JOURNAL OF PURE AND APPLIED MICROBIOLOGY, Dec. 2017.

Vol. 11(4), p. 1909-1917

Bio-management of Cucumber wilt Complex Caused by

Root-knot nematode, Meloidogyne incognita and Fusarium
oxysporum f. sp. cucumerinum in Polyhouse under Protected
Cultivation
Jaydeep Patil*1, S R Goel2 and Saroj Yadav2
1
Department of Plant Protection, School of Agriculture LPU, Phagwara, Punjab - 144401, India.
2
Department of Nematology, College of Agriculture,CCS HAU Hisar 125004 Haryana, India.

doi:

(Received: 15 November 2017; accepted: 01 December 2017)

A polyhouse study was conducted to determine the potential of biocontrol agents

viz. Trichoderma viride, Pseudomonas fluorescence, Purpureocillium lilacinum (Paecilomyces
lilacinus) @ 0.3 and 0.5 g/kg soil and liquid formulation of bioagents, (T. viride + P. fluorescence
+ P. lilacinus) @ 0.5 and 1 ml/ kg soil, against root-knot nematode, Meloidogyne incognita and
Fusarium oxysporum f. sp. cucumerinum disease complex on cucumber. Three main treatments,
viz., nematode alone, fungus alone and both inoculated simultaneously were taken. Chemical
checks with Bavistin @ 2 g/ l water and carbofuran @ 0.1 mg/ kg soil as well as untreated check
were also maintained. Fungus was grown on sand maize meal medium. Soil was autoclaved
and infested with root-knot nematode (1000 J2/ kg soil) and fungus (50 g/ kg soil). The bio-agents
were mixed with the potted soil treatment wise. A waiting period of three days was given for
multiplication of bioagents on the organic matter before sowing. All the treatments significantly
improved the plant growth parameter, viz., shoot length (SL), root length (RL), fresh shoot
weight (FSW), fresh root weight (FRW), dry shoot weight (DSW) and dry root weight (DRW) as
compared to untreated check. However, maximum improvement in plant growth parameter was
recorded in case of carbofuran @ 0.1 mg /kg soil followed by higher dose liquid formulation
of bioagents. Bavistin was least effective among all the treatments against root-knot nematode,
Meloidogyne incognita and Fusarium oxysporum f. sp. cucumerinum disease complex.

Keywords: Meloidogyne incognita, Fusarium oxysporum f. sp. cucumerinum, shoot length,

Root length, Fresh shoot weight, and fresh root weight, shoot weight, dry root weight.

