8. section-6 individual and interactive effect of...

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8. SECTION-6

Individual and interactive effect of different plant symbionts, bio-

organic wastes and antagonistic fungi in the management of root-

knot nematode, Meloidogyne incognita infecting chickpea var.

Avrodhi

8.1 INTRODUCTION

Chickpea (Cicer arietinum L.) is an important pulse crop of India and an

important source of protein in the vegetarian diet. Among the various constraints to

chickpea production, the plant-parasitic nematodes are one of the major threat (Sikora

and Greco, 1990). Although large number of nematodes have been associated with

chickpea, root-knot nematode Meloidogyne incognita (Kofoid and White) Chitwood

is highly prevalent.

Rhizosphere microorganisms utilize compounds and materials released from

the crop roots and provide microorganisms with nutrition. Consequently, the

rhizosphere supports large and active microbial population capable of exerting

beneficial, neutral or detrimental effects on plant growth. Certain species of

nodulating bacteria, such as Bradyrhizobium japonicum or other Rhizobium spp. are

applied as a form of soil enrichment to enhance chickpea production. However, in soil

where there are nematode infestations, the effectiveness of the soil enrichment with B.

japonicum may not be fully realized, if complimentary control measures such as

nematode antagonists or suppressants are not used (Azcon-de-aquilar et al., 1979;

Hillocks, 2001). The use of biological agents rather than harmful synthetic

nematicides may have added advantage when certain nodulating bacteria are used in

soil enrichment.

Chemicals that are being used for controlling plant-parasitic nematodes are

costly and hazardous in nature. Researchers all over the world are engaged in

standardizing the nematode management strategies by following non-chemical and

eco-friendly approaches such as soil amendments with botanicals (Sukul et al, 2001;

Rajendra and Saritha, 2005), organic soil amendments (Singh et al., 1990; Vedhera et

al, 1998; Nagesh and Reddy, 1997), biological control agents (Babu et al., 2000; Nico

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et al., 2004; Kantharaju et al., 2005; Sumathi et al., 2006) and judicious use of

nematicides (Taylor and Sasser, 1978) to stabilize crop production.

Arbuscular mycorrhizal fungi (AMF) are being widely used as biocontrol

agents as they can enhance growth and production of crops and also provide

protection against plant-parasitic nematodes (Azcon-de-Aguilar et al., 1979; Hussey

and Roncadori, 1982; Jeffries, 1987; Schonbeck et al., 1994; Mahaveer et al., 1994;

Dugassa et al., 1996; Hillocks, 2001; Gernns et al., 2001; Jothi and Sundarababu,

2002; Oyekanmi et al., 2007; Shreenivasa et al., 2007). Investigations carried out, so

far, had been mostly on the management of root-knot nematodes by utilizing AM

fungi indigenous isolates (Mishra and Shukla, 1997; Kantharaju et al., 2005). Both

AMF and root-knot nematodes are biotrophic and metabolic sink-inducing entophytic

organisms living on the same resources. Due to this fact, interaction between the

symbionts and the parasites are taken in the study.

Of the rhizosphere organisms, antagonistic fungi have great potential against

plant pathogens (Papavizas, 1985). Different Trichoderma spp. have also long been

recognized for their potential as biocontrol agents against soilborne, foliar and

postharvest phytopathogenic fungal pathogens and root-knot nematodes (Chet, 1987,

1990; Calvet et al., 1990; De et al., 1996; Reddy et al., 2000; Sharon et al., 2001;

Bandyopadhyay and Cardwell, 2003). These fungi may also promote plant growth

(Inbar et al., 1994) and have the ability to colonize root surfaces and the cortex

(Kleifeld and Chet, 1992; Yedidia et al., 1999). Application of Trichoderma resulted

increase in the yield of several crops infected by plant pathogens (Lewis and

Papavizas, 1980; Siddiqui and Mahmood, 1996).

Application of organic amendments to soil is considered a good management

practice in many agriculture production system because it stimulate soil microbial

growth and activity, with subsequent mineralization of plant nutrients (Eriksen, 2005),

and therefore increase soil fertility and quality (Doran and Smith, 1987). Soil

amendments, especially with those with high nitrogen (%), have been reported to

inhibit nematicidal and fungicidal activity, mainly through the release of ammonia

from the amendment during their decomposition in the soil or through increased

population of antagonistic microorganisms (Rodriguez-Kabana, 1986; Rodriguez-

Kabana et al., 1987; Spiegel et al., 1987; Oka et al., 1993).

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For the management of plant-parasitic nematodes, the application of a

combination of two or more beneficial microbes in a biocontrol preparation has been

recommended (Meyer and Roberts, 2002) in order to maximum the potential benefits

of the various agents. In general a single biocontrol agent is used for biocontrol of

plants disease against a single pathogen (Wilson and Backman, 1999). This may

sometimes account for the inconsistent performance because a single biocontrol agent

is not active in all soil environments or against all pathogens that attack the host plant.

On the other hand, mixture of biocontrol agents with different plant colonization

patterns may be useful for biocontrol of different plant pathogens via different

mechanism of disease suppression (Akhtar and Siddiqui, 2007). Moreover, mixture of

biocontrol agents with taxonomically different organisms that require different

optimum temperature, pH and moisture conditions may colonize roots more

aggressively, improve plant growth and efficacy of biocontrol (Siddiqui, 2006).

Presence of rhizobia in the rhizosphere protect the host root from damage

caused by pathogens (Siddiqui and Husain, 1992; Siddiqui and Mahmood, 1995).

AMF have been shown to reduce root-knot nematode population densities on tomato

and other plants (Sikora and Schonbeck, 1975; Sikora, 1990, 1992). Similarly,

Trichoderma spp. are known to reduce root-knot nematode infections (Windham et

al., 1989; Spiegel and Chet, 1998) by producing nematicidal substances (Sharon et al.,

2001). The combined application of microorganisms with different antagonistic

mechanisms is an approach that could possibly improve biocontrol efficacy. However,

interference and competition between antagonists must be considered.

All the microorganisms that have been applied as biocontrol agents against

nematodes were tested singly and there is not much information available about the

efficacy of mixing the bioagents. We suggest that a combination of rhizospheric

microorganisms with antagonistic fungi and organic wastes could be more likely to

have a greater variety of traits responsible for suppression of root-knot nematode over

a wide range of environmental conditions. Therefore, the aim of the present study is to

increase the efficacy of AM fungus, Glomus fasciculatum in the biocontrol of

nematodes by integrating it with an antagonistic fungus T. harzianum and root-nodule

bacterium Mesorhizobium ciceri (as these organisms are generally important

components of rhizophere soil) in combination with decomposed organic wastes (A.

sativa straw, C. album leaves and poultry manure) to serve as replacements or

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alternatives to the synthetic nematicides. All of them were tested alone and in various

combinations in the absence as well as in the presence of root-knot nematode, M.

incognita, for their efficacy on plant growth, mycorrhization and disease development

in chickpea plant.

8.2 MATERIALS AND METHODS

8.2.1 Preparation and sterilization of soil mixture

Soil, river sand and organic manure were mixed in a ratio of 3:1:1 (v/v/v),

divided and kept in jute bags. Little water was poured into each bag to wet the soil

after transferring them to an autoclave for sterilization at 137.9 KPa for 20 minutes.

Sterilized soil was allowed to cool down at room temperature before filling 15cm

diameter clay pots with 1kg of sterilized soil.

The study was carried out in three parts:

8.2.2 EXPERIMENT 8A: With straw of A. sativa

8.2.2.1 Growth and maintenance of test plant

Seeds of chickpea (Cicer arietinum L.) cv. Avrodhi were surface sterilized

with 0.01% mercuric chloride for 2min and then washed three times with distilled

water. Five sterilized seeds of chickpea were then sown in 15cm diameter earthen pots

containing 1kg sterilized soil and later thinned to one seedling per pot.

8.2.2.2 Plant straw

Ten grams of decomposed plant straw of Avena sativa L. was added around

each seedling in the pots. Prior to use, straw has been allowed to decompose in

separate containers for 6 months, with sufficient water being added at ten days

intervals.

8.2.2.3 AM fungus (G. fasciculatum) inoculum

The inoculum of Glomus fasciculatum was maintained as mentioned in

Section-2. The inoculum of G. fasciculatum was raised and maintained on Chloris

gayana Kunth (Rhode‟s grass) grown in the greenhouse of the department. The

proportion of G. fasciculatum in the inoculum was assessed by the most probable

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number method (Porter, 1979). It was found that 1g of soil contained approximately

16 propagules of G. fasciculatum, so to inoculate 800 infective propagules of G.

fasciculatum per pot, 50g of soil with inoculum was added around each seedling with

AM fungus treatment.

8.2.2.4 Mesorhizobium ciceri inoculum

Chickpea strain of Rhizobium (Charcoal-based culture), Mesorhizobium ciceri

was obtained from Indian Agricultural Research Institute (IARI), New Delhi, India.

100g of charcoal culture of M. ciceri was dissolved in 1L distilled water. 10mL of this

suspension, containing 1g culture was inoculated per plant.

8.2.2.5 Trichoderma harzianum inoculums

Trichoderma harzianum was obtained from Institute of Microbial Technology

(IMTECH), Chandigarh and was cultured on potato dextrose agar (PDA, 20% potato,

2.0% dextrose, 2.0% agar, pH =7.0). Seven days after incubation at 27 ºC, the culture

broth was filtered and the filtrate containing fungal spores served as stock solution for

screening nematicidal activity. Inoculum was prepared by maintaining 106 spores of

T. harzianum per mL of the solution and was inoculated per plant.

8.2.2.6 Nematode inoculum

Root-knot nematode, Meloidogyne incognita culture was maintained as

mentioned in Section-3. Second stage juveniles (J2) Meloidogyne incognita race 2

were used as inoculum. Large number of M. incognita egg masses were handpicked,

using sterilized forceps, from heavily infected tomato roots on which a pure culture of

the nematode was maintained. These egg masses were washed in distilled water and

then placed in 10cm diameter, 15-mesh coarse sieves containing crossed layers of

tissue paper, placed in Petri plates containing water just enough to contact the egg

masses and were kept in an incubator running at 25 °C. The hatched juveniles (J2)

were collected from the Petri plates every 24h and fresh water was added. The

concentration of second stage juveniles (J2) of M. incognita in the water suspension

was adjusted so that each milliliter contained 200±5 nematodes. 10ml of this

suspension containing 2000 freshly hatched juveniles were added to each pot

containing a chickpea seedling.

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8.2.2.7 Inoculation technique

For the addition of decomposed straw of A. sativa and inoculation of G.

fasciculatum, M. ciceri, T. harzianum and M. incognita, soil around the roots was

carefully removed without damaging the roots. The inoculum suspension or soil with

inoculum was poured or placed around the roots and the soil was replaced. All the

treatments were given concomitantly 15days prior to the nematode inoculation. An

equal volume of sterile water was added to the control treatments.

8.2.2.8 Experimental design

A. sativa, G. fasciculatum, M. ciceri and T. harzianum were applied around the

seedlings of chickpea in the presence as well as in the absence of nematode in each of

the possible (single, dual, triple and quadrate) combinations. The experiment was set

up in a completely randomized block design with sixteen experimental

variables/treatments: 1) Control (C); 2) G. fasciculatum (GF); 3) M. ciceri (MC); 4) A.

sativa (AS); 5) T. harzianum (TH); 6) GF+MC; 7) GF+AS; 8) GF+TH; 9) MC+AS;

10) MC+TH; 11) AS+TH; 12) GF+MC+AS; 13) GF+MC+TH; 14) GF+AS+TH; 15)

MC+AS+TH and 16) GF+MC+AS+TH with and without M. incognita (MI). Each

treatment was replicated five times. The pots were watered upto the soil capacity and

kept on a glasshouse bench with air temperature ranging from 22±3 oC.

8.2.2.9 Observations

The chickpea plants were terminated 90 days after the nematode inoculation

for determining the plant growth, chlorophyll content, nutrient status, mycorrhization

and nematode-related parameters (as discussed earlier in Section-2 and 3). The plants

of each treatment were taken out from the pots and soil particles adhering to roots

were removed by washing under tap water and properly labelled. Length of the plants

was measured by measuring tape and fresh as well as dry weight of the plants were

with the help of a physical balance. Excess water was removed by blotting paper

before weighing shoots and roots separately. For dry weight determination, shoots and

roots were kept in labelled envelopes and dried in a hot air oven running at 60 °C for

24-48 h before weighing. Chlorophyll content was estimated by the method of Arnon

(1949). Nutrient contents (N, P & K) were also estimated per 1g of fresh leaf weight.

Nitrogen content of the shoot was estimated by the method of Lindner (1944), while

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phosphorus and potassium contents were estimated by the methods of Fiske and

Subbarow (1925) and flame photometer, respectively.

8.2.2.10 Parameters studied

After termination of the experiment, the following parameters were

determined for each treatment:

Plant length (cm)

Plant fresh weight (g)

Plant dry weight (g)

Pods plant-1

Nodules plant-1

Chlorophyll content (mg g-1

fresh leaves)

Nutrient contents (mg g-1

fresh leaves)

Mycorrhization parameters

Nematode-related parameters

8.2.2.11 Plant growth and chemical parameters

Plant growth and chemical parameters were studied by the methods mentioned

in Section-2.

8.2.2.12 Mycorrhization parameters

Mycorrhization was recorded in terms of external colonization (%), internal

colonization (%), per cent arbuscules, number of chlamydospores in 1cm root

segment and number of chlamydospores recovered from 100g rhizosphere soil as in

Section-2.

8.2.2.13 Nematode-related parameters

Nematode related parameters in terms of nematode population (both in soil

and root); number of galls root system-1

; number of eggmasses root system-1

; number

of eggs eggmass-1

; root-knot index (0-5) and reproduction factor (pf/pi) were studied

by the methods mentioned in Section-3.

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8.2.3 EXPERIMENT 8B: With green leaves of botanical C. album


Seedlings of chickpea (Cicer arietinum L.) cv. Avrodhi were raised and one

seedling per pot was maintained as described above in Experiment 8A.

8.2.3.2 C. album green leaves

Ten grams of decomposed green leaves of C. album were added around each

seedling in the pots. Prior to use, leaves of the botanical has been allowed to

decompose in separate pots for 15 days, with sufficient water being added at 3 days

interval.


The inoculum of Glomus fasciculatum was maintained as mentioned above in

Experiment 8A and in Section-2.



was maintained as mentioned above in Experiment 8A and in Section-4.

8.2.3.5 Trichoderma harzianum inoculum

The inoculum of Trichoderma harzianum was maintained as mentioned above

in Experiment 8A.

8.2.3.6 Preparation of nematode inoculum


mentioned above in Experiment 8A and in Section-3.


For the addition of decomposed green leaves of C. album and inoculation of

G. fasciculatum, M. ciceri, T. harzianum and M. incognita, soil around the roots was



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C. album, G. fasciculatum, M. ciceri and T. harzianum were applied around

the seedlings of chickpea in the presence as well as in the absence of nematode. The

experiment was set up in a completely randomized block design with sixteen

experimental variables/treatments: 1) Control (C); 2) G. fasciculatum (GF); 3) M.

ciceri (MC); 4) C. album (CA); 5) T. harzianum (TH); 6) GF+MC; 7) GF+CA; 8)

GF+TH; 9) MC+CA; 10) MC+TH; 11) CA+TH; 12) GF+MC+CA; 13) GF+MC+TH;

14) GF+CA+TH; 15) MC+CA+TH and 16) GF+MC+CA+TH with and without M.

incognita (MI). Each treatment was replicated five times. The pots were watered upto

the soil capacity and kept on a glasshouse bench with air temperature ranging from

22±3 oC.




and nematode-related parameters (as discussed earlier in Section-2 and 3).


After termination of the experiment, the above parameters (as mentioned in

Experiment 8A) were determined for each treatment.



in Section-2.





Section-2.

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; number

of eggs eggmass-1



8.2.4 EXPERIMENT 8C: With poultry manure


Seedlings of chickpea (Cicer arietinum L.) cv. Avrodhi were raised and one

seedling per pot was maintained as described above in Experiment 8A.

8.2.4.2 Poultry manure

Ten grams of decomposed poultry manure was added around each seedling in

the pots. Prior to use, manure has been allowed to decompose in separate containers

for 6 months, with sufficient water being added at ten days intervals.


The inoculum of Glomus fasciculatum was maintained as mentioned above in

Experiment 8A and in Section-2.



was maintained as mentioned above in Experiment 8A and in Section-4.

8.2.4.5 Trichoderma harzianum inoculum

The inoculum of Trichoderma harzianum was maintained as mentioned above

in Experiment 8A.

8.2.4.6 Preparation of nematode inoculum

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mentioned above in Experiment 8A and in Section-3.


