Indian Phytopath.

50 (2) : 229-234 (1997)

Evaluating methods of application of biocontrol agent

in the control of mungbean root rot
T. RAGUCHANDER,

K. RAJAPPAN

Department of Plant Pathology.

and R. SAMIAPPAN

Tamil Nadu Agricultural

University. Coimbatore

641 003

ABSTRACT : Dry root rot of mungbean caused by Macrophomina phaseolina was reduced by seed pelleting of
Trichoderma viride isolates when talc was used as carrier. Among the isolates, T yiride-III supported higher plant
growth, better native Rhizobium nodulation and grain yield. Sclerotial number and root rot incidence were greatly
reduced in seed pelleting of antagonists as compared to row application. Rhizosphere soil had a higher number of
Trichoderma chlamydospores in seed pelleting treatment.
Keywords : Root rot, Macrophomina

phaseolina,

biocontrol,

Macrophomina phaseolina (Tassi) Goid. causes seedling rot/dry root rot in mungbean (Vigna
radiata (Roxb.) Wilczek).
Chemical control
achieved by treatment with broad spectrum fungicide showed greater variation between the effectiveness of fungicides (Hooda and Grover, 1983;
Grewal, 1988; Muthukrishnan, 1989). Moreover
chemical measures may establish imbalances in
the microbiological community unfavourable for
activity of beneficial
organisms.
However
biocontrol potential of Trichoderma spp. has been
indicated in a number of reports (Kehri and
Chandra, 1991; Elad et ai., 1986; Jeyarajan et ai.,
1991). In this study, we investigated the efficacy
of seed pelleting
and row application
of
Trichoderma spp. on the control of Macrophomina
rootrot in mung bean and the results are presented herein.
MATERIALS AND METHODS
A field trial was conducted at National Pulses
Research Centre, Vamban during KHARIF 1991
Received for publication

August, 6, 1994.

Trichoderma

viride, seed pelleting

under rainfed condition to see the effect of

Trichoderma harzianum Rifai and Trichoderma
vir ide Pers: Fr (isolates I, II and III) used as seed
pelleting and row application on seed germination, plant biometrics, native nodulation and root
rot incidence. The study was conducted in well
maintained sick plot having Macrophomina population at the rate of 22 sclerotia per g of soil. The
experiment was laid out in randomised block design with three replications in 5x4m plots using
the cultivar C03 mungbean with a spacing of
30xlOcm.
T viride isolate I was obtained from Commonwealth Mycological Institute, London. Isolate
II was from Netherlands while Isolate III and T.
harzianum were from Dept. of Plant Pathology,
TNAU, Coimbatore. For soil application, mass
multiplication of biocontrol agents was done on
wheat bran medium adopting the method of
Kousalya-Gangadharan and Jeyarajan (1990). Sterilization of wheat bran was done at 126C for 2
hours for two successive days in polypropylene
bags. Inoculation was done with 5 mm disc of T
viride isolates I, II and III; and T harzianum

230

[Vol. 50(2) 1997]

Indian Phytopathology

Table I. Effect of Trichoderma

application

on germination

of C03 mung bean

SJ.No.

Treatment

I:

Trichoderma

2.

T. harzianum

3.

T. vir ide I - RA

82.7
(65.65)ab

4.

T. viride I - SP

87.2
(69.09)"

5.

T. viride II - RA

85.4
(67.62)b

6.

T. viride II - SP

84.9
(67.24)ab

7.

T. viride III - RA

85.6
(66.26)'b

8.

T. viride III - SP

89.5
(71.25)'

9.

Seed treatment with carbendazim

(2g1kg of seed)

85.8
(67.86)ob

10.

Control

83.7
(66.26)'b

Germination
harzianum

- RA

77.8
(62.03)b

- SP

83.3
(65.90)ab

SE

1.84

CV (%)

4.7

*Figures in parentheses

are transformed

values.

X = Values followed by the same letter do not differ significantly

range test (DMRT).
RA = Row application

(%)

(P=0.05) according to Duncan's

multiple

at 50g!5m row.

