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Indian Journal of Pure & Applied Biosciences (IJPAB)
Year : 2020, Volume : 8, Issue : 4
First page : (66) Last page : (75)
Article doi: : http://dx.doi.org/10.18782/2582-2845.7968

Use of Plant Products, Bio Control Agents, Chemicals and Organic Amendments for Integrated Management of Rhizoctonia solani in Cowpea

Tribikram Sahoo¹, Abhisek Tripathy^2*, Shriram Ratan Pradhan³ and Anirudha Tarai⁴
College of Agriculture, Odisha University of Agriculture and Technology, Bhubaneswar-751003
*Corresponding Author E-mail: abhi.plantpathology@gmail.com
Received: 23.02.2020  |  Revised: 29.03.2020   |  Accepted: 8.04.2020 

 ABSTRACT

For integrated disease management of root and stem rot of cowpea caused by Rhizoctonia solani experiments were conducted using different plant products, bio control agents, chemicals and organic amendments during Kharif of 2014-15. Out of seven phytoextracts tested against Rhizoctonia solani, the maximum inhibition was recorded in garlic(100%) followed by turmeric(34.08%) at 10 % concentration, whereas maximum inhibition 100% was recorded in both 15% and 20% concentration in garlic and calotropis respectively. Out of four biological agents were tested against Rhizoctonia solani, Bacillus subtilis was recorded maximum growth inhibition of 72.04% as compared to Trichoderma hamatum (49.78%) and Trichoderma harzianum (46.00%).  In vitro studies of 13 fungicides against Rhizoctonia solani revealed that Carboxin 37.5% + Thiram 37.5%, Hexaconazole 5%, Difenoconazole 25%, Tebuconazole 25% and Tebuconazole 50% + Trifloxystrobin 25% recorded maximum growth inhibition (100%). In vivo studies of 13 fungicides maximum mortality was recorded from Carbendazim 50% followed by Propineb 70% (82% and 65.97%) respectively. Among the organic products tested against Rhizoctonia solani, the minimum mortality was recorded from spent mushroom substrate + Cowdung + Vermicompost followed by Vermicompost with 10% and 15% mortality respectively. The maximum mortality was recorded from Poultry manure (66.67%) as compared to rest of the treatments and control.

Keywords: Phytoextract, Rhizoctonia solani, Biological agents, Bacillus subtilis, Cowpea

Full Text : PDF; Journal doi : http://dx.doi.org/10.18782

Cite this article: Sahoo, T., Tripathy, A., Pradhan, S.R., & Tarai, A. (2020). Use of Plant Products, Bio Control Agents, Chemicals and Organic Amendments for Integrated Management of Rhizoctonia solani in Cowpea, Ind. J. Pure App. Biosci. 8(4), 66-75. doi: http://dx.doi.org/10.18782/2582-2845.7968

INTRODUCTION

MATERIALS AND METHODS

The research  was  carried out both in the laboratory and  field  during  2014-16.The  field and laboratory experiment was conducted at the AICRP on vegetable crops, Orissa  University  of  Agriculture  and  Technology, Bhubaneswar  and  Research facilities of  Department  of  Plant  Pathology, OUAT, Bhubaneswar.
In vitro evaluation of plant extracts for fungal growth inhibition by poison food technique-
To  study  the  antifungal  mechanism  of  plant  extracts, the  poisoned  food  technique  was  used (Nene and Thapliyal, 1973) in vitro.
Different plant extracts used in these experiments are enlisted below.

Table 1: List of plant extracts used in In vitro evaluation against R. solani

Sl. No	Common name	Botanical name	Plant part used
1	Neem	Azadirachta indica A.	Leaf
2	Garlic	Allium sataivum L.	Clove
3	Turmeric	Curcuma  longa	Rhizome
4	Karanja	Pongamia glabra	Leaf
5	Begunia	Begunia nirgundi	Leaf
6	Datura	Datura stramonium	Leaf
7	Milkweed	Calotropis spp.	Leaf

The per cent  inhibition  over  control  was  calculated  according  to  formula   given  by  Vincent (1947) as  follows.
                                      
 I = Per cent inhibition of mycelium
C = Growth of mycelium in control
T = Growth of mycelium in treatment

In vitro evaluation of bioagents by Dual culture technique
The antagonistic  microorganisms  like  Trichoderma viride, Trichoderma harzianum, Trichoderma hamatum and Bacillus subtilis were  maintained  in  medium potato  dextrose  agar  and  evaluated  for  their  antagonistic  effect  under  in vitro  conditions  against  Rhizoctonia solani  by  dual  culture  technique. In  dual  culture  technique, twenty  ml  of  sterilized  and  cooled  potato  dextrose  agar  was  poured  into  sterile  Petri dishes  and  allowed  to  solidfy. Fungal  antagonistics  inoculating  the  pathogen  at  one  side  of  Petridish  and  the  antagonist  inoculated  at  exactly  opposite  side  of  the  same  plate  by  leaving  3-4 cm  gap. For this, actively growing cultures were used. Each treatment was replicated 2 times. After required period of incubation i.e.  after  control  plate  reached  growth  of  90  mm  diameter, the  radial  growth  of  pathogen  was  measured. Per cent  inhibition  over  control  was  worked  out  according  to  formulae  given  by Vincent (1947).
The  fungicides  were  tested  initially  under  in vitro  condition by  poison  food  technique (Nene & Thapliyal,1973)  at  desired  concentration  in in  vivo condition by  pot  culture  method. Per cent  inhibition of mycelial  growth  over  control  was  calculated  by  using  the  formulae  given  below  by  Vincent (1947).
In vivo evaluation of fungicides
Soil was collected from the research plot of Department of Plant Pathology.    The soil was sterilized in autoclave at 1210C at 15 psi for 2 hours. Sterilized soils were put into polythene bag mixing with inoculum of actively growing culture of R. solani. Then it was incubated for 10days. After that seeds of cowpea at the rate of ten seeds per pot were sown. After thirteen days, thinning was done. Then  the  required  concentrations  of  chemicals  were  prepared  and  incorporated  into pot. Three replications were maintained for each treatment.

Table 2: List of different fungicides tested for efficacy in R.solani

Sl. No	Common names	Chemical  name	Conc. (%)
1	Bavistin	Carbendazim 50%WP	0.2
2	Vitavax power	Carboxin 37.5% + Thiram 37.5%	0.2
3	Sheathmar 3L	Validmycin 3%L	0.1
4	Onestar	Azoxystrobin 23% SC	0.1
5	Sixer	Mancozeb 63%WP + Carbendazim 12% WP	0.2
6	Curzate	Cymoxanil 8% WP+Mancozeb 64% WP	0.15
7	Roko	Thiophanate Methyl 70% WP	0.15
8	Antracol	Propineb 70% WP	0.2
9	Contaf	Hexaconazole 5% EC	0.05
10	Score	Difenconazole 25% EC	0.05
11	Folicur	Tebuconazole 25%EC	0.05
12	Tilt	Propiconazole 25% EC	0.05
13	Nativo	Tebuconazole50%WP+ Trifloxystrobin25%WP	0.05

In vivo efficacy of organic products against R. solani
Different organic products were taken to evaluate their efficacy in field condition for controlling the Rhizoctonia solani infecting cowpea. Organic products were selected for experimental purpose as per their local availability viz.  Mustard oil cake, Karanj oil cake, Neem oil cake, Mahua oil cake, Poultry manure, Goat manure, VAM, Vermicompost, Spent Musroom Substrate and fresh cow dung. Each treatment was replicated thrice. Soil was sterilized at 15 psi at temperature of 1210C for 20 minutes.The soil was inoculated with the culture of Rhizoctonia solani. The poly pots were closed with rubber bands and kept under shade. The organic products were amended thoroughly with previously incubated soil at the rate of 10gms per kg of soil. After mixing the organic products, the poly pots were closed with rubber bands and kept for seven days. Ten cowpea seeds were sown in each pot and watering was carried out thrice a week to promote germination and growth of the plants. The regular observation was recorded with respect to mortality percentage and plant height.

