STUDY ON EFFECT OF NEEM FORMULATIONS ...

7/18/2017

STUDY ON EFFECT OF NEEM FORMULATIONS, FUNGICIDES AND INSECTCIDES ON GROWTH OF Pseud monas fluorescens | Ashutosh…

STUDY ON EFFECT OF NEEM FORMULATIONS, FUNGICIDES AND INSECTCIDES ON GROWTH OF Pseud monas fluorescens UPLOADED BY

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Progressive Research – An International Journal Print ISSN : 0973-6417, Online ISSN : 2454-6003 Volume 10 (Special-IV) : 2015-2019 (2015)

Society for Scientific Development in Agriculture and Technology Meerut (U.P.) INDIA

STUDY ON EFFECT OF NEEM FORMULATIONS, FUNGICIDES AND INSECTCIDES ON GROWTH OF Pseudomonas fluorescens M. Geethu, Ashutosh Gautam Division of Crop Improvement, Indian Cardamom Research Institute, Myladumpara, Idukki Dist Kerala-685553 E-mail : [email protected]

ABSTRACT Pseudomonas fluorescens

is a commonly used bio-control agent against several crop diseases. A study was undertaken to evaluate the compatibility of selected neem formulations, fungicides and insecticides with Pseudomonas fluorescens . Three neem formulations, nine fungicides and seven insecticides were tested along with Pseudomonas fluorescens , of which all the insecticides are compatible with Pseudomonas fluorescens . Econeem plus, Copper oxychloride and wettable sulphur inhibited the growth of Pseudomonas fluorescens .All other chemicals are compatible with Pseudomonas fluorescens. Key words :

Compatibility, Bio-control agent, Neem formulations, Fungicides, Insecticides, Pseudomonas fluorescens

Agriculture is increasingly dependent on the use of chemical fertilizers, growth regulators and pesticides to increase yield. This dependency is associated with problems such as environmental pollution, health hazards, interruption of natural ecological nutrient cycling and destruction of biological communities that otherwise support crop production. Hence, crop improvement and disease management have to be achieved in shorter intervals of time with fewer detrimental inputs. Therefore, the concepts of integrated pest management and organic farming have been accepted in Indian agriculture. Biological control forms an integral part of integrated Pest Management (IPM) for sustainable agriculture and in organic agriculture. Pseudomonas fluorescens is commonly used bio control agent against several crop diseases. Pseudomonas spp . is aerobic, gram-negative bacteria, ubiquitous in agricultural soils, and are well adapted to growing in the rhizosphere. Pseudomonades possess many traits that make them well suited as bio control and growth-promoting agents. Fluorescent pseudomonades are gram negative rod shaped, chaemoheterotrophic bacteria with polar flagella and are characterized by the yellow green iron chelating low molecular weight siderophores called pyoverdines or pseudobactins that fluoresce under UV light (O’Sullivan and O’ Gara, 1992). Pseudomonas fluorescens are common inhabitants of rhizosphere and are the most studied group within the genus Pseudomonas . They can be visually distinguished from the other Pseudomonas species by their ability to produce water soluble yellow green pigments. Pseudomonas fluorescens have noticeably been observed at the root surface, often forming microcolonies or discontinued biofilms in the grooves between epidermal cells. Certain strains are also capable of

endophytic colonization. Within root tissues, they are mostly found in the intracellular spaces of the epidermis and the cortex (Duijff et al, 1997). They are effective at utilizing seed and root exudates for growth and colonize the rhizosphere aggressively. The major mechanism of action of Pseudomonas fluorescens includes root surface colonization, the competition with root pathogens for nutrients, antibiosis through the production of antibiotics, siderophores and other metabolites. The antimicrobial compounds includes, 2,4-diacetylphloroglucinol (DAPG), phenazines, pyrrolnitrin, pyoluteorin, hydrogen cyanide (HCN) and surfactants. The main objective of this study is to test the compatibility of selected neem formulations, fungicides and insecticides used in rice and vegetables with Pseudomonas fluorescens.

MATERIALS AND METHODS The study on “Effect of neem formulations, fungicides and insecticides on the growth of Pseudomonas fluorescens has been carried out during 2014-15 at the ICRI, Myladumpara. The materials used and methods followed are given hereafter. Maintenance of pure culture of Pseudomonas fluorescens : The pure culture of Pseudomonas fluorescens available at the Department of Plant Pathology, Regional Agricultural Research Station (RARS), Pattambi was utilized for the study. This culture is being developed and being used in KAU for the mass production. The culture was maintained on King’s B medium. Composition of King’s B medium : Peptone-20g Glycerol-10 ml K2HPO4-1.5g

MgSO4.7H2O -1.5g Agar-15g

Distilled water-1000 ml

pH-7.2 Preparation of media : Water and peptone were mixed

142

Geethu et. al.,

Table-1 : Effect of Neem formulations on Pseudomonas fluorescