Effect of Some Bacterial and Fungal Bioagents for Management of Root-knot Nematode and Soil Born Fungus in Cucumber under Protected Conditions

doi:10.21203/rs.3.rs-280942/v1

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Research Article

Effect of Some Bacterial and Fungal Bioagents for Management of Root-knot Nematode and Soil Born Fungus in Cucumber under Protected Conditions

https://doi.org/10.21203/rs.3.rs-280942/v1

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Background: Complex disease caused by root-knot nematode and Fusarium fungus in cucumber is the most destructive disease under polyhouse conditions. The study was conducted to determine the potential of biocontrol agents viz. Trichoderma viride, Pseudomonas fluorescence, Purpureocillium lilacinum against root-knot nematode, Meloidogyne incognita and Fusarium oxysporum f. sp. cucumerinum disease complex on cucumber.

Results: All bioagents were taken @ 0.3 and 0.5 g/kg soil and liquid formulation of bioagents, @ 0.5 and 1 ml/ kg soil, Chemical checks as well as untreated check were also maintained. The bio-agents were mixed with the potted soil treatment wise. All the treatments significantly improved the plant growth parameter, viz., shoot length, root length, fresh shoot weight, fresh root weight, dry shoot weight and dry root weight as compared to untreated check. However, maximum improvement in plant growth parameter was recorded in case of carbofuran followed by higher dose of liquid formulation of bioagents. Among bioagents, liquid formulation of bioagents was significantly improved the plant growth parameter, viz., shoot length (147.3), fresh shoot weight (55.6), dry shoot weight (22.51) and dry root weight (4.50) and most effective in suppressing root knot nematode galling (43) and final population in soil (131) and fungus wilt incidence (25 %) at 30^th day of after germination followed by P. lilacinum T. viride and P. fluorescence.

Conclusion: The results suggest that the liquid formulation of bioagents was more effective in suppression of root-knot nematode and fungus complex disease than the powder formulations of bioagents. More studies should be needed in future to evaluate the efficacy of bioagents as seed treatments and soil applications under field conditions.

General Microbiology

Seed treatment

bio-agents

Meloidogyne incognita

disease complex

management

cucumber

and polyhouse.

Protected agriculture is considered an important means of increasing the productivity and quality of most vegetable crops (Hanafi, and Papasolomontos,1999). Recently, there has been an increase in interest in protected agriculture in India. Vegetable crops are grown worldwide as a source of nutrients and fiber in the human diet (Punja, and Utkhede, 2003). Restricted air exchange results in the atmospheric humidity being much higher inside insulated greenhouses than conventional ones which encourage several plant diseases and cause physiological disorders (El-Mougy et al., 2012). Root knot nematode, Meloidogyne incognita and Fusarium oxysporum f. sp. cucumerinum disease complex is recognized as foremost agricultural pathogens and are notorious to attack plants and cause crop losses throughout the world (Akhtar et al., 2005; Hadian et al., 2011). Internationally, root knot nematode has been causes severe yield losses in abundant crops due to their ability to invade several of crop species (Williamson and Hussey, 1996; Kayani et al. 2017). Likewise, it encourages typically root galling, yellowing of leaves, defoliation, stunting, and wilting of infested plants (Sasser et al., 1983). Management of root knot nematodes and fungus is a major worldwide challenge for polyhouse growers all over the world. Numerous agricultural practices have been tested against root knot nematodes (Collange et al., 2011). But they are not much effective against both pathogens. An actual approach toward management of root knot nematode and fungus is given by the use of nematicides (Giacometti et al., 2010; Nicolopoulou-Stamati et al., 2016). One possible potential and supportive substitute to nematicides is use of bio-control agents (Dutta and Thakur, 2017). Among the biological control agents that have been evaluated are egg-parasitic fungi, antagonist’s fungi, antagonist’s bacteria, and predatory nematodes has played an important role to suppress growth of plant pathogens and enhance growth of agricultural crops (Rao et al., 1998; Kiewnick and Sikora, 2005; Abdelmoneim, 2006: Al-Shammari et al., 2013).