Table 3: List of organic products used against Rhizoctonia soalni

Sl. No	Name	Quantity(gm)/Kg of soil
1	Mustard oil cake	10
2	Karanja oil cake	10
3	Poultry manure	10
4	Goat manure	10
5	Vescicular arbuschular mycorrihiza(VAM)	10
6	Vermi compost	10
7	Spent mushroom substrate (SMS)	10
8	SMS + Cow dung	10
9	SMS + Vermi compost	10
10	SMS + Vermicompost +Cow dung	10
11	Neem oil cake	10
12	Mahua oil cake	10

Statistical analysis was carried out by following the standard procedures (Panse & Sukhatme, 1967). Data in percentage were transformed to angular values before analysis.

RESULTS

Efficacy of plant extracts in inhibiting the growth of the fungus
Seven plant extracts, which are easily available in the locality were evaluated for their fungitoxicity activity against the growth of Rhizoctonia solani at  10,15 and 20% concentration as per the procedure described above. The radial growth of the fungal colony on plant extract mixed medium were measured after seven days of incubation and the data obtained were presented in Table  4.
All the plant extracts under study inhibited mycelial growth of Rhizoctonia solani at 10, 15 and 20 per cent concentrations and were significantly superior over control. The results indicated that, the maximum inhibition was recorded in Garlic (100%) followed by Turmeric (34.08) at 10% concentration. At 15% concentration, a total inhibition was noticed in Garlic and Calotropis (100%) followed by Turmeric (41.11%). At 20% concentration, a total inhibition was noticed in Garlic and Calotropis (100%) followed by Turmeric (68.33%). The least reduction of growth was observed in Begunia (0.38%) at 10% concentration and in Karanja 5.56% and 9.82% at 15% and 20% concentration respectively. In general all the leaf extracts tested, significantly inhibited the mycelia growth of Rhizoctonia solani above 1% at 10% concentration, 5% at 15% concentration and 9% at 20% concentration except Begunia and Karanja.

Table 4: Efficacy of plant extracts against Rhizoctonia solani in in vitro

Sl. No.	Common name	% inhibition at 10% conc.	% inhibition at 15% conc.	% inhibition at 20% conc.
1	Garlic (Allium sataivum L.)	*100 (90.02)	100 (90.02)**	100 (90.02)
2	Neem (Azadirachta indica A.)	1.11 (6.05)	17.41 (24.67)	21.11 (27.36)
3	Begunia (Vitex nirgundi)	0.38 (3.52)	6.67 (14.97)	20.87 (27.20)
4	Datura (Datura stramonium)	30.93 (33.80)	33.33 (35.27)	41.67 (40.21)
5	Karanja (Pongamia glabra)	9.82 (18.27)	5.56 (13.64)	9.82 (18.27)
6	Turmeric (Curcuma  longa)	34.08 (35.72)	41.11 (39.89)	68.33 (55.77)
7	Milkweed (Calotropis spp.)	7.78 (16.20)	100 (90.02)	100 (90.02)
8	Control	0	0	0
	SE(m)±	0.79	0.66	0.55
	CD(0.05)	2.36	1.88	1.65