In India, growing of horticultural crops in cultivated plant in the gourd family, Cucurbitaceae.
polyhouses under protected cultivation is becoming It is a creeping vine that bears cylindrical fruits
very popular among the farmers throughout the that are used as culinary vegetables. There are
country. Large numbers of polyhouses are being three main varieties of cucumber: slicing, pickling,
erected in Haryana under the ages of National and burp less. Within these varieties, several
Horticulture Mission to grow short duration different cultivars have emerged. The cucumber
crops. Cucumber (Cucumis sativus L.) is a widely is originally from South Asia, but now grows on
most continents. Cucumber is an edible cucurbit
popular throughout the world due to a good
* To whom all correspondence should be addressed.
E-mail: rajhau99@gmail.com source of vitamins, minerals, fiber and roughages.
It having crisps texture and taste. Cucumber is
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truly a versatile vegetable because of wide range with a mixture of autoclaved sandy loam soil (sand
of uses from salads to pickles and digestive aids 70%, silt 22% and clay 8%, pH 7.5). Autoclaved
to beauty products. the caloric and nutritional soil would be infested with root-knot nematode @
value of cucumber is very low but it is a primary 1000 J2/kg soil and fungus (50 g/kg soil) as per the
source of vitamins, mineral and fiber for human treatment. The experiment was conducted in pots
body (Keoprapari,1997). The annual production (1 kg capacity) containing infested soil. Inoculum
of cucumber in India is 698000 MT from 45000 of root knot nematode was obtained from the
ha area with productivity of 15.5 per ha only nematode infected cucumber at farmer’s field in
during 2012-13 (Anonymous, 2014). Polyhouse Hisar Haryana, India. Root knot nematode females
cultivation involves intensive cultivation of crops, collected from the cucumber roots were processed
optimum use of fertilizers and frequent use of for perineal pattern to confirm the species of root
irrigation, but continuous growing of the same crop knot nematode associated with the plant. The pure
with high day temperature and relative humidity culture was prepared in steamed sterilized soil in
within the greenhouse, polyhouse and low tunnel pots and Inoculum was used for experimentation.
along with poor plant hygienic conditions inside Pure culture of F. oxysporum isolated
and outside the greenhouse increase problem of soil from the infested plants during random survey
borne pests and diseases including plant parasitic of polyhouses was maintained on PDA (Potato
nematodes (Minuto et al., 2006) which results in Dextrose Agar) in petriplates at (27±5) °C in order
the availability of ideal conditions for the growth to mass-produce pure culture of the Fungus was
and multiplication of these pests. grown on sand maize meal medium (700gm sand
Under polyhouse cultivation crops, + maize meal 300gm + 150ml distilled water).
are attacked by a number of pests and diseases The flasks were incubated in a BOD (Biological
including nematodes which interfere with the Oxygen Demand) incubator at a temperature of
successful cultivation under protected conditions. (27±1) °C for 15 days. During incubation, the flasks
Among the nematodes, root-knot nematode were shaken three times in a day, to ensure proper
(Meloidogyne spp.) is the most damaging under growth of the fungal mycelium on the sand maize
polyhouse conditions, parasitizing almost all the meal medium.
polyhouses crops. The damage becomes very Root-knot nematode and fungus was
severe in association with fungi. Though, yield also inoculated carefully adding the homogenous
loss due to this nematode is difficult to predict, suspension of the two pathogens at the root
approximate yield loss due to this nematode has zone of the plants, as per treatment. Each pot
been predicted by many authors in various crops. would be infested with root-knot nematode
Another important biotic stress to which the crop (1000 J2/kg soil) and fungus (50g /kg soil) and
exposed is the fungus, Fusarium oxysporum f. treated with (carbofuran at 1 mg a.i./kg soil,
sp. cucumerinum. Considering the soil health, Bavistin at 1 mg a.i./kg soil, Trichoderma viride,
environmental safety and the long term hazards Pseudomonas fluorescence, Purpureocillium
posed by the indiscriminate use of pesticides, lilacinum (Paecilomyces lilacinus) @ 0.3 and
bioagents promise to be the next best alternative 0.5 g/kg soil and liquid formulation of bioagents,
for nematode management. With this aim, a (T. viride + P. fluorescence + P. lilacinus) @ 0.5
study was conducted in a polyhouse to test the and 1 ml/ kg soil were incorporated to the potted
efficacy of certain easily available bioagents (P. soil as per treatment. Also waiting period of three
lilacinus, T. viride and P. fluorescence) against days was given for multiplication of bioagents
Meloidogyne incognita and Fusarium oxysporum on the organic matter before sowing. After seven
f. sp. cucumerinum disease complex on cucumber. days each pot would be sown with cucumber
cv Sania @ 5 seeds as per treatment and also
MATERIALS AND METHODS maintain untreated check. Uninoculated pots and
nematode + fungus inoculated pots served as
Experiment was conducted in polyhouse controls. One plant per pot was retained after 30
(26.7±3) °C, 73.5±11% Relative Humidity and days. Each treatment was replicated four times in
0.918 kPa) in earthen pots (18 cm diameter) filled a completely randomized block design during the

