EVALUATION THE ACTIVITY OF HIRSUTELLA SP. FUNGUS AND THE NEMATICIDE VEROX TO CONTROL ROOT KNOTS NEMATODE MELOIDOGYNE SPP . ON FIG SEEDLINGS

This study was conducted in the plastic house of the College of Agricultural Engineering Sciences at the University of Baghdad to evaluate the activity of Hirsutella sp. fungus and Nematode Verox against Meloidogyne spp. causative agent of root knots in Fig seedlings. Treatment of Meloidogyne spp. eggs with Hirsutella sp. at 20ml/L and with Verox at 2g/L caused significant inhibition in eggs hatching after 3,5 and 7 d .The number of non-hatching eggs were 96.70, 54.00and 29.30 eggs respectively in petri plates treated with Hirsutella sp., 106.70, 60.00and 46.00 eggs respectively in petri plates treated with Verox compared with 116.70, 111.00and96.00 eggs respectively in control. The treatment caused significant inhibition of juvenile-2 at the same periods . The number of juvenile-2 were 123.30, 114.00and 109.30 petri plates treated with Hirsutella sp at 20ml/L110.00, 63.00 and 26.70 in seedlings treated with Verox at 2g/

It has been reported that Fig trees are subjected to infection with many pathogens. Of these pathogens root knots Nematode is the most important that affect quality and quantity of Fig production (Martinuz, 2012). In addition to high losses caused by root knots Nematode, it caused breakdown of plant resistance to other pathogens and formed disease complex with other pathogen making its control more difficult (Qiao et al., 2013). The control of root knots Nematode was restricted for long time on highly toxic Nematicides. Due to enormous problems to ecosystem and human caused by Nematicide the

Samples from Fig seedlings root infected with
Meloidogyne spp. showing root knots were collected. The infected roots were cut to small pieces, 2 cm length, and dipped in 1% sodium hypochlorite solution to 4 min in flask with agitation. The hypochlorite solution with the root pieces was transferred to 300, 150and 25μm sieves respectively. The pieces were subjected to light flow water to eliminate NaOCl. The eggs were collected from 25μm sieve by light flow water in flask, 125 mL, observed under microscope at 40× and the eggs with juvenile-2 were counted in 1 mL of the total volume. The eggs suspension was poured in dark petri plates, 20 cm diam, with small volume of distilled water and maintained at 25°C for 1-3 d to obtain juvenile-2. Evaluation the effect of Hirsutella sp. fungus and Verox Nematicide on Meloidogyne spp. eggs hatching and Juvenile-2 viability.
One ml of Meloidogyne spp. inoculum containing 250± 5 eggs was placed in each of petri plates. Hirsutella sp. was added into the plates at 5, 10and20 ml/L. One mL of Verox Nematicide at 1, 1.5and 2 g/L was added to other plates containing the inoculum. The plates were incubated at 25± 5°C and the number of eggs and dead juvenile-2 were counted after 3, 5and7 d of treatment (AL-Ubaidy, 1985). Distilled water was added to other plates containing the inoculum for control, in three replicates for each concentration.The plates were distributed in Complete Randomized Design. Seedlings height were measured from soil surface to higher part of growing point befor treatment and at the end of experiment. Leaf area was determined by taking 3 leaves from each experiment unit by Al-Zaidi method as described by (Sadik et al., 2011) .

Estimation of nitrogen and potassium e in treated Fig seedlings.
Nitrogen percentage was determined in the leaver after 3, 6and9 d of treatment. The leaves were dried. in oven at 50°C and ground. The leaves powder was added to a mixture of sulfuric and perchloric (1:2) for digestion. The nitrogen in the solution was determined by kjeldhal system (Haynes, 1980). The potassium was determined by flame photometer as described by (Haynes,1980). The data was statistical analysis using Gen State 12 and compared by LSD 0.05. The treatment of Meloidogyne spp. eggs with Hirsutella sp. at concentrations 5,10and20 ml/L and with Verox at 1,1.5and 2 g/L caused significant inhibition in eggs hatching compared with control after 3,5,7 d (  Results of this study indicated that the fungus Hirsutella sp. at 20ml/L and the Nematicide Verox at 2g/L were of high activity to inhibit eggs hatching of Meloidogyne spp. parasitized the eggs producing secondary metabolites toxic to eggs. It was reported that Hirsutella sp. caus reduction in eggs hatching of Meloidogyne incognita to Laboratory conditions (Hallmann et al., 2019; Abokora,2021). Verox at 2g/L showed high activity against eggs of Meloidogyne compared with control. Verox is composed of plant growth promoting rhizobacteria (PGPR), I produce secondary metabolites (enzymes and toxins) including Chitinase that decompose chitin in eggs and toxin affecting eggs hatching (Siddiqui & Mahmood, 1999; Chauhan et al., 2015).
The treatment of juvenile -2 of Meloidogyne spp. with Hirsutella sp. fungus at 5, 10and20 ml/L and with Verox at, 1,1.5and 2 g/L induced significant inhibition of juvenile-2 compared with control after 3, 5and7 d. The results (