Thus, the purpose of the present study is to assess the potential antagonistic effect of biocontrol agents viz., Trichoderma viride, Pseudomonas fluorescence, Purpureocillium lilacinum and combination of liquid formulation of bio-agents (T. viride + P. fluorescence + P. lilacinum) against Meloidogyne incognita and Fusarium oxysporum f. sp. cucumerinum their effect on growth development of cucumber under polyhouse conditions.

Experimental site

The experiment was conducted during summer season (2015-16) in the polyhouse of the Department of Horticulture, authors institute, Hisar, Haryana (India).

Collection, isolation and maintenance of root-knot nematode and fungus

Cucumber roots showing galls were collected from the naturally infested polyhouses during survey and brought to the laboratory. Nematode and fungus were isolated, identified by Patil et. al., (2018) and pure culture of both the organisms were maintained. Fungus was grown on sand maize meal medium for mass production and nematode culture was maintained in screen house on brinjal. Nematode juveniles were collected by sodium hypochlorite method for inoculation.

Source of bioagents

Bioagents Trichoderma viride (10⁷ cfu/g), Pseudomonas fluorescence ((10⁸cfu/g)), Purpureocillium lilacinum ((10⁶ cfu/g) and liquid formulation of bio-agents (T. viride + P. fluorescence + P. lilacinum) were procured from IIHR, Bangalore were used in this study.

Seed treatment with bioagents

Carboxy methyl cellulose (CMC) was used as an adhesive for treating cucumber seeds with fungal and bacterial bioagents (1x10⁸ cfu/ml.). For preparing 1% (v/w) adhesive solution, adhesive was added to fungal and bacterial bioagents. Now required amount of seeds was taken in a petriplate and the fungal as well as bacterial bioagent with the adhesive was added on the seeds stirring continuously and stopped when all the seeds got smeared with suspension. After treating, the seeds were dried in shade for 6 hours and used for sowing.

Extraction of nematodes from soil

Nematode extraction from soil was done by Cobb’s sieving and decanting method (Cobb, 1918) followed by modified Baermann funnel method (Whitehead & Hemming, 1965).

Experimental setup

This experiment was conducted in polyhouse ((24± 2 º C)) to evaluate the effect of seed treatment of bio-agents in cucumber for management of root-knot nematode and fungus (Fusarium oxysporum f. sp. cucumerinum). Experiment was conducted in pots (1 kg capacity). The soil of each pot (1500 g) was a mixture of sand, sandy loam and peat moss (2:1:1), which was previously steam-sterilized (15 Psi at 121 º C) with an autoclave for 30 min. In previous in vitro test, we tested bio-agents against M. incognita. Based on the results of these tests (un-published data), two promising doses namely: T. viride, P. fluorescence, P. lilacinum 10 and 20 g/kg seed and liquid formulation of bio-agents (T. viride + P. fluorescence + P. lilacinum) 10 ml/ kg and 15 ml/ kg seed were selected for the present study. Seed treatment of bio-agents were tested against nematode alone, fungus alone and nematode and fungus simultaneously inoculated. Treatments were arranged in completely randomized design with four replications. Cucumber seeds (cv. Sania) were treated with bio-agents for 2 hours dried in shade for one hour and then sown to each pot treatment wise @ 5 seeds per pot. One plant per pot was maintained. Sterilized soil was inoculated with 1000 J₂/ kg soil of root-knot nematode and fungus 50g /kg soil.

Maintenance of plants

General care and maintenance of plants were done as recommended by CCS Haryana Agricultural University, Hisar. Hoagland solution was applied at 10 days’ interval. Hoeing, watering other practices were done as per recommendation.

Observations

After crop maturity at 60 days after sowing, plants were harvested and 200 cc soil was collected for estimation of nematode population. Observations viz. plant growth parameters (shoot and root length, fresh and dry shoot and root weight), number of galls per plant, number of egg masses per plant, number of eggs/egg mass, final nematode population (Cobb, 1918), pre-emergence damping off (after 15 and 30 days) were taken.

