Screening of entomopathogenic nematodes for the management ... · PDF fileScreening of entomopathogenic nematodes for the management ... determine the effect of Entomopathogenic nematodes

19 Raza et al.

Int. J. Biosci. 2015

RESEARCH PAPER OPEN ACCESS

Screening of entomopathogenic nematodes for the management

of Meloidogyne incognita in Brinjal

Muhammad Subtain Raza1, Muhammad Imran2, Tahira Yasmin1, Muhammad Azeem3 ,

Hira Manzoor4, Muhammad Awais5*

1Department of Environmental Sciences, GC University, Faisalabad, Pakistan

2Directorate of Agriculture, Pest Warning & Quality Control of Pesticides, Punjab, Lahore,

Pakistan

3Department of Soil Science, Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi, Pakistan

4Department of Plant pathology, University of Agriculture, Faisalabad, Pakistan

5Department of Plant Breeding & Genetic, university of Agriculture, Faisalabad,Pakistan

Key words: Brinjal, Entomopathogenic Nematodes , Plant Parasitic Nematodes,Screening.

http://dx.doi.org/10.12692/ijb/6.11.19-31 Article published on June 08, 2015

Abstract

Nematodes cause severe damage to economically important vegetable crops. Meloidogyne spp. an important

group of nematodes that reduce both the quantity and quality of vegetables. The present study was conducted to

determine the effect of Entomopathogenic nematodes for the management of Meloidogyne incognita. Effect of

Entomopathogenic nematodes native and already available (Heterorhbiditis indica, H. bactereophora,

Steinernema krussi and S. feltiae) was evaluated for invasion and development of M. incognita in brinjal. The

results revealed that S. felti was most effective in reducing numbers of galls, females, egg masses and

reproduction factor in screening experiment. Findings of this investigation may suggest that EPN can be

successfully used for the management of root knot nematodes.

* Corresponding Author: Muhammad Awais [email protected]

International Journal of Biosciences | IJB |

ISSN: 2220-6655 (Print), 2222-5234 (Online)

http://www.innspub.net

Vol. 6, No. 11, p. 19-31, 2015

http://dx.doi.org/10.12692/ijb/6.11.19-5

http://www.innspub.net/

20 Raza et al.


Introduction

Brinjal or eggplant (Solanum melongena L.) is a low

priced and one of the most common grown vegetable

crop in Pakistan. It contains sufficient amount of

vitamin A and B. (FAO, 2009). There are various

restrictive factors in the production of crop but root

knot nematodes (Meloidogyne spp.) are the most

destructive one that tremendously reduce both

quality and quantity of crop produce. Meloidogyne

spp. is an important group of nematodes and results

in reducing both the quantity and quality of products

in Punjab as a result of intensive agriculture (Anwar

and Mckenry, 2010). Nematodes cause severe damage

to economically important vegetable crops. In

Pakistan Meloidogyne spp is found with proportion

e.g., M. incognita 58 %, M. javanica 31 %, M.

arenaria 8 %, M. halpa 7 % and other species are

about 2% (Maqbool, 1986). Sasser (1980) reported

that out of the 80 Meloidogyne species, M. incognita,

47%; Meloidogyne javanica, 40%; M. arenaria, 7%

and M. hapla, 6% is are important regarding their

devastating impacts . The warmer climates allow a

longer growing season with more susceptible crops

being grown per year and this together with shorter

life cycle can lead to high nematode reproduction

rates and severe crop losses. Sikora and Fernandez

(2005) described that root knot nematodes have high

reproductive potential that’s why it is difficult to

manage.

Microbial control agents (i.e. viruses, bacteria, fungi,

protozoa and nematodes) have also become major

contributors in biological control of pests.

Entomopathogenic nematodes are categorized into

two families Steinernematidae and

Heterorhabditidae. Steinernematidae is

characterizeed by the genera Steinernema and

Neoste inernema and Heterorhabditidae is

denoted by the genus Heteror habdit is (Kaya

and Gaugler, 1993; Burnell and Stock, 2000).

Entomopathogenic Nematodes are correlated with

same bacteria in the genus Xenor habdus for

Steinernematidae and Photor habdus for

Heterorhabditidae (Boemare, 2002). Amongst the

various biological control microorganisms are

utilized Pasturia penetrans, egg-parasitic pathogens

fungi, nematode-trapping fungi, bacteria, and

polyphagous predatory nematodes are generally

employed (Kerry, 1988; Kiewnick and Sikora, 2005).

EPN also worn as bioinsecticides (Georgis and

Manweiler, 1994). Inundative pathogens can b

produced by crop plants and other microorganisms in

which proteinaceous toxins can be induced

(Somasekhar et al., 2002). Bird and Bird (1986)

conducted several experiments to find the way of

reduction of parasitic nematodes by the using

entomopathogenic nematodes. They proposed three

mechanisms: i. Crowding effect of nematodes with

the roots that force away the plant parasitic

nematodes. ii. Antagonistic effect of

entomopathogenic nematodes due to massive doses

that leads to the reduction of parasitic nematodes in

the soil. iii. Allelochemicals that are manufactured by

entomopathogenic nematodes, repel or intoxicate the

effect of plant parasitic nematodes. Ishibashi and

Kondo (1986) made experiments and observed that

the soil in sealed containers having

entomopathogenic nematodes had lesser plant

parasitic nematodes than the control soil. They made

various green house experiments on different tomato

cultivars and concluded that by the application of

Steinernema glaseri the population of Meloidogyne

javanica was suppressed. They also observed that the

population of plant nematodes was reduced with the

application of Steinernerna glaseri and Steinernerna

feltiae to non- sterilized soil or bark compost but it

increases the Rhabditoid nematodes. Akhurst (1990)

suggested that the symbiotic bacteria of

entomopathogenic nematodes releases substances

with high antibiotic activity that protects the cadavers

from the invasion of plant parasitic nematodes into

the roots. Georgis and Manweiler (1994) reported the

bacteria associated with entomopathogenic

nematodes Steinernema and Heterorhabditis are

Xenorhabdus spp. are commercially available to

control soil insect pest. They studied that

entomopathogenic nematodes are adaptive to soil and

are efficient as a biological management against a

number of insect pests. Verschoor and De Goede

(2000) reported that entomopathogenic nematodes

21 Raza et al.


can be detected by placing nematode susceptible

insect larvae into the soil samples. The nematodes

present in the soil infect and kill the larvae. After that

nematodes can be isolated from the insect cadavers.

Perez and Lewis (2002) recently has been

demonstrated that EPNs can affect the number of

root knot nematode infecting plants, or the number of

eggs produced when they are apply near the root

system. However the application of the bacterium has

not shown a consistent suppression of root knot

nematode in some studies. Entomopathogenic

nematodes have been used as biological management

against different insect pests. These nematodes are

soil-dwelling organisms and are obligate parasites of

insects. The only stage which lives freely on the soil is

the infective juvenile (J3) and to complete their life

cycles, those J3s must find a suitable host. When they

locate the host, penetrate into them through the body

natural openings (Poinar, 1979) or also through the

cuticle in Heterorhabditidae (Bedding and Molyneux,

1982), reaching the hemocoel and releasing a

symbiotic bacterium (Xenorhabdus in Steinernema

and Photorhabdus in Heterorhabditis) which starts

reproducing and finally kills the insect by septicaemia

between 24 to 72 hours. Some species of EPN such S.

glaseri, S. carpocapsae and H. megidis have been

reported on many vegetable root plants (Bird and

Bird, 1986; Kanagy and Kaya, 1996) or their exudates

when insect consume them (Rasmann et al., 2005),

probably as a result of a defensive strategy used by

plants to protect themselves from insect attacks.

Recently it has been demonstrated that EPNs can

affect the number of RKN infecting plants, or the

number of eggs produced when they are apply near

the root system (Fallon et al., 2004; Perez and

Lewis,2004). The explanation of this effect has been

theorized as a result of an allelopathic response

produced by the symbiotic bacteria of EPNs which is

repellent to Root knot nematode (Grewal et al., 1999),

however the application of the bacterium has not

shown a consistent suppression of nematode in some

studies.

