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8
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
9
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).
10
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
11
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
12
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.
13
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
14
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.
15
8.2.3 EXPERIMENT 8B: With green leaves of botanical C. album
8.2.3.1 Growth and maintenance of test plant
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.
8.2.3.3 AM fungus (G. fasciculatum) inoculum
The inoculum of Glomus fasciculatum was maintained as mentioned above in
Experiment 8A and in Section-2.
8.2.3.4 Mesorhizobium ciceri inoculum
Chickpea strain of Rhizobium (Charcoal-based culture), Mesorhizobium ciceri
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
Root-knot nematode, Meloidogyne incognita culture was maintained as
mentioned above in Experiment 8A and in Section-3.
8.2.3.7 Inoculation technique
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
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
16
treatments were given concomitantly 15days prior to the nematode inoculation. An
equal volume of sterile water was added to the control treatments.
8.2.3.8 Experimental design
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.
8.2.3.9 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).
8.2.3.10 Parameters studied
After termination of the experiment, the above parameters (as mentioned in
Experiment 8A) were determined for each treatment.
8.2.3.11 Plant growth and chemical parameters
Plant growth and chemical parameters were studied by the methods mentioned
in Section-2.
8.2.3.12 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.
17
8.2.3.13 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.
8.2.4 EXPERIMENT 8C: With poultry manure
8.2.4.1 Growth and maintenance of test plant
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.
8.2.4.3 AM fungus (G. fasciculatum) inoculum
The inoculum of Glomus fasciculatum was maintained as mentioned above in
Experiment 8A and in Section-2.
8.2.4.4 Mesorhizobium ciceri inoculum
Chickpea strain of Rhizobium (Charcoal-based culture), Mesorhizobium ciceri
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
18
Root-knot nematode, Meloidogyne incognita culture was maintained as
mentioned above in Experiment 8A and in Section-3.
8.2.4.7 Inoculation technique
For the addition of decomposed poultry manure 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.
8.2.4.8 Experimental design
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.
8.2.4.9 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).
8.2.4.10 Parameters studied
After termination of the experiment, the above parameters (as mentioned in
Experiment 8A) were determined for each treatment.
19
8.2.4.11 Plant growth and chemical parameters
Plant growth and chemical parameters were studied by the methods mentioned
in Section-2.
8.2.4.12 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.
8.2.4.13 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.
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
20
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).
1
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
1
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.
8.3.1.2.1 Plant length (cm)
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
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
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
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
Pla
nt
length
(c
m)
0
20
40
60
80
100
120
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
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
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
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).
8.3.1.2.2 Plant fresh weight (g)
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).
8.3.1.2.3 Plant dry weight (g)
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
Table 28. 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 infected with root-knot
nematode, Meloidogyne incognita
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
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
Data mean±SD of five replicates
GF = Glomus fasciculatum; MC = Mesorhizobium ciceri; AS = Avena sativa; TH = Trichoderma harzianum; MI = Meloidogyne incognita
Mean values with different letters within the column are significantly different at P = 0.05
1
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
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
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
Pla
nt
length
(c
m)
0
20
40
60
80
100
120
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
Pla
nt
fres
h w
eight
(g)
0
20
40
60
80
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
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
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
Fig. 23 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 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).
8.3.1.2.5 Nodules plant-1
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).
8.3.1.2.6 Chlorophyll content (mg g-1
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).
8.3.1.2.7 Nutrient contents (N, P & K) (mg g-1
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
Table 29. 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 infected with root-knot nematode Meloidogyne incognita
Data mean±SD of five replicates
GF = Glomus fasciculatum; MC = Mesorhizobium ciceri; AS = Avena sativa; TH = Trichoderma harzianum; MI = Meloidogyne incognita
Mean values with different letters within the column are significantly different at P = 0.05
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
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
8.3.1.2.8 Mycorrhization parameters
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
Data mean±SD of five replicates
GF = Glomus fasciculatum; MC = Mesorhizobium ciceri; AS = Avena sativa; TH = Trichoderma harzianum; MI = Meloidogyne incognita
Mean values with different letters within the column are significantly different at P = 0.05
1
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
No.