Statistical analysis

Data were analyzed in a factorial completely randomized design with four replications. Data were subjected to analysis of variance (ANOVA), and the means were compared with a critical difference. A significance level of P ≤ 0.05 was used in all analyses. Nematode data were transformed, using square root transformation to homogenize error variances. All calculations were performed with the software available online at the website www.hau.ac.in.

Effect of bio-control agents on plant growth performance

Generally, the data (Table 1) indicated that shoot and root length in all the treatments was significantly improved over untreated inoculated checks viz., nematode alone, fungus alone and nematode + fungus simultaneously. Among the various treatments, highest shoot and root length was detected in seed treatment with liquid formulation of bio-agents 15 ml/ kg seed trailed by P. lilacinum 20 g per kg seed nevertheless of whether nematode or fungus inoculated individually or concomitantly. In plants inoculated with both the pathogens concomitantly, extreme shoot length of both the parameters was in case of liquid formulation of bio-agents followed by P. lilacinum as compared to untreated inoculated check. Data (Table 2) displayed that fresh shoot and root weight in altogether the treatments was significantly improved over untreated inoculated checks viz., nematode alone, fungus alone and nematode + fungus simultaneously. Among the various treatments, extreme fresh shoot and root weight was observed in liquid formulation of bio-agents (T. viride + P. fluorescence + P. lilacinum) 15 ml per kg seed followed by P. lilacinum 20 g per kg seed irrespective of whether nematode or fungus inoculated individually or concomitantly. Though, in plants inoculated with nematode alone, fresh shoot and root weight was extreme in case of liquid formulation of bio-agents followed by P. lilacinum as compared to untreated inoculated check.

The data (Table 3) articulated that dry shoot and root weight in all the treatments was significantly better over untreated inoculated checks viz., nematode alone, fungus alone and nematode + fungus simultaneously. Among the various treatments, maximum fresh shoot and root weight was observed in liquid formulation of bio-agents (T. viride + P. fluorescence + P. lilacinum) 15 ml per kg seed trailed by P. lilacinum 20 g per kg seed irrespective of whether nematode or fungus inoculated individually or concomitantly. Though, in plants inoculated with nematode alone, fresh shoot and root weight was maximum in case of liquid formulation of bio-agents followed by P. lilacinum 20 g per kg seed as compared to untreated inoculated check. In plants inoculated with fungus alone, fresh shoot and root weight was maximum in case of liquid formulation of bio-agents followed by T. viride 20 g per kg seed as compared to untreated inoculated check. In plants inoculated with nematode and fungus concomitantly, fresh shoot and root weight was maximum in case of liquid formulation of bio-agents followed by P. lilacinum as compared to untreated inoculated check. In general, fresh shoot and root weight was significantly reduced in all the treatments compared to untreated uninoculated check notwithstanding of whether inoculated individually or concomitantly with nematode and fungus.

Effect of bio-agents on nematode reproduction

The data (Table 4) indicated that nematode reproduction in all the treatments was significantly reduced over untreated inoculated checks viz., nematode alone and nematode + fungus simultaneously. Among the various treatments, minimum nematode reproduction was observed in liquid formulation of bio-agents 15 ml per kg seed followed by P. lilacinum 20 g per kg seed irrespective of whether nematode inoculated individually or concomitantly. However, in plants inoculated with nematode alone, nematode reproduction was minimum in case of liquid formulation of bio-agents followed by P. lilacinum as compared to untreated inoculated check. In case of plants inoculated with nematode and fungus concomitantly, nematode reproduction was minimum in case of liquid formulation of bio-agents followed by P. lilacinum as compared to untreated inoculated check. Maximum reduction in nematode reproduction was observed in the presence of both nematode and fungus followed by nematode alone. On the additional, the obtained results from experiment indicated that soil juvenile population (Fig. 1), developing stages, no. of eggs/egg mass, no. of egg masses, number of females in root and no. of galls were significantly decreased in plant treated with bio-agents as compared with untreated check.