Effect of Bio control Agents on Mycelial Growth of Rhizoctonia solani.
An experiment was conducted to explore the capabilities of four bio-agents against the test pathogen. The antagonistic nature of these bioagents against the test fungus was studied adopting dual culture technique and data obtained are presented in Table 5.
Antagonistic studies revealed that, growth of pathogen was significantly checked over control by the antagonistic nature of all the antagonists tested. The antagonists also restricted the growth of the pathogen and didn’t allow it to grow further.                            The lowest growth of the pathogen was observed in Bacillus subtilis (25.16mm) followed by Trichoderma hamatum (45.20mm), Trichoderma harzianum (48.60mm), Trichoderma viride (50.60mm). However, the growth of Rhizoctonia solani in Trichoderma viride was maximum.
          Regarding the degree of growth inhibition, Bacillus subtilis inhibited the maximum growth of the pathogen (72.04%) followed by Trichoderma hamatum, Trichoderma harzianum but such inhibition of the growth of the pathogen with  Trichoderma viride was relatively least among all the antagonists. The range of growth inhibition was from 43.78% to 72.04% which undoubtly positively establishes a check in the growth of pathogen without treatment with fungicides.

Table 5: In vitro bio assay of bio control agent

Sl.no.	Name of antagonist	Mean diameter of Rhizoctonia solani (mm)	*Mean diameter in dual culture (mm)	Growth inhibition (%)
1	Trichoderma hamatum	90	45.20	49.78
2	Trichoderma viride	90	50.60	43.78
3	Trichoderma harzianum	90	48.60	46.00
4	Bacillus subtilis	90	25.16	72.04
5	Control	90	90.00	0.00

Efficacy of fungicides on growth of Rhizoctonia solani in in vitro
In order to evaluate the efficacy of some selected fungicides on growth of Rhizoctonia solani, the experiment was conducted as per the procedure mentioned earlier following posion food technique. The colony diameter and per cent growth inhibition have been presented in Table 6. It was observed from the data that, all the fungicides tested in solid medium significantly reduced the fungal colony in comparision to control.
It was revealed that fungicides namely Carboxin 37.5% + Thiram 37.5% along with Hexaconazole 5% EC, Difenoconazole 25% EC, Tebuconazole 25% EC and Tebuconazole 50% + Trifloxystrobin 25% proved efficacious in inhibiting the mycelial growth with 100 per cent inhibition. The next effective fungicides were Cymoxanil 8% WP+Mancozeb 64% WP  and Propiconazole 25% EC which recorded  79.81 and 77.57 per cent inhibition respectively. However Validamycin, Mancozeb 63%WP + Carbendazim 12% WP Azoxystrobin 23% SC and Propineb 70 WP registered 54.63, 52.78 and 5.07 respectively.

Table 6: Efficacy of  fungicides on growth of  Rhizoctonia solani in in-vitro

Sl. No.	Chemical  name	Conc.(%)	% growth inhibition
1	Carbendazim 50%WP	0.2	0.00* (0)
2	Carboxin 37.5% WP + Thiram 37.5% WP	0.2	100 (90.02)
3	Validamycin 3%L	0.1	54.63 (47.67)
4	Azoxystrobin 23% SC	0.1	8.37 (16.82)
5	Mancozeb 63%WP + Carbendazim 12% WP	0.2	52.78 (46.60)
6	Cymoxanil 8% WP+ Mancozeb 64% WP	0.15	79.81 (63.31)**
7	Thiophanate Methyl 70% WP	0.15	1.26 (6.44)
8	Propineb 70% WP	0.2	5.07 (13.02)
9	Hexaconazole 5% EC	0.05	100 (90.02)
10	Difenoconazole 25% EC	0.05	100 (90.02)
11	Tebuconazole 25% EC	0.05	100 (90.02)
12	Propiconazole 25% EC	0.05	77.57 (61.74)
13	Tebuconazole50% WP+ Trifloxystrobin25%WP	0.05	100 (90.02)
	SE (m)±		0.90
	CD(0.05)		2.71

