‘




 
To compare the effectiveness of the bioagents against the wilt and
root-knot diseases, separate treatments with efficacious fungicide
(carbendazim) and nematicides (fenamiphos) were maintained.
Carbendazim (50%w.p) @1.25 kg/h was applied in broadcast manner a
applied to soil @ 6kg/h and for seed treatment the doze was 2g/kg seed.
Half dose of carbendazim was mixed with the half dose of fenamiphos to
get the required dose of the combination.
For soil application dose of the comination was maintained as 40
g/microplot (50 kg/ha). To achieve this 20 g of %.
 cultured on
baggasse-soil-molasses mixture was mixed with the 20 g leaf litter
colonized by

  and /or 20ml nutrient broth culture of
P. flusorescens. The mixture was applied in rows where seeds were to be
sown in a microplot. Fo seed application 2.5 g of sorghum seeds
colonized with the bioagent was mixed with the 2.5 g of other bioagent or
2.5ml nutrient broth to get the total dose of 5 g/kg seed.
Æ  
Ýeeds of chickpea, Æ  . var. BG-256 were sown in
rows (57 seeds/row, 4 rows/microplot) where antagonist(s) had already
been applied. One week after sowing irrigation was done. Weeds were
removed manually three times (monthly) and any chemical fertilizer or
pesticide was not applied in the fiel. Plants were grown for four months.
During this period they were regularly observed for any symptom.
Ý  

‘


Ýoil population of the wilt fungi and bio-agents was estimated
monthly using dilution plate method. Ýoil was collected from the
rhizosphere of five plants in each microplot and was mixed to make a
composite sample. The soil was sieved through a coarse sieve. One gram
of the soil was taken in a conical flask to which 10ml sterile water was
added. The soil-water mixture was stirred over a magnetic stirrer for 5
minutes. One mol of this suspension was transferred into a 9 ml sterile
water in a test tube blank. One ml samples were then transferred
immediately through successive 9ml sterile water blanks until the desired
final dilution of 1:10000(for fungi) and 1:100000(for bacteria) was
achieved.
For fungi, 1 ml of the desired dilution was aseptically transferred
(under Laminar flow) to the surface of the hardened potato dextrose agar
medium in Petri dishes and 3 such plates were maintained for each
dilution. The transferred suspension was spread over the agar surface
with a glass spreader. The agar plates were prepared four days previously
to ensure that the medium in the plate was free from contamination.
The plates were then incubated at 25 -27oC for 7-10 days to get the
colonies. To determine the population of bacteria, 0.3 ml of the final
dilution was spread over hardened surface of nutrient broth in a Petri
plate and incubated at 37 oC (
) or 30oC (
) for 3
days. After incubation, the plates were examined under a colony counter
to determine soil population of the desired microorganism.
ñ 


 
Population of M. incognita was determined by Cob¶s decanting the
sieving method. A composite soil sample was made by collecting the soil
from the rhizosphere of five plants in each microplot. The sool was sifted
through a coarse sieve. The soil (1 kg) was mixed in 5 liters of water ina
plastic bucket. The soil-water mixture was stirred and then allowed to
stand for 30 seconds. The liquid was then decanted over a combination of
3 sieves (60, 200, and 500 mesh). The catch from the final sieve was
carefully washed and transferred to a beaker.
A small coarse sieve with two layers of wet paper towels was kept
in a Baermann funnel filled with water. The nematode suspension from
the beaker was poured onto the sieve and allowed to stand overnight. The
nematodes move actively and migrate throught he paper towel into the
water and aggregate in the bottom of the rubber tubing of Baermann
funnel. The nematode suspension recovered from the Baermann funnel
was taken into a counting dish for examination under microscope.

Visual observations were made on five and two and a half months
old plants of pigeonpea and chickpea, respectively to determine wil t
incidence and severity according to the following formulae
Number of plants showing wilt symptoms in a microplot
Wilt incidence = ×100
Total number of plants in a microplot

Number of branches/twigs showing wilt symptom

Wilt severity = ×100
Total number of branches /twigs of plant
ñ 
 

Two months old plants were randomly uprootd from each
microplot (5 plants/microplot) to count root nodules. Poink and healthy
nodules were recognized as a functional nodules. Were as dark brown and
degenerated ones as nonfunctional nodules.


At matuarity in April, nine and four months of sowing of
pigeonpea and chickpea, respectively, ten plants from each microplot
were randomly uprooted and the following parameters were determined.
1.m Plant length
2.m Dry weight of plants
3.m Number of pods/plant
4.m Number of seeds/plants
5.m Weight of seeds/plant
6.m Root-knot severity
7.m Functional nodules/root system
8.m Total nodules/root system
9.m Non-functional nodules/root system
10.mNematode reproduction
11.mRhizosphere population of pathogens and bioagents
ñ 

Plants were carefully uprooted to avoid root loss. Roots were
gently washed in slow stream of water. The roots were visually observed
to count galls which are out growth/swellings of root parts. To cou nt
eggmass, roots were treated with phloxine B solution (0.159g/L) which
gave a blue stain to eggmasses.
Ý