Per cent disease incidence

The data (Table 5) exposed that in general, all the treatments significantly reduced incidence of fungus in fungus alone and nematode and fungus concomitantly on cucumber as compared to untreated inoculated check. Data was recorded 15 and 30 days after sowing. At 15 days after sowing, disease incidence was minimum in case of for liquid formulation of bio-agents 15 ml per kg seed or T. viride 20 g per kg seed as compared to untreated inoculated check. At 30 days after sowing, disease incidence was minimum in case of liquid formulation of bio-agents followed by in T. viride as compared to untreated inoculated check.

Significantly higher plant growth parameters were obtained in seed treatment with liquid formulation of bio-agents followed by P. lilacinus irrespective of whether nematode or fungus were inoculated individually or concomitantly. This is in line with studies done by Amer-Zareen (2001) who reported that seed/soil drench with P. lilacinus, Trichoderma harzianum and T. flavus recorded higher plant growth compared to control. Minimum plant growth as parameters were recorded in case of P. fluorescence treated plants. Nematode reproduction in all the treatments was significantly reduced over untreated inoculated checks. However, in plants inoculated with nematode alone, number of galls per plant was minimum in case of liquid formulation of bio-agents followed by P. lilacinus as compared to untreated inoculated check. Lowest number of eggs per egg mass was observed in liquid formulation of bio-agents followed by P. lilacinus irrespective of whether nematodes were inoculated individually or concomitantly. Our findings are in agreement with Dhawan et al. (2004) who reported that P. lilacinus tested against M. incognita on okra as seed treatment at 10, 15 and 20 g / kg seed improved plant growth as well as reduced in number of galls, egg masses and eggs per egg mass. Lamovsek et al. (2013) also reported that the use of commercial product of Purpureocillium lilacinum and Bacillus firmus as drenching and seed treatment on tobacco and soybean, improved plants growth characters and reduced of Meloidogyne spp. population under field condition.

Similarly, Rao et al. (1997) reported seed treatment with P. lilacinus, resulted in lowest root-knot index, reduced final population of M. incognita and increased fruit yield of cucumber. The reason for reduction in nematode population might be due to parasitic activity of P. lilacinus on eggs and all stages of nematodes. Spores of the P. lilacinus also adhere to the cuticle of vermiform stages of the nematodes as they migrate through the soil. The spores of P. lilacinus penetrate the cuticle and engulf the nematode. The hyphae of the P. lilacinus could have entered the nematode through body openings, such as the anus and vulva. The developing P. lilacinus killed the nematode by feeding on its body contents. As a whole, P. lilacinus acted as a parasite on the nematode. Maximum reduction in nematode population J₂/ 200 cc soil was observed in the presence of both nematode and fungus followed by nematode alone. All the treatments significantly reduced incidence of fungus on cucumber as compared to untreated inoculated check. Disease incidence was minimum (10 %) in case of liquid formulation of bio-agents followed by 15 % in case of T. viride as compared to untreated inoculated of both pathogens concomitantly. These finding also was similar to Druzhinina et al. (2011), Hallman et al. (2009), Robab et al. (2012) and Vos et al. (2012).

Table 1

Effect of seed treatment with bio-agents on shoot and root length of cucumber infested with *M. incognita* and fungus
Treatments	Shoot length (cm)				Root length (cm)
Treatments	Nematode alone	Fungus alone	Nematode + fungus	Pooled Mean	Nematode alone	Fungus alone	Nematode + fungus	Pooled Mean
T. viride 10 g/ kg seed	136.8	130.3	129.9	132.3	37.0	37.1	31.9	35.3
T. viride 20 g/ kg seed	142.8	139.9	135.8	139.5	42.0	47.1	37.0	42.0
P. fluorescence 10 g/ kg seed	138.3	131.3	130.2	133.3	38.6	38.5	32.8	36.6
P. fluorescence 20 g/ kg seed	144.8	138.2	138.3	140.4	43.0	46.2	38.1	42.4
P. lilacinum 10 g/ kg seed	139.4	132.5	131.2	134.3	39.3	39.9	34.1	37.8
P. lilacinum 20 g/ kg seed	147.4	135.8	139.4	140.9	46.6	43.9	38.7	43.0
LFB 10 ml/ kg seed	140.3	134.5	133.3	136.0	39.9	42.8	35.6	39.4
LFB 15 ml/ kg seed	152.3	144.0	142.6	146.3	54.1	48.4	42.4	48.2
Carbosulfan 3% v/w	176.1	149.6	164.5	163.4	64.6	54.0	60.1	59.5
Untreated check (inoculated)	83.3	86.4	85.3	85.0	23.3	25.1	24.6	24.3
Untreated check (uninoculated)	111.8	111.3	112.8	111.9	28.2	29.5	28.9	28.8
Pooled Mean	137.6	130.3	131.2		41.5	41.1	36.7
CD at 5%	Pathogen: 0.6; Bio-agents: 1.2; Pathogen X Bio-agents: 2.1				Pathogen: 0.5; Bio-agents: 1.0; Pathogen X Bio-agents: 1.8
LFB = liquid formulation of bio-agents, Data are mean of four replications