‫املستهلك‬ ‫ية‬ ‫ومحا‬ ‫لسىق‬ ‫ا‬ ‫لبحىث‬ ‫لعراقية‬ ‫ا‬ ‫اجمللة‬
Results of this study indicated that the fungus Hirsutella sp. at 20ml/L and the Nematicide Verox at 2g/L were of high activity to juvenile 2 viability of Meloidogyne spp. parasitized juvenile-2 producing secondary metabolites toxic to juvenile-2. Hirsutella rhossiliensis reported to be internal parasite produce stick substances to attract juvenile-2, induced reduction in juvenile-2 viability up to 95% (Hallmann et al., 2019; Wang et al.,  2007). Verox at 2g/L showed high activity against Juvenile of Meloidogyne compared with control. Verox is composed of plant growth promoting rhizobacteria (PGPR), I produce secondary metabolites (enzymes and toxins) including Chitinase that decompose chitin in eggs and toxin affecting juvenile-2 viability (Siddiqui & Mahmood, 1999; Chauhan et al., 2015).

Effect of treatment Fig seedlings infected with Meloidogyne spp on disease index and severity.
Results ( Table-3) showed that treatment of Fig seedling with Hirsutella sp. fungus and Verox and inoculating with Meloidogyne spp. induced high significant reduction in both disease index and disease severity. The disease index and severity were found to be 2.00 and 1.33 m seedlings treated with Hirsutella sp. and Verox respectively compart with 3.33 and 83.5 in contest respectively. Table ( High reduction in root knots index and disease severity were observed upon treatment big seedlings with Hirsutella sp. and Verox. This may come from reduction. the number of juvenile-2 penetrated the roof Lending To reduction in knot formation on the root. It has been reported that Hirsutella sp. was active in reduction Juvenile-2 of Meloidogyne incognita, number of knots and Nematode population (Abokora, 2021), The treatment with PGPR caused reduction in root knots number and disease severity (Liu et al., 2022).

Effect of treatment Fig seedlings with Hirsutella sp. and Verox on phenylalanine Ammonia-Lyase (PAL) and Chitinase enzymes activity.
Results (Table-4) showed that treatment Fig seedling with Hirsutella sp. fungus and vortex caused significant increase in PAL activity compared with control. PAL activity attained to 17.43, 19.23and 18.85 mg cinnamic acid/ h/g fresh weight in seedling treated with Hirsutella sp., 16.76, 18.56and18.05 mg cinnamic acid /h/g fresh weight in seedling treated with Verox compared with 15.33, 17.13and15.74 mg cinnamic acid/h/ g fresh weight in control after 3, 6and 9 d of treatment respectively. The significant differences in PAL activity were found more obvious at 3 d of treatment. No significant difference in PAL activity between treatments offer bod of d of treatments has been observed.  The effect of Hirsutella sp. and Verox on Meloidogyne spp. may be indirectly through induction systemic resistance in big seedlings against Meloidogyne spp. as proved by increase in PAL and Chitinase activity. It was reported that Nematophagous lungus Arthrobotrys oligospore induced increase, enzymes activity related with resistance including PAL associated with reduction in Nematode number in tomato compared. with plant, inoculated with M. incognita (Mostafanezhad et al., 2014).

Effect of treatment Fig seedlings with Hirsutella sp. and Verox on Loves content of Nitrogen and potassium.
Results (Table-5) indicated to significant increase in nitrogen and potassium percentages in the leaves of seedlings treated with Hirsutella sp. and Verox compared with control. (Treated with Meloidogyne spp.). Nitrogen and potassium percentage were attained to 3.617% and 2.120% in seedlings treated. with Hirsutella sp., 3.558% and 2.080% in seedlings treated with Verox compared with 2.858% and 1.820% respectively in control. Table ( (Table-6). Significant increases in seedlings height treated with the two agents were observed, 2.667 cm and 3.833 cm respectively compared with 0.667 cm in seedlings inoculated with Meloidogyne spp. (control).

CONCLUSION
The restriction of Meloidogyne spp. development was found associated with promotion of thy seedlings. This may come directly from the inhibition of juvenile-2 penetration into the cost and indirectly from the secondary metabolites producing by Hirsutella sp. and PGPR that provide the plants with nutrient elements and make other more available.