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months of April to June, 2015 in the polyhouse bio-agents (147.9 cm), followed by P. lilacinus
under protected conditions and watered daily so (146.9 cm) as compared to untreated inoculated
that each pot as per requirement. check (83.9 cm). In general, shoot length was
Evaluations were performed 60 days after significantly less in all the treatments compared
sowing. Measurements were made on the plant to untreated uninoculated check irrespective of
growth parameters (shoot length, fresh and dry whether inoculated individually or concomitantly
shoot and root weight) observations were made on both pathogens. Maximum reduction in shoot
the root population of nematode viz., Number of length was observed in the presence of nematode
galls per plant, Number of egg masses per plant, and fungus followed by fungus alone while
Number of eggs per egg mass, Final nematode minimum in case of nematode alone.
population per pot. Nematode population in Fresh shoot weight in all the treatments
soil was processed as per the sieving method of was significantly better over untreated inoculated
Cobb´s sieving and decanting technique followed checks viz., nematode alone (23.2 g), fungus alone
by Modified Baermann´s funnel technique for (23.4 g) and nematode + fungus simultaneously
estimation of nematode population in soil. Per cent (22.90 g). Among the various treatments,
wilt incidence due to fungus was assessed using maximum fresh shoot weight was observed in
number of wilt infected plants /total number of liquid formulation of bio-agents (T. viride + P.
plants taken for observation. fluorescence + P. lilacinus) @ 15 ml per kg soil
Statistical analysis (59.1 g), followed by Paecilomyces lilacinus @
Data were analysed using analysis of 0.5 g per kg soil (55.7 g) irrespective of whether
variance (ANOVA). Treatment means were nematode or fungus inoculated individually or
compared and critical differences (CD) was concomitantly. However, in plants inoculated with
calculated at P=0.05 to test for significant nematode alone, fresh shoot weight was maximum
differences between treatments (T) in case of liquid formulation of bio-agents (64.7 g),
followed by P. lilacinus @ 0.5 g per kg soil (64.7
RESULTS g) as compared to untreated inoculated check (23.2
g). Plants inoculated with fungus alone, fresh shoot
Data indicated that shoot length in all the weight was maximum in case of liquid formulation
treatments was significantly better over untreated (57.2 g), followed by T. viride @ 0.5 g per kg soil
inoculated checks viz., nematode alone (87.5 cm), (54.1 g) as compared to untreated inoculated check
fungus alone (85.6 cm) and nematode + fungus (23.4). Plants inoculated with nematode and fungus
simultaneously (83.9 cm). Among the various concomitantly, fresh shoot weight was maximum
treatments, maximum shoot length was observed in case of liquid formulation of bio-agents (55.2
in liquid formulation of bio-agents (T. viride + P. g), followed by P. lilacinus (5.6 g) as compared to
fluorescence + P. lilacinus) @ 15 ml per kg soil untreated inoculated check. In general, fresh shoot
(151.1 cm), followed by Paecilomyces lilacinus @ weight was significantly less in all the treatments
0.5 g per kg soil (145.7 cm) irrespective of whether compared to untreated uninoculated check
nematode inoculated individually or concomitantly. irrespective of whether inoculated individually
However, in plants inoculated with nematode or concomitantly with nematode and fungus.
alone, shoot length was maximum in case of liquid Maximum reduction in fresh shoot weight was
formulation of bio-agents (155.8 cm), followed by observed in the presence of both nematode and
P .lilacinus (151.7 cm) as compared to untreated fungus followed by fungus alone and minimum in
inoculated check (87.5 cm). Plants inoculated case of nematode alone.
with fungus alone, shoot length was maximum Dry root weight in all the treatments
in case of liquid formulation of bio-agents (149.6 was significantly better over untreated inoculated
cm), followed by Trichoderma viride @ 0.5 g checks viz., nematode alone (2.19 g), fungus alone
per kg soil (143.9 cm) as compared to untreated (2.00 g ) and nematode + fungus simultaneously
inoculated check (85.6 cm). Plants inoculated with (1.76 g). Among the various treatments, maximum
nematode and fungus concomitantly, shoot length dry root weight was observed in liquid formulation
was maximum in case of liquid formulation of of bio-agents (T. viride + P. fluorescence + P.

J PURE APPL MICROBIO, 11(4), DECEMBER 2017.

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lilacinus) @ 15 ml per kg soil (6.57 g), followed in plants inoculated with nematode alone, dry root
by Paecilomyces lilacinus @ 0.5 g per kg soil (5.56 weight was maximum in case of liquid formulation
g) irrespective of whether nematode or fungus of bio-agents (7.27 g), followed by P. lilacinus
inoculated individually or concomitantly. However, (6.7 g) as compared to untreated inoculated check

Table 1. Effect of soil treatment with bio-agents on shoot

length (cm) of cucumber infested with M. incognita and fungus

Treatments Nematode Fungus Nematode Mean

alone alone + fungus

T1: Trichoderma viride @ 0.3 g/pot 136.5 131.8 126.6 131.6

T2: Trichoderma viride @ 0.5 g/pot 145.6 143.9 137.9 142.4
T3: Pseudomonas fluorescence @ 0.3 g/pot 138.1 128.6 130.8 132.5
T4: Pseudomonas fluorescence @ 0.5 g/pot 149.1 141.1 140.9 143.7
T5: Paecilomyces lilacinus @ 0.3 g/pot 140.4 127.7 134.6 134.2
T6: Paecilomyces lilacinus @ 0.5 g/pot 151.7 138.6 146.9 145.7
T7: Liquid formulation of bio-agents (T. viride + 142.6 135.9 134.8 137.8
P. fluorescence + P. lilacinus) @ 10 gm/ pot
T8: Liquid formulation of bio-agents @ 15 gm/ pot 155.8 149.6 147.9 151.1
T9: Carbofuran @ 0.1 g/ pot 159.2 121.7 155.9 145.6
T10: Drenching with Bavistin @ 2 g/l water 119.6 154.2 122.2 132.0
T11: Untreated check (inoculated) 87.5 85.6 83.9 85.7
T12: Untreated check (uninoculated) 164.8 165.4 166.1 165.4
Mean 140.9 135.3 135.7