Table 2

Effect of seed treatment with bio-agents on fresh shoot weight and root weight of cucumber infested with *M. incognita* and fungus
Treatments	Shoot weight (g)				Root weight (g)
Treatments	Nematode alone	Fungus alone	Nematode + fungus	Pooled Mean	Nematode alone	Fungus alone	Nematode + fungus	Pooled Mean
T. viride 10 g/ kg seed	44.05	30.90	36.06	37.00	15.25	12.74	13.57	13.85
T. viride 20 g/ kg seed	51.30	40.00	42.08	44.46	18.30	18.64	17.80	18.24
P. fluorescence 10 g/ kg seed	45.05	32.65	37.11	38.26	15.50	13.75	15.25	14.82
P. fluorescence 20 g/ kg seed	53.60	38.80	43.18	45.19	18.80	17.50	18.39	18.22
P. lilacinum 10 g/ kg seed	48.08	33.98	38.78	40.27	16.09	14.25	16.00	15.44
P. lilacinum 20 g/ kg seed	56.68	37.05	44.55	46.1	19.93	16.41	19.74	18.69
LFB 10 ml/ kg seed	49.20	35.50	40.83	41.84	17.16	14.99	16.90	16.54
LFB 15 ml/ kg seed	63.23	41.93	46.05	50.40	21.49	19.25	20.74	19.07
Carbosulfan 3% v/w	75.82	47.78	63.04	62.21	22.10	23.08	21.74	21.24
Untreated check (inoculated)	23.91	25.05	23.60	24.18	6.76	6.74	6.35	10.24
Untreated check (uninoculated)	30.12	28.93	29.81	29.61	10.29	11.76	10.65	13.84
Pooled Mean	49.18	35.68	40.46		16.51	15.37	16.86
CD at 5%	Pathogen: 0.5; Bio-agents: 1.1; Pathogen X Bio-agents: 1.9				Pathogen: 0.4; Bio-agents: 0.8; Pathogen X Bio-agents: 1.4
LFB = liquid formulation of bio-agents, Data are mean of four replications