of
eggs
eggm
ass-1
020406080
100120140160180
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
No.
of
eggm
asse
s
root
syst
em-1
0
20
40
60
80
100
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
Nem
atode
popula
tion
0
200
400
600
800
1000
1200
1400Soil (×10)
Root
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
No.
of
gal
ls
root
syst
em-1
0
50
100
150
200
250
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
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
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
Fig. 24 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
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
8.3.2.1 In the absence of M. incognita
8.3.2.1.1 Plant length (cm)
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).
8.3.2.1.2 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 (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).
8.3.2.1.3 Plant dry weight (g)
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).
8.3.2.1.5 Nodules plant-1
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
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 43.64g±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
Data mean±SD of five replicates
GF = Glomus fasciculatum; MC = Mesorhizobium ciceri; CA = Chenopodium album; TH = Trichoderma harzianum
Mean values with different letters within the column are significantly different at P = 0.05
1
8.3.2.1.6 Chlorophyll content (mg g-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).
8.3.2.1.7 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 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).
8.3.2.1.8 Mycorrhization parameters
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).
8.3.2.2 In the presence of M. incognita
Inoculation with M. incognita caused a significant reduction in all the growth
parameters compared with uninoculated control.
8.3.2.2.1 Plant length (cm)
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 32. 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 chlorophyll content, nutrient status and
mycorrhization parameters of chickpea plant
Data mean±SD of five replicates
GF = Glomus fasciculatum; MC = Mesorhizobium ciceri; CA = Chenopodium album; TH = Trichoderma harzianum
Mean values with different letters within the column are significantly different at P = 0.05
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 2.402h±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
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
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
Pla
nt
length
(c
m)
0
20
40
60
80
100
120
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
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
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
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
Fig. 25 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, chlorophyll and nutrient contents of chickpea
plant
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 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).
8.3.2.2.2 Plant fresh weight (g)
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. 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).
8.3.2.2.3 Plant dry weight (g)
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. 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
8.3.2.2.4 Pods plant-1
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).
8.3.2.2.5 Nodules plant-1
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).
8.3.2.2.6 Chlorophyll content (mg g-1
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, 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).
8.3.2.2.7 Nutrient contents (N, P & K) (mg g-1
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
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
Table 33. 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
infected with root-knot nematode, Meloidogyne incognita
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 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
Data mean±SD of five replicates
GF = Glomus fasciculatum; MC = Mesorhizobium ciceri; CA = Chenopodium album; TH = Trichoderma harzianum; MI = Meloidogyne incognita
Mean values with different letters within the column are significantly different at P = 0.05
2
Table 34. 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 chlorophyll content, nutrient status and
mycorrhization parameters of chickpea plant infected with root-knot nematode, Meloidogyne incognita
Data mean±SD of five replicates
GF = Glomus fasciculatum; MC = Mesorhizobium ciceri; CA = Chenopodium album; TH = Trichoderma harzianum; MI = Meloidogyne incognita
Mean values with different letters within the column are significantly different at P = 0.05
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 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
Fig. 26 Individual and interactive effect of AM fungus Glomus fasciculatum,
root-nodule bacterium Rhizobium, leaves of Chenopodium album and an
antagonistic fungi Trichoderma harzianum on the growth parameters,
chlorophyll and nutrient contents of chickpea plant infected with
Meloidogyne incognita
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
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
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
Pla
nt
length
(c
m)
0
20
40
60
80
100
120
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
Pla
nt
fres
h w
eight
(g)
0
20
40
60
80
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
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
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.
8.3.2.2.8 Mycorrhization parameters
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. 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.
8.3.2.2.9 Root-knot development
8.3.2.2.9a Nematode population
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.