For the addition of decomposed poultry manure and inoculation of G.

fasciculatum, M. ciceri, T. harzianum and M. incognita, soil around the roots was






Poultry manure, G. fasciculatum, M. ciceri and T. harzianum were applied

around the seedlings of chickpea in the presence as well as in the absence of

nematode. The experiment was set up in a completely randomized block design with

sixteen experimental variables/treatments: 1) Control (C); 2) G. fasciculatum (GF); 3)

M. ciceri (MC); 4) Poultry manure (PM); 5) T. harzianum (TH); 6) GF+MC; 7)

GF+PM; 8) GF+TH; 9) MC+PM; 10) MC+TH; 11) PM+TH; 12) GF+MC+PM; 13)

GF+MC+TH; 14) GF+PM+TH; 15) MC+PM+TH and 16) GF+MC+PM+TH both in

the presence and absence of M. incognita (MI). Each treatment was replicated five

times. The pots were watered upto the soil capacity and kept on a glasshouse bench

with air temperature ranging from 22±3 oC.




and nematode-related parameters (as discussed earlier in Section-2 and 3).


After termination of the experiment, the above parameters (as mentioned in

Experiment 8A) were determined for each treatment.

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in Section-2.





Section-2.





; number

of eggs eggmass-1



8.2.5 Statistical analysis

All the data in Experiment 8A, 8B and 8C were analyzed statistically by the

method of Panse and Sukhatme (1985). Minimum difference required for significance

(C.D.) at P=0.01 and P=0.05 was calculated by the ANOVA model 4 given in

Appendix. Duncan‟s multiple range test was employed to test for significant

differences between treatments at P = 0.05.

8.3 RESULTS

8.3.1 EXPERIMENT 8A: With straw of A. sativa

8.3.1.1 In the absence of M. incognita

8.3.1.1.1 Plant length (cm)

Inoculation of the plant symbionts (G. fasciculatum, M. ciceri), antagonistic

fungi, T. harzianum and A. sativa straw in all (single, dual, triple and quadrate)

combinations caused a significant increase in plant length (shoot, root and total

length) over control except M. ciceri individual treatment, which failed to cause a

significant increase. Treatments GF+MC+AS, GF+AS+TH, MC+AS+TH and

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GF+MC+AS+TH were not statistically different from each other. However, the

highest increase in plant length (63.46%) was observed while inoculation of all

biocontrol agents and A. sativa straw. Individually, G. fasciculatum is the most

effective of all the biocontrol agents in promoting the plant length (Table 26 and Fig.

22).

8.3.1.1.2 Plant fresh weight (g)

Individual inoculation of M. ciceri did not cause a significant increase in plant

fresh weight in terms of shoot, root and total weight over control plants. Highest

increase was observed (66.30%) while combined inoculation of GF+MC+AS+TH.

Individual inoculation of A. sativa straw resulted in highest fresh weight (26.2%)

followed by individual inoculation of G. fasciculatum (25%), T. harzianum (23.5%)

and M. ciceri (10.9%). Treatments GF+AS, GF+TH, AS+TH and GF+MC+TH were

statistically similar to each other (Table 26 and Fig. 22).

8.3.1.1.3 Plant dry weight (g)

Plant dry weight (shoot, root and total dry weight) was significantly increased

by the inoculation of all biocontrol agents and A. sativa straw but the increase was not

significant in case of individual inoculation of M. ciceri (Table 26). However, the

highest increase was observed (67.8%) while inoculating all the plant symbionts and

A. sativa straw together (Fig. 22).

8.3.1.1.4 Pods plant -1

Significant increase in pods number was observed in case of all the treatments.

Highest pod number was recorded in the treatment GF+MC+AS+TH (87) and lowest

while inoculation with M. ciceri (43) alone (Table 26).

8.3.1.1.5 Nodules plant-1

Nodules number increased to a greater extent while inoculating with root-

nodulating bacteria, M. ciceri. 9, 10 and 8 nodules were recorded in plants inoculated

individually with G. fasciculatum, A. sativa and T. harzianum respectively which

were comparatively very less as compared to the nodules number in plants treated

individually with M. ciceri (56) (Table 26).

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Table 26. Individual and interactive effect of AM fungus Glomus fasciculatum, root-nodule bacterium Mesorhizobium ciceri, straw of

Avena sativa and an antagonistic fungi Trichoderma harzianum on the growth parameters of chickpea plant

Treatments Plant length (cm) Plant fresh weight (g) Plant dry weight (g) Pods

plant-1 Nodules

plant-1 Shoot Root Total Shoot Root Total Shoot Root Total

Control 43.64f±2.18 21.82g±1.09 65.46g±3.27 43.36f±2.17 10.84g±0.54 54.20g±2.71 6.51h±0.33 1.62g±0.08 8.13h±0.41 31.0k±1.55 4.0j±0.20

GF 53.67c±2.68 26.84f±1.34 80.51f±4.03 54.18d±2.71 13.57f±0.68 67.75e±3.39 8.27de±0.41 2.06f±0.10 10.33ef±0.52 50.0i±2.50 9.0i±0.45

MC 46.85ef±2.34 23.45g±1.17 70.30g±3.52 48.13e±2.41 12.03g±0.60 60.16f±3.01 7.19g±0.36 1.91f±0.10 9.10g±0.46 43.0j±2.15 56.0f±2.80

AS 50.02de±2.50 31.15d±1.56 81.17ef±4.06 51.13de±2.56 17.27cd±0.86 68.40de±3.42 7.58f±0.38 2.85d±0.14 10.43e±0.52 51.0i±2.55 10.0i±0.50

TH 48.14e±2.41 31.07de±1.55 79.21f±3.96 50.32e±2.52 16.61de±0.83 66.93e±3.35 7.51fg±0.38 2.63e±0.13 10.14f±0.51 47.0ij±2.35 8.0ij±0.40

GF+MC 61.21a±3.06 30.67e±1.53 91.88cd±4.59 62.49b±3.12 15.62e±0.78 78.11bc±3.91 9.34bc±0.47 2.46e±0.12 11.80cd±0.59 64.0fg±3.20 75.0d±3.75

GF+AS 59.59a±2.98 38.26bc±1.91 97.85b±4.89 60.91bc±3.05 21.75a±1.09 82.66b±4.13 9.12c±0.46 3.38a±0.17 12.50b±0.63 70.0de±3.50 23.0h±1.15

GF+TH 58.45b±2.92 36.80c±1.84 95.25bc±4.76 60.37c±3.02 20.00b±1.00 80.37b±4.02 8.85cd±0.44 3.27a±0.16 12.12bc±0.61 68.0e±3.40 22.0h±1.10

MC +AS 54.21c±2.71 33.11d±1.66 87.32de±4.37 55.18d±2.76 18.31c±0.92 73.49cd±3.67 7.97e±0.40 3.17bc±0.16 11.14de±0.56 61.0gh±3.05 72.0de±3.60

MC +TH 52.77cd±2.64 32.45d±1.62 85.22e±4.26 53.99d±2.70 18.09c±0.90 72.08d±3.60 7.91ef±0.40 3.00cd±0.15 10.91e±0.55 59.0h±2.95 70.0e±3.50

AS+TH 59.23ab±2.96 37.00c±1.85 96.23b±4.81 60.41c±3.02 20.34b±1.02 80.75b±4.04 8.88c±0.44 3.31a±0.17 12.19b±0.61 68.0e±3.40 20.0h±1.00

GF+ MC +AS 62.85a±3.14 40.11b±2.01 102.96a±5.15 64.58ab±3.23 21.97a±1.10 86.55a±4.33 9.84ab±0.49 3.33a±0.17 13.17a±0.66 80.0b±4.00 86.0b±4.30

GF+ MC +TH 61.35a±3.07 38.54b±1.93 99.89b±4.99 61.67b±3.08 21.80a±1.09 83.47b±4.17 9.52b±0.48 3.19b±0.16 12.71b±0.64 73.0cd±3.65 83.0bc±4.15

GF+AS+TH 61.45a±3.07 44.26a±2.21 105.71a±5.29 66.64a±3.33 22.14a±1.11 88.78a±4.44 10.06a±0.50 3.40a±0.17 13.46a±0.67 84.0ab±4.20 30.0g±1.50

MC +AS+TH 61.93a±3.10 39.00b±1.95 100.93ab±5.05 62.97b±3.15 21.85a±1.09 84.82a±4.24 9.61b±0.48 3.22ab±0.16 12.83ab±0.64 75.0c±3.75 80.0c±4.00

GF+ MC+AS+TH 62.36a±3.12 44.64a±2.23 107.00a±5.35 67.40a±3.37 22.73a±1.14 90.13a±4.51 10.23a±0.51 3.42a±0.17 13.65a±0.68 87.0a±4.35 95.0a±4.75

C.D. (P = 0.05) 4.00 2.43 6.42 4.11 1.31 5.41 0.62 0.20 0.82 4.56 4.16

C.D. (P = 0.01) 5.38 3.28 8.64 5.53 1.76 7.28 0.83 0.27 1.10 6.14 5.60

Data mean±SD of five replicates

GF = Glomus fasciculatum; MC = Mesorhizobium ciceri; AS = Avena sativa; TH = Trichoderma harzianum

Mean values with different letters within the column are significantly different at P = 0.05

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8.3.1.1.6 Chlorophyll content (mg g-1

) fresh leaves

Single, double, triple and quadrate combinations of G. fasciculatum, M. ciceri,

A. sativa and T. harzianum significantly increase the chlorophyll content of plants.

However, the highest increase (60%) was observed in GF+MC+AS+TH treated

plants. Chlorophyll content in plants with treatments GF+MC+AS, GF+MC+TH,

GF+AS+TH, MC+AS+TH and GF+MC+AS+TH were not significantly different

from each other (Table 27 and Fig. 22).

8.3.1.1.7 Nutrient contents (N, P & K) (mg g-1

) fresh leaves

Nutrient contents in terms of N, P and K were significantly increased in the

individual and interactive treatments. Nutrient contents in plants with treatments

GF+MC+AS, GF+MC+TH, GF+AS+TH, MC+AS+TH and GF+MC+AS+TH were at

par. Maximum nutrients (72.8%N, 69.2%P and 64.1%K) were recorded in plants

while inoculating with GF+ MC+AS+TH (Table 27 and Fig. 22).

8.3.1.1.8 Mycorrhization parameters

Inoculation of M. ciceri and straw of A. sativa brings about an increase in the

mycorrhization parameters (external and internal colonization, per cent arbuscules,

no. of chlamydospores in 1cm root segment and no. of chlamydospores recovered

from 100g rhizosphere soil) but the increase was not significant. Increase in

mycorrhization was comparatively more in plants treated with M. ciceri. Antagonistic

fungi, T. harzianum suppress the mycorrhizal colonization of G. fasciculatum,

although the reduction was not significant. Combined inoculation of GF+MC+AS and

GF+MC+AS+TH brings about a significant increase in mycorrhization. Moreover, all

the parameters in both the treatments were at par (Table 27).

8.3.1.2 In the presence of M. incognita

Inoculation with M. incognita caused a significant reduction in all the growth

parameters compared with uninoculated control.


The inoculation of M. incognita caused a significant reduction in plant length

(shoot, root and total length) of all the treatments over nematode-uninoculated

1

Table 27. Individual and interactive effect of AM fungus Glomus fasciculatum, root-nodule bacterium Mesorhizobium ciceri, straw of Avena

sativa and an antagonistic fungi Trichoderma harzianum on the chlorophyll content, nutrient status and mycorrhization

parameters of chickpea plant


GF = Glomus fasciculatum MC = Mesorhizobium ciceri; AS = Avena sativa; TH = Trichoderma harzianum


Treatments Chlorophyll

content

(mg g-1)

Nutrient contents (mg g-1) External

Colonization

(%)

Internal

Colonization

(%)

Per cent

arbuscules No. of

chlamydospores in

1cm root segment

No. of

chlamydospores

recovered from 100 g

rhizosphere soil N P K

Control 2.402f±0.120 2.68f±0.13 0.240f±0.012 1.92e±0.096 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0d±0.00

GF 2.876e±0.144 3.61de±0.18 0.329de±0.016 2.33d±0.117 62.7c±3.14 61.7bc±3.09 59.3c±2.97 58.0bc±2.90 893.0c±44.65

MC 3.135d±0.157 3.82cd±0.19 0.308e±0.015 2.29d±0.114 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0d±0.00

AS 2.894e±0.145 3.63d±0.18 0.313e±0.016 2.35d±0.118 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0d±0.00

TH 2.821e±0.141 3.54e±0.18 0.311e±0.016 2.30d±0.115 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0d±0.00

GF+MC 3.411c±0.171 4.35ab±0.22 0.373b±0.019 2.89b±0.145 67.8a±3.39 66.5a±3.32 64.0a±3.20 63.0a±3.15 960.0a±48.00

GF+AS 3.242cd±0.162 4.18b±0.21 0.379b±0.019 2.93b±0.146 63.6bc±3.18 62.4b±3.12 59.9bc±2.99 59.0b±2.95 906.0b±45.30

GF+TH 3.194d±0.160 4.11bc±0.21 0.361bc±0.018 2.81bc±0.141 61.5c±3.07 60.8c±3.04 58.3c±2.91 57.0c±2.85 880.0c±44.00

MC +AS 3.466bc±0.173 4.40a±0.22 0.355c±0.018 2.77c±0.138 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0d±0.00

MC +TH 3.283c±0.164 4.27b±0.21 0.354cd±0.018 2.75c±0.138 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0d±0.00

AS+TH 3.259c±0.163 4.21b±0.21 0.357c±0.018 2.91b±0.146 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0d±0.00

GF+ MC +AS 3.807a±0.190 4.59a±0.23 0.399a±0.020 3.08a±0.154 69.3a±3.46 68.5a±3.42 65.7a±3.28 64.0a±3.20 987.0a±49.35

GF+ MC +TH 3.766a±0.188 4.53a±0.23 0.393a±0.020 2.99ab±0.150 66.8ab±3.34 65.4ab±3.27 63.0ab±3.15 61.0ab±3.05 948.0ab±47.40

GF+AS+TH 3.578b±0.179 4.42a±0.22 0.403a±0.020 3.11a±0.155 62.8c±3.14 61.8b±3.09 59.4c±2.97 0.0d±0.00 895.0bc±44.75

MC +AS+TH 3.665ab±0.183 4.48a±0.22 0.384ab±0.019 3.00a±0.150 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0d±0.00

GF+ MC+AS+TH 3.843a±0.192 4.63a±0.23 0.406a±0.020 3.15a±0.158 68.3a±3.42 66.9a±3.35 64.3a±3.22 0.0d±0.00 971.0a±48.55

C.D. (P=0.05) 0.234 0.291 0.025 0.193 3.769 3.706 3.561 3.352 53.64

C.D. (P=0.01) 0.315 0.392 0.034 0.260 5.076 4.991 4.795 4.514 72.24

1

T1 = Control; T2 = G. fasciculatum (GF); T3 = M. ciceri (MC); T4 = A. sativa (AS); T5 = T. harzianum (TH); T6 = GF+MC;

T7 = GF+AS; T8 = GF+TH; T9 MC+AS; T10 = MC+TH; T11 = AS+TH; T12 = GF+MC+AS; T13 = GF+MC+TH; T14 = GF+AS+TH; T15 = MC+AS+TH; T16 = GF+MC+AS+TH

Fig. 22 Individual and interactive effect of AM fungus Glomus fasciculatum,

root-nodule bacterium Mesorhizobium ciceri, straw of Avena sativa and an

antagonistic fungi Trichoderma harzianum on the growth parameters,

chlorophyll and nutrient contents of chickpea plant

T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16

Chlo

rop

hyll

co

nte

nt

(mg g

-1)

0

1

2

3

4

5T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16

Pla

nt

dry

wei

ght

(g)

02468

10121416


Pla

nt

length

(c

m)

0

20

40

60

80

100

120


Pla

nt

fres

h w

eight

(g)

0

20

40

60

80

100

T1 = Control; T2 = G. fasciculatum; T3 = Rhizobium; T4 = A. sativa; T5 = T. harzianum; T6 = G. fasciculatum+Rhizobium;

T7 = G. fasciculatum+A. sativa; T8 = G. fasciculatum+T. harzianum; T9 = Rhizobium+A. sativa; T10 = Rhizobium+

T. harzianum; T11 = A. sativa+T. harzianum; T12 = G. fasciculatum+Rhizobium+A. sativa; T13 = G. fasciculatum+

Rhizobium+T. harzianum; T14 = G. fasciculatum+A. sativa+T. harzianum; T15 = Rhizobium+A. sativa+T. harzianum;

T16 = G. fasciculatum+Rhizobium+A. sativa+T. harzianum

Treatments


Nutr

ient

conte

nts

(mg g

-1)

0.0

1.0

2.0

3.0

4.0

5.0

6.0N P K

2

untreated control plants. However, the lowest reduction by nematode occurred in the

plants inoculated with T. harzianum (9.63%) as compared to other plant symbionts

(G. fasciculatum, M. ciceri), and A. sativa straw. All the biocontrol agents and A.

sativa in various combinations reduce the deleterious effect of nematode and caused a

significant increase in plant length (shoot, root and total length). Individually, T.

harzianum with M. incognita caused the maximum enhancement of plant length

(26%) over nematode-inoculated untreated plants. In combined treatments,

GF+AS+TH+MI and GF+MC+AS+TH+MI were statistically similar. However, the

increase was highest in GF+MC+AS+TH+MI (68.4%) over all other treatments and

nematode-inoculated untreated control (Table 28 and Fig. 23).