CD @ 5% level
Treatment: 1.4
Sub treatment: 2.8
Treatment X Sub treatment: 4.9

Table 2. Effect of soil treatment with bio-agents on dry

shoot weight (g) of cucumber infested with M. incognita and fungus

Treatments Nematode Fungus Nematode Mean

alone alone + fungus

T1: Trichoderma viride @ 0.3 g/pot 12.49 11.7 10.50 13.24
T2: Trichoderma viride @ 0.5 g/pot 19.47 17.6 16.24 17.85
T3: Pseudomonas fluorescence @ 0.3 g/pot 14.49 13.7 12.49 13.91
T4: Pseudomonas fluorescence @ 0.5 g/pot 19.39 18.3 16.99 18.10
T5: Paecilomyces lilacinus @ 0.3 g/pot 15.74 14.7 13.75 14.08
T6: Paecilomyces lilacinus @ 0.5 g/pot 20.24 19.0 17.50 18.86
T7: Liquid formulation of bio-agents (T. viride + 18.65 16.9 15.49 16.55
P. fluorescence + P. lilacinus) @ 10 gm/ pot
T8: Liquid formulation of bio-agents @ 15 gm/ pot 21.94 20.7 19.07 21.13
T9: Carbofuran @ 0.1 g/ pot 23.50 22.5 21.24 22.66
T10: Drenching with Bavistin @ 2 g/l water 11.75 24.0 12.74 16.16
T11: Untreated check (inoculated) 5.82 5.6 4.76 5.56
T12: Untreated check (uninoculated) 24.75 25.0 24.49 24.75
Mean 17.35 17.48 15.44

CD @ 5% level
Treatment: 0.74
Sub treatment: 1.49
Treatment X Sub treatment: 2.59

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(2.19 g). Plants inoculated with fungus alone, check (2.0 g). Plants inoculated with nematode
dry root weight was maximum in case of liquid and fungus concomitantly, dry root weight was
formulation of bio-agents (6.70 g), followed by T. maximum in case of liquid formulation of bio-
viride (6.30 g) as compared to untreated inoculated agents (5.78 g), followed by P. lilacinus (4.99 g)
Table 3. Effect of soil treatment with bio-agents on dry root
weight (g) of cucumber infested with M. incognita and fungus

Treatments Nematode Fungus Nematode Mean

alone alone + fungus

T1: Trichoderma viride @ 0.3 g/pot 3.86 4.2 3.20 3.75

T2: Trichoderma viride @ 0.5 g/pot 5.04 6.3 4.26 5.19
T3: Pseudomonas fluorescence @ 0.3 g/pot 4.04 3.8 3.45 3.75
T4: Pseudomonas fluorescence @ 0.5 g/pot 5.43 5.2 4.74 5.11
T5: Paecilomyces lilacinus @ 0.3 g/pot 4.42 3.5 3.67 3.86
T6: Paecilomyces lilacinus @ 0.5 g/pot 6.75 4.9 4.99 5.56
T7: Liquid formulation of bio-agents (T. viride + 4.72 4.5 3.93 4.36
P. fluorescence + P. lilacinus) @ 10 gm/ pot
T8: Liquid formulation of bio-agents @ 15 gm/ pot 7.27 6.7 5.78 6.57
T9: Carbofuran @ 0.1 g/ pot 8.12 3.6 7.81 6.50
T10: Drenching with Bavistin @ 2 g/l water 4.09 7.3 3.72 5.01
T11: Untreated check (inoculated) 2.19 2.0 1.76 1.99
T12: Untreated check (uninoculated) 9.01 8.8 8.50 8.75
Mean 5.40 5.05 4.65

CD @ 5% level
Treatment: 0.43
Sub treatment: 0.87
Treatment X Sub treatment: 1.51