Various strategies have been used extensively over the

years in Pakistan to manage nematodes and use of

bio-products in Integrated Pest Management (IPM) is

now accepted as an ecologically sound and

economically viable alternative to chemical pest

control. Thus, the present Study was planned to

Screening of entamopathogenic nematode for the

management of M. incognita in Brinjal plant and

evaluation of different species of EPN for their effect

against invasion, development and reproduction of

M. incognita infection.

Material and methods

Preparation of nematode inoculums

Preparation and sterilization of Soil

The soil was thoroughly mixed and air dried by

spreading in a thin layer on woody bench, covered

with plastic sheath. After drying the large stones and

plant debris was removed. After this, mechanical

analysis of the soil was performed. The sterilization

of sandy loam soil (70% sand, 22% silt, 8% clay) was

accomplished by applying formalin (1:20). Diluted

formalin was poured in the small heap of soil and

covered with polythene sheet to stop the fumes

completely. This process continued for a week. After a

week the soil was exposed to get rid of residual

formalin. The soil was mixed l thoroughly and then

filled the empty pots.

Generating and Transplanting of egg plant seedlings

Seedling of eggplant cv Dilnasheen was raised in

sterilized sandy loam soil in 20-cm diameter. earthen

pots. They were allowed to grow for 30-days.The

small earthen pots of 13-cm diameter were filled with

soil. One day before transplanting the seedlings, the

soil was mixed by sand and clay in ratio 75:25 and

watered to settle the soil. Seedlings were removed

from pots carefully and then adhering soil was

removed gently by shaking and then transplanted

singly in pots.

Single egg mass culture of M. incognita

A single egg mass of M. incognita was picked with a

fine forceps from galled egg plant root and then

inoculated at the base of each egg plant by making 3-

cm deep hole one week after transplanting. The holes

were covered with soil to prevent drying and

22 Raza et al.


immediately watered.

Identification of Meloidogyne spp.

A piece of fresh galled root from single egg mass

culture was harvested and placed in petri dish. The

root tissue was teased apart with forceps to remove

adult females and collected into a drop of 45% lactic

acid in another petri dish. A fully developed female

was placed over a clean glass slide and its posterior

half of the body was cut off using a surgical scalpel.

The lower piece of the cuticle having perineal patterns

was further trimmed to square shaped and inner

tissue was completely removed by flexible bristle. The

perineal pattern bearing portion was transferred to

drop of glycerin on a slide so that the anterior surface

was in contact with the glass and the anus was

oriented upwards. The glass cover slip was gently

placed and sealed with paraffin and labeled. The

pattern was examined under a research microscope

(Eisenback et al., 1981). Root knot nematodes species

were identified on the bases of perineal pattern

(Hartmann and sasser, 1985).

Inoculation of plants with M. incognita

Three holes (3-cm deep) around each plant were

made with the help of pointed wood. Approximately

2000 eggs of M. incognita were pipetted in these

holes held in a small volume of egg suspension. The

holes were covered with soil to prevent drying. One

day after inoculation these pots were watered

carefully to prevent loss of eggs through leaching or

excess drying.

Extraction of M. incognita eggs from infected

seedling

The roots were cut into 2-3-cm segments and shaken

vigorously (manually) for 3-4 minutes in a coffee jar

(1 liter) with a tightly fitted lid, containing 200 ml of

1% sodium hypochlorite (NaOCl) solution (Chlorax)

to dissolve the gelatinous matrix and to release the

eggs from the egg masses (Hussy and Barker, 1973).

This suspension was quickly passed through 200-

mesh (75 µm) sieve nested over 500-mesh (25 µm)

sieve to collect root fragments on the former and

freed eggs on the latter. The eggs collected on the

500-mesh sieve were rinsed with tap water to remove

the residual NaOCl. Rinsing of eggs was done for

several minutes. Then these freed eggs were collected

in a beaker. This process was repeated twice, for

removing additional eggs and getting rid of residual

Chlorax.

Screening of entomopathogenic nematodes for the

management of M. incognita in brinjal

Brinjal plants were inoculated with different

available and native species of EPN at density

1000/pot in brinjal plants at various times of

applications (5 days before, at the same time, and

after 5 days) of application of RKN (500/plant) was

studied. There were destructive harvest at 35 days to

assess the reproduction (No. of galls, galling index,

No. of egg masses, No. of females and reproductive

rate, final population / initial population) of M.

incognita in experimental plants.

Data Analysis

The data collected were analyzed under Complete

Randomized Design (CRD). The analysis of variance

(ANOVA) and the differences among means was

analyzed by applying LSD test at 5% level of

probability.(Steel et al., 1997).