Plant fresh weight (shoot, root and total) declined significantly in the presence

of M. incognita. Inoculation of plant symbionts, antagonistic fungi and organic waste

alone as well as in combinations overcome the loss caused by the nematode and

brings about a significant improvement in the fresh weight of chickpea plants over

nematode-inoculated untreated plants (Table 28). Highest increase in fresh weight

(70%) was reported while combined inoculation of all the microorganisms i.e.

GF+MC+AS+TH+MI. Reduction by M. incognita was lowest in plants treated with T.

harzianum (17.97%) as compared to other plant symbionts and A. sativa straw (Fig.

23).


Plant dry weight (shoot, root and total dry weight) depicted almost a similar

trend of results as in case of plant fresh weight. T. harzianum proved to be most

effective in suppressing M. incognita and cause a highest increase (27.3%) over other

individual treatments of G. fasciculatum (25.1%), A. sativa (25.6%) and M. ciceri

(17.7%) in the presence of nematode. Combined inoculation of all biocontrol agents

with M. incognita promoted highest increase (71.7%) over nematode-inoculated

untreated plants (Table 28 and Fig. 23).

8.3.1.2.4 Pods plant-1

Number of pods were significantly reduced by the presence of root-knot

nematode, M. incognita. Lowest reduction occurred in T. harzianum (6.38%)

1


sativa and an antagonistic fungi Trichoderma harzianum on the growth parameters of chickpea plant infected with root-knot

nematode, Meloidogyne incognita


plant-1 Nodules


MI 37.85i±1.89 18.96h±0.95 56.81i±2.84 34.66g±1.73 8.67g±0.43 43.33h±2.17 5.18g±0.26 1.29i±0.06 6.47g±0.32 18.0i±0.90 0.0k±0.00

GF+MI 46.65gh±2.33 23.22g±1.16 69.87g±3.49 43.19ef±2.16 10.75f±0.54 53.94fg±2.70 6.08f±0.30 2.02fg±0.10 8.10f±0.41 38.0g±1.90 7.0j±0.35

MC+MI 43.08h±2.15 20.04h±1.00 63.12h±3.16 40.10f±2.01 10.24f±0.51 50.34g±2.52 6.12f±0.31 1.50h±0.08 7.62f±0.38 26.0h±1.30 47.0f±2.35

AS+MI 46.81g±2.34 23.35g±1.17 70.16g±3.51 43.38e±2.17 10.80f±0.54 54.18f±2.71 6.13f±0.31 2.00g±0.10 8.13f±0.41 40.0fg±2.00 8.0j±0.40

TH+MI 47.76fg±2.39 23.82fg±1.19 71.58fg±3.58 44.03e±2.20 10.87f±0.54 54.90f±2.75 6.16f±0.31 2.08f±0.10 8.24f±0.41 44.0f±2.20 10.0j±0.50

GF+MC+MI 51.42e±2.57 25.61ef±1.28 77.03ef±3.85 47.56d±2.38 11.81e±0.59 59.37e±2.97 6.75e±0.34 2.18ef±0.11 8.93e±0.45 55.0e±2.75 60.0e±3.00

GF+AS+MI 55.40cd±2.77 27.76cd±1.39 83.16cd±4.16 51.67c±2.58 12.87cd±0.64 64.54cd±3.23 7.34cd±0.37 2.39cd±0.12 9.73cd±0.49 60.0d±3.00 18.0i±0.90

GF+TH+MI 56.86c±2.84 28.35c±1.42 85.21c±4.26 52.65bc±2.63 13.21c±0.66 65.86bc±3.29 7.41c±0.37 2.50c±0.13 9.91bc±0.50 65.0c±3.25 21.0hi±1.05

MC+AS+MI 51.26ef±2.56 25.83e±1.29 77.09e±3.85 47.96d±2.40 11.92e±0.60 59.88e±2.99 6.83e±0.34 2.20e±0.11 9.03e±0.45 56.0de±2.80 62.0e±3.10

MC+TH+MI 52.94de±2.65 26.37de±1.32 79.31de±3.97 49.19cd±2.46 12.25de±0.61 61.44de±3.07 6.94de±0.35 2.27de±0.11 9.21de±0.46 60.0d±3.00 68.0d±3.40

AS+TH+MI 57.23c±2.86 28.55c±1.43 85.78c±4.29 52.91b±2.65 13.30c±0.66 66.21b±3.31 7.45bc±0.37 2.53bc±0.13 9.98b±0.50 66.0c±3.30 22.0h±1.10

GF+MC+AS+MI 58.20bc±2.91 29.12bc±1.46 87.32bc±4.37 53.74b±2.69 13.42bc±0.67 67.16b±3.36 7.53b±0.38 2.56b±0.13 10.09b±0.50 68.0bc±3.40 70.0cd±3.50

GF+MC+TH+MI 61.64ab±3.08 30.66ab±1.53 92.30ab±4.62 56.68a±2.83 14.25ab±0.71 70.93a±3.55 8.04a±0.40 2.67ab±0.13 10.71a±0.54 71.0ab±3.55 73.0bc±3.65

GF+AS+TH+MI 62.82a±3.14 31.50a±1.58 94.32a±4.72 58.32a±2.92 14.56a±0.73 72.88a±3.64 8.32a±0.42 2.70a±0.13 11.02a±0.55 73.0a±3.65 27.0g±1.35

MC+AS+TH+MI 59.67b±2.98 29.64b±1.48 89.31b±4.47 55.57ab±2.78 13.77b±0.69 69.34ab±3.47 7.91ab±0.40 2.61b±0.13 10.52ab±0.53 69.0b±3.45 76.0ab±3.80

GF+MC+AS+TH+MI 63.86a±3.19 31.80a±1.59 95.66a±4.78 58.91a±2.95 14.75a±0.74 73.66a±3.68 8.33a±0.42 2.78a±0.14 11.11a±0.56 75.0a±3.75 79.0a±3.95

C.D. (P=0.05) 3.78 1.88 5.66 3.50 0.87 4.37 0.50 0.16 0.66 4.02 3.61

C.D. (P=0.01) 5.09 2.53 7.62 4.71 1.18 5.89 0.67 0.22 0.89 5.41 4.86


GF = Glomus fasciculatum; MC = Mesorhizobium ciceri; AS = Avena sativa; TH = Trichoderma harzianum; MI = Meloidogyne incognita


1


Chlo

rophyll

co

nte

nt

(mg g

-1)

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16

Pla

nt

dry

wei

ght

(g)

0

2

4

6

8

10

12


Pla

nt

length

(c

m)

0

20

40

60

80

100

120


Pla

nt

fres

h w

eight

(g)

0

20

40

60

80






Treatments


Nutr

ient

conte

nts

(mg g

-1)

0.0

1.0

2.0

3.0

4.0

5.0N P K

T1 = M. incognita (MI); T2 = G. fasciculatum (GF) + M. incognita (MI); T3 = M. ciceri (MC) + M. incognita (MI); T4 = A.

sativa (AS)+ M. incognita (MI); T5 = T. harzianum (TH) + M. incognita (MI); T6 = GF+MC+MI; T7 = GF+AS+MI; T8 =

GF+TH+MI; T9 = MC+AS+MI; T10 = MC+TH+MI; T11 = AS+TH+MI; T12 = GF+MC+AS+MI; T13 = GF+MC+TH+MI; T14 = GF+AS+TH+MI; T15 = MC+AS+TH+MI; T16 = GF+MC+AS+TH




chlorophyll and nutrient contents of chickpea plant infected with root-

knot nematode, Meloidogyne incognita

2

compared to other agents applied with nematode. Highest pod number was observed

while combined inoculation of all the biocontrol agents, GF+MC+AS+TH+MI (75)

and lowest in the M. ciceri (26) treated plants (Table 28).


No nodules were observed while inoculating with M. incognita in untreated

control. M. ciceri inhibits the effect of nematode and increase the nodule number to a

greater extent as compared to other microorganisms and straw. In individual

treatments, highest nodule number (47) was observed in M. ciceri followed by T.

harzianum (10), A. sativa (8) and G. fasciculatum (7) with M. incognita. In the

presence of nematode, all the plant symbionts and straw together resulting highest

nodule number (79) as compared to all other single, double and triple combinations

(Table 28).


fresh leaves)

M. incognita reduced the chlorophyll content in all the plants as compared to

the chickpea plants in its absence. However, the lowest reduction (18.4%) was

reported in case of plants treated with T. harzianum. G. fasciculatum, M. ciceri, A.

sativa and T. harzianum overcome the loss caused by the nematode and highest

increase (74.6%) was observed in plants inoculated with all of them in combination

over nematode-inoculated untreated control plants (Table 29 and Fig. 23).


fresh leaves)

Lowest nutrient contents (N, P and K) were observed in plants inoculated with

M. incognita as compared to uninoculated control. All the plant symbionts and straw

were effective in suppressing the deleterious effect of nematode and significantly

increase the nutrient contents. Individually, M. ciceri resulted a highest increase in N

content (35.2%) and G. fasciculatum cause a highest increase in P content (31.8%) as

compared to other microorganisms and plant straw in nematode-inoculated plants.

Combined inoculations (dual, triple and quadrate) proved much better than individual

appplication. GF+MC+AS+TH+MI caused highest increase in nutrients (85.6%N,

80%P and 71.4%K) over nematode-inoculated-untreated plants (Table 29 and Fig.

23).

1


sativa and an antagonistic fungi Trichoderma harzianum on the chlorophyll content, nutrient status and mycorrhization

parameters of chickpea plant infected with root-knot nematode Meloidogyne incognita





content (mg g-1)


Colonization

(%)

Internal

Colonization

(%)

Per cent

arbuscules No. of

chlamydospores

in 1cm root

segment

No. of

chlamydospores

recovered from

100 g

rhizosphere soil

N P K

MI 1.825e±0.091 2.30g±0.12 0.160e±0.008 1.54f±0.08 0.00d±0.00 0.00d±0.00 0.00e±0.00 0.0d±0.00 0.0d±0.00

GF+MI 2.208d±0.110 2.75f±0.14 0.211d±0.011 1.85e±0.09 59.10c±2.95 59.20bc±2.96 57.05cd±2.85 56.0bc±2.80 858.0bc±42.90

MC+MI 2.376d±0.119 3.11e±0.16 0.195d±0.010 1.81e±0.09 0.00d±0.00 0.00d±0.00 0.00e±0.00 0.0d±0.00 0.0d±0.00

AS+MI 2.253d±0.113 2.89ef±0.14 0.198d±0.010 1.86e±0.09 0.00d±0.00 0.00d±0.00 0.00e±0.00 0.0d±0.00 0.0d±0.00

TH+MI 2.300d±0.115 2.96e±0.15 0.204d±0.010 1.88e±0.09 0.00d±0.00 0.00d±0.00 0.00e±0.00 0.0d±0.00 0.0d±0.00

GF+MC+MI 2.875b±0.144 3.83bc±0.19 0.262b±0.013 2.25cd±0.11 65.20a±3.26 64.90a±3.25 62.30a±3.12 61.0a±3.05 938.0a±46.90

GF+AS+MI 2.737c±0.137 3.52d±0.18 0.264b±0.013 2.29c±0.11 61.80b±3.09 61.08b±3.05 58.58bc±2.93 57.0b±2.85 885.0b±44.25

GF+TH+MI 2.839bc±0.142 3.65cd±0.18 0.270b±0.014 2.40bc±0.12 58.30c±2.92 57.07c±2.85 54.67d±2.73 54.0c±2.70 824.0c±41.20

MC+AS+MI 2.902b±0.145 3.88b±0.19 0.250c±0.013 2.26c±0.11 0.00d±0.00 0.00d±0.00 0.00e±0.00 0.0d±0.00 0.0d±0.00

MC+TH+MI 2.956b±0.148 3.98b±0.20 0.257bc±0.013 2.28c±0.11 0.00d±0.00 0.00d±0.00 0.00e±0.00 0.0d±0.00 0.0d±0.00

AS+TH+MI 2.887b±0.144 3.84b±0.19 0.259b±0.013 2.36c±0.12 0.00d±0.00 0.00d±0.00 0.00e±0.00 0.0d±0.00 0.0d±0.00

GF+MC+AS+MI 3.042a±0.152 4.09a±0.20 0.276a±0.014 2.13d±0.11 66.46a±3.32 65.71a±3.29 63.03a±3.15 62.0a±3.10 947.0a±47.35

GF+MC+TH+MI 3.095a±0.155 4.15a±0.21 0.280a±0.014 2.48b±0.12 63.64ab±3.18 62.74ab±3.14 59.89ab±2.99 59.0ab±2.95 905.0ab±45.25

GF+AS+TH+MI 2.993ab±0.150 4.04ab±0.20 0.284a±0.014 2.59a±0.13 60.06bc±3.00 59.23b±2.96 57.07c±2.85 0.0d±0.00 929.0a±46.45

MC+AS+TH+MI 3.148a±0.157 4.22a±0.21 0.272ab±0.014 2.53ab±0.13 0.00d±0.00 0.00d±0.00 0.00e±0.00 0.0d±0.00 0.0d±0.00

GF+MC+AS+TH+MI 3.186a±0.159 4.27a±0.21 0.288a±0.014 2.64a±0.13 66.15a±3.31 64.98a±3.25 62.45a±3.12 0.0d±0.00 942.0a±47.10

C.D. (P=0.05) 0.193 0.254 0.017 0.155 3.61 3.57 3.43 3.37 52.07

C.D. (P=0.01) 0.260 0.342 0.024 0.209 4.86 4.81 4.62 4.53 70.13

1


Presence of M. incognita and T. harzianum adversely affected mycorrhization

parameters. M. incognita was more detrimental to mycorrhization of G. fasciculatum

as compared to T. harzianum. Combined inoculation of M. incognita and T.

harzianum significantly reduced the colonization. Treatments GF+MC+MI,

GF+MC+AS+MI and GF+MC+AS+TH+MI were not significantly different from

each other. However, lowest reduction by nematode was observed in case of plants

inoculated with GF+AS (2.9% external and 2.12% internal colonization, 2.18%

arbuscules, 3.4% chlamydospores in 1cm root segment and 2.3% chlamydospores

recovered from 100g rhizosphere soil). Highest mycorrhization in M. incognita

inoculated plants was observed when GF+MC+AS were together inoculated (Table

29).

8.3.1.2.9 Root-knot development

8.3.1.2.9a Nematode population

The population of M. incognita in soil as well as in root was significantly

reduced in the presence of either of the plant symbionts, fungi or straw. Inoculation of

T. harzianum resulted in an acute reduction in soil and root population of nematode

i.e. 65% and 67.5% respectively compared to other agents (Table 30 and Fig. 24).

8.3.1.2.9b Number of galls root system-1

Highest galling (210) was observed in plants which were inoculated with M.

incognita alone. All the treatments significantly decrease the number of galls in root

system. Lowest number of galls (10) was observed while combined inoculation of

GF+MC+AS+TH+MI (Table 30 and Fig. 24).

8.3.1.2.9c Number of eggmasses root system-1

No. of eggmasses root system-1

were highest when M. incognita alone was

inoculated. Presence of G. fasciculatum, M. ciceri, A. sativa and T. harzianum caused

significant reduction in this number. Maximum reduction (95%) was observed when

all of them were inoculated together and this reduction was significantly higher than

the reduction caused by either of them alone (Table 30 and Fig. 24).