Table 4. Effect of soil treatment with bio-agents on number

of galls/plant of cucumber infested with M. incognita and fungus

Treatments Nematode Nematode Mean

alone + fungus

T1: Trichoderma viride @ 0.3 g/pot 212 (14.6) 206 (14.4) 209 (14.5)
T2: Trichoderma viride @ 0.5 g/pot 179 (13.4) 172 (13.2) 175.5 (13.3)
T3: Pseudomonas fluorescence @ 0.3 g/pot 205 (14.4) 194 (13.9) 199.5 (14.1)
T4: Pseudomonas fluorescence @ 0.5 g/pot 174 (13.3) 166 (12.9) 170 (13.1)
T5: Paecilomyces lilacinus @ 0.3 g/pot 192 (13.9) 185 (13.6) 188.5 (13.8)
T6: Paecilomyces lilacinus @ 0.5 g/pot 167 (13.0) 161 (12.7) 164 (12.8)
T7: Liquid formulation of bio-agents (T. viride + 186 (13.7) 180 (13.4) 183 (13.6)
P. fluorescence + P. lilacinus) @ 10 gm/ pot
T8: Liquid formulation of bio-agents @ 15 gm/ pot 160 (12.7) 153 (12.4) 156.5 (12.6)
T9: Carbofuran @ 0.1 g/ pot 153 (12.4) 145 (12.1) 149 (12.3)
T10: Drenching with Bavistin @ 2 g/l water 217 (14.8) 208 (14.4) 212.5 (14.6)
T11: Untreated check (inoculated) 313 (17.7) 307 (17.6) 310 (17.6)
T12: Untreated check (uninoculated) 0 (1.0) 0 (1.0) 0.0 (1.0)
Mean 12.9 12.6

Data in parenthesis are the square root (“n+1) transformed values of respective data
CD @ 5% level
Treatment: 0.04
Sub treatment: 0.09
Treatment X Sub treatment: 0.14

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as compared to untreated inoculated check (1.76 nematode alone. In general, dry root weight was
g). Maximum reduction in dry root weight was significantly lesser in all the treatments compared
observed in the presence of nematode and fungus to untreated uninoculated check irrespective of
followed by fungus alone and minimum in case of
Table 5. Effect of soil treatment with bio-agents on number of
egg masses/plant of cucumber infested with M. incognita and fungus

Treatments Nematode Nematode Mean

alone + fungus

T1: Trichoderma viride @ 0.3 g/pot 222 (14.9) 204 (14.3) 213 (14.6)
T2: Trichoderma viride @ 0.5 g/pot 173 (13.2) 164 (12.9) 168.5 (13.0)
T3: Pseudomonas fluorescence @ 0.3 g/pot 215 (14.7) 195 (14.0) 205 (14.4)
T4: Pseudomonas fluorescence @ 0.5 g/pot 162 (12.8) 154 (12.4) 158 (12.6)
T5: Paecilomyces lilacinus @ 0.3 g/pot 195 (14.0) 186 (13.7) 190.5 (13.8)
T6: Paecilomyces lilacinus @ 0.5 g/pot 156 (12.5) 149 (12.2) 152.5 (12.4)
T7: Liquid formulation of bio-agents (T. viride + 183 (13.6) 173 (13.2) 178 (13.4)
P. fluorescence + P. lilacinus) @ 10 gm/ pot
T8: Liquid formulation of bio-agents @ 15 gm/ pot 149 (12.2) 144 (12.1) 146.5 (12.1)
T9: Carbofuran @ 0.1 g/ pot 128 (11.3) 122 (11.1) 125 (11.2)
T10: Drenching with Bavistin @ 2 g/l water 345 (18.6) 342 (18.5) 343.5 (18.5)
T11: Untreated check (inoculated) 454 (21.3) 448 (21.2) 451 (21.2)
T12: Untreated check (uninoculated) 0.0 (1.0) 0.0 (1.0) 0.0 (1.0)
Mean 13.3 13.1

Data in parenthesis are the square root (“n+1) transformed values of respective data
CD @ 5% level
Treatment: 0.04
Sub treatment: 0.11
Treatment X Sub treatment: 0.16