Table 3

Effect of seed treatment with bio-agents on dry shoot weight and dry root weight of cucumber infested with *M. incognita* and fungus
Treatments	Dry shoot weight (g)				Dry root weight (g)
Treatments	Nematode alone	Fungus alone	Nematode + fungus	Pooled Mean	Nematode alone	Fungus alone	Nematode + fungus	Pooled Mean
T. viride 10 g/ kg seed	14.50	14.23	14.25	14.32	3.59	3.51	3.89	3.66
T. viride 20 g/ kg seed	18.25	19.99	17.65	18.63	4.72	4.90	4.98	4.89
P. fluorescence 10 g/ kg seed	15.48	15.24	14.74	15.15	3.94	3.70	3.90	3.84
P. fluorescence 20 g/ kg seed	18.99	19.24	19.23	19.15	5.00	4.60	5.75	5.11
P. lilacinum 10 g/ kg seed	16.82	15.75	15.50	16.02	4.02	4.01	4.33	4.11
P. lilacinum 20 g/ kg seed	19.74	18.74	19.73	19.40	5.76	4.48	6.26	5.5
LFB 10 ml/ kg seed	17.75	16.99	16.90	17.21	4.33	3.99	4.53	4.28
LFB 15 ml/ kg seed	20.97	20.23	20.31	20.50	6.08	5.50	6.51	6.02
Carbosulfan 3% v/w	23.33	21.92	22.75	22.66	8.18	6.00	7.36	7.17
Untreated check (inoculated)	6.28	7.01	7.27	6.85	2.20	2.10	1.77	2.02
Untreated check (uninoculated)	9.74	9.25	10.47	9.82	3.27	3.00	3.25	3.17
Pooled Mean	16.43	16.07	9.22		4.64	4.16	4.77
CD at 5%	Pathogen: 1.2; Bio-agents: 2.4; Pathogen X Bio-agents: 4.2				Pathogen: 0.22; Bio-agents: 0.42; Pathogen X Bio-agents: 0.73
LFB = liquid formulation of bio-agents, Data are mean of four replications

Table 4

Effect of seed treatment with bio-agents on nematode reproduction in cucumber infested with *M. incognita* and fungus
Treatments	Numbers of galls/plant			Numbers of egg masses/plant			FNP / 200cc soil
Treatments	N alone	N + F	Pooled Mean	N alone	N + F	Pooled Mean	N alone	N + F	Pooled Mean
T. viride 10 g/ kg seed	53 (7.3)	65 (8.1)	59 (7.7)	63 (7.9)	64 (8.0)	63.5 (8.0)	185 (13.6)	189 (13.8)	187 (13.7)
T. viride 20 g/ kg seed	47 (6.8)	55 (7.5)	51 (7.2)	53 (7.3)	52 (7.2)	52.5 (7.3)	168 (12.9)	176 (13.3)	172 (13.1)
P. fluorescence 10 g/ kg seed	51 (7.2)	63 (7.9)	57 (7.6)	61 (7.9)	63 (8.0)	62 (8.0)	182 (13.5)	186 (13.7)	184 (13.6)
P. fluorescence 20 g/ kg seed	46 (6.8)	52 (7.3)	49 (7.0)	51 (7.2)	53 (7.3)	52 (7.3)	165 (12.9)	174 (13.2)	169.5 (13.0)
P. lilacinum 10 g/ kg seed	51 (7.1)	58 (7.6)	54.5 (7.4)	59 (7.7)	61 (7.8)	60 (7.8)	177 (13.3)	180 (13.4)	178.5 (13.4)
P. lilacinum 20 g/ kg seed	46 (6.8)	50 (7.1)	48 (7.0)	47 (6.9)	50 (7.1)	48.5 (7.0)	162 (12.8)	166 (12.9)	164 (12.8)
LFB 10 ml/ kg seed	48 (6.9)	57 (7.6)	52.5 (7.3)	55 (7.4)	53 (7.3)	54 (7.4)	173 (13.2)	178 (13.3)	175.5 (13.3)
LFB 15 ml/ kg seed	45 (6.7)	48 (6.9)	46.5 (6.9)	45 (6.8)	48 (6.9)	46.5 (6.9)	157 (12.5)	164 (12.8)	160.5 (12.7)
Carbosulfan 3% v/w	43 (6.6)	45 (6.8)	44 (6.7)	43 (6.6)	47 (6.9)	45 (6.8)	149 (12.2)	159 (12.6)	154 (12.4)
Untreated check (inoculated)	448 (21.2)	453 (21.3)	450.5 (21.2)	250 (15.8)	249 (15.8)	249.5 (15.8)	820 (28.6)	825 (28.7)	822.5 (28.7)
Untreated check (uninoculated)	0.0 (1.0)	0.0 (1.0)	0.0 (1.0)	0.0 (1.0)	0.0 (1.0)	0.0 (1.0)	0.0 (1.0)	0.0 (1.0)	0.0 (1.0)
Pooled Mean	7.7	8.1		7.5	7.6		13.3	13.5
CD at 5%	Pathogen: 0.04; Bio-agents: 0.09; Pathogen X Bio-agents: 0.13			Pathogen: 0.1; Bio-agents: 0.09; Pathogen X Bio-agents :0.04			Pathogen: 0.02; Bio-agents: 0.06; Pathogen X Bio-agents: 0.09
N = nematode; N + F = nematode + Fungus; FNP = final nematode population; LFB = liquid formulation of bio-agents