1

Table 30. Individual and interactive effect of AM fungus Glomus fasciculatum, root-nodule bacterium Mesorhizobium ciceri, straw of

Avena sativa and an antagonistic fungi Trichoderma harzianum on the root-knot development of Meloidogyne incognita in

chickpea plant

Treatments Nematode population No. of galls root

system-1 No. of eggmasses

root system-1 No. of eggs

eggmass-1 Root-knot index

(0-5)

Reproduction factor

(pf/pi) Soil Root

MI 11,915.0a±595.8 243.0a±12.1 210.0a±10.5 84.0a±4.2 154.0a±7.7 4.0a 12.20a±0.61

GF+MI 9,008.0c±450.4 176.0c±8.8 148.0c±7.4 60.0c±3.0 107.0c±5.4 2.0c 8.80c±0.44

MC+MI 10,052.0b±502.6 202.0b±10.1 173.0b±8.7 70.0b±3.5 128.0b±6.4 3.0b 10.11b±0.51

AS+MI 7,089.0d±354.4 141.0e±7.0 122.0d±6.1 48.0d±2.4 89.0d±4.4 1.0d 7.05e±0.35

TH+MI 4,170.0e±208.5 79.0f±3.9 69.0e±3.5 28.0e±1.4 52.0e±2.6 0.0 3.92f±0.20

GF+MC+MI 7,445.0d±372.3 152.0d±7.6 124.0d±6.2 51.0d±2.5 94.0d±4.7 1.0d 7.60d±0.38

GF+AS+MI 3,395.0f±169.8 67.0g±3.3 57.0f±2.8 24.0f±1.2 42.0f±2.1 0.0 3.33g±0.17

GF+TH+MI 1,787.0h±89.4 33.0i±1.7 30.0h±1.5 12.0h±0.6 22.0h±1.1 0.0 1.69i±0.08

MC+AS+MI 3,872.0e±193.6 75.0fg±3.8 66.0e±3.3 26.0ef±1.3 48.0e±2.4 0.0 3.72fg±0.19

MC+TH+MI 2,383.0g±119.2 45.0h±2.3 40.0g±2.0 16.0g±0.8 29.0g±1.4 0.0 2.25h±0.11

AS+TH+MI 1,489.0hi±74.5 32.0i±1.6 27.0hi±1.4 11.0hi±0.6 19.0hi±1.0 0.0 1.64i±0.08

GF+MC+AS+MI 1,740.0h±87.0 35.0i±1.8 31.0h±1.5 13.0gh±0.7 23.0h±1.2 0.0 1.74i±0.09

GF+MC+TH+MI 1,310.0i±65.5 26.0ij±1.3 22.0ij±1.1 9.0i±0.4 17.0ij±0.8 0.0 1.30ij±0.06

GF+AS+TH+MI 953.0j±47.6 19.0jk±1.0 15.0jk±0.8 6.0jk±0.3 11.0kl±0.6 0.0 0.95jk±0.05

MC+AS+TH+MI 1,062.0ij±53.1 21.0j±1.1 18.0j±0.9 8.0ij±0.4 13.0jk±0.7 0.0 1.07j±0.05

GF+MC+AS+TH+MI 655.0j±32.8 12.0k±0.6 10.0k±0.5 4.0k±0.2 7.0l±0.4 0.0 0.70k±0.04

C.D. (P=0.05) 457.4 9.2 7.8 3.15 5.75 0.458

C.D. (P=0.01) 616.0 12.3 10.5 4.24 7.74 0.617




1


No.

of

eggs

eggm

ass-1

020406080

100120140160180


No.

of

eggm

asse

s

root

syst

em-1

0

20

40

60

80

100


Nem

atode

popula

tion

0

200

400

600

800

1000

1200

1400Soil (×10)

Root


No.

of

gal

ls

root

syst

em-1

0

50

100

150

200

250






Treatments


Rep

roduct

ion f

acto

r (p

f/pi)

0.0

2.0

4.0

6.0

8.0

10.0

12.0

14.0

T1 = M. incognita (MI); T2 = G. fasciculatum (GF) + M. incognita (MI); T3 = M. ciceri (MC) + M. incognita (MI); T4 = A.

sativa (AS) + M. incognita (MI); T5 = T. harzianum (TH) + M. incognita (MI); T6 = GF+MC+MI; T7 = GF+AS+MI; T8 =

GF+TH+MI; T9 = MC+AS+MI; T10 = MC+TH+MI; T11 = AS+TH+MI; T12 = GF+MC+AS+MI; T13 = GF+MC+TH+MI;

T14 = GF+AS+TH+MI; T15 = MC+AS+TH+MI; T16 = GF+MC+AS+TH+MI



antagonistic fungi Trichoderma harzianum on the root-knot development

of Meloidogyne incognita in chickpea plant

2

8.3.1.2.9d Number of eggs eggmass-1

Significant reduction in fecundity was observed when plants were inoculated

with either G. fasciculatum, M. ciceri, A. sativa or T. harzianum. Combination of all

these biocontrol agents resulted a maximum decline (95.4%) in fecundity. T.

harzianum alone caused significantly less number of eggs eggmass-1

(52) as compared

to other biocontrol agents viz., G. fasciculatum (107), M. ciceri (128) and A. sativa

(89) (Table 30 and Fig. 24).

8.3.1.2.9e Root-knot index (0-5)

Highest root-knot index (4) was observed in plants inoculated with M.

incognita alone. Inoculation of plant symbionts, antagonistic fungi and straw resulted

in a significant reduction in root-knot index (RKI). No infection was observed in the

roots of plants with combined inoculation of the above organisms and straw (Table

30).

8.3.1.2.9f Reproduction factor (pf/pi)

Plants inoculated with M. incognita showed the highest reproduction rate i.e.

12.20. Addition of G. fasciculatum, M. ciceri, A. sativa and T. harzianum brings about

a significant reduction in reproduction of nematode and the reduction was more

pronounced in the plants having all the biocontrol agents with M. incognita (0.70)

(Table 30 and Fig. 24).

8.3.2 EXPERIMENT 8B: With leaves of C. album



Plant length in terms of shoot, root and total length increased to a significant

level by the inoculation of the plant symbionts (G. fasciculatum, M. ciceri),

antagonistic fungi, T. harzianum and C. album in various combinations and in

individual treatments over control plants. The difference lies that individually M.

ciceri failed to cause a significant increase. Length of plants receiving treatments

GF+MC+CA, GF+CA+TH, MC+CA+TH and GF+MC+CA+TH were statistically

similar. However, the highest increase in plant length (67%) was observed while

inoculation of all biocontrol agents and C. album leaves. Individually, C. album is the

3

most effective in promoting the plant length (27%) followed by G. fasciculatum, T.

harzianum and M. ciceri (Table 31 and Fig. 25).




increase was observed (70.5%) while combined inoculation of GF+MC+CA+TH.

Inoculation of C. album leaves resulted in highest fresh weight (29.15%) as compared

to individual inoculation of A. sativa straw (26.2%) in the previous experiment 8A.

Treatments GF+MC+CA, GF+MC+TH, GF+CA+TH and GF+MC+CA+TH were at

par (Table 31 and Fig. 25).


Dry weight (shoot, root and total weight) of chickpea plants got improved by

inoculation of all biocontrol agents and C. album leaves in all treatments but the

increase was not significant in case of individual inoculation of M. ciceri (Table 31).

Combined treatments proved more efficient than individual treatments of either agent.

However, the highest increase was observed (72.4%) while inoculating all the plant

symbionts and C. album leaves together. Lowest dry weight (9.1) was observed in

plants treated with M. ciceri (Fig. 25).

8.3.2.1.4 Pods plant -1

Number of pods were increased significantly in case of all the treatments.

Highest pod number was recorded in the combined treatment GF+MC+CA+TH (92)

and lowest (43) while individual inoculation with M. ciceri (Table 31).


Nodulation increased to a greater extent while inoculating with root-

nodulating bacteria, M. ciceri. Individually, highest number of nodules (56) were

recorded in plants treated with M. ciceri followed by G. fasciculatum (9), C. album

(12) and T. harzianum (8). Leaves of botanical proved more effective than the straw

tested in experiment 8A (Table 31).

1

Table 31. Individual and interactive effect of AM fungus Glomus fasciculatum, root-nodule bacterium Mesorhizobium ciceri, leaves of

botanical Chenopodium album and an antagonistic fungi Trichoderma harzianum on the growth parameters of chickpea plant


plant-1 Nodules


Control 43.64g±2.18 21.82g±1.09 65.46g±3.27 43.36g±2.17 10.84i±0.54 54.20i±2.71 6.51h±0.33 1.62g±0.08 8.13h±0.41 31.0m±1.55 4.0j±0.20

GF 53.67d±2.68 26.84f±1.34 80.51f±4.03 54.18d±2.71 13.57h±0.68 67.75g±3.39 8.27de±0.41 2.06f±0.10 10.33f±0.52 50.0ij±2.50 9.0i±0.45

MC 46.85fg±2.34 23.45g±1.17 70.30g±3.52 48.13f±2.41 12.03i±0.60 60.16i±3.01 7.19g±0.36 1.91f±0.10 9.10g±0.46 43.0l±2.15 56.0f±2.80

CA 51.19de±2.56 31.94de±1.60 83.13ef±4.16 52.40de±2.62 17.60ef±0.88 70.00fg±3.50 7.57f±0.38 2.91d±0.15 10.48f±0.52 53.0i±2.65 12.0i±0.60

TH 48.14ef±2.41 31.07e±1.55 79.21f±3.96 50.32ef±2.52 16.61fg±0.83 66.93h±3.35 7.51fg±0.38 2.63e±0.13 10.14f±0.51 47.0jk±2.35 8.0ij±0.40

GF+MC 61.21a±3.06 30.67e±1.53 91.88cd±4.59 62.49b±3.12 15.62g±0.78 78.11de±3.91 9.34bc±0.47 2.46e±0.12 11.80cd±0.59 64.0fg±3.20 75.0d±3.75

GF+CA 60.73ab±3.04 38.01bc±1.90 98.74b±4.94 62.30bc±3.12 20.73bc±1.04 83.03bc±4.15 9.14c±0.46 3.42a±0.17 12.56bc±0.63 71.0d±3.55 24.0h±1.20

GF+TH 58.45bc±2.92 36.80c±1.84 95.25c±4.76 60.37c±3.02 20.00cd±1.00 80.37cd±4.02 8.85cd±0.44 3.27b±0.16 12.12c±0.61 68.0ef±3.40 22.0h±1.10

MC+CA 55.06cd±2.75 33.63d±1.68 88.69de±4.43 55.91d±2.80 18.85de±0.94 74.76e±3.74 8.22e±0.41 3.16bc±0.16 11.38de±0.57 63.0gh±3.15 74.0de±3.70

MC+TH 52.77d±2.64 32.45d±1.62 85.22e±4.26 53.99d±2.70 18.09e±0.90 72.08ef±3.60 7.91ef±0.40 3.00cd±0.15 10.91ef±0.55 59.0h±2.95 70.0e±3.50

CA+TH 59.68b±2.98 37.39c±1.87 97.07bc±4.85 61.14c±3.06 20.32c±1.02 81.46c±4.07 8.98c±0.45 3.36a±0.17 12.34c±0.62 69.0de±3.45 26.0h±1.30

GF+MC+CA 64.71a±3.24 40.35b±2.02 105.06a±5.25 65.98ab±3.30 22.09a±1.10 88.07a±4.40 9.93a±0.50 3.33ab±0.17 13.26ab±0.66 85.0bc±4.25 89.0b±4.45

GF+MC+TH 61.35a±3.07 38.54b±1.93 99.89b±4.99 61.67c±3.08 21.80ab±1.09 83.47ab±4.17 9.52b±0.48 3.19b±0.16 12.71b±0.64 73.0d±3.65 83.0c±4.15

GF+CA+TH 62.81a±3.14 45.00a±2.25 107.81a±5.39 67.81a±3.39 22.48a±1.12 90.29a±4.51 10.37a±0.52 3.41a±0.17 13.78a±0.69 89.0ab±4.45 34.0g±1.70

MC+CA+TH 63.80a±3.19 39.15b±1.96 102.95ab±5.1

5 64.21b±3.21 21.96a±1.10 86.17a±4.31 9.87ab±0.49 3.25b±0.16 13.12b±0.66 81.0c±4.05 82.0c±4.10

GF+MC+CA+TH 62.90a±3.15 46.40a±2.32 109.30a±5.47 69.31a±3.47 23.10a±1.15 92.41a±4.62 10.49a±0.52 3.53a±0.18 14.02a±0.70 92.0a±4.60 98.0a±4.90

C.D. (P=0.05) 4.04 2.46 6.48 4.15 1.31 5.45 0.62 0.21 0.83 4.69 4.23

C.D. (P=0.01) 5.44 3.31 8.73 5.59 1.76 7.34 0.84 0.28 1.11 6.32 5.70


GF = Glomus fasciculatum; MC = Mesorhizobium ciceri; CA = Chenopodium album; TH = Trichoderma harzianum


1


) fresh leaves

All combinations of G. fasciculatum, M. ciceri, C. album and T. harzianum

significantly increase the chlorophyll content of plants. However, the highest increase

(64%) was observed in GF+MC+CA+TH treated plants. Chlorophyll content in plants

with treatments GF+MC+CA, GF+MC+TH, GF+CA+TH, MC+CA+TH and

GF+MC+CA+TH were not significantly different from each other (Table 32 and Fig.

25).


) fresh leaves


individual and interactive treatments of all the microorganisms and leaves of

botanical. Nutrient contents in plants recieving treatments GF+MC+CA,

GF+MC+TH, GF+CA+TH, MC+CA+TH and GF+MC+CA+TH were at par.

Maximum nutrients (76.5%N, 74.2%P and 70.3%K) were recorded in plants while

inoculating with GF+ MC+CA+TH (Table 32 and Fig. 25).


Inoculation of M. ciceri and leaves of C. album brings about an increase in the

mycorrhization parameters (external and internal colonization, per cent arbuscules,

no. of chlamydospores in 1cm root segment and no. of chlamydospores recovered

from 100g rhizosphere soil) but the increase was not significant. Increase in

mycorrhization was comparatively more in plants treated with M. ciceri. Antagonistic

fungi, T. harzianum suppress the mycorrhizal colonization of G. fasciculatum,

although the reduction was not significant. Combined inoculation of GF+MC+CA and

GF+MC+CA+TH brings about a significant increase in mycorrhization. Moreover, all

the parameters in both the treatments were at par (Table 32).







1


botanical Chenopodium album and an antagonistic fungi Trichoderma harzianum on the chlorophyll content, nutrient status and

mycorrhization parameters of chickpea plant


GF = Glomus fasciculatum; MC = Mesorhizobium ciceri; CA = Chenopodium album; TH = Trichoderma harzianum



content

(mg g-1)


Colonization

(%)

Internal

Colonization

Per cent

arbuscules No. of

chlamydospores in

1cm root segment

No. of

chlamydospores

recovered from 100

g rhizosphere soil N P K

Control 2.402h±0.120 2.68f±0.13 0.240f±0.012 1.92e±0.10 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0e±0.00 0.0d±0.00

GF 2.876g±0.144 3.61e±0.18 0.329de±0.016 2.33d±0.12 62.7c±3.14 61.7bc±3.09 59.3c±2.97 58.0cd±2.90 893.0c±44.65

MC 3.135ef±0.157 3.82de±0.19 0.308e±0.015 2.29d±0.11 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0e±0.00 0.0d±0.00

CA 2.962fg±0.148 3.70e±0.18 0.315e±0.016 2.37d±0.12 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0e±0.00 0.0d±0.00

TH 2.821g±0.141 3.54e±0.18 0.311e±0.016 2.30d±0.12 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0e±0.00 0.0d±0.00

GF+MC 3.411d±0.171 4.35b±0.22 0.373bc±0.019 2.89c±0.14 67.8a±3.39 66.5a±3.32 64.0a±3.20 63.0ab±3.15 960.0a±48.00

GF+CA 3.315d±0.166 4.23bc±0.21 0.386b±0.019 2.97b±0.15 64.3b±3.21 62.9b±3.15 60.6b±3.03 59.0c±2.95 909.0b±45.45

GF+TH 3.194e±0.160 4.11cd±0.21 0.361c±0.018 2.81c±0.14 61.5c±3.07 60.8c±3.04 58.3c±2.91 57.0d±2.85 880.0c±44.00

MC+CA 3.518cd±0.176 4.46ab±0.22 0.356c±0.018 2.78c±0.14 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0e±0.00 0.0d±0.00

MC+TH 3.283de±0.164 4.27b±0.21 0.354cd±0.018 2.75c±0.14 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0e±0.00 0.0d±0.00

CA+TH 3.374d±0.169 4.25b±0.21 0.358c±0.018 2.93bc±0.15 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0e±0.00 0.0d±0.00

GF+MC+CA 3.877a±0.194 4.66a±0.23 0.402a±0.020 3.12ab±0.16 69.9a±3.50 68.5a±3.42 65.7a±3.28 65.0a±3.25 988.0a±49.40

GF+MC+TH 3.766ab±0.188 4.53a±0.23 0.393ab±0.020 2.99b±0.15 66.8ab±3.34 65.4ab±3.27 63.0ab±3.15 61.0bc±3.05 948.0ab±47.40

GF+CA+TH 3.663bc±0.183 4.49a±0.22 0.411a±0.021 3.18a±0.16 63.3bc±3.17 62.3b±3.11 60.0bc±3.00 59.0c±2.95 904.0bc±45.20

MC+CA+TH 3.790a±0.190 4.62a±0.23 0.389b±0.019 3.05b±0.15 0.0d±0.00 0.0d±0.00 0.0d±0.00 0.0e±0.00 0.0d±0.00

GF+MC+CA+TH 3.939a±0.197 4.73a±0.24 0.418a±0.021 3.27a±0.16 69.3a±3.46 67.6a±3.38 65.1a±3.26 64.0a±3.20 982.0a±49.10

C.D. (P=0.05) 0.237 0.293 0.025 0.195 3.79 3.72 3.58 3.50 53.80

C.D. (P=0.01) 0.319 0.395 0.034 0.262 5.10 5.01 4.82 4.72 72.46

1


Chlo

rop

hyll

co

nte

nt

(mg g

-1)

0

1

2

3

4


Pla

nt

dry

wei

ght

(g)

02468

10121416


Pla

nt

length

(c

m)

0

20

40

60

80

100

120


Pla

nt

fres

h w

eight

(g)

0

20

40

60

80

100






Treatments


Nutr

ient

conte

nts

(mg g

-1)

0.0

1.0

2.0

3.0

4.0

5.0

6.0N P K

T1 = Control; T2 = G. fasciculatum (GF); T3 = M. ciceri (MC); T4 = C. album (CA); T5 = T. harzianum (TH); T6 = GF+MC; T7 = GF+CA; T8 = GF+TH; T9 = MC+CA; T10 = MC+TH; T11 = CA+TH; T12 = GF+MC+CA; T13 =

GF+MC+TH; T14 = GF+CA+TH; T15 = MC+CA+TH; T16 = GF+MC+CA+TH


root-nodule bacterium Mesorhizobium ciceri, leaves of botanical

Chenopodium album and an antagonistic fungi Trichoderma harzianum on

the growth parameters, chlorophyll and nutrient contents of chickpea

plant

2



(G. fasciculatum, M. ciceri), and C. album leaves. All the biocontrol agents with

botanical in various combinations reduce the deleterious effect of nematode and

caused a significant increase in plant length. Individually, T. harzianum with M.

incognita caused the maximum enhancement of plant length (26%) over nematode-

inoculated untreated plants. In combined treatments, GF+CA+TH+MI and

GF+MC+CA+TH+MI were statistically similar. However, treatment

GF+MC+CA+TH+MI was most effective in reducing the nematode and resulted

highest increase (73.8%) over all other treatments and nematode-inoculated untreated

control (Table 33 and Fig. 26).



of M. incognita. Inoculation of plant symbionts, antagonistic fungi and organic waste

alone as well as in combinations overcome the loss caused by the nematode and

brings about a significant improvement in the fresh weight of chickpea plants over

nematode-inoculated untreated plants. Highest increase in fresh weight (75.6%) was

reported while combined inoculation of all the microorganisms i.e.