SP = Seed pelleting at 4g1kg of seed.

maintained on PDA and further incubated at room

temperature for 20 days. Fifty gram of wheat bran
medium containing I x 106 CFU/g of substrate
was applied to the soil in rows of 5 m length
during sowing.
For seed pelleting, eight-day old culture of T
viride isolates and T harzianum maintained in
250 ml Erlenmeyer flask containing 70 ml of yeast
molasses broth were mixed including the mycelial
mat and metabolites using a mixie. Further, it was

mixed with taIc powder at the rate of 1:2 (v/w)

and shade dried for 2 days. Carboxymethyl cellulose (lOg) was added as sticky material for 1 kg
of talc powder. The talc based Trichoderma formulation was used for treating the seeds at 4g!kg
of seeds. This taIc based formulation contains I x
107 CFU/g of talc on serial dilution (Ramakrishnan
et al., 1994). For comparison, carbendazim was
used for seed treatment at 2g!kg of seed. One
control without the addition of wheat bran or

(Vol. 50(2) 1997]

Indian Phytopathology

Table 2. Effect of Trichoderma

SI.No.

treatments on plant biometrics at 30 DAS

Treatments

Root
weight
(rng),

Shoot
weight
(rng),

population
per 100g of soil,

Trichoderma

Sclerotialg
of soil x

I.

T harzianum

- RA

261.661>0

2380.0

9x 103e

26.00,b

2.

T harzianum

- SP

338.33'b

4116.7ab

22.6x 1030

12.00do

3.

T vir ide I - RA

263.331>0

3179.01>0

17.6x103d

22.331>0

4.

T viride I - SP

403.33"

3715.0,b

27x 103b

15.66cd

5.

T viride II - RA

268.331>0

3669.7'b

9x I03e

26.33'b

6.

T vir ide II - SP

316.66'bc

4101.7,b

22.3x103c

16.00cd

7.

T viride III - RA

345.66'b

4029.7,b

7xlQ3f

18.331

8.

T viride III - SP

355.00'b

4859.3'

30x 103a

6.33'

9.

Carbendazim
(2g1kg - ST)

329.00ab

40J3.3ab

Ox103a

21.33bo

Control

215.00

4044.3ab

Oxl03g

30.33'

S.E.

33.99

378.33

180.37

2.07

CV (%)

19.13

17.19

35.87

18.47

10.

231

X - Values followed by the same letter do not differ significantly (P

range test.

.,

i>f

...
=

0.05) according to Duncan's multiple

SP - Seed pelleting at 4g1kg of seed. RA - Row application at 50 g!5m row.

antagonists was also maintained. Observations at
30 and 60 days after sowings (DAS) on seed
germination (%), root and shoot length (mm), root
and shoot weight (mg), Trichoderma per 100 g of
soil (Elad and Chet, 1983), scIerotial number of
M. phaseolina (per g of soil), number of nodules
(per plant), root rot incidence (%), number of
pods (per plant) and grain yield (kglha) were also
recorded.
RESULTS AND DISCUSSION
The germination percentage of mungbean presented in Table 1 showed that maximum germinationof 89.51 % was obtained in seed pelleting of T
viride III followed by T viride 1 applied as seed
pelleting. The influence of carbendazim,
T
harzianum, T viride II as seed treatment, and T
viride isolates I, II and III as row application on

seed germination was at par with control (Table I).

The biometrical observations on plant growth
at 30 days after sowing (BAS) indicated that T
viride III as seed pelleting recorded highest root
length of 112.9mm followed by T viride II as
seed pelleting (107.7 mm). However, seed pelleting
treatments of T viride and T harzianum differed
significantly, from row application treatments.
Control recorded 93.8 mm root length. The shoot
length was, however, not influenced by any of the
treatments. T viride III as seed pelleting had the
highest shoot weight of 4859.3 mg as against
control (4044.3 mg) which was at par with other
treatments (Table 2).
The survival of Trichoderma estimated from
rhizosphere soil indicated that seed pelleting of T
vir ide III recorded higher CFU (colony forming

232

Indian Phytopathology

Table 3. Effect of Trichoderma

SI.No.

Treatments

I.

T harzianum

2.

T harzianum

3.