Table 6. Effect of soil treatment with bio-agents on final nematode

population/200 cc soil of cucumber infested with M. incognita and fungus

Treatments Nematode Nematode Mean

alone + fungus

T1: Trichoderma viride @ 0.3 g/pot 226 (15.0) 216 (14.7) 221 (14.9)
T2: Trichoderma viride @ 0.5 g/pot 189 (13.8) 182 (13.5) 185.5 (13.6)
T3: Pseudomonas fluorescence @ 0.3 g/pot 216 (14.7) 205 (14.4) 210.5 (14.5)
T4: Pseudomonas fluorescence @ 0.5 g/pot 185 (13.6) 179 (13.4) 182 (13.5)
T5: Paecilomyces lilacinus @ 0.3 g/pot 205 (14.4) 198 (14.1) 201.5 (14.2)
T6: Paecilomyces lilacinus @ 0.5 g/pot 180 (13.4) 176 (13.3) 178 (13.4)
T7: Liquid formulation of bio-agents (T. viride + 194 (13.9) 188 (13.7) 191 (13.8)
P. fluorescence + P. lilacinus) @ 10 gm/ pot
T8: Liquid formulation of bio-agents @ 15 gm/ pot 175 (13.2) 168 (13.0) 171.5 (13.1)
T9: Carbofuran @ 0.1 g/ pot 166 (12.9) 154 (12.4) 160 (12.7)
T10: Drenching with Bavistin @ 2 g/l water 466 (21.6) 452 (21.3) 459 (21.4)
T11: Untreated check (inoculated) 644 (25.4) 637 (25.3) 640.5 (25.3)
T12: Untreated check (uninoculated) 0 (1.0) 0 (1.0) 0.0 (1.0)
Mean 14.4 14.1

Data in parenthesis are the square root (“n+1) transformed values of respective data
CD @ 5% level
Treatment: 0.03
Sub treatment: 0.09, Treatment X Sub treatment: 0.13

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whether inoculated individually or concomitantly inoculated checks viz., nematode alone (313) and
with nematode and fungus. nematode + fungus simultaneously (307). Among
Number of galls per plant in all the the various treatments, minimum number of galls
treatments was significantly reduced over untreated per plant was observed in liquid formulation

Table 7. Effect of soil treatment with bio-agents on fungus

incidence (%) of cucumber infested with M. incognita and fungus

Treatments After 15 days After 30 days Mean

T1: Trichoderma viride @ 0.3 g/pot 30 (33.4) 35 (36.5) 32.5 (35.0)

T2: Trichoderma viride @ 0.5 g/pot 20 (26.9) 25 (30.3) 27.5 (28.6)
T3: Pseudomonas fluorescence @ 0.3 g/pot 25 (30.2) 30 (33.4) 27.5 (31.8)
T4: Pseudomonas fluorescence @ 0.5 g/pot 20 (26.9) 25 (30.2) 22.5 (28.5)
T5: Paecilomyces lilacinus @ 0.3 g/pot 25 (30.2) 33 (34.9) 29 (32.5)
T6: Paecilomyces lilacinus @ 0.5 g/pot 15 (23.0) 20 (26.9) 17.5 (25.0)
T7: Liquid formulation of bio-agents (T. viride + 25 (30.2) 25 (30.2) 25 (30.2)
P. fluorescence + P. lilacinus) @ 10 gm/ pot
T8: Liquid formulation of bio-agents @ 15 gm/ pot 15 (23.0) 15 (23.0) 15 (23.0)
T9: Carbofuran @ 0.1 g/ pot 30 (33.4) 35 (36.4) 32.5 (34.9)
T10: Drenching with Bavistin @ 2 g/l water 0 (4.1) 10 (18.6) 5 (11.3)
T11: Untreated check (inoculated) 60 (51.1) 75 (60.4) 67.5 (55.7)
T12: Untreated check (uninoculated) 0 (4.1) 0 (4.1) 0.0 (4.1)
Mean 26.4 30.4

Data in parenthesis are the angular transformed values

CD @ 5% level
Treatment: 1.5
Sub treatment: 3.7
Treatment X Sub treatment: 5.2

Table 8. Effect of soil treatment with bio-agents on nematode +

fungus incidence (%) of cucumber infested with M. incognita and fungus

Treatments After 15 days After 30 days Mean

T1: Trichoderma viride @ 0.3 g/pot 35 (36.5) 40 (39.5) 37.5 (38.0)