GF+MC+AS+TH+MI which was comparatively more than combined treatment with

A. sativa straw (70%) in previous experiment. Reduction by M. incognita was lowest

in plants treated with T. harzianum (17.97%) as compared to other plant symbionts

and botanical (Table 33 and Fig. 26).



trend of results as in case of plant fresh weight. T. harzianum proved to be most

effective in suppressing M. incognita. Combined inoculation of all biocontrol agents

and botanical overcome the loss caused by the nematode and the most effective is the

GF+MC+CA+TH+MI treatment with lowest reduction and thus promoted highest

increase with M. incognita (76.0%) over nematode-inoculated untreated plants (Fig.

26). Treatments CA+TH+MI and GF+MC+CA+MI were at par in suppressing

nematode activity and resulted in dry weight which was not significantly different

(Table 33).

3


Root-knot nematode, M. incognita significantly reduced the number of pods

over uninoculated untreated control plants. Lowest reduction occurred in T.

harzianum (6.38%) compared to other agents applied with nematode. Highest pod

number was observed while combined inoculation of all the biocontrol agents,

GF+MC+CA+TH+MI (81) and lowest in the M. ciceri (26) treated plants (Table 33).


It was evident from the table 33 that no nodules were observed while

inoculating the untreated control plants with M. incognita. M. ciceri inhibits the effect

of nematode and increase the nodule number to a greater extent as compared to other

microorganisms and botanical. In individual treatments, highest nodule number (47)

was observed in M. ciceri followed by T. harzianum (10), C. album (9) and G.

fasciculatum (7) with M. incognita. In the presence of nematode, all the plant

symbionts and botanical together resulting highest nodule number (85) as compared

to all other single, double and triple combinations (Table 33).


fresh leaves)



reported in case of plants treated with T. harzianum. G. fasciculatum, M. ciceri, C.

album and T. harzianum overcome the loss caused by the nematode and highest

increase (78%) was observed in plants inoculated with all of them in combination

over nematode-inoculated untreated control plants (Table 34 and Fig. 26). Plants with

treatments MC+CA+MI, MC+TH+MI, CA+TH+MI and GF+CA+TH+MI were

statistically same in inhibiting the nematode activity (Fig. 26).


fresh leaves)


M. incognita as compared to uninoculated control. All the plant symbionts and

botanical were effective in suppressing the deleterious effect of nematode and

significantly increase the nutrient contents. Individually, M. ciceri resulted a highest

increase in N content (35.2%) and G. fasciculatum cause a highest increase in P

1


botanical Chenopodium album and an antagonistic fungi Trichoderma harzianum on the growth parameters of chickpea plant

infected with root-knot nematode, Meloidogyne incognita

Treatments Plant length (cm) Plant fresh weight (g) Plant dry weight (g)

Pods plant-1 Nodules


Control 37.85j±1.89 18.96i±0.95 56.81j±2.84 34.66h±1.73 8.67h±0.43 43.33i±2.17 5.18g±0.26 1.29j±0.06 6.47h±0.32 18.0l±0.90 0.0l±0.00

GF+MI 46.65hi±2.33 23.22h±1.16 69.87h±3.49 43.19fg±2.16 10.75g±0.54 53.94gh±2.70 6.08f±0.30 2.02h±0.10 8.10g±0.41 38.0j±1.90 7.0k±0.35

MC+MI 43.08i±2.15 20.04i±1.00 63.12i±3.16 40.10g±2.01 10.24g±0.51 50.34h±2.52 6.12f±0.31 1.50i±0.08 7.62g±0.38 26.0k±1.30 47.0g±2.35

CA+MI 47.23h±2.36 23.66h±1.18 70.89h±3.54 43.66f±2.18 10.83g±0.54 54.49g±2.72 6.15f±0.31 2.00h±0.10 8.15g±0.41 43.0i±2.15 9.0k±0.45

TH+MI 47.76gh±2.39 23.82gh±1.19 71.58gh±3.58 44.03f±2.20 10.87g±0.54 54.90g±2.75 6.16f±0.31 2.08gh±0.10 8.24g±0.41 44.0i±2.20 10.0k±0.50

GF+MC+MI 51.42fg±2.57 25.61fg±1.28 77.03fg±3.85 47.56e±2.38 11.81f±0.59 59.37f±2.97 6.75e±0.34 2.18fg±0.11 8.93f±0.45 55.0h±2.75 60.0f±3.00

GF+CA+MI 56.23de±2.81 28.08de±1.40 84.31de±4.22 52.00d±2.60 13.00de±0.65 65.00de±3.25 7.35cd±0.37 2.43de±0.12 9.78de±0.49 62.0ef±3.10 20.0j±1.00

GF+TH+MI 56.86d±2.84 28.35d±1.42 85.21d±4.26 52.65cd±2.63 13.21cd±0.66 65.86cd±3.29 7.41c±0.37 2.50cd±0.13 9.91cd±0.50 65.0de±3.25 21.0ij±1.05

MC+CA+MI 52.00f±2.60 26.00f±1.30 78.00f±3.90 48.14e±2.41 12.10f±0.61 60.24f±3.01 6.84e±0.34 2.23f±0.11 9.07f±0.45 57.0gh±2.85 64.0e±3.20

MC+TH+MI 52.94ef±2.65 26.37ef±1.32 79.31ef±3.97 49.19de±2.46 12.25ef±0.61 61.44ef±3.07 6.94de±0.35 2.27ef±0.11 9.21ef±0.46 60.0fg±3.00 68.0d±3.40

CA+TH+MI 57.55d±2.88 28.80cd±1.44 86.35d±4.32 53.29c±2.66 13.44c±0.67 66.73c±3.34 7.45c±0.37 2.60c±0.13 10.05c±0.50 67.0cd±3.35 24.0i±1.20

GF+MC+CA+MI 58.86cd±2.94 29.48c±1.47 88.34cd±4.42 54.81c±2.74 13.56c±0.68 68.37c±3.42 7.59bc±0.38 2.63bc±0.13 10.22c±0.51 70.0c±3.50 74.0c±3.70

GF+MC+TH+MI 61.64bc±3.08 30.66bc±1.53 92.30bc±4.62 56.68b±2.83 14.25b±0.71 70.93b±3.55 8.04ab±0.40 2.67b±0.13 10.71b±0.54 71.0bc±3.55 73.0c±3.65

GF+CA+TH+MI 63.86ab±3.19 31.98ab±1.60 95.84ab±4.79 58.85ab±2.94 14.81ab±0.74 73.66ab±3.68 8.34a±0.42 2.79ab±0.14 11.13ab±0.56 75.0b±3.75 30.0h±1.50

MC+CA+TH+MI 59.91c±3.00 30.02c±1.50 89.93c±4.50 55.80bc±2.79 14.05bc±0.70 69.85bc±3.49 7.90b±0.39 2.65b±0.13 10.55bc±0.53 69.0c±3.45 79.0b±3.95

GF+MC+CA+TH+MI 65.93a±3.30 32.85a±1.64 98.78a±4.94 60.76a±3.04 15.32a±0.77 76.08a±3.80 8.53a±0.43 2.86a±0.14 11.39a±0.57 81.0a±4.05 85.0a±4.25

C.D. (P=0.05) 3.81 1.90 5.70 3.52 0.88 4.40 0.50 0.16 0.66 4.10 3.72

C.D. (P=0.01) 5.13 2.55 7.68 4.74 1.19 5.93 0.67 0.22 0.89 5.53 5.00


GF = Glomus fasciculatum; MC = Mesorhizobium ciceri; CA = Chenopodium album; TH = Trichoderma harzianum; MI = Meloidogyne incognita


2


botanical Chenopodium album and an antagonistic fungi Trichoderma harzianum on the chlorophyll content, nutrient status and

mycorrhization parameters of chickpea plant infected with root-knot nematode, Meloidogyne incognita





content (mg g-1)


Colonization

(%)

Internal

Colonization

Per cent

arbuscules No. of

chlamydospores

in 1cm root

segment

No. of

chlamydospores

recovered from

100 g

rhizosphere soil

N P K

Control 1.825e±0.091 2.30g±0.12 0.160e±0.008 1.54f±0.08 0.0e±0.0 0.0e±0.0 0.0e±0.0 0.0e±0.00 0.0e±0.0

GF+MI 2.208d±0.110 2.75f±0.14 0.211d±0.011 1.85e±0.09 59.1cd±3.0 59.2cd±3.0 57.1cd±2.9 56.0c±2.80 858.0cd±42.9

MC+MI 2.376d±0.119 3.11e±0.16 0.195d±0.010 1.81e±0.09 0.0e±0.0 0.0e±0.0 0.0e±0.0 0.0e±0.00 0.0e±0.0

CA+MI 2.290d±0.114 2.94ef±0.15 0.200d±0.010 1.87e±0.09 0.0e±0.0 0.0e±0.0 0.0e±0.0 0.0e±0.00 0.0e±0.0

TH+MI 2.300d±0.115 2.96e±0.15 0.204d±0.010 1.88e±0.09 0.0e±0.0 0.0e±0.0 0.0e±0.0 0.0e±0.00 0.0e±0.0

GF+MC+MI 2.875bc±0.144 3.83c±0.19 0.262bc±0.013 2.25d±0.11 65.2ab±3.3 64.9ab±3.2 62.3ab±3.1 61.0ab±3.05 938.0ab±46.9

GF+CA+MI 2.792c±0.140 3.54d±0.18 0.266b±0.013 2.32cd±0.12 62.1bc±3.1 61.1c±3.1 58.9bc±2.9 58.0bc±2.90 850.0d±42.5

GF+TH+MI 2.839c±0.142 3.65cd±0.18 0.270b±0.014 2.40c±0.12 58.3d±2.9 57.1d±2.9 54.7d±2.7 54.0d±2.70 824.0d±41.2

MC+CA+MI 2.931b±0.147 3.96b±0.20 0.253c±0.013 2.26d±0.11 0.0e±0.0 0.0e±0.0 0.0e±0.0 0.0e±0.00 0.0e±0.0

MC+TH+MI 2.956b±0.148 3.98b±0.20 0.257c±0.013 2.28d±0.11 0.0e±0.0 0.0e±0.0 0.0e±0.0 0.0e±0.00 0.0e±0.0

CA+TH+MI 2.898b±0.145 3.90bc±0.20 0.264b±0.013 2.47b±0.12 0.0e±0.0 0.0e±0.0 0.0e±0.0 0.0e±0.00 0.0e±0.0

GF+MC+CA+MI 3.051ab±0.153 4.12ab±0.21 0.277ab±0.014 2.47b±0.12 68.0a±3.4 66.6a±3.3 64.2a±3.2 63.0a±3.15 968.0a±48.4

GF+MC+TH+MI 3.095a±0.155 4.15a±0.21 0.280a±0.014 2.48b±0.12 63.6b±3.2 62.7bc±3.1 59.9b±3.0 59.0b±2.95 905.0bc±45.3

GF+CA+TH+MI 3.017b±0.151 4.06b±0.20 0.288a±0.014 2.64a±0.13 60.8c±3.0 59.5c±3.0 57.6c±2.9 56.0c±2.80 864.0c±43.2

MC+CA+TH+MI 3.205a±0.160 4.29a±0.21 0.274b±0.014 2.56ab±0.13 0.0e±0.0 0.0e±0.0 0.0e±0.0 0.0e±0.00 0.0e±0.0

GF+MC+CA+TH+MI 3.241a±0.162 4.32a±0.22 0.293a±0.015 2.70a±0.13 67.4a±3.4 66.0a±3.3 63.6a±3.2 62.0a±3.10 957.0a±47.9

C.D. (P=0.05) 0.194 0.256 0.018 0.158 3.64 3.59 3.45 3.38 51.67

C.D. (P=0.01) 0.262 0.345 0.024 0.213 4.90 4.83 4.64 4.56 69.59

1

T1 = M. incognita (MI); T2 = G. fasciculatum (GF) + M. incognita (MI); T3 = M. ciceri (MC) + M. incognita (MI); T4 = C.

album (CA) + M. incognita (MI); T5 = T. harzianum (TH) + M. incognita (MI) ; T6 = GF+MC+MI; T7 = GF+CA+MI; T8 = GF+TH+MI; T9 = MC+CA+MI; T10 = MC+TH+MI; T11 = CA+TH+MI; T12 = GF+MC+CA+MI; T13 = GF+MC+TH+MI;

T14 = GF+CA+TH+MI; T15 = MC+CA+TH+MI; T16 = GF+MC+CA+TH+MI


root-nodule bacterium Rhizobium, leaves of Chenopodium album and an


chlorophyll and nutrient contents of chickpea plant infected with

Meloidogyne incognita


Chlo

rophyll

co

nte

nt

(mg g

-1)

0.0

0.5

1.0

1.5

2.0

2.5

3.0


Pla

nt

dry

wei

ght

(g)

0

2

4

6

8

10

12


Pla

nt

length

(c

m)

0

20

40

60

80

100

120


Pla

nt

fres

h w

eight

(g)

0

20

40

60

80






Treatments


Nutr

ient

conte

nts

(mg g

-1)

0.0

1.0

2.0

3.0

4.0

5.0N P K

2

content (31.8%) as compared to other microorganisms and C. album leaves in

nematode-inoculated plants. Combined inoculations (dual, triple and quadrate) proved

much better than individual appplication. GF+MC+CA+TH+MI proved to be most

effective in reducing the loss caused by nematode, thus resulted highest increase in

nutrients (87.8%N, 83.1%P and 75.3%K) over nematode-inoculated-untreated plants

(Table 34 and Fig. 26) and were comparatively more than A. straw combined

treatment (85.6%N, 80%P and 71.4%K) in experiment 8A.





harzianum significantly reduced the colonization. However, lowest reduction by

nematode was observed in case of plants inoculated with GF+R+CA and thus resulted

in highest mycorrhization values (67.96% external colonization, 66.63% internal

colonization, 64.16% arbuscules, 63 chlamydospores in 1cm root segment and 968

chlamydospores in 100g rhizosphere soil) in M. incognita inoculated plants(Table 34).

Botanical prove more effective in increasing the mycorrhization over straw.



AM fungi, root-nodule bacteria, botanical and antagonistic fungi significantly

reduced the population of M. incognita in soil as well as in root. Inoculation of T.

harzianum resulted in an acute reduction in soil and root population of nematode i.e.

65% and 67.5% respectively compared to other agents (Table 35 and Fig. 27).

Reduction in nematode population in the plants with treatments CA+TH+MI,

GF+MC+TH+MI, GF+CA+TH+MI and MC+CA+TH+MI were at par.


