut palms in Egypt. showed that Dbx-1003 containing nematophagus, D. Intrnational Journal of Nematology 16(1) 56-69. brochopaga suppressed M. incognita larvae in soil Ahman, J., T. Johansson, M.Olsson, P.J. Punt, C. Hondel and root galls formation. These results are in and A. Tunlid (2002). Improving the pathogenicity of a agreement with those obtained by Aboul-Eid et al. nematode trapping fungus by genetic engineering of a (2006). Generally, the percentage nematode subtilisin with nematotoxic activity. Appl. Environ. reduction increased as the rate and peroid after Microbiol. 68(7): 3408-3415. application increased. Deleij, F.A.A.M., K.G. Davies and B.R. Kerr (1992). The use of Verticillium chlamydosporium Goddard and Also ADX-1004 containing nematophagus, Arthrobotrys Pasteuria penetrans (Thorne) Sayre & Starr alone and dactyloides suppressed the root-knot nematode. The in combination to control Meloidogyne incognita on reduction percents to the juveniles in mid season tomato plants. Fundamental and Applied Nematology 15, 235-242. were 74% with liquid form and 79% with granules form at the high dosge these percents increased at Duncan, D.B. (1955). Multiple Range and Multiple F. Tests. final season to reach 89% and 94%. Similar results Biometrics 11, 1-42. were obtained with the fungus A. dactyloides has Duponnois, R., T. Mateille and A. Ba (1997). Potential shown high potential as a biological control agent effects of Sahelian nematophagous fungi against against M. javanica in soil microcosms (Stirling and Meloidogyne mayaguensis on tobacco, Annales due Mani, 1995). Tabac setion-2, No. 29, 61-70. Hertz, B.N. and C. Dackman (1992). Conidial traps-a new Predation of nematodes by nematophagous fungi is survival structure of the nematode trapping fungus presumably confined to those J2 exposed on the root Arthrobotrys oligospora, Mycological Research 96, surface or in the soil. Therefore, for Dbx-1003 to be 194-198. effective at the initial invasion period during which the Kerry, B.R. (1989). Fungi as biological control agents for J2 are active in the soil, need to be increased (De Leij plant parasitic nematodes. In Biotechnology of fungi et al., 1992). This is presumably to enable the fungus for improving plant growth (Eds. Whipps, J.M. and R.D to capture J2 before they invade the roots. Lumsden).

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