8.3.2.2.9b Number of galls root system-1
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
Table 35. 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 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
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
Data mean±SD of five replicates
GF = Glomus fasciculatum; MC = Mesorhizobium ciceri; CA = Chenopodium album; TH = Trichoderma harzianum; MI = Meloidogyne incognita
Mean values with different letters within the column are significantly different at P = 0.05
1
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
Ch
loro
ph
yll
con
ten
t
(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
gh
t
(g)
02468
10121416
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
Pla
nt
len
gth
(cm
)
0
20
40
60
80
100
120
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
Pla
nt
fres
h w
eigh
t
(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
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
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
Fig. 27 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 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).
8.3.2.2.9c Number of eggmasses root system-1
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).
8.3.2.2.9d Number of eggs eggmass-1
Significant reduction in fecundity was observed when plants were inoculated
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).
8.3.2.2.9e Root-knot index (0-5)
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).
8.3.2.2.9f Reproduction factor (pf/pi)
Plants inoculated with M. incognita showed the highest reproduction rate i.e.
12.20. Addition of G. fasciculatum, M. ciceri, 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
8.3.3.1 In the absence of M. incognita
8.3.3.1.1 Plant length (cm)
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.
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 (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).
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 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
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 21.82l±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
Data mean±SD of five replicates
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).
8.3.1.1.5 Nodules plant-1
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).
8.3.1.1.6 Chlorophyll content (mg g-1
) 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).
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+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).
8.3.1.1.8 Mycorrhization parameters
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
Table 37. 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 chlorophyll content, nutrient status and
mycorrhization parameters of chickpea plant
Data mean±SD of five replicates
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
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 2.402g±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
Fig. 28 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, chlorophyll and nutrient contents of chickpea plant
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
Chlo
rophyll
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
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(g)
02468
10121416
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
Pla
nt
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(c
m)
0
20
40
60
80
100
120
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
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
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
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).
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.
8.3.1.2.1 Plant length (cm)
The inoculation of M. incognita caused a significant reduction in plant length
(shoot, root and total length) of all the treatments over nematode-uninoculated
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
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).
8.3.1.2.2 Plant fresh weight (g)
Plant fresh weight (shoot, root and total) declined significantly in the presence
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).
8.3.1.2.3 Plant dry weight (g)
Plant dry weight (shoot, root and total dry weight) depicted almost a similar
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).
8.3.1.2.4 Pods plant-1
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).
8.3.1.2.5 Nodules plant-1
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
Table 38. 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 infected with
root-knot nematode, Meloidogyne incognita
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
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
Data mean±SD of five replicates
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
8.3.1.2.6 Chlorophyll content (mg g-1
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,
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).
8.3.1.2.7 Nutrient contents (N, P & K) (mg g-1
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 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).
8.3.1.2.8 Mycorrhization parameters
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+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
Table 39. 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 chlorophyll content, nutrient status and
mycorrhization parameters of chickpea plant infected with root-knot nematode, Meloidogyne incognita
Data mean±SD of five replicates
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
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
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
Fig. 29 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, chlorophyll and nutrient contents of chickpea plant infected
with root-knot nematode, Meloidogyne incognita
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
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
14
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
Pla
nt
length
(c
m)
0
20
40
60
80
100
120
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
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
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
Nutr
ient
conte
nts
(mg g
-1)
0.0
1.0
2.0
3.0
4.0
5.0N P K
2
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, 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).
8.3.1.2.9b Number of galls root system-1
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).
8.3.1.2.9c Number of eggmasses root system-1
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).
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, 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
Table 40. 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 on the root-knot development of, Meloidogyne
incognita in chickpea plant
Treatments Nematode population No. of galls root
system-1 No. of egg- masses
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 12.2a±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
Data mean±SD of five replicates
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
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
No.
of
eggs
eggm
ass-1
020406080
100120140160180
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
No.
of
eggm
asse
s
root
syst
em-1
0
20
40
60
80
100
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
Nem
atode
popula
tion
0
200
400
600
800
1000
1200
1400Soil (×10)
Root
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
No.
of
gal
ls
root
syst
em-1
0
50
100
150
200
250
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
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16
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
Fig. 30 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 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).
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 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).
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, 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.