Highest number of galls (210) were observed in plants which were inoculated

with M. incognita alone. All the treatments significantly decrease the number of galls

in root system. Lowest number of galls (10) was observed while combined

1


botanical Chenopodium album and an antagonistic fungi, Trichoderma harzianum on the root-knot development of Meloidogyne

incognita in chickpea plant Treatments Nematode population No. of galls root

system-1 No. of eggmasses



(0-5)

Reproduction factor

(pf/pi) Soil Root

Control 11,915.0a±595.8 243.0a±12.1 210.0a±10.5 84.0a±4.2 154.0a±7.7 4.0a 12.2a±0.61

GF+MI 9,008.0c±450.4 176.0c±8.8 148.0c±7.4 60.0c±3.0 107.0c±5.4 2.0c 8.8c±0.44

MC+MI 10,052.0b±502.6 202.0b±10.1 173.0b±8.7 70.0b±3.5 128.0b±6.4 3.0b 10.1b±0.51

CA+MI 4,863.0e±243.1 95.0e±4.8 82.0e±4.1 32.0e±1.6 61.0e±3.0 1.0d 4.8e±0.24

TH+MI 4,170.0f±208.5 79.0f±3.9 69.0f±3.5 28.0f±1.4 52.0f±2.6 0.0 3.9f±0.20

GF+MC+MI 7,445.0d±372.3 152.0d±7.6 124.0d±6.2 51.0d±2.5 94.0d±4.7 1.0d 7.6d±0.38

GF+CA+MI 2,562.0h±128.1 51.0gh±2.5 43.0h±2.1 18.0gh±0.9 33.0h±1.7 0.0 2.6gh±0.13

GF+TH+MI 1,787.0i±89.4 33.0i±1.7 30.0i±1.5 12.0i±0.6 22.0i±1.1 0.0 1.7i±0.08

MC+CA+MI 3,098.0g±154.9 58.0g±2.9 52.0g±2.6 21.0g±1.1 39.0g±1.9 1.0d 2.9g±0.15

MC+TH+MI 2,383.0h±119.2 45.0h±2.3 40.0h±2.0 16.0h±0.8 29.0h±1.4 0.0 2.3h±0.11

CA+TH+MI 1,192.0jk±59.6 24.0jk±1.2 20.0jk±1.0 8.0jk±0.4 15.0jk±0.8 0.0 1.2jk±0.06

GF+MC+CA+MI 1,548.0ij±77.4 30.0ij±1.5 27.0ij±1.4 11.0ij±0.6 20.0ij±1.0 0.0 1.5ij±0.08

GF+MC+TH+MI 1,072.0k±53.6 21.0k±1.0 18.0k±0.9 7.0k±0.4 14.0k±0.7 0.0 1.0k±0.05

GF+CA+TH+MI 774.0lm±38.7 16.0kl±0.8 14.0kl±0.7 6.0kl±0.3 10.0k±0.5 0.0 0.7l±0.04

MC+CA+TH+MI 953.0kl±47.6 18.0k±0.9 16.0k±0.8 7.0k±0.4 12.0k±0.6 0.0 0.9kl±0.04

GF+MC+CA+TH+MI 540.0m±27.0 11.0l±0.6 10.0l±0.5 4.0l±0.2 11.0k±0.6 0.0 0.6l±0.03

C.D. (P=0.05) 441.37 8.83 7.53 3.04 5.55 0.442

C.D. (P=0.01) 594.40 11.89 10.14 4.09 7.47 0.596




1


Ch

loro

ph

yll

con

ten

t

(mg g

-1)

0

1

2

3

4


Pla

nt

dry

wei

gh

t

(g)

02468

10121416


Pla

nt

len

gth

(cm

)

0

20

40

60

80

100

120


Pla

nt

fres

h w

eigh

t

(g)

0

20

40

60

80

100






Treatments


Nutr

ien

t co

nte

nts

(mg g

-1)

0.0

1.0

2.0

3.0

4.0

5.0

6.0N P K

T1 = M. incognita (MI); T2 = G. fasciculatum (GF) + M. incognita (MI); T3 = M. ciceri (MC) + M. incognita (MI); T4 = C.

album (CA) + M. incognita (MI); T5 = T. harzianum (TH) + M. incognita (MI) ; T6 = GF+MC+MI; T7 = GF+CA+MI; T8 = GF+TH+MI; T9 = MC+CA+MI; T10 = MC+TH+MI; T11 = CA+TH+MI; T12 = GF+MC+CA+MI; T13 = GF+MC+TH+MI;

T14 = GF+CA+TH+MI; T15 = MC+CA+TH+MI; T16 = GF+MC+CA+TH+MI


root-nodule bacterium Mesorhizobium ciceri, leaves of botanical

Chenopodium album and an antagonistic fungi Trichoderma harzianum

on the root-knot development of chickpea plant infected with

Meloidogyne incognita

2

inoculation of GF+MC+CA+TH+MI and the same number of galls were reported in

the previous experiment in combination with straw (Table 35 and Fig. 27).


Number of eggmasses root system-1

were highest when M. incognita alone

was inoculated. Presence of G. fasciculatum, M. ciceri, C. album and T. harzianum

caused significant reduction in this number. Maximum reduction (95%) was observed

when all of them were inoculated together and this reduction was significantly higher

than the reduction caused by either of them alone (Table 35 and Fig. 27).



with either of the plant symbionts and botanical. Combination of all these biocontrol

agents resulted a maximum decline (95.4%) in fecundity (Table 35 and Fig. 27). T.

harzianum alone caused significantly less number of eggs eggmass-1

(52) as compared

to other biocontrol agents viz., G. fasciculatum (107), M. ciceri (128) and C. album

(61).


Inoculation of plant symbionts, antagonistic fungi and botanical resulted in a

significant reduction in root-knot index (RKI). Highest root-knot index (4) was

observed in plants inoculated with M. incognita alone. No infection was observed in

the roots of plants with combined inoculation of the above organisms and botanical

(Table 35).



12.20. Addition of G. fasciculatum, M. ciceri, C. album and T. harzianum brings

about a significant reduction in reproduction of nematode and the reduction was more

pronounced in the plants having all the biocontrol agents with M. incognita (0.60)

(Table 35 and Fig. 27). C. album inhibits the nematode reproductive ability over A.

sativa straw used earlier in the experiment 8A.

3

8.3.3 EXPERIMENT 8C: With poultry manure



It was evident from the table 36 that inoculation of the AM fungus G.

fasciculatum, root-nodulating bacteria M. ciceri, antagonistic fungi, T. harzianum and

poultry manure in all possible combinations caused a significant increase in plant

length (shoot, root and total length) over control except M. ciceri individual treatment,

which failed to cause a significant increase. Individually, poultry manure is the most

effective of all the plant symbionts in promoting the plant length (Fig. 28). Treatments

GF+MC+PM, GF+PM+TH, MC+PM+TH and GF+MC+PM+TH were not

statistically different from each other. However, the highest increase in plant length

(71%) was observed while inoculation of all biocontrol agents with poultry manure as

compared to A. sativa straw (63.46%) in experiment 8A and C. album leaves (67%) in

experiment 8B.




increase was observed (73.5%) while combined inoculation of GF+MC+PM+TH.

Individual inoculation of poultry manure resulted in highest fresh weight (32.3%)

followed by individual inoculation of G. fasciculatum (25%), T. harzianum (23.5%)

and M. ciceri (10.9%). Fresh weight of plants with treatments GF+MC+PM,

GF+PM+TH, MC+PM+TH and GF+MC+PM+TH were statistically similar.

However, GF+MC+PM+TH caused highest value of fresh weight (94.03) (Table 36

and Fig. 28).


Plant dry weight (shoot, root and total dry weight) was significantly increased

by the inoculation of all biocontrol agents and poultry manure but the increase was

not significant in case of individual inoculation of M. ciceri. However, the highest

increase was observed (74.0%) while inoculating all the plant symbionts and poultry

manure together which is comparatively more than the dry weight of plants inoculated

with straw and botanical in previous experiments (Table 36 and Fig. 28).

1

Table 36. Individual and interactive effect of AM fungus Glomus fasciculatum, root-nodule bacterium Mesorhizobium ciceri, animal waste

as poultry manure and an antagonistic fungi Trichoderma harzianum on the growth parameters of chickpea plant


plant-1 Nodules


Control 43.64f±2.18 21.82l±1.09 65.46h±3.27 43.36h±2.17 10.84h±0.54 54.20h±2.71 6.51g±0.33 1.62h±0.08 8.13h±0.41 31.0j±1.55 4.0l±0.20

GF 53.67d±2.68 26.84k±1.34 80.51g±4.03 54.18e±2.71 13.57g±0.68 67.75f±3.39 8.27cd±0.41 2.06g±0.10 10.33f±0.52 50.0h±2.50 9.0k±0.45

MC 46.85ef±2.34 23.45l±1.17 70.30h±3.52 48.13g±2.41 12.03h±0.60 60.16g±3.01 7.19f±0.36 1.91g±0.10 9.10g±0.46 43.0i±2.15 56.0f±2.80

PM 55.60cd±2.78 31.72j±1.59 87.32ef±4.37 53.40ef±2.67 18.31e±0.92 71.71f±3.59 7.91d±0.40 2.88e±0.14 10.79f±0.54 57.0g±2.85 16.0j±0.80

TH 48.14e±2.41 31.07j±1.55 79.21g±3.96 50.32fg±2.52 16.61f±0.83 66.93f±3.35 7.51ef±0.38 2.63f±0.13 10.14f±0.51 47.0hi±2.35 8.0kl±0.40

GF+MC 61.21ab±3.06 30.67j±1.53 91.88de±4.59 62.49c±3.12 15.62f±0.78 78.11d±3.91 9.34ab±0.47 2.46f±0.12 11.80d±0.59 64.0f±3.20 75.0d±3.75

GF+PM 62.58a±3.13 42.16de±2.11 104.74b±5.24 65.99b±3.30 22.08b±1.10 88.07b±4.40 9.10b±0.46 4.15a±0.21 13.25b±0.66 75.0d±3.75 30.0h±1.50

GF+TH 58.45b±2.92 36.80gh±1.84 95.25cd±4.76 60.37cd±3.02 20.00d±1.00 80.37cd±4.02 8.85bc±0.44 3.27c±0.16 12.12cd±0.61 68.0ef±3.40 22.0i±1.10

MC+PM 58.15bc±2.91 34.67hi±1.73 92.82d±4.64 56.28de±2.81 21.23cd±1.06 77.51de±3.88 8.42c±0.42 3.25c±0.16 11.67de±0.58 66.0f±3.30 78.0d±3.90

MC+TH 52.77d±2.64 32.45ij±1.62 85.22fg±4.26 53.99e±2.70 18.09e±0.90 72.08ef±3.60 7.91d±0.40 3.00de±0.15 10.91ef±0.55 59.0g±2.95 70.0e±3.50

PM+TH 61.83a±3.09 40.68ef±2.03 102.51b±5.13 64.05bc±3.20 21.31c±1.07 85.36bc±4.27 9.07b±0.45 3.80b±0.19 12.87bc±0.64 72.0de±3.60 26.0hi±1.30

GF+MC+PM 62.84a±3.14 45.50bc±2.27 108.34a±5.42 67.72ab±3.39 23.12a±1.16 90.84a±4.54 9.55a±0.48 4.21a±0.21 13.76a±0.69 91.0b±4.55 97.0b±4.85

GF+MC+TH 61.35a±3.07 38.54fg±1.93 99.89bc±4.99 61.67c±3.08 21.80bc±1.09 83.47c±4.17 9.52a±0.48 3.19cd±0.16 12.71c±0.64 73.0d±3.65 83.0c±4.15

GF+PM+TH 63.00a±3.15 47.82ab±2.39 110.82a±5.54 68.84a±3.44 23.90a±1.20 92.74a±4.64 9.77a±0.49 4.28a±0.21 14.05a±0.70 96.0a±4.80 38.0g±1.90

MC+PM+TH 62.27a±3.11 43.90cd±2.20 106.17ab±5.31 66.21b±3.31 22.95ab±1.15 89.16ab±4.46 9.41a±0.47 4.17a±0.21 13.58ab±0.68 85.0c±4.25 87.0c±4.35

GF+MC+PM+TH 63.10a±3.15 48.84a±2.44 111.94a±5.60 70.47a±3.52 23.56a±1.18 94.03a±4.70 9.83a±0.49 4.32a±0.22 14.15a±0.71 98.0a±4.90 107.0a±5.35

C.D. (P=0.05) 4.07 2.57 6.62 4.21 1.35 5.56 0.62 0.23 0.84 4.88 4.44

C.D. (P=0.01) 5.49 3.46 8.92 5.67 1.82 7.48 0.83 0.31 1.13 6.58 5.98


GF = Glomus fasciculatum; MC = Mesorhizobium ciceri; PM = Poultry manure; TH = Trichoderma harzianum Mean values with different letters within the column are significantly different at P = 0.05

1

8.3.1.1.4 Pods plant -1

Significant increase in pods number was observed in case of all the treatments.

Highest pod number was recorded in the treatment GF+MC+PM+TH (98).

Individually 57 pods were reported in poultry manure treated plants followed by G.

fasciculatum (50), T. harzianum (47) and M. ciceri (43) (Table 36).


Nodules number increased to a greater extent while inoculating with root-

nodulating bacteria, M. ciceri. 9, 16 and 8 nodules were recorded in plants inoculated

individually with G. fasciculatum, poultry manure and T. harzianum respectively

which were comparatively very less as compared to the nodules number in plants

treated individually with M. ciceri (56). Pods number in plants with treatments

GF+PM+TH and GF+MC+PM+TH were at par (Table 36).


) fresh leaves

Single, double, triple and quadrate combinations of G. fasciculatum, M. ciceri,

poultry manure and T. harzianum significantly increase the chlorophyll content of

plants. Chlorophyll content in plants with treatments GF+MC+PM, GF+MC+TH,

MC+ PM +TH and GF+MC+ PM +TH were not significantly different from each

other (Table ). However, the highest increase (66%) was observed in

GF+MC+PM+TH treated plants over other treatments (Table 37 and Fig. 28).


) fresh leaves


individual and interactive treatments. Nutrient contents in plants with treatments

GF+MC+PM, GF+PM+TH and GF+MC+PM+TH were at par. Maximum nutrients

(80%N, 78.3%P and 74.5%K) were recorded in plants while inoculating with GF+

MC+PM+TH as compared to other treatments (Table 37). Comparatively less

nutrients were recorded in earlier experiments with straws and botanicals (Fig. 28).


M. ciceri and poultry manure brings about an increase in the mycorrhization

parameters in terms of external and internal colonization, per cent arbuscules, no. of

1


as poultry manure and an antagonistic fungi Trichoderma harzianum on the chlorophyll content, nutrient status and

mycorrhization parameters of chickpea plant


GF = Glomus fasciculatum; MC = Mesorhizobium ciceri; PM = Poultry manure; TH = Trichoderma harzianum Mean values with different letters within the column are significantly different at P = 0.05


content

(mg g-1)


Colonization

(%)

Internal

Colonization

(%)

Per cent

arbuscules No. of

chlamydospores in

1cm root segment

No. of

chlamydospores

recovered from 100

g rhizosphere soil N P K

Control 2.402g±0.120 2.68f±0.13 0.240g±0.012 1.92f±0.10 0.00e±0.00 0.00e±0.00 0.00e±0.00 0.0d±0.00 0.0e±0.00

GF 2.876ef±0.144 3.61e±0.18 0.329ef±0.016 2.33e±0.12 62.70d±3.14 61.70d±3.09 59.30d±2.97 58.0c±2.90 893.0d±44.65

MC 3.135d±0.157 3.82de±0.19 0.308f±0.015 2.29e±0.11 0.00e±0.00 0.00e±0.00 0.00e±0.00 0.0d±0.00 0.0e±0.00

PM 3.093de±0.155 3.76e±0.19 0.323f±0.016 2.43e±0.12 0.00e±0.00 0.00e±0.00 0.00e±0.00 0.0d±0.00 0.0e±0.00

TH 2.821f±0.141 3.54e±0.18 0.311f±0.016 2.30e±0.12 0.00e±0.00 0.00e±0.00 0.00e±0.00 0.0d±0.00 0.0e±0.00

GF+MC 3.411c±0.171 4.35b±0.22 0.373c±0.019 2.89c±0.14 67.80bc±3.39 66.50bc±3.32 64.00bc±3.20 63.0ab±3.15 960.0bc±48.00

GF+PM 3.242c±0.162 4.22c±0.21 0.384bc±0.019 2.97bc±0.15 66.14c±3.31 64.78c±3.24 62.21c±3.11 61.0b±3.05 942.0c±47.10

GF+TH 3.194cd±0.160 4.11cd±0.21 0.361cd±0.018 2.81cd±0.14 61.45d±3.07 60.77d±3.04 58.29d±2.91 57.0c±2.85 880.0d±44.00

MC+PM 3.338c±0.167 4.30b±0.21 0.356d±0.018 2.94c±0.15 0.00e±0.00 0.00e±0.00 0.00e±0.00 0.0d±0.00 0.0e±0.00

MC+TH 3.283c±0.164 4.27bc±0.21 0.354de±0.018 2.75d±0.14 0.00e±0.00 0.00e±0.00 0.00e±0.00 0.0d±0.00 0.0e±0.00

PM+TH 3.218c±0.161 4.17c±0.21 0.379c±0.019 3.02b±0.15 0.00e±0.00 0.00e±0.00 0.00e±0.00 0.0d±0.00 0.0e±0.00

GF+MC+PM 3.927a±0.196 4.77a±0.24 0.407ab±0.020 3.16ab±0.16 71.79a±3.59 70.34a±3.52 67.72a±3.39 66.0a±3.30 1,022.0a±51.10

GF+MC+TH 3.766ab±0.188 4.53ab±0.23 0.393b±0.020 2.99b±0.15 66.77c±3.34 65.40c±3.27 62.97c±3.15 61.0b±3.05 948.0c±47.40

GF+PM+TH 3.706b±0.185 4.50b±0.23 0.421a±0.021 3.29a±0.16 65.21cd±3.26 63.85cd±3.19 61.49cd±3.07 60.0bc±3.00 929.0cd±46.45

MC+PM+TH 3.835a±0.192 4.67a±0.23 0.392b±0.020 3.07b±0.15 0.00e±0.00 0.00e±0.00 0.00e±0.00 0.0d±0.00 0.0e±0.00

GF+MC+PM+TH 3.987a±0.199 4.82a±0.24 0.428a±0.021 3.35a±0.17 70.85ab±3.54 69.41ab±3.47 66.65ab±3.33 65.0a±3.25 1,009.0ab±50.45

C.D. (P=0.05) 0.237 0.294 0.026 0.197 3.84 3.77 3.62 3.54 54.64

C.D. (P=0.01) 0.319 0.396 0.035 0.266 5.17 5.07 4.88 4.76 73.59

1

T1 = Control; T2 = G. fasciculatum (GF); T3 = M. ciceri (MI); T4 = Poultry manure (PM); T5 = T. harzianum (TH); T6 =

GF+MC; T7 = GF+PM; T8 = GF+TH; T9 = MC+PM; T10 = MC+TH; T11 = PM+TH; T12 = GF+MC+PM; T13 = GF+MC+TH;

T14 = GF+PM+TH; T15 = MC+PM+TH; T16 = GF+MC+PM+TH


root-nodule bacterium Mesorhizobium ciceri, animal waste as poultry

manure and an antagonistic fungi Trichoderma harzianum on the growth

parameters, chlorophyll and nutrient contents of chickpea plant


Chlo

rophyll

co

nte

nt

(mg g

-1)

0

1

2

3

4


Pla

nt

dry

wei

ght

(g)

02468

10121416


Pla

nt

length

(c

m)

0

20

40

60

80

100

120


Pla

nt

fres

h w

eight

(g)

0

20

40

60

80

100






Treatments


Nutr

ient

conte

nts

(mg g

-1)

0.0

1.0

2.0

3.0

4.0

5.0

6.0N P K

2

chlamydospores in 1cm root segment and no. of chlamydospores recovered from 100g

rhizosphere soil but the increase was not significant. Increase in mycorrhization was

comparatively more in plants treated with M. ciceri. Antagonistic fungi, T. harzianum

suppress the mycorrhizal colonization by G. fasciculatum, although the reduction was

not significant. Combined inoculation of GF+MC+PM and GF+MC+PM+TH brings

about a significant increase in mycorrhization. Moreover, all the parameters in both

the treatments were at par (Table 37).









and poultry manure. All the biocontrol agents and poultry manure in various

combinations reduce the deleterious effect of nematode and caused a significant

increase in plant length (shoot, root and total length). Individually, T. harzianum with

M. incognita caused the maximum enhancement of plant length (26%) over

nematode-inoculated untreated plants. Poultry manure was more effective than A.

sativa straw and C. album leaves in suppressing the nematode. In combined

treatments, GF+PM+TH+MI and GF+MC+PM+TH+MI were statistically similar.