T2: Trichoderma viride @ 0.5 g/pot 25 (30.3) 30 (33.5) 27.5 (31.9)
T3: Pseudomonas fluorescence @ 0.3 g/pot 35 (36.5) 40 (39.5) 37.5 (38.0)
T4: Pseudomonas fluorescence @ 0.5 g/pot 25 (30.3) 25 (30.3) 25 (30.3)
T5: Paecilomyces lilacinus @ 0.3 g/pot 30 (33.4) 35 (36.5) 32.5 (35.0)
T6: Paecilomyces lilacinus @ 0.5 g/pot 15 (23.0) 20 (26.9) 17.5 (25.0)
T7: Liquid formulation of bio-agents (T. viride + 30 (33.4) 30 (33.5) 30 (33.5)
P. fluorescence + P. lilacinus) @ 10 gm/ pot
T8: Liquid formulation of bio-agents @ 15 gm/ pot 15 (23.0) 20 (26.9) 17.5 (25.0)
T9: Carbofuran @ 0.1 g/ pot 10 (18.6) 10 (18.6) 10 (18.6)
T10: Drenching with Bavistin @ 2 g/l water 40 (39.5) 45 (42.4) 42.5 (41.0)
T11: Untreated check (inoculated) 65 (54.1) 85 (68.2) 75 (61.2)
T12: Untreated check (uninoculated) 0 (4.1) 0 (4.1) 0.0 (4.1)
Mean 30.2 33.3

Data in parenthesis are the angular transformed values

CD @ 5% level
Treatment: 1.2
Sub treatment: 3.0
Treatment X Sub treatment: 4.2

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of bio-agents (T. viride + P. fluorescence + P. compared to untreated inoculated check (65 %).
lilacinus) @ 15 ml per kg soil (156), followed by At 30 days after sowing, disease incidence was
Paecilomyces lilacinus @ 0.5 g per kg soil (164) minimum (15 %) in case of soil treated with liquid
irrespective of whether nematode inoculated formulation of bio-agents followed by 20 % in case
individually or concomitantly. However, in of P. lilacinus) @ 0.5 g per kg soil as compared to
plants inoculated with nematode alone, number untreated inoculated check (85 %).
of galls per plant was minimum in case of liquid
formulation of bio-agents (160), followed by P. DISCUSSION
lilacinus (167) as compared to untreated inoculated
check (313). Plants inoculated with nematode and Cucumber is highly susceptible to M.
fungus concomitantly, number of galls per plant incognita and F. oxysporum disease complex as
was minimum in case of liquid formulation of indicated by severity in root-knot development,
bio-agents (153), followed by P. lilacinus (161) nematode population densities, root colonization
as compared to untreated inoculated check (307). by fungus and plant growth suppression in the
Maximum reduction in number of galls per plant inoculated controls. Our results indicated that
was observed in the presence of both nematode and carbofuran is most effective among the treatments
fungus followed by nematode alone. in improving plant growth and reducing M.
Nematode population J2/200 cc soil in incognita population densities in soil. Carbofuran
all the treatments was significantly reduced over impairs nematode neuromuscular activity by
untreated inoculated checks viz., nematode alone inhibiting the function of the enzyme acetyl
(644) and nematode + fungus simultaneously (637). cholinesterase resulting in reduced movement
Among the various treatments, minimum final and ability of invasion and multiplication (Evans,
nematode population J2/200 cc soil was observed 1973; Wright, 1981). The nematodes may also be
in liquid formulation of bio-agents (T. viride + P. killed while feeding on root tissues by the systemic
fluorescence + P. lilacinus) @ 15 ml per kg soil action of these nematicides when they are absorbed
(171), followed by Paecilomyces lilacinus @ 0.5 g by the plant roots and translocated in the plant
per kg soil (178) irrespective of whether nematode system (van Berkum and Hoestra, 1979). Abuzar
inoculated individually or concomitantly. However, (2003) found similar effectiveness of carbofuran
in plants inoculated with nematode alone, final in suppressing M. incognita on Abelmoschus
nematode population J2/200 cc soil was minimum esculetus. Bavistin was found most effective in
in case of liquid formulation of bio-agents (175), controlling root colonization by fungus. It inhibits
followed by P. lilacinus (168) as compared to the nuclear division of fungi by inactivating the
untreated inoculated check (644). Plants inoculated spindle, which is composed of microtubules.
with nematode and fungus concomitantly, final Bavistin as an important control measure
nematode population J2/200 cc soil was minimum against F. oxysporum (Prasad et al., 2000; Haseeb
in case of liquid formulation of bio-agents (180), and Shukla, 2002; Abuzar, 2003, Haseeb et at.,
followed by P. lilacinus (176) as compared to 2006). To maintain a low inoculum load by
untreated inoculated check (637). Maximum continuous application of systemic fungicide
reduction in nematode population J2/200 cc soil alone is not practical for the control of wilt
was observed in the presence of both nematode disease. To cope with this, A. indica seed powder
and fungus followed by nematode alone. may be applied. It is clear from the results that
In general, all the treatments significantly besides chemicals A. indica seed powder were
reduced incidence of nematode and fungus sufficiently effective against both the pathogens,
concomitantly on cucumber as compared to this may be due to presence of active principles
untreated inoculated check. Data were recorded 15 and toxic chemicals in A. indica cake (Abuzar and
and 30 days after sowing. At 15 days after sowing, Haseeb, 2009; Abuzar and Haseeb, 2010). Initial
disease incidence was minimum (15 %) in case of investigations on antagonistic rhizobacteria against
soil treated with liquid formulation of bio-agents root-knot nematodes; include work by Kloepper
(T. viride + P. fluorescence + P. lilacinus) @ 15 ml et al. (1992). P. fluorescens was found not only
per kg soil or P. lilacinus) @ 0.5 g per kg soil as effective against M. incognita but also against wilt