Figures in parenthesis are the square root (n + 1) transformed values, Data are mean of four replications

Table 5

Effect of seed treatment with bio-agents on fungal incidence (%) in cucumber infested with *M. incognita* and *F. oxysporum* f. sp. *cucumerinum*
Treatments	Fungus alone			Nematode + Fungus
Treatments	Pre emergence damping off (after 15 days)	Pre emergence damping off (after 30 days)	Pooled Mean	Pre emergence damping off (after 15 days)	Pre emergence damping off (after 30 days)	Pooled Mean
T. viride 10 g/ kg seed	15 (23.0)	25 (30.3)	20 (26.7)	20 (26.8)	30 (33.4)	25 (30.1)
T. viride 20 g/ kg seed	5 (13.6)	10 (18.4)	7.5 (16.0)	10 (18.6)	15 (23.0)	12.5 (20.8)
P. fluorescence 10 g/ kg seed	15 (23.0)	20 (26.9)	17.5 (25.0)	20 (26.9)	30 (33.4)	25 (30.2)
P. fluorescence 20 g/ kg seed	10 (18.6)	15 (23.0)	12.5 (20.8)	15 (23.0)	20 (26.8)	17.5 (24.9)
P. lilacinum 10 g/ kg seed	15 (23.0)	20 (26.9)	17.5 (25.0)	20 (26.8)	25 (30.3)	22.5 (28.5)
P. lilacinum 20 g/ kg seed	10 (18.6)	20 (26.9)	15 (22.75)	15 (23.0)	15 (23.0)	15 (23.0)
LFB 10 ml/ kg seed	15 (23.0)	20 (26.9)	17.5 (25.0)	15 (23.0)	20 (26.8)	17.5 (24.9)
LFB 15 ml/ kg seed	5 (13.6)	5 (13.6)	5 (13.6)	5 (13.6)	10 (18.6)	7.5 (16.1)
Carbosulfan 3% v/w	5 (13.6)	5 (13.6)	5 (13.6)	5 (13.6)	10 (18.6)	7.5 (16.1)
Untreated check (inoculated)	55 (48.2)	65 (54.1)	60 (51.2)	60 (51.2)	65 (54.1)	60 (51.2)
Untreated check (uninoculated)	0 (4.1)	0 (4.1)	0.0 (4.1)	0 (4.1)	0 (4.1)	0.0 (4.1)
Pooled Mean	20.2	24.0		22.7	26.3
CD at 5%	Pathogen 1.3; Bio-agents: 3.2; Pathogen X Bio-agents: N.S.			Pathogen: 1.5; Bio-agents: 3.7; Pathogen X Bio-agents: N.S.
LFB = liquid formulation of bio-agents

Figures in parenthesis are the angular transformed values, Data are mean of four replications

All combinations of bioagents were effective in suppressing nematode and fungus disease in all combinations (nematode alone, fungus alone and nematode and fungus inoculated simultaneously) as well as improving plant growth. Among all the bioagents, liquid formulations of all bioagents were more effective than the powder formulations in all possible combinations. The results of this study seem to indicate that the liquid formulations of bioagents could be a potential candidate in organic farming, where a few/less options are available to manage phytonematodes and fungus disease complexes.

M. incognita: Meloidogyne incognita; F. oxysporum f. sp. cucumerinum: Fusarium oxysporum f. sp. cucumerinum; T. viride: Trichoderma viride; P. fluorescence: Pseudomonas fluorescence; P. lilacinum: Purpureocillium lilacinum

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Effect of Some Bacterial and Fungal Bioagents for Management of Root-knot Nematode and Soil Born Fungus in Cucumber under Protected Conditions

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