[Vol. 50(2) 1997]

treatments on plant biometrics at 60 DAS
Root
weight
(mg)x

Shoot
weight
(mg),

Trichoderma
population
per 100g of soil,

Sclerotialg
of soil x

- RA

403.3"

6430.0"

4x 103e

26.33"

- SP

471.6"

6470.0"

20.0x J03b

4.66ed

T vir ide I - RA

351.6"

5448.3'

5.8x 103e

15.33b

4.

T viride I - SP

373.3"

4583.3"

18.8x 103b

a.ss=

5.

T viride II - RA

391.0"

5475.6"

4.8x 103e

23.33"

6.

T viride II - SP

410.0

6490.0"

19.3x 103b

s.oo=

7.

T viride III - RA

400.0'

5466.6"

5.5x 103e

8.

T viride III - SP

496.6'

6695.0'

27x J03a

3.33d

9.

Carbendazim

411.6'

6055.'

Ox J03d

26.66'

10.

Control

428.3"

5705.0"

Ox 103d

25.00"

S.E.

47.43

874.55

1103.71

2.24

CV (%)

19.80

25.75

37.0

25.64

(2g1kg) - ST

X - Values followed by the same letter do not differ significantly

range test.

units) production (30.0 x 103) followed by T. viride

II (27 x 103) as seed pelleting. Row application
recorded lower population of Trichoderma ranging from 17.6 x 103 to 7 x 103 Further, the sclerotial population of the pathogen was least (6.33
per g of soil) in seed pelleting of T. viride 1II.
Control recorded highest sclerotial population of
30.33 per g of soil. Carbendazim as seed treatment recorded 21.33 sclerotial population of the
pathogen which is at par with row application of
antagonistic organisms. The trend was same at 60
DAS (Table 3).
The number of nodules per plant on 30 and
60 DAS along with root rot incidence are presented in Table 4. Though all the treatments -are at
par with respect to number of nodules per plant,
seed pelleting with isolate III recorded maximum
number of nodules at both intervals (58.6 and
54.8 respectively). This clearly indicates that T.
viride isolates did not inhibit native nodulation in

(P

11.33bc

0.05) according to Duncan's

multiple

mungbean. This finding was similar to that of

Uma Maheswari (\ 991) and Sridhar et al. (\ 992).
The root rot incidence was least in the treatment
of T. viride III (27.7%) which was on par with
isolate II (31.0%) applied as seed pelleting. Seed
treatment with carbendazim (2g1kg of seed) recorded 39.03% root rot incidence which was at
par with other treatments.
The effect of root rot incidence on yield components presented in Table 4 indicated that the
number of pods was more in T. viride seed pelleting
of isolate III and recorded highest yield of 323.6
kglha, followed by seed pel1eting of isolate II
(279.16 kglha). Control recorded lowest yield of
255.5 kg/ha. The results of this experiment clearly indicated that the survival of T. vir ide isolates
was higher in seed pelleting treatment as compared to row application under rainfed condition
(Table 2 and 3). In seed pelleting treatment, the
antagonistic organism was able to multiply with

[Vol. 50(2) 1997]

Indian Phytopathology

Table 4. Effect of Trichoderma

treatments

SI.

Number of
nodules!
plant on
30th day.

Number of
nodules!
plant on
60th. day

Root rot
incidence
(%)

Number of
pods!plant

Grain yield
(kg/ha)

Treatments

No.

on native nodulation,

root rot incidence and yield components

I.

T harzianum

- RA

47.2liaY

36.26aY

39.2Z
(38.I3)ab

18.85a

263.88a

2.

T harzianum - SP

48.26a

43.96a

43.63
(41.23)b

20.80a

273.60a

3.

T viride I - RA

45.86a

50.4a

35.0
. (36.12)ab

21.05a

265.27a

4.

T viride I - SP

50.5a

44.5a

32.54
(34.72)ab

21.95a

226.38a

5.

T viride II - RA

52.33a

52.5a

45.46
(42.25)a

18.01a

263.88a

6.

T viride II - SP

47.53a

53.46a

31.0
(33.01)a

18.15a

279.16a

7.

T viride III - RA

51.6a

38.66a

44.76ab
(41.79)

2055a

277.77a

8.

T viride III - SP

58.6a

54.8a

27.7
(31.38)a

23.03a

323.6a

9.

Carbendazim

54.2a

50.83a

39.03
(38.I3)ab

16.75a

268.05a

10.