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 1917

causing fungi. Results show that the Meloidogyne 4. Evans A A F. Mode of action of nematicides.
incognita- Fusarium oxysporum disease complex Annals of Applied Biology, 1973; 75:469-473.
can cause severe yield losses in V. radiata as in 5. Haseeb A and Shukla P K. Management of
other crops. Although chemicals viz. carbofuran wilt disease of chickpea by the application of
chemicals, bio-pesticides and bioagents under
and Bavistin showed a significant effect in increase
field conditions. Current Nematology, 2002;
of growth parameters and in suppression of the 13:61-63.
disease complex, these can be replaced to some 6. Haseeb A, Sharma A and Shukla P K Effect of
extent by A. indica neem cake avoid the hazards Different Initial Inoculum Levels of Fusarium
of chemicals. oxysporum on Vigna radiata. 2005; Proceedings
of 7th Indian Agricultural Scientists and Farmer’s
ACKNOWLEDGEMENT Congress, Meerut, p.1.
7. Kloepper J W, Rodriguez-kabana R, M C
The financial assistance provided by Inroy, J A and Young R W. 1992. Rhizospheric
bacteria antagonistic to soybean cyst (Heterodera
Department of Science and Technology (DST),
glycines) and root-knot (Meloidogyne incognita)
government of India, New Delhi, in the form of nematodes: Identification by fatty acid analysis
INSPIRE Fellowship to carry out this research and frequency of biological control activity. Plant
work is greatly acknowledged. I acknowledged to and Soil, 2005; 139:75-84.
Miss Saroj for assisting me during the experiment. 8. Oostendrop M and Sikora R A. Utilization of
antagonistic rhizobacteria as a seed treatment for
REFERENCES the biological control of Heterodera schachtii in
sugarbeet. Revue de Nematology, 1989; 12:77-
1. Abuzar, S. Studies on the interactive effects of 83.
root-knot nematode (Meloidogyne incognita) 9. Prasad K V V and Rao, O.P, Agrawal, S.C.
and wilt fungus (Fusarium oxysporum f. sp. Control of seed borne Fusarium spp. in lentil.
vasinfectum) on the growth of okra and its Journal of Mycology and Plant Pathology, 2000;
management. 2003; M.Sc. Dissertation submitted 30:256.
in Department of Plant Protection, A.M.U. 10. Van berkum J A and Hoestra H. Practical Aspects
Aligarh India. of the Chemical Control of Nematicides in Soil.
2. Abuzar S and Haseeb A. Bio-Management of In: Mulde, D. (Ed.), Soil Disinfestation. Elsevier,
Plant-Parasitic Nematodes in Pigeon Pea Field Amsterdam, 1979; p.53-134.
Crop Using Neem-Based Products and Manurial 11. Wright D J. Nematicides: Mode of Action and
Treatments. World Applied Sciences Journal, New Approaches to Chemical Control. In:
2009; 7(7): 881-884. Zukerman, B.M., Rhode, R.A. (Eds.), Plant
3. Abuzar S and Haseeb A. Plant Growth and Parasitic Nematodes. Vol. 3, Academic Press,
Plant Parasitic Nematodes in Response to Soil New York & London, 1981; p.421-449.
Amendments with Plant Growth Promoting 12. Minuto A, Gullino M L, Lamberti F D, Adabbo
Rhizobacteria and Inorganic Fertilizer in Pigeon T, Tescari E and Ajwa, H. Garibaldi. Application
Pea, Cajanus cajan L.World Applied Sciences of emulsifiable mixture of 1,3 Dichloropropene
Journal. 2010; 8 (4): 411-413. and chloropicrin against root knot nematode and
soil fungi for green house tomato in Italy. Crop
Protection, 2006; 25, 1244-1252.

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