However, the increase was highest in GF+MC+PM+TH+MI (80%) over all other

treatments and nematode-inoculated untreated control (Table 38 and Fig. 29).



of M. incognita. Inoculation of plant symbionts, antagonistic fungi and organic

manure alone as well as in combinations overcome the loss caused by the nematode

and brings about a significant improvement in the fresh weight of chickpea plants

over nematode-inoculated untreated plants. Highest increase in fresh weight (81%)

was reported while combined inoculation of all the microorganisms and poultry

3

manure i.e. GF+MC+PM+TH+MI. Reduction by M. incognita was lowest in plants

treated with T. harzianum (17.97%) (Table 38 and Fig. 29).



trend of results as of plant fresh weight. T. harzianum proved to be most effective in

suppressing M. incognita and cause a highest increase (27.3%) over other individual

treatments of G. fasciculatum (25.1%), poultry manure (26.4%) and M. ciceri (17.7%)

in the presence of nematode. Combined inoculation of all biocontrol agents and

poultry manure with M. incognita promoted highest increase (81.7%) over nematode-

inoculated untreated plants which is comparatively more than the combined

inoculation with A. sativa straw (71.7%) and C. album (76%) leaves with M.

incognita in previous experiments (Table 38 and Fig. 29).


Number of pods were significantly reduced by the presence of root-knot

nematode, M. incognita in untreated control. Lowest reduction occurred in T.

harzianum (6.38%) compared to other agents applied with nematode. Highest pod

number was observed while combined inoculation of all the biocontrol agents and

poultry manure, GF+MC+PM+TH+MI (85) and lowest pods (26) in the plants

inoculated alone with M. ciceri (Table 38).


No nodules were observed while inoculating with M. incognita. M. ciceri

inhibits the effect of nematode and increase the nodule number to a greater extent as

compared to other microorganisms and manure. In individual treatments, highest

nodule number (47) was observed in M. ciceri followed by Poultry manure (12), T.

harzianum (10) and G. fasciculatum (7) with M. incognita. In the presence of

nematode, all the plant symbionts and manure together resulted highest nodule

number (90) as compared to all other single, double, triple and quadrate combinations

(Table 38).

1


as poultry manure and an antagonistic fungi Trichoderma harzianum on the growth parameters of chickpea plant infected with

root-knot nematode, Meloidogyne incognita


plant-1 Nodules


MI 37.85j±1.89 18.96g±0.95 56.81j±2.84 34.66h±1.73 8.67j±0.43 43.33i±2.17 5.18h±0.26 1.29h±0.06 6.47h±0.32 18.0l±0.90 0.0l±0.00

GF+MI 46.65hi±2.33 23.22f±1.16 69.87h±3.49 43.19fg±2.16 10.75i±0.54 53.94gh±2.70 6.08g±0.30 2.02f±0.10 8.10g±0.41 38.0j±1.90 7.0k±0.35

MC+MI 43.08i±2.15 20.00g±1.00 63.12i±3.16 40.10g±2.01 10.24i±0.51 50.34h±2.52 6.12g±0.31 1.50g±0.08 7.62g±0.38 26.0k±1.30 47.0f±2.35

PM+MI 47.43h±2.37 23.75ef±1.19 71.18h±3.56 43.67f±2.18 10.92hi±0.55 54.59g±2.73 6.55fg±0.33 1.63g±0.08 8.18g±0.41 47.0i±2.35 12.0j±0.60

TH+MI 47.76gh±2.39 23.82e±1.19 71.58gh±3.58 44.03ef±2.20 10.87i±0.54 54.90g±2.75 6.16g±0.31 2.08f±0.10 8.24g±0.41 44.0i±2.20 10.0jk±0.50

GF+MC+MI 51.42fg±2.57 25.61e±1.28 77.03fg±3.85 47.56de±2.38 11.81gh±0.59 59.37f±2.97 6.75f±0.34 2.18ef±0.11 8.93f±0.45 55.0h±2.75 60.0e±3.00

GF+PM+MI 56.44de±2.82 28.21cd±1.41 84.65de±4.23 52.51bc±2.63 13.05ef±0.65 65.56de±3.28 7.37de±0.37 2.48d±0.12 9.85de±0.49 64.0ef±3.20 20.0i±1.00

GF+TH+MI 56.86d±2.84 28.35c±1.42 85.21d±4.26 52.65b±2.63 13.21e±0.66 65.86d±3.29 7.41d±0.37 2.50d±0.13 9.91d±0.50 65.0e±3.25 21.0i±1.05

MC+PM+MI 52.36f±2.62 26.32de±1.32 78.68f±3.93 48.87d±2.44 12.00g±0.60 60.87f±3.04 6.92ef±0.35 2.22e±0.11 9.14f±0.46 58.0gh±2.90 66.0d±3.30

MC+TH+MI 52.94ef±2.65 26.37d±1.32 79.31ef±3.97 49.19cd±2.46 12.25fg±0.61 61.44ef±3.07 6.94e±0.35 2.27e±0.11 9.21ef±0.46 60.0fg±3.00 68.0d±3.40

PM+TH+MI 58.48cd±2.92 29.29bc±1.46 87.77cd±4.39 53.84b±2.69 13.32de±0.67 67.16d±3.36 7.56d±0.38 2.54cd±0.13 10.10d±0.50 70.0d±3.50 27.0h±1.35

GF+MC+PM+MI 60.09c±3.00 29.84b±1.49 89.93c±4.50 55.39ab±2.77 13.85d±0.69 69.24cd±3.46 7.85cd±0.39 2.60c±0.13 10.45cd±0.52 70.0d±3.50 72.0c±3.60

GF+MC+TH+MI 61.64bc±3.08 30.66b±1.53 92.30c±4.62 56.68a±2.83 14.25cd±0.71 70.93c±3.55 8.04c±0.40 2.67bc±0.13 10.71c±0.54 71.0cd±3.55 73.0c±3.65

GF+PM+TH+MI 64.03ab±3.20 34.25a±1.71 98.28ab±4.91 58.11a±2.91 17.72b±0.89 75.83ab±3.79 8.67ab±0.43 2.81b±0.14 11.48ab±0.57 80.0b±4.00 35.0g±1.75

MC+PM+TH+MI 62.09b±3.10 30.85b±1.54 92.94bc±4.65 57.35a±2.87 15.10c±0.76 72.45bc±3.62 8.20bc±0.41 2.73b±0.14 10.93bc±0.55 75.0c±3.75 86.0b±4.30

GF+MC+PM+TH+MI 67.25a±3.36 35.00a±1.75 102.25a±5.11 58.72a±2.94 19.71a±0.99 78.43a±3.92 8.78a±0.44 2.98a±0.15 11.76a±0.59 85.0a±4.25 90.0a±4.50

C.D. (P=0.05) 3.84 1.93 5.76 3.52 0.93 4.45 0.51 0.16 0.67 4.22 3.81

C.D. (P=0.01) 5.17 2.60 7.76 4.75 1.25 5.99 0.68 0.22 0.90 5.68 5.13


GF = Glomus fasciculatum; MC = Mesorhizobium ciceri; PM = Poultry manure; TH = Trichoderma harzianum; MI = Meloidogyne incognita


1


fresh leaves)



reported in case of plants treated with T. harzianum. G. fasciculatum, M. ciceri,

poultry manure and T. harzianum overcome the loss caused by the nematode and

highest increase in chlorophyll content (81.6%) was observed in plants inoculated

with all of them in combination over nematode-inoculated untreated control plants

(Table 39). Chlorophyll content in pots receiving treatments MC+PM+MI,

MC+TH+MI, GF+MC+PM+MI and GF+PM+TH+MI were not significantly different

from each other. Highest increase (81.6%) occurred in plants treated with

GF+MC+PM+TH in the presence of M. incognita (Fig. 29).


fresh leaves)


M. incognita as compared to uninoculated control. All the plant symbionts and poultry

manure were effective in suppressing the deleterious effect of nematode and

significantly increase the nutrient contents. Combined inoculations (dual, triple and

quadrate) proved much better than individual appplication. However,

GF+MC+PM+TH+MI caused highest increase in nutrients (90.4%N, 87.5%P and

80.5%K) over nematode-inoculated-untreated plants (Table 39). Poultry manure

proved to be more effective than straw and botanical used in experiment 8A and 8B in

inhibiting the nematode activity (Fig. 29).





harzianum significantly reduced the colonization. Treatments GF+MC+PM+MI and

GF+MC+PM+TH+MI were statistically similar. However, highest mycorrhization

(9.2%) in M. incognita inoculated plants was observed when GF+MC+PM were

together inoculated (Table 39).

1


as poultry manure and an antagonistic fungi Trichoderma harzianum on the chlorophyll content, nutrient status and

mycorrhization parameters of chickpea plant infected with root-knot nematode, Meloidogyne incognita


GF = Glomus fasciculatum; MC = Mesorhizobium ciceri; PM = Poultry manure; TH = Trichoderma harzianum; MI = Meloidogyne incognita



content (mg g-1)


Colonization

(%)

Internal

Colonization

(%)

Per cent

arbuscules No. of

chlamydospores

in 1cm root

segment

No. of

chlamydospores

recovered from

100 g

rhizosphere soil N P K

MI 1.825f±0.091 2.30g±0.12 0.160f±0.008 1.54f±0.08 0.00e±0.00 0.00e±0.00 0.00e±0.00 0.0e±0.00 0.0e±0.0

GF+MI 2.208e±0.110 2.75f±0.14 0.211e±0.011 1.85e±0.09 59.10cd±2.95 59.20cd±2.96 57.05cd±2.85 56.0cd±2.80 858.0cd±42.9

MC+MI 2.376e±0.119 3.11e±0.16 0.195e±0.010 1.81e±0.09 0.00e±0.00 0.00e±0.00 0.00e±0.00 0.0e±0.00 0.0e±0.0

PM+MI 2.294e±0.115 2.95ef±0.15 0.202e±0.010 1.90e±0.10 0.00e±0.00 0.00e±0.00 0.00e±0.00 0.0e±0.00 0.0e±0.0

TH+MI 2.300e±0.115 2.96e±0.15 0.204e±0.010 1.88e±0.09 0.00e±0.00 0.00e±0.00 0.00e±0.00 0.0e±0.00 0.0e±0.0

GF+MC+MI 2.875d±0.144 3.83cd±0.19 0.262cd±0.013 2.25d±0.11 65.20ab±3.26 64.90ab±3.25 62.30ab±3.12 61.0ab±3.05 938.0ab±46.9

GF+PM+MI 2.819d±0.141 3.59d±0.18 0.267c±0.013 2.36cd±0.12 63.64b±3.18 62.32bc±3.12 60.01b±3.00 58.0bc±2.90 906.0b±45.3

GF+TH+MI 2.839d±0.142 3.65d±0.18 0.270c±0.014 2.40c±0.12 58.30d±2.92 57.07d±2.85 54.67d±2.73 54.0d±2.70 824.0d±41.2

MC+PM+MI 2.947c±0.147 3.97b±0.20 0.254d±0.013 2.27d±0.11 0.00e±0.00 0.00e±0.00 0.00e±0.00 0.0e±0.00 0.0e±0.0

MC+TH+MI 2.956c±0.148 3.98b±0.20 0.257d±0.013 2.28d±0.11 0.00e±0.00 0.00e±0.00 0.00e±0.00 0.0e±0.00 0.0e±0.0

PM+TH+MI 2.929cd±0.146 3.94bc±0.20 0.265c±0.013 2.51bc±0.13 0.00e±0.00 0.00e±0.00 0.00e±0.00 0.0e±0.00 0.0e±0.0

GF+MC+PM+MI 3.039c±0.152 4.11b±0.21 0.277bc±0.014 2.53b±0.13 68.65a±3.43 67.25a±3.36 64.75a±3.24 64.0a±3.20 977.0a±48.8

GF+MC+TH+MI 3.095bc±0.155 4.15ab±0.21 0.280b±0.014 2.48c±0.12 63.64b±3.18 62.74b±3.14 59.89bc±2.99 59.0b±2.95 905.0b±45.3

GF+PM+TH+MI 2.998c±0.150 4.05b±0.20 0.293ab±0.015 2.67ab±0.13 62.07bc±3.10 60.89c±3.04 58.58c±2.93 57.0c±2.85 887.0bc±44.3

MC+PM+TH+MI 3.255ab±0.163 4.32a±0.22 0.272c±0.014 2.59b±0.13 0.00e±0.00 0.00e±0.00 0.00e±0.00 0.0e±0.00 0.0e±0.0

GF+MC+PM+TH+MI 3.314a±0.166 4.38a±0.22 0.300a±0.015 2.78a±0.14 68.03a±3.40 66.63a±3.33 63.92a±3.20 63.0a±3.15 968.0a±48.4

C.D. (P=0.05) 0.195 0.257 0.018 0.160 3.66 3.61 3.47 3.40 52.37

C.D. (P=0.01) 0.263 0.346 0.024 0.215 4.94 4.86 4.67 4.58 70.53

1

T1 = M. incognita (MI); T2 = G. fasciculatum (GF) + M. incognita (MI); T3 = M. ciceri (MC) + M. incognita (MI); T4 = Poultry manure (PM) + M. incognita (MI); T5 = T. harzianum (TH) + M. incognita (MI); T6 = GF+MC+MI; T7 = GF+PM+MI;

T8 = GF+TH+MI; T9 = MC+PM+MI; T10 = MC+TH+MI; T11 = PM+TH+MI; T12 = GF+MC+PM+MI; T13 = GF+MC+TH+MI; T14 = GF+PM+TH+MI; T15 = MC+PM+TH+MI; T16 = GF+MC+PM+TH+MI



manure and an antagonistic fungi Trichoderma harzianum on the growth

parameters, chlorophyll and nutrient contents of chickpea plant infected

with root-knot nematode, Meloidogyne incognita


Chlo

rophyll

co

nte

nt

(mg g

-1)

0.0

0.5

1.0

1.5

2.0

2.5

3.0


Pla

nt

dry

wei

ght

(g)

0

2

4

6

8

10

12

14


Pla

nt

length

(c

m)

0

20

40

60

80

100

120


Pla

nt

fres

h w

eight

(g)

0

20

40

60

80

100






Treatments


Nutr

ient

conte

nts

(mg g

-1)

0.0

1.0

2.0

3.0

4.0

5.0N P K

2



The population of M. incognita in soil as well as in root was significantly

reduced in the presence of either of the plant symbionts, antagonistic fungi or poultry

manure. Inoculation of T. harzianum resulted in an acute reduction in soil and root

population of nematode i.e. 65% and 67.5% respectively compared to other agents

(Table 40). Reduction in nematode population by T. harzianum and poultry manure

were at par (Fig. 30).