Results

Effect on plant growth parameters when EPN

applied before 5 days of RKN

Root length

The effect of four EPN species was assessed on the

basis of the development of roots length. All the

treatments were statistically significant. Effects of

the entire four spp. were significantly different from

each other (Table 1.1). Steinernema feltiae was

found most effective as the maximum root length

(18.40A) was found in the plant inoculated with this

spp. Minimum root length (15.15D) was checked in

the plant inoculated with S. glaseri as compared to

control. The control has minimum root length

(11.84E) due to the RKN. Heterorhabditis

bacteriophora and H. indica were intermediate of

these two (Table 1.1).

23 Raza et al.


Root weight

Effects of four EPN species were evaluated on the

basis of roots weight when applied 5 days before the

application of RKN. All the treatments were

statistically significant (Table 4.1). The maximum

root weight was observed in the plant inoculated

with RKN only (control). Minimum root weight

(2.30E) was checked in the plant inoculated with S.

feltiae (Table 4.1) as compared to control. The least

effective spp. Found was S. glaseri as the maximum

root weight was observed in plant inoculated with this

spp. as compared to control (Table 1). The control has

maximum root weight (3.77A) due to the attack of

RKN. Heterorhabditis bacterophora (2.43D) and H.

indica (4.26C) were intermediate of these two (Table

1.1).

Table 1.1. Screening of different EPN spp on plant growth responses when EPN applied before 5 days

application of RKN.

Entomopathogenic nematodes

species

Plant growth parameters (EPN applied before 5 days application of RKN)

Root length Root weight Shoot length Shoot weight

S. glaseri 15.15 D* 2.74B 23.7 D 6.14D

S. feltiae 18.4A 2.3E 27.3 A 8.24A

H.bacteriophora 15.76 C 2.43 D 24.6 C 6.75 C

H. indica 17.3B 4 2.6 C 25.52B 4 7.5B

Control 11.8E 3.77A 15.4 E 5.2E

Means with in a column sharing the same letter are not significantly different from each other at P = 0.05

according to Least Significant Difference Test.

Table 1.2. Screening of different EPN spp on plant growth responses EPN and RKN applied at same time.

Entomopathogenic nematodes species Plant growth parameters (EPN and RKN applied at same time)


S. glaseri 14.2D* 2.9B 22.8D 5.15D

S. feltiae 17.3A 2.24D 26.16A 7.16A

H.bacteriophora 14.7C 2.50CD 24.1C 5.68C

H. indica 16.04B 2.70BC

24.954B

6.50B

Control 10.9E 3.94A 11.248E 4.12E



Shoot weight

Four EPN species were applied 5 day defore the

application of RKN and their effect on the

development of shoot weight were evaluated. All the

treatments were statistically significant. Effects of

the entire four spp. were significantly different from

each other (Table 4.1). S. feltiae was found most

effective, as the maximum shoot weight. Minimum

shoot weight was checked in the plant inoculated

with S. glassri as compared to control. The control

has minimum shoot weight due to the attack of

RKN. Shoot weight of the plants inoculated with

H.bacteriophora and H. indica were intermediate of

S. feltiae and S. glaseri (Table 1.1).

Shoot Length

Effects of four EPN species were evaluated on the

basis of shoots length. Effects of the entire four spp.

were significantly different from each other (Table

1.1). All the treatments were statistically significant.

Steinernema feltiae was found most effective as the

maximum shoot length was found in the plant

24 Raza et al.


inoculated with this spp. Minimum shoot length was

observed in the plant inoculated with S. glassri as

compared to control. The control has minimum

shoot weight due to the assault of RKN. Effect of H.

bacterophora and H. indica on the development

shoot weight were intermediate of these two (Table

1.1).

Table 1.3. Screening of different EPN spp on plant growth responses EPN applied after 5 days application of

RKN.

Entomopathogenic nematodes

species

Plant growth parameters (EPN applied after 5 days application of RKN)


S. glaseri 12.96D 3.06B 4.43D 21.78C

S. feltiae 16.26A 2.6D 6.52B 25.1A

H.bacteriophora 13.62C 2.75C 5.12C 22.9B

H. indica 15.18B 2.94B 8.92A 23.14B

Control 9.86E 4.16A 3.72E 10.58D



Table 1. 4. Screening of different EPN spp on nematode reproduction parameters EPN applied before 5 days

application of RKN.

