Microbial control of root-knot nematode which infested Eggplant (Solanum melongena) by using the nematode-trapping fungus Dactylaria brochopaga & Verticilium chlamydosporium as a combined with yeast, molasses and vermiculate (F1+F2AVYM) is reported under field condition study. The results revealed that the highest percentage reduction in number of nematode larvae per 1kg soil was achieved when applying the fungi D. brochopaga & V. chlamydosporium together combined with yeast, molasses and vermiculate. Also the highest percentage reduction in number of root-galls per plant (93.1 %) was achieved by using the fungi D. brochopaga & V. chlamydosporium together combined with yeast, molasses and vermiculate. The data revealed that the weight of fruits per plant were significantly (P≤0.05 and/or P≤ 0.01) increased in all nematode-trapping fungus D. brochopaga treatments compared to the untreated check treatment
Keywords: Microbial control; Dactylaria brochopaga; Verticilium chlamydosporium; Meloidogyne incognita; Yeast; Molasses; Vermiculite; Eggplant
Most research on microbial agents that attack nematodes in soil has concerned fungi, especially those that form traps to ensnare their prey. The nematode-trapping fungi; the first nematode- trapping fungus, the ubiquitous Arthrobotrys oligospora, was described by Fersenius, [1] Zopf [2] observed that the fungus could capture motile nematodes and parasitize them. Mai and Chen [3] used the nematode trapping fungi (N.T.F.) Arthrobotrys superba, A. dactyloides, A. arthrobotryoides, and Dactylella doedycoides to reduce the penetration of alfaalfa roots by Pratylenchus penetrans in sterilized soil. Niblack and Hussey [4] evaluated (N.T.F.), A. amerospora, combined in 3 commercial preparations with Rhizobium japanicum inoculums for controlling of Heterodera glycines on soybean (Glycine max) in the field and greenhouse. In a greenhouse experiment conducted to evaluate the effect of pigeon droppings and A. oligospora as a soil amendment on the root- knot nematodes Meloidogyne incognita on Muskmelon, Ali [5] indicated that there was a trend towards nematode population diminution with greater efficiency when the fungus has been added soil 2 weeks before to nematode inoculation and planting. Inoculums density of A. oligospora was positively correlated with number of Dactylaria brochopaga & Verticilium chlamydosporium as The fungus, Verticillium chlamydosporium parasitizes eggs of root-knot and cyst nematodes [6]. Also, it infects nematode eggs and sedentary females of cyst nematodes by hyphae produced on actively growing mycelium [7]. V. chlamydosporium colonizes the rhizosphere, which facilitates the infection of egg masses protruding from female root-knot nematodes on infected roots [8]. The fungi A. dactyloides, A. oligosora, Macrosporium ellipsosporium, and M. cionopagum, parasites on most of the Pratylenchus penetrans adults and juveniles which added to the fungus cultures [9]. Anter et al. [10] in a greenhouse experiment, A. conoides and A. oligospora showed the highest effect on reducing M. incognita numbers during the first 4 weeks. A. conoides and A. oligospora showed highly influential on the activity J2 of M. incognita compared to the control treatment [11]. Arthrobotrys oligospora, A. conoides, Arthrobotrys sp., Dactylaria shelensis, and Dactylaria sp. and Monacrosporium were tested by Duponnois et al., [12] for their trapping ability against Meloidogyne mayaguensis. One Dactylaria strain and most of the Arthrobotrys strains reduced the population of nematodes. An Egyptian population of D. brochopaga the effect on nematodes has a great impact through production of traps which capture the larvae and dissolve nematode outer cuticle and digest the inner content of the victim [13].