Control

45.06a

41.93a

51.65
(45.97)b

20.la

235.55a

SE

7.56

7.76

3.43

2.04

49.43

CV (%)

26.67

28.76

18.22

17.81

31.74

x - Values

2g!kg-ST

233

followed by the same letter do not differ significantly

(P = 0.05) according

to Duncan's

multiple

range test.
Y - Nodulation was due to native rhizobia, Z - Figures in parentheses

available moisture in root region during crop

growth under rainfed condition, thereby supporting the crop growth, yield and reduction of root
rot pathogen. The result was in concordance with
various workers (Elad et aI., 1986; Kehri and
Chandra, 1991; Jeyarajan et aI., 1991).
Further, the control of Macrophomina root
rot was also better by seed pelleting as compared
to row application. In row application of T viride

are transformed

values.

isolates, the biocontrol agent at first have to multiply in the introduced environment and then these
have to move near the root region to prevent the
entry of the pathogen. In seed pelleting treatment,
the antagonisitc organism readily multiplies on
seed surface which, in turn, prevents the entry of
the pathogen. Similarly, Harman and Taylor
(1988) reported that the use of solid matrix priming as seed treatment effectively controlled the

234

Indian Phytopathology

[Vol.

Pythium ultimum and Fusarium graminearum. Further, Uma Maheswari (1991) supported seed treatment of T. longibrachiatum which recorded
less
incidence of root rot as compared to furrow application in groundnut.
Hence, from this study, it is
clear that seed treatment of antagonistic
organism
is effective in controlling
root rot as compared to
row application

under

rainfed

condition.

REFERENCES
Elad, Y. and Chet, I. (1983). Improved selective media
for isolation of Trichoderma spp. or Fusarium spp.
Phytoparasitica 11: 55-58.
Elad, Y., Zvieli, Y. and Chet, I. (1986). Biological
control of Macrophomina phaseolina (Tassi) Goid
by Trichoderma harzianum. Crop Protection 5:
288-292.

50(2) 1997]

Jeyarajan,
R., Ramakrishnan,
G., Rajamanickam,
B. and Sangeetha, P. (1991). Field demonstrations of efficacy of Trichoderma as biocontrol agent
for root rot disease of grain legumes and oilseeds.
Petria 1: 143.
Kehri, H.K. and Chandra, S. (1991). Antagonism of
Trichoderma viride to Macrophomina phaseolina
in the control of dry root-rot of mung. Indian
Phytopath. 44: 60-63.
Kousalya Gangadharan
and Jeyarajan,
Mass multiplication of Trichoderma
logical Control 4: 70-71.

R. (1990).
spp. 1. Bio-

Muthukrishnan,
K. (1989). Studies on Macrophomina
root rot in rice/allow urd bean (Vigna mungo (L.)
Hepper.) M.Sc. (Ag.) thesis, Tamil Nadu Agricultural University, Coimbatore. 76 p.

Grewal, J.S. (1988). Disease of pulse crops - An overview. Indian Phytopath. 41: \-14.

Ramakrishnan,
G., Jeyarajan,
R. and Dinakaran,
D. (1994). Talc based formulation of Trichoderma
viride for biocontrol of Macrophomina phaseolina.
1. Biological Control 8: 41-44.

Harman, G.E. and Taylor, A.G. (1988). Improved

seedling performance by integration of biological
control agents at favourable pH levels with solid
matrix priming. Phytopathology 78: 520-525.

Sridhar, R., Ramakrishnan,

G., Dinakaran,
D. and
Jeyarajan,
R. (1992). Studies on the compatibility of antagonists with Rhizobium in groundnut. 1.
Oilseeds Research 9: 348-350.

Hooda, I. and Grover, R.K. (\983). Comparative antifungal activity of fungitoxicants against Rhizoctonia bataticola causing seedling rot and foliage
blight of mungbean. Indian 1. Plant Pathol. 1: 7582.
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Uma Maheswari, C. (1991). Biological control of dry

root rot 0/ groundnut (Arachis hypogaea L.) caused
by Macrophomina phaseolina (Tassi) Goid. M.Sc.
(Ag.) Thesis, Tamil Nadu Agricultural University,
Coimbatore. pp. 90.