Entomopathogenic

nematodes species

Nematode reproduction parameters (EPN applied before 5 days application of RKN)

Root galls Egg masses No of females Final population Rate of reproduction1

S. glaseri 91B 72 B 97 B 1260 B 2.52B

S. feltiae 30 E 24 E 35E 671 E 1.34 E

H.bacteriophora 83 C 59 C 92.2C 1022 C 2.04C

H. indica 57 D 43D 63 D 940 D 1.88D

Control 158 A 122 A 163 A 2280 A 4.56 A

1 Rate of reproduction = Pf/Pi (Final Population / Initial Population)

2 Means with in a column sharing the same letter are not significantly different from each other at P = 0.05


Effect on plant growth parameters when EPN and

RKN applied at the same time

Comparison of four EPN species on the basis of root

length

Effect of four EPN species were assess on the basis

of the development of roots length when applied

with RKN at the same time. All the treatments were

statistically significant. Effects of the entire four spp.

were significantly different from each other (Table

1.2). Steinernema feltaei was found most effective as

the maximum root length (17.30A) was found in the

plant vaccinated with this spp. Minimum root length

(14.20D) was observed in the plant inoculated with

S. glaseri as contrast to control. The control has

maximum root length (10.90E) due to the attack of

root knot nematode. H. bacterophora and H. indica

were intermediate of these two (Table 1.2).

Comparison of four EPN species on the basis of root

weight

Effect of four EPN species were evaluated on the

basis of roots weight when applied with RKN at the

same time. All the treatments were statistically

significant (Table 1.2). The maximum root weight

was checked in the plant inoculated with RKN only

(Control). Minimum root weight (2.24D) was

25 Raza et al.


observed in the plant inoculated with S. feltiae

(Table 1.2) as compared to control. The least effective

spp. was found S. glaseri as the maximum root

weight was observed in plant inoculated with this spp.

as compared to control (Table 1.2). The control has

maximum root weight (3.94A) due to the result of

RKN. Heterorhabditis. bacteriophora (2.50CD) and

H. indica (2.70BC) were intermediate of these two

(Table 1.2).

Table 1.5. Screening of different EPN spp on nematode reproduction parameters EPN and RKN applied at same

time.

Entomopathogenic

nematodes species

Nematode reproduction parameters (EPN and RKN applied at same time)


S. glaseri 100 B 80 B 109.4 B 1280 B 2.16 B

S. feltiae 40 E 32D 50.8 E 680E 1.36 C

H.bacteriophora 90 C 75 B 94 C 1031C 2.06 B

H. indica 65D 52 C 72 D 948D 1.89 B

Control 164.8 A 129.8 A 169.4 A 2285 A 4.57 A


2 Means with in a column sharing the same letter are not significantly different from each other at P = 0.05


Table 1.6. Screening of different EPN spp on nematode reproduction parameters EPN applied after 5 days

application of RKN.

Entomopathogenic

nematodes species

Nematode reproduction parameters (EPN applied after 5 days application of RKN)


S. glaseri 150 B2 100 B 160.40 B 1303.4 B 2.6 B

S. feltiae 60 E 60 D 71.800 E 708E 1.41E

H. bacteriophora 100 C 90C 107.80 C 1051.2 C 2.10 C

H. indica 70.D 65D 80.2 D 991D 1.98 D

Control 204.8 A 160 A 215.6 A 2320 A 4.64 A




Comparison of four EPN species on the basis of

Shoot Length

Effect of four EPN species was evaluated on the

basis of shoots length when applied with RKN at the

same time. Effect of the entire four spp. was

significantly dissimilar from each other (Table 1.2).

All the treatments were statistically significant.

Steinernemaa feltiae was found most effective as the

maximum shoot length (26.16A) was found in the

plant inoculated with this spp. Minimum shoot

length (22.80D) was checked in the plant inoculated

with S. glaseri as compared to control. The control

has minimum shoot lenth (11.24E) due to the

development of RKN. Effect of H. bacteriophora and

H. indica on the development shoot weight were

intermediate of these two (Table 1.2).