Highest number of galls (210) was observed in plants which were inoculated

with M. incognita alone. All the treatments significantly decrease the number of galls

in root system. Lowest number of galls (5) were observed while combined inoculation

of GF+MC+PM+TH+MI (Table ). Galls were less in combined inoculation with

poultry manure as compared to combined inoculation with straw (10) and botanical

(10) (Table 40 and Fig. 30).


Number of eggmasses root system-1

were highest (84) when M. incognita

alone was inoculated. Presence of G. fasciculatum, M. ciceri, poultry manure and T.

harzianum caused significant reduction in this number. Maximum reduction (97.4%)

was observed when all of them were inoculated together and this reduction was

significantly higher than the reduction caused by either of them alone (Table 40 and

Fig. 30).



with either G. fasciculatum, M. ciceri, poultry manure or T. harzianum. Combination

of all these biocontrol agents resulted a maximum decline (96.7%) in fecundity (Table

40). T. harzianum alone caused significantly less number of eggs eggmass-1

(52) as

compared to other biocontrol agents viz., G. fasciculatum (107), M. ciceri (128) and

1


as poultry manure and an antagonistic fungi Trichoderma harzianum on the on the root-knot development of, Meloidogyne

incognita in chickpea plant

Treatments Nematode population No. of galls root

system-1 No. of eggmasses



(0-5)

Reproduction factor

(pf/pi) Soil Root

MI 11,915.0a±595.8 243.0a±12.1 210.0a±10.5 84.0a±4.2 154.0a±7.7 4.0a 12.2a±0.61

GF+MI 9,008.0c±450.4 176.0c±8.8 148.0c±7.4 60.0c±3.0 107.0c±5.4 2.0c 8.8c±0.44

MC+MI 10,052.0b±502.6 202.0b±10.1 173.0b±8.7 70.0b±3.5 128.0b±6.4 3.0b 10.1b±0.51

PM+MI 4,527.0e±226.3 89.0e±4.4 76.0e±3.8 31.0e±1.5 56.0e±2.8 0.0 4.4e±0.22

TH+MI 4,170.0e±208.5 79.0f±3.9 69.0e±3.5 28.0e±1.4 52.0e±2.6 0.0 3.9f±0.20

GF+MC+MI 7,445.0d±372.3 152.0d±7.6 124.0d±6.2 51.0d±2.5 94.0d±4.7 1.0d 7.6d±0.38

GF+PM+MI 1,728.0g±86.4 35.0h±1.8 29.0g±1.4 12.0g±0.6 22.0h±1.1 0.0 1.7h±0.09

GF+TH+MI 1,787.0g±89.4 33.0h±1.7 30.0g±1.5 12.0g±0.6 22.0h±1.1 0.0 1.7h±0.08

MC+PM+MI 2,681.0f±134.0 53.0g±2.6 47.0f±2.4 19.0f±1.0 35.0f±1.8 0.0 2.6g±0.13

MC+TH+MI 2,383.0f±119.2 45.0g±2.3 40.0f±2.0 16.0f±0.8 29.0g±1.4 0.0 2.3g±0.11

PM+TH+MI 882.0h±44.1 19.0i±1.0 16.0h±0.8 6.0h±0.3 12.0i±0.6 0.0 1.0i±0.05

GF+MC+PM+MI 1,192.0h±59.6 23.0i±1.2 20.0h±1.0 8.0h±0.4 15.0i±0.8 0.0 1.1i±0.06

GF+MC+TH+MI 810.0hi±40.5 16.0ij±0.8 15.0hi±0.8 6.0h±0.3 11.0ij±0.6 0.0 0.8i±0.04

GF+PM+TH+MI 476.0i±23.8 9.0jk±0.5 7.0j±0.4 3.0i±0.2 6.0jk±0.3 0.0 0.4j±0.02

MC+PM+TH+MI 715.0i±35.8 14.0j±0.7 12.0ij±0.6 5.0hi±0.3 9.0j±0.5 0.0 0.7ij±0.04

GF+MC+PM+TH+MI 266.0j±13.3 6.0k±0.3 5.0j±0.3 2.0i±0.1 5.0k±0.3 0.0 0.3j±0.02

C.D. (P=0.05) 437.9 8.8 7.47 3.02 5.50 0.44

C.D. (P=0.01) 589.7 11.8 10.07 4.07 7.41 0.59


GF = Glomus fasciculatum; MC = Mesorhizobium ciceri; PM = Poultry manure; TH = Trichoderma harzianum; MI = Meloidogyne incognita Mean values with different letters within the column are significantly different at P = 0.05

1


No.

of

eggs

eggm

ass-1

020406080

100120140160180


No.

of

eggm

asse

s

root

syst

em-1

0

20

40

60

80

100


Nem

atode

popula

tion

0

200

400

600

800

1000

1200

1400Soil (×10)

Root


No.

of

gal

ls

root

syst

em-1

0

50

100

150

200

250






Treatments


Rep

roduct

ion f

acto

r (p

f/pi)

0.0

2.0

4.0

6.0

8.0

10.0

12.0

14.0

T1 = M. incognita (MI); T2 = G. fasciculatum (GF) + M. incognita (MI); T3 = M. ciceri (MC) + M. incognita (MI); T4 =

Poultry manure (PM) + M. incognita (MI); T5 = T. harzianum (TH) + M. incognita (MI); T6 = GF+MC+MI; T7 = GF+PM+MI;

T8 = GF+TH+MI; T9 = MC+PM+MI; T10 = MC+TH+MI; T11 = PM+TH+MI; T12 = GF+MC+PM+MI; T13 =

GF+MC+TH+MI; T14 = GF+PM+TH+MI; T15 = MC+PM+TH+MI; T16 = GF+MC+PM+TH+MI



manure and an antagonistic fungi Trichoderma harzianum on the on the

root-knot development of Meloidogyne incognita in chickpea plant

2

poultry manure (56). Treatments GF+PM+MI and GF+TH+MI possess same number

of eggs eggmass-1

i.e. 22 (Fig. 30).


Highest root-knot index (4) was observed in plants inoculated with M.

incognita alone. Inoculation of plant symbionts, antagonistic fungi and poultry

manure resulted in a significant reduction in root-knot index (RKI). No infection was

observed in the roots of plants inoculated with either T. harzianum and poultry

manure as well as in the combined treatments of all the biocontrol agents (Table 40).



12.20. Addition of G. fasciculatum, M. ciceri, poultry manure and T. harzianum

brings about a significant reduction in reproduction of nematode and the reduction

was more pronounced in the plants having all the biocontrol agents with poultry

manure in the presence of M. incognita (0.30). Reproduction rate of M. incognita was

less in poultry manure (4.4) as compared to A. sativa straw (7.05) and C. album leaves

(4.8) in the earlier experiment (Table 40 and Fig. 30).

8.4 DISCUSSION

All the application of bio-organic fertilizers and plant symbionts generally

resulted better plant growth, however, the combined application showed more growth

than uninoculated control plants. The increase in growth of chickpea could also be

due to nutrient supplementation among the inoculated organisms, which might have

enhanced their efficiencies like N fixation by Rhizobium and P-uptake by G.

fasciculatum and effective pathogen suppression by T. harzianum. This was profound

for our study and other workers (Singh et al., 2000; Cakmakci et al., 2001; Singh,

2001; Rudresh et al., 2005; Gomaa et al., 2002; Srivastava et al., 2002; Patidar and

Mali, 2004; Uyanoz, 2007) also got similar results. Different organic manures, plant

symbionts and antagonistic fungi significantly affected the growth parameters and

improved nutrient content of crop. As compared to single inoculation of biological

fertilizers, the growth parameters, chlorophyll and nutrient contents increased more

when organic fertilizers and plant symbionts were applied in combinations, which

indicates that organic wastes and plant symbionts supply nutrients to plants

3

(Cakmacki et al., 2001; Patidar and Mali, 2004). Organic fertilizers as plant straw,

green leaves and poultry manure are biologically decomposed by the microbes and on

the other hand, plant symbionts such as AM fungi and N2 fixing organisms as

Rhizobium are important for soil productivity and plant nutrition.

Mycorrhization parameters were increased by the inoculation of Rhizobium

spp. and organic wastes/manures while T. harzianum reduced the mycorrhizal

colonization by Glomus fasciculatum. Akhtar and Siddiqui (2008) and Serfoji et al.

(2010) concluded the same in their study. In the present study, negative interaction

was observed between G. fasciculatum and T. harzianum, the same findings were also

obtained by Rousseau et al. (1996) and Green et al. (1998).

Meloidogyne incognita infestation was suppressed in all the combination of

the microorganisms and organic wastes applications, leading to improved growth and

yield of the chickpea plant. A similar result was obtained by Rao et al. (1995). The

present experiment indicate that application of T. harzianum is superior to G.

fasciculatum, Rhizobium and organic wastes (A. sativa straw, C. album leaves and

poultry manure) in reducing nematode population in soil as well as in roots, root-

galling, number of eggmasses, fecundity, root-knot index and reproduction rate of M.

incognita.

Application of G. fasciculatum together with T. harzianum to the seedlings

were found to be highly useful for reducing the total nematode population as well as

for enhancement of plant growth parameters, chlorophyll and nutrient contents.

However, the extent of nematode reduction depends on the qualitative and

quantitative application of the biocontrol agent. This may be due to the altered host

physiology (Saleh and Sikaro, 1984; Sikora, 1978; Hussey and Roncadori, 1982). AM

fungal association with the root system may possibly secrete certain

metabolites/compounds that restrict the multiplication of M. incognita in the

rhizosphere (Sikora and Sitaramaiah, 1995; Nagesh and Reddy, 2004).

The current study indicated that the application of Trichoderma harzianum

significantly reduce root and soil population, root-galling, egg mass formation and

egg production of M. incognita. Trichoderma harzianum is a well established

biocontrol fungus which possess nematicidal properties (Windham et al., 1993, Rao et

al., 1996, Spiegel and Chet, 1998; Sharon et al., 2001; Suarez et al., 2004) and also

4

colonizes eggs and infests second stage juveniles (Safiuallah and Thomas, 1996;

Sharon et al., 2001). Trichoderma spp. are thoroughly investigated for their biocontrol

potential against root-knot nematodes on a range of crops like tomato, okra,

mungbean and bell pepper (Meyer et al., 2001; Siddiqui and Shaukat, 2004; Siddiqui

et al., 2001). It is still premature to pinpoint the exact mechanism of management of

nematode by the addition of a bioagent like T. harzianum but it is likely that a

reduction of the nematode population may be due to the production of nematode toxic

metabolites in the root rhizosphere. A number of studies has demonstrated that

Trichoderma spp. frequently enhances root colonization and development, crop

productivity, resistance to abiotic stresses and uptake and use of nutrients (Spiegel

and Chet, 1998, Harman et al., 2004; Sharon et al., 2001; Siddiqui et al., 2001;

Affokpon et al., 2011).

There is overall increase in the biomass of chickpea plants when decomposed

wastes were added. The results were more pronounced when these wastes were added

in combinations. This may be due to the formation of better soil structure, build up of

antagonistic organisms and supply of nutrients. Increase in plant growth and reduction

in nematode population by the use of organic fertilizers may be attributed to the above

mentioned reasons (Southey, 1978). The nematicidal effects of the amendments is

mainly due to ammonia toxicity (Rodriguez-Kabana et al., 1986; Ben-Yep-Het et al.,

2005; Oka et al., 2006). Another feasible mechanism that has been suggested involves

the antagonistic effect of the increased microbial population in the soil (Kaplan et al.,

1992; Riegel et al., 1996). In brief, the results obtained in this study suggest that

application of nitrogen-rich organic amendments to soil is effective in control of root-

knot nematodes in organic greenhouse farming systems.

Nematode reproduction rate was also significantly lower in G. fasciculatum +

Rhizobium + T. harzianum + organic wastes inoculated plants. Vedhera et al. (1998)

reported that besides increasing the yield, application of different organic manures

also inhibited reproduction in females and penetration of second stage juveniles into

ginger roots. Goswami and Vijayalakshmi (1981); Rajendran and Saritha (2005) and

Kantharaju et al. (2005) also showed the same results in tomato.

Various mechanisms have been suggested for the biocontrol activity of

Trihoderma spp. against phytopathogenic fungi: antibiosis, competition,

mycoparsitism and enzymatic hydrolysis (Elad, 1995; Sivan and Chet 1992.)

5

Trichoderma spp. overgrew the pathogen colony and inhibited its growth. This

showed that Trichoderma used mycoparasitism as mechanism of control.

Mycoparasitism occurs when intimate association exist between the pathogen and the

biocontrol and involves coiling of hyphae around the pathogen, penetration,

production of haustoria and lysis of hyphae. Trichoderma spp. inhibited the colony

growth of the pathogen without establishment of contact. This may indicate the

involvement of volatile compounds such as acetaldehyde ethylene, acetone and

carbon dioxide (Tamini and Hutchinson, 1975) and some non-volatile antibiotics such

as viridin (Dennis and Webster, 1971) produced by Trichoderma.

Maximum root nodule numbers were recorded with poultry manure +

Rhizobium + G. fasciculatum and T. harzianum treatments. Same results were shown

by Gomaa et al. (2002) in case of vetch (a leguminous forage crop). The data indicate

that the combined inoculums associated with either of the organic waste (straw,

botanical or manure) may be feasible to get reasonable productivity when the bio-

organic farming was taken into consideration. Thus, legume cultivation under

application of organic fertilizers has the potential to increase crop production and soil

sustainable. In addition, legume plants improve the fertility of the soil via providing a

substantial input of N2 fixation.

The present study also suggests that greater emphasis on the development of

mixtures of biocontrol agents is needed, because they may better adapt to the

environmental changes that occur throughout the growing season and protect against a

broader range of pathogens. Pierson and Weller (1994) also found that mixture of

microorganisms may increase the genetic diversity of biocontrol systems that may

persist longer in the rhizosphere and utilize a wider array of biocontrol mechanisms.

Combinations of biocontrol agents have been recommended to cope with the often

highly variable conditions common in farmer‟s fields and optimize the potential

benefits of the various agents (Meyer and Roberts, 2002).

Data revealed that combined application of poultry manure and T. harzianum

suppress the nematode population and resulted significant increase in plant growth

parameters. Organic amendments stimulate increased activity of biological

antagonists of nematodes, which in turn reduced the loss caused by the nematode to a

higher extent. Other researchers also demonstrated a relationship between organic

6

amendments and microbial activity, especially for disease supression (Bailey and

Lazarovits, 2003; Serfoji et al., 2010).

Thus, it can be concluded that T. harzianum, G. fasciculatum, M. ciceri and

organic wastes have prospect of enhancing nematode management through nematode

density reduction and enhanced crop improvement. These selected beneficial plant

symbionts along with organic waste could be effective alternatives to the toxic

nematicides and inorganic fertilizers used in crop protection.

8.5 CONCLUSION

In the present study, antagonistic fungi, T. harzianum alongwith AM fungus,

G. fasciculatum, root-nodule bacterium, Mesorhizobium ciceri and organic wastes

(straw of Avena sativa, green leaves of Chenopodium album and poultry manure)

were tested for managing root-knot nematode, M. incognita in chickpea plants. The

results demonstrated the potential benefits of applying microorganisms in the

presence of nematode. In general, combined treatments responded better than the

individual treatments. T. harzianum resulted in highest reduction in root-knot

development compared to other plant symbionts and organic wastes. Among different

treatments, all plant symbionts together with poultry manure resulted in the highest

plant growth, chlorophyll and nutrient content and brings about a significant decline

in the nematode-related parameters in chickpea var. Avrodhi.

8.6 SUMMARY

6. A study was conducted to carried out the individual and interactive effect of

different plant symbionts (Glomus fasciculatum, Mesorhizobium ciceri), bio-

organic wastes (Avena sativa straw, Chenopodium album leaves, poultry

manure) and antagonistic fungi, Trichoderma harzianum in all possible

combinations for biocontrol of root-knot nematode, Meloidogyne incognita

infecting chickpea var. Avrodhi.

7. Biocontrol agents and organic wastes were evaluated for their efficacy in

terms of growth characteristics, chlorophyll content, nutrients status,

mycorrhization and nematode-related parameters.

7

8. Combined treatments resulted in higher plant growth, biomass, chlorophyll

and nutrient status (N, P & K). Wastes with high N content i.e. poultry manure

(6.3%N) content caused greater reduction in nematode population in chickpea

plant over straw (1.05%N) and botanical (3.78%N).

9. Antagonistic fungi, Trichoderma harzianum @ 106

spores plant-1

proved to be

most effective of all the plant symbionts (G. fasciculatum, M. ciceri) and

organic wastes in suppressing the nematode-related parameters in chickpea

plants.

10. Of all the treatments, combined inoculation of AM fungus (Glomus

fasciculatum @ 800 spores plant-1

) + root-nodulating bacteria (Mesorhizobium

ciceri @ 1g plant-1

) + organic waste (poultry manure @ 10g plant-1

) and

antagonistic fungi (Trichoderma harzianum @ 106

spores plant-1

) was found to

be the most efficient in improving the plant growth parameters and decreasing

the root-knot development of M. incognita in chickpea plants.

8. section-6 individual and interactive effect of...

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