population in the check plots before application ………………………………………………………............................……………..................................... X 100 effect of V. chlamydosporium combined with some organic manure on Meloidogyne incognita and other soil micro-organisms on tomato under field condition decreased the counts of Meloidogyne incognita juveniles in soil, as well as, gall formation on roots. Noweer and Aboul-Eid [15] obtained that the biological control of root-knot nematode Meloidogyne incognita infesting cucumber Cucumis sativus L. cvs. Alfa by (N.T.F.) D. brochopaga under field conditions. They showed that the D. brochopaga alone or in combination with yeast, molasses and vermiculite reduced the juvenile-M. incognita-population density per 1 kg soil and number of root-galls per 1 gm roots. Aboul-Eid et al. [16] studied the impact of the nematode-trapping fungus, Dactylaria brochopaga as a biocontrol agent against Meloidogyne incognita infesting Superior grapevine. They found that all treatments significantly reduced M. incognita J in soil 2 and number of root galls compared with the untreated control. Significant yield increases have been observed with all treatments compared with the untreated control. Spores suspension twice applications gave the highest yield production. Noweer and Mona E.M. Al-Shalaby [17] evaluated nematophagous fungi Dactylaria brochopaga and Arthrobotrys dactyloides against Meloidogyne incognita infesting peanut plants under field conditions. They found 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. Noweer [18] found that the effects of some nematode-trapping fungi on the root-knot nematode Meloidogyne sp. infesting white bean Phaseolus vulgaris and sugar beet Beta vulgaris sp.vulgaris under field conditions. He showed that the D. brochopaga affected the development and reproduction of M. incognita on sugar beet and white bean under field conditions. Noweer., [19] investigated the effect of the nematode-trapping fungus Dactylaria brochopaga and the nematode egg parasitic fungus Verticilium chlamydosporium in Controlling Citrus Nematode Infesting Mandarin, and Interrelationship with the Co inhabitant Fungi. He showed that the mixed compound treatment greatly affected the citrus nematode (Tylenchulus semipnetrans) numbers both in soil and roots, in comparing with those of Vydate or induced by mixed compound was 97% and 70%; respectively in soil and roots. In the present study, the effect of the nematode-trapping fungus D. brochopaga & Verticilium chlamydosporium as a combined with yeast, molasses and vermiculate on the development and reproduction of J2population in the treated plots after application x J2 Re % = 1- M. incognita infested Eggplant plants is reported under field condition.
The experiment was carried out through the period from 20 January 2018 to 27 May 2018 on an area almost ten karats' cultivated to Eggplant, Solanum melongena, variety Balady in Abdel-samad village, Imbaba, Giza. The soil of the experimental site is sand with a high level of natural infestation of root-knot nematodes Meloidogyne incognita. The seedlings of Eggplant were planted in January 20, 2018. Pre-treatment soil samples were taken, to determine the level of root-knot nematode infestation and contained from three simple samples (65 gm/sample) from each three rows per treatment as one composite soil sample (250 gm). A modified technique of Christie and Perry [20] methods had been chosen for nematode extraction because it illustrates a simpler technique requiring only minimum equipment for sandy soil processing. The sieves used in nematode processing were 2 mm hole diameter sieve and two 325 mesh sieves. Trapped Nematode suspensions washed from the last two sieves were transferred directly into glass beakers without using a centrifugal flotation technique step. Examination and counting of nematodes were made under a research microscope and root- knot nematode juveniles were identified by referring to the identification of ten rootknot females attached to the naturally- infested plant roots. Root – knot nematode identification was achieved according to Jepson Susan [21].
The fungus (Every one gm contain 5000 spores from the nematode-trapping fungus Dactylaria brochopaga collecting from the surface of Petri dishes contains the fungus grows on water agar medium, this fungus which has constricting rings responsible for nematode capturing through trapping mechanism with 5000 spores from the nematode egg parasitic fungus Verticilium chlamydosporium collecting from the slants contains the fungus grows on PDA medium this fungus which feeding on the nematode egg-masses was introduced into soil by different ways see table 1. The use of the two fungi together for different feeding method on the nematodes, whether nematode egg-masses or larvae that hatches from the nematode egg-masses. Compared with the nematicide Furidan treatment and an untreated treatment as control (Table 1). Percentage reduction in Root-knot second stage juvenile population in soil were determined according to the formula of Handerson & Tilton (Puntener) [22] where % Re= % reduction (% efficiency).
Verticilium chlamydosporium A=Agar, V= Vermiculite, Y= Yeast, M= Molasses, FUR= Furidan Data on population densities of M. incognita in both soil and root samples taken from the three replicates per treatment on 20/1, and 25/3 and 27/5/2018. The yield of
*With planting F1= Dactylaria brochopaga, F2=
Effects of the nematode-trapping fungus Dactylaria brochopaga & Verticilium chlamydosporium as a combined with yeast, molasses and vermiculate (F1+F2AVYM) on reproduction of root-knot nematode Meloidogyne incognita infesting Eggplant plants variety Balady are shown in tables (2, 3). Data in table 2 reveals that the highest decrease in the number of larvae per one kg soil was shown with treatment by (F1+F2AVYM). On March 25, 2018: The number of larvae per kg soil was decreased in almost all treatments especially it was 263 larvae per kg for the (F1+F2AVYM) compared to the untreated control 995. On May 27, 2018: The number of larvae per kg soil was decreased in almost all treatments especially it was 235 larvae per kg for the (F1+F2AVYM) compared to the untreated control 1739.
The percentage reduction of the number of larvae per one kg soil was 84.7% for (F1+F2AVYM) on March 25, 2018, while it was 88.6% for (F1+F2AVYM) on May 27, 2018. Data in table (3) reveals that the highest decrease in the number of galls per 5gm roots was noted with treatment by the (F1+F2AVYM). On March 25, 2018:
The number of galls per 5gm roots was decreased in almost all treatments especially it was 10 galls for (F1+F2AVYM) compared to the untreated control 36. On May 27, 2018: The number of galls per 5gm roots was decreased in almost all treatments especially it was 11 galls for (F1+F2AVYM) compared to the untreated control 94.