Comparison of four EPN species on the basis of

shoot weight

Four EPN species were applied at the same time

with RKN and their effect on the development of

shoot weight were evaluated. All the treatments

were shown significant results. Effect of the entire

four spp. was significantly dissimilar from each

26 Raza et al.


other (Table 1). Steinernema feltiae was found most

effective, as the maximum shoot weight (7.16A) was

found in the plant inoculated with RKN and S. felti.

Minimum shoot weigth (5.15) were checked in the

plant inoculated with S. glassri as compared to

control. The control has minimum shoot weight

(4.12E) due to the result of RKN. Shoot weight of the

plants inoculated with H.bacterophora and H.

indica were intermediate of S. feltiae and S. glaseri

(Table 1.2).

Fig. 1. Screening of different EPN spp on plant

growth responses when EPN applied before 5 days

application of RKN.

Effect on plant growth parameters when EPN

applied after 5 days of RKN

Root weight

Effect of EPNs spp. on the development of root

weight was noted (Table 1.3). All the treatments

were statistically significant. Minimum root weight

was checked in plants treated with S. feltiae. The

control has maximum root weight due to the attack

of RKN.

Root length

Effect of four EPN species was evaluated on the

basis of the development of roots length. S. feltiae

was found most effective as the maximum root

length was found in the plant inoculated with this

spp. Minimum root length was observed in the plant

inoculated with S. glaseri as compared to control.

The control has minimum root length due to the

assault of RKN. (Table 1.3).

Shoot weight

Maximum shoot weight was observed in treatment

with S. feltiae followed by H. bacteriophora and H.

indica (Table 1.3).

Shoot Length

Effect of EPN on the development of shoot length

was noted. Maximum shoot length was checked in

plant treated with S. feltiae (Table 4.3).

Effect on nematode reproduction parameters when

EPN applied before 5 days of RKN

Number of galls per root system

Number of galls was more significant (P = 0.05) on

control plant, S. felti was significant as compared to

other treatment. Comparison of treatment means

indicated that S. feltiae was more effective as

compared with other treatments (Table 1.4).

Number of egg mass

Minimum number of egg masses (24E) was observed

in S. feltiae followed by H. indica (43D).Maximum

number of egg masses were observed in control

treatment (Table 4.4).

Number of females per root system

Control showed highest number of females due to

pathogenic effect of nematodes and S. felti

treatment showed lowest number of females as

compared to other treatment. Significant number of

females was formed in control treatments (Table

1.4).


growth responses EPN and RKN applied at same

time.

Final Population

Effect of all the EPN spp. was statistically

significantly in reducing the final population of RKN

(Table 1.4). The minimum number of nematodes

27 Raza et al.


(671E) was checked in the plant treated with S.

feltiae as compared to control. Final population in

the plant treated with other three spp. S. glaseri, H.

bacteriophora and H. indica, was 1260B, 1022 C and

940 D respectively. The hightes final population was

found in the control (Table 1.4).

Rate of reproduction

When the EPNs were evaluated on the basis’s rate of

reproduction, all the treatment was found statistically

significantly. Highest rate of reproduction was

observed in the control plant to fallow this was plants

treated with S. glaseri , H. bacteriophora and H.

indica as these has statistically same rate of

reproduction. Minimum rate of reproduction (1.36)

was found in the plant treated with S. feltiae (Table

1.4).


growth responses EPN applied after 5 days

application of RKN.


EPN and RKN applied with at same time


Each EPN was found effective in reducing the

number of gall as compared to control (Table 1.5).


control plant. Minimum number of galls (40E) was

found in the plant treated with S. feltiae as

compared to control. Maximum number of galls

(100B) was found on plant treated with S. glassri as

compared to control. Evaluation of treatment

indicated that S. feltiae was more effective as

compared with other treatments. (Table 1.5).


Control showed maximum number of females

(169A) due to pathogenic effect of nematodes and

plant with S. felti showed the minimum number of

females as compared to other treatments. Significant

number of females was formed in other treatments

(Table 1.5).

Number of egg mass

Minimum number of egg masses (32D) was

observed in S. feltiae treatment followed by H.

indica (52C). Development of egg masses were

statistically same in the plant treated with S. glaseri

and H. bacteriophora. Highest number of egg masses

(129.8A) were checked in the plants treated with RKN

alone (control).