The observations taken from ten plants from a microplot were
averaged and considered as one replicate. Ýince three microplots were
maintained for each treatment, there were three replicates. The data on
plant growth and yield was subjected to a two -factor analysis of variance.
Pathogens were considered as one factor, whereas bioagent treatments as
second factor. The experiment on the relative effectiv eness of bioagents
was conducted during two consecutive years. The data on these
experiments was analyzed separately. The data on wilt incidence,
severity, root-knot, soil population etc. were analyzed for single factor
ANOVA. Wilt incidence and severity was angularly transformed before
the analysis. Least Ýignificance Difference (L.Ý.D) was calculated at  =
0.05 for all the variables to compare individual treatments. Histograms
have been prepared for most of the data and bars are presented with
percent variance in comparison to control and an star for significance.
ANOVA tables have been presented as Appendix after Reference
Ýection.
ñ 
Æ  
‘‘Æ
Ý ‘ Ý Æ
    Ý
ñ

Ý


 
Ý  
 
 ñ
 
  

ÝÝÆ 
Ý



Plants grown in the plots inoculated with ‘
  f. sp. 
developed characteristic symptoms of wilt disease. Ýome plants at
seedling stage exhibited drooping that led to their mortality. At later stage
branches or twigs shoed wilting even in the condition of adequate soil
moister. Leaf of such twigs later turned yellowish. At the advanced stage
of plant growth, the whole plant, branch or twig became brown and
subsequently dried and died (fig.). On concomitantly inoculated plants
severity of the wilt was increased (86%) but gall formation was decreased
(13%). Transverse section of the roots of infected plants showed
mycelium of the wilt fungus especially in vascular tissue. Incidence of the
wilt disease was 38% and its severity was 3.4 on 0-5 scale. Application of
the bioagents decreased the severity of wilt. Ýoil application of %

resulted to the decrease in wilt index to 2.3, followed by 2.1, 2.0 and 2.1
by %

, 
 and 

Ýeed treatment was found
3-7% less effective in controlling the disease. Disease incidence
remained statistically uninfluenced.
ñ 

Nematode infected plants showed stunted growth and vigour with
pale green foliage. On roots, characteristic galls or knots were developed
(Fig). Nematode inoculation @ 2000 juveniles of Meloidogyne incognita
per kg of soil caused considerable galling on chickpea roots (Fig. **).
Application of various bioagents through soil application of various
bioagents through soil application or seed treatment inf luent the gall
formation to some extent, and the decrease in the number of gall per root
system was, however, significant for only 
 and
fenamiphos, where 9-11% decrease was recorded. Reproduction of
nematode in terms of egg mass production was significantly decreased
only due to application of nematicide through soil application or seed
treatment.

!
Incidence of the wilt and its severity were significantly increased
on the plants grown in the plots which were inoculated with ‘
 
f sp.  and    concomitantly. The
incidence was increased by 15%, whereas the disease severity by 29% in
comparison to ‘ alone. Application of various treatments
checked the wilt disease to a varied extents. There was no discernible
impact of treatments on the incidence of wilt as it remained statistically
unchanged. Lowest wilting i.e., 28% was recorded due to soil application
of mixture of carbendazim and fenamiphos, fo llowed by 31-32% wilting
with %
 spp., 35% with 
 against 86% wilt in the
concomitantly inoculated control.
Ýeverity of root-knot caused by the 
 was, however,
decreasedby 13% and egg massproduction by 20%in the prescence of
FUsarium. Ýome treatments also checked the root-knot disease.
Application of the mixture of fungicide+nematicide resulted to 15%
decrease in the galling following by 9-10% decrease due to nematicide
above,

  or P. . These treatments also
caused to a significant decline in the egg mass production. Other
treatments, however did not provided a significant control of the disease.
Ýeed treatments were found some 2-6% less effective in decreasing the
severity of wilt or root-knot disease.


 
Ý  

Application of 
 resulted to a significant increase in
the plant length (12.2%) and dry weight (10.6%) of chickpea compared to
uninoculated control. Inoculations with ‘
  f. sp.  and 

 singly or concomitantly significantly decreased the plant

growth. The pathogenic fungus caused 17.6 and 23.7% decrease in the
plant length and its dry weight, respectively; corresponding values for
root-knot nematode were 12.6 and 10.4%. Concomitant inoculations with
the fungus and nematode caused greater suppression in plant length
(38.2%) and dry weight (42.3%) compared to uninoculated control.
Application of bioagents checked the suppressive effects of the
pathogens that led to increase in the plant growth variables considered.
The bioagents except

  significantly increased the
plant length and dry weight. Greatest increase in plant length of ‘
inoculated plants was recorded due to application of T. virens (2.3% )
whereas, in dry weight with %

 or 
 (31%)
compared to inoculated control. Application of carbendazim resulted to
21.1 and 15.9% increase in the plant length and dry weight. Plant length
of nematode inoculated plants was significant ly increased due to
application of 
 (11.4%) or fenamiphos (11.9%). The
nematicide also significantly increased the dry weight of chickpea plants.
Bioagents treatments were also effective in increasing the growth of the
plants inoculated concomitantly with ‘
 
f. sp.  and 

. Greatest increase i.e., 42.1 and 39.4% in the growth variables
occurred due to application of 
‘ followed by 
 T.
harzianum and %
 compared to concomitantly inoculated control. A
treatment with the mixture of carbendazim and fenamiphos resulted to 26
and 24% increase I the plant length and dry weight of chickpea.
Ý


Application of bioagents through seed treatment was relatively less
effective compared to soil application. Plant length of chickpea was
significantly increased due to application of 
 compared soil
application. Plant length of chickpea was significantly increased due to
application P. fluorescens compared to inoculated control. Greatest
increase in the plant length of Fusarium inoculated plants was recorded
due to %
 (13.6%) followed by %

 (12.1%) and P.
 (11.2%) against 21.6% due to carbendazim. dry