Table 7: Efficacy of  fungicides on Rhizoctonia solani in in-vivo

Sl. No.	Chemical  name	Conc. (%)	*Mortality (%)
1	Carbendazim 50%WP	0.2	82.00*
2	Carboxin 37.5%WP + Thiram 37.5%WP	0.2	2.60
3	Validamycin 3%L	0.1	8.93
4	Azoxystrobin 23% SC	0.1	48.77
5	Mancozeb 63%WP + Carbendazim 12% WP	0.2	7.73
6	Cymoxanil 8% WP+ Mancozeb 64% WP	0.15	1.83
7	Thiophanate Methyl 70% WP	0.15	60.07
8	Propineb 70% WP	0.2	65.97
9	Hexaconazole 5% EC	0.05	2.67
10	Difenoconazole 25% EC	0.05	2.00
11	Tebuconazole 25% EC	0.05	12.63
12	Propiconazole 25% EC	0.05	12.10
13	Tebuconazole50%WP+ Trifloxystrobin25%WP	0.05	3.47
14	Control		99.07
	SE (m)±		1.51
	CD(0.05)		4.54

Efficacy of fungicides on growth of R. solani  in in-vivo
The fungicides which were studied in laboratory, they were further studied under pot culture experiment as per the procedure described under ‘Materials and Methods’ and presented in table 7.
        This study reaveled that least mortality was recorded from Cymoxanil 8% WP+mancozeb 64% WP (1.83%) followed by  Difenoconazole 25% EC (2.00%) and Carboxin 37.5% + Thiram 37.5% (2.60%). The maximum mortality  was recorded in Carbendazim 50%WP(82.00%) followed by Propineb 70% WP (65.97%)and Azoxystrobin 23% SC (60.07%). However the control pots recorded as high as 99.07% mortality.
Effect of organic amendment on growth of Rhizoctonia solani
This study revealed that, least mortality was recorded from spent mushroom substrate+ cowdung+ vermicompost (10.00%) followed by Vermicompost (15.00%). The  mortality was recorded in poultry manure amendment(66.67%) followed by mustard oil cake (58.33%). Fifty per cent mortality was recorded in Spent mushroom substrate +vermicompost, Karanja oil cake and Mahua oil cake amended treatments. However control pots recorded as high as 91.66% mortality. So far as plant height was concerned the result was obtained in the same trend. Spent mushroom substrate + cowdung +vermicompost has recorded the maximum plant height (89.33cm) followed by vermicompost (81.67cm). However control pot recorded only (11.00cm).

Table 8:  In vivo efficacy of organic products against  Rhizoctonia solani

Sl.No.	Treatment	Mortality %	Plant Height(cm)
1	Control	91.66*	11.00*
2	Mustard oil cake	58.33	3.33
3	Karanja oil cake	50.00	58.33
4	Poultry manure	66.67	47.33
5	Goat manure	41.67	47.00
6	Vescicular arbuschular mycorrhiza (VAM)	41.67	58.33
7	Vermicompost	16.00	81.67
8	Spent mushroom substrate (SMS)	50.00	51.67
9	SMS + Cowdung	41.67	79.00
10	SMS + vermicompost	50.00	64.33
11	SMS+Cowdung+Vermicompost	10.00	89.33
12	Neem oil cake	41.67	31.33
13	Mahua oil cake	50.00	37.33
	SE(m)±	9.67	11.67
	CD(0.05)	28.40	34.27