Eggplant was taken throw the period 25/3 to 27/5/2018 at the end of experiment (table 4).
The percentage reduction of the number of galls per 5 gm roots were 92.9% for (F1+F2AVYM) on March 25, 2018, while it was 93.1% for (F1+F2AVYM) on May 27, 2018.
The highest mean production of Eggplant variety Balady plants per one plant in the different treatments was 18 kg/plant for the (F1+F2AVYM), and it was 2160 kg per one plot (kirate). While the mean production of Eggplant variety Balady plants per one plant in the untreated control treatment was 5 k g/plant, and it was 600 kg per one plot (kirate). The length period of collection of the eggplant fruits explained that the use of difference in the quantity of production to show the efficiency of different treatments.
The present results indicate that the fungus Dactylaria brochopaga or the fungus Verticilium chlamydosporium was affected on the development and reproduction of Meloidogyne incognita on Eggplant plants variety Balady under field conditions especially for the (F1+F2AVYM). This was indicated by the lower numbers of juveniles in soil, lower numbers of galls per 5gm roots, the % reduction in population density of soil larvae, in treatment of the nematophagous A. dactyloides. Aboul-Eid et al., [13] reported that Dactylaria brochopaga has constricting rings responsible for nematode capturing through trapping mechanism.
* Data with the same letter(s) within a column are not significantly different according ** % Red. = % reduction (% efficiency) according to Anderson & Tilton formula.
*Data with the same letter(s) within a column are not significantly different according to Duncan’s a new multiple range test ** % Red. = % reduction (% efficiency) according to Anderson & Tilton formula.
*Data with the same letter(s) within a column are not significantly different according to Duncan’s a new multiple range test
***XR=Mean reduction due to treatments. F1= Dactylaria brochopaga A=Agar, V= Vermiculite, Y= Yeast, M= Molasses, FUR= Furidan
***XR=Mean reduction due to treatments.
F1= Dactylaria brochopaga, F2= Verticilium chlamydosporium.
A=Agar, V= Vermiculite, Y= Yeast, M= Molasses, FUR= Furidan
F1= Dactylaria brochopaga, F2= Verticilium chlamydosporium.
A=Agar, V= Vermiculite, Y= Yeast, M= Molasses, FUR= Furidan.
to Duncan’s a new multiple range test.
, F2= Verticilium chlamydosporium.
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The two fungi proved to be more effective nematodeantagonist and may have been affecting nematode population larvae through production of traps which capture the larvae and dissolve the nematode outer cuticle digest the inner content of the victim and parasitic on nematode eggs [23]. This information explains the results of this work. Moreover, Mankau [24] stated that the nematode- destroying fungi play a major role in recycling the carbon, nitrogen, and other important elements from the rather substantial of nematodes which browse on microbial primary decomposers. Certain fungal agents gave similar results in controlling citrus nematode in citrus groves and orchards [25]. The results of work with similar formulations of the nematode-trapping fungus Arthrobotrys dactyloides also has constricting rings responsible for nematode capturing through trapping mechanism has shown potential as a biological control agent against Meloidogyne javanica in soil microcosms [26].
Finally the (F1+F2AVYM) could be of great impact on the future of biotic and/or organic farming approach especially for the exported crops and other important foodstuff agricultural commodities especially for the two Fungi Dactylaria brochopaga & Verticilium chlamydosporium spores and broadcasted mycelia carried on granules of vermiculite, yeast, and molass.
Microbial control of root-knot nematode which infested Eggplant (Solanum melongena) by using the nematodetrapping fungus Dactylaria brochopaga & Verticilium chlamydosporium as a combined with yeast, molasses and vermiculate (F1+F2AVYM) is reported under field condition study. The results revealed that the highest percentage reduction in number of nematode larvae per 1kg soil was achieved when applying the fungi D. brochopaga & V. chlamydosporium together combined with yeast, molasses and vermiculate. The highest percentage reduction in number of root-galls per plant (93.1 %) was achieved by using the fungi D. brochopaga & V. chlamydosporium together combined with yeast, molasses and vermiculate. The weight of fruits per plant were significantly (P≤0.05 and/or P≤ 0.01) increased in all nematode-trapping fungus D. brochopaga treatments compared to the untreated check treatment. The use of the two Fungi Dactylaria brochopaga & Verticilium chlamydosporium spores and broadcasted mycelia carried on granules of vermiculite, yeast, and molass could be of great impact on the future of biotic and/or organic farming approach especially for the exported crops and other important foodstuff agricultural commodities.