Final population




(680E) was checked in the plant treated with S. felti

as compared to control. Final population in the plant

treated with other three spp S. glaseri, H.

bacteriophora and H. indica, was 1280B, 1031C and

948D respectively. The highest final population was


Fig. 4. Screening of different EPN spp on nematode

reproduction parameters EPN applied before 5 days

application of RKN.



reproduction, all the treatment were found

statistically significantly. Highest rate of reproduction

(4.57A) was checked in the control plant to fallow this

was plants treated with (2.16B), H. bacterophora

(2.06B) and H. indica (1.89B), as these has

statistically same rate of reproduction. Minimum rate

28 Raza et al.


of reproduction (1.36C) was found in the plant treated

with S. glaseri.


EPN applied after 5 days of RKN



control plant. S. felti as protective was significant as

compared to other treatment. Comparison of

treatment means indicated that S. felti as curative

was more effective as compared with other

treatments. (Table 1.6).


Control showed highest number of females due to

pathogenic effect of nematodes (215.6A) and

application of S. feltiae showed lowest number of

females as compared to other treatment. Significant

higher number of females was formed in other

treatments as compared to control.


reproduction parameters EPN and RKN applied at

same time.

Number of egg mass

Minimum number of egg masses was observed in

plant treated with S. feltiae while higher no of egg

mass was shown by S. glaseri as compared to

control (100B).

Final reproduction




was observed in the plant treated with S. feltiae as

compared to control. Final population in the plant

treated with other three spp S. glaseri, H.

bacteriophora and H. indica, was 1303B, 1051 C and

991D respectively. The highest final population was




reproduction, all the treatment was found statistically

significantly. Highest rate of reproduction was

checked in the control plant to fallow this was plants

treated with S. glaseri, H. bacterophora and H.

indica. Minimum rate of reproduction (1.41E) was

found in the plant treated with S. feltiae (Table 1.6)

Discussion

Meloidogyne spp., are the important plant parasitic

nematodes associated with vegetable crops. The areas

having soil type sandy loam and loam has more

population of RKN. The soil was important factor

affecting nematode movement, rate of growth,

reproduction (Starr et al., 1993; Ogbuji, 2004) and

host suitability (Anwar et al., 2007). The four

entomopathogenic nematodes i.e., S, glaseri, S.

feltiae, H. indica and H. bacteriophora showed

variable response against M. incognita. Plant growth

parameters as well as nematode reproduction

parameters varied depending upon EPN Spp. applied

at three times of interval. Entomopathogenic

nematodes had a delaying result on the maturity of

RKN due to crowding effect, release of toxins by

mutualistic bacteria The involved mechanism of the

increase in root weight in affected plants might be

due to the larger amount of growth substances, more

tryptophan and other amino acids than un-inoculated

plants (Setty and Wheeler, 1968) and had inverse

impact on shoot length. There was inverse

relationship between shoot and root weight at all level

of M. incognita but these findings did not agree with

the hypothesis that root and shoot are mutually

dependent upon each other for exchanging nutrients,

carbohydrates, growth substances and are

physiologically in equilibrium and any reduction in

root growth limit the shoot growth or vice versa. So

these observations suggested that root weight was not

a good parameter for the assessment of plant growth.

It was being observed that an increase in the

inoculum level causes a progressive increase in the

29 Raza et al.


host infection as indicated by number of galls, gall

index and egg masses per root system. The statistical

analysis indicated that there was a direct relationship

between root gall and production of egg masses. Gall

index and total plant fresh weight showed inverse

relationship. The nematode multiplication was the

maximum at initial inoculum level and then started

decreasing at higher inoculum levels. It might be due

to the intra specific competition among nematodes

for food (Seinhorst, 1962).


reproduction parameters EPN applied after 5 days

application of RKN.

Conclusion

Effect of entomopathogenic nematodes native and

already available (Heterorhbiditis indica,

Heterorhbditis bactereophora, Steinernema krussi

and Steinernema feltiae) was evaluated for invasion

and development of M. incognita in brinjal. Invasion

and development of M. incognita was reduced when

Steinernema feltiae @ 2000/pot were applied to soil.

Steinernema feltiae was most effective in reducing no

of galls, no of females, no of egg masses and

reproduction factor in screening experiment. These

plants also had minimum juveniles per root system.

From the current investigation it may be concluded

that EPN can be successfully employed for the control

of root knot nematodes.

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