DISCUSSIONS

All the phytoextracts under study inhibited mycelial growth of Rhizoctonia solani at 10%, 15% and 20% concentration as compared to control. The maximum inhibition was recorded in garlic (100%) at 10% concentration. Garlic and calotropis were recorded the maximum per cent growth inhibition (100%) in both 15% and 20% concentration respectively. Garlic, a potential phytoextract effective against a wide range of diseases including R. solani has been reported earlier by Shashidhara et al. (2008) and Sehajpal et al. (2009) which supports our findings. Dawar et al.(2010) reported potentiality of Datura alba against R.solani as efficacious one which also corroborating the present investigation.
In the present study the antifungal characterstics of bio-agents viz. Trichoderma viride, Trichoderma hamatum, Trichoderma  harzianum and Bacillus subtilis were tested in vitro to study the effectiveness against R.solani  employing dual culture technique. Maximum growth inhibition was found in Bacillus subtilis (72.04%) followed by Trichoderma hamatum (49.78%) and Trichoderma harzianum (46.00%). The effectiveness of Trichoderma spp. in field of plant disease management  was confirmed earlier by various workers like Elad et al.(1983), Chang and Choi (1990), Nagarajkumar et al.(2004), Sharma et al. (2009), Bhat et al. (2009) and Panwar et al. (2012). The antagonistic nature of Bacillus subtilis also supports the findings of Schmiedeknecht (1993), who reported that B. subtilis is a suitable antagonists for biological control of Rhizoctonia disease on potato.
              In vitro bio assay of fungicides against Rhizoctonia solani revealed that, Carboxin 37.5% + Thiram 37.5% @ 0.2%, Hexaconazole 5% @ 0.05%, Difenoconazole 25% @ 0.05%, Tebuconazole 25% @ 0.05% and Tebuconazole 50% + Trifloxystrobin 25% @ 0.05%   recorded maximum growth inhibition however Validamycin 3% @ 0.1%, Cymoxanil 8% + Mancozeb 64% @ 0.15%, Mancozeb 63% + Carbendazim 12% @ 0.2%, Propiconazole 25% @ 0.05% concentration also checked the growth of Rhizoctonia solani ranging 52.78% to 91.81%  inhibition. It was supported by earlier report of Sahu (1986), who reported that Carboxin performed best against R.solani in Rapeseed and mustard.
In in vivo study with chemicals in pot culture experiment, the minimum mortality was recorded in Cymoxanil 8% + Mancozeb 64% @ 0.15% followed by Difenoconazole 25% @ 0.05% and Carboxin 37.5% + Thiram 37.5% @ 0.2 % concentration registered the mortality as 1.83%, 2.0% and 2.60% respectively. The maximum mortality was recorded in Carbendazim 50% @ 0.2 % followed by Propineb 70% @ 0.2% recording mortality as with 82.0% and 65.97% respectively. The effectiveness of Carbendazim was found to be less against R.solani, but Sunder et al. (2009) reported that Carbendazim was proved effective against R. solani in mung bean, which contradicts the present findings.
In pot culture experiment, the effectiveness of organic products were explored  on the  growth of Rhizoctonia solani  which revealed that, the minimum mortality was recorded from Spent mushroom substrate + Cowdung + Vermicompost followed by Vermicompost with 10% and 15% respectively. The maximum mortality was recorded from Poultry manure 66.67%  as compared to rest of the treatments including control. With regard to the plant height the result was also obtained in same trend. Spent mushroom substrate + Cowdung + Vermicompost recorded maximum plant height followed by Vermicompost with 89.33cm and 81.67cm respectively. Such findings are also in agreement with the findings of El-Mohamedy et al. (2006), who amended soil with Trichoderma hazianum. Ashlesha et al. (2009) amended soil with dry powder of cowdung and fresh cowdung urine and milk and reported effective against soil borne Rhizoctonia and Fusarium pathogens. The efficacy of Vermicompost against R.solani has been reported earlier by Sinha et al. (2010) which is also in agreement with the present finding.

CONCLUSION

Therefore the phytoextracts and organic products may be included in integrated disease management strategy against Rhizoctonia root and stem rot management in cowpea to the farming community of Odisha. Use of phyto extracts as a means of non-chemical crop disease management is found imperative in view of its eco-friendly nature. However use of chemicals in integrated disease management schedule has been the last resort, so far as crop disease management is concerned. But exploration of antagonists and organic products found to be efficacious and economic and non-hazardous to environment. The farming community of Odisha may be educated to use indigenous products like botanicals and organic products in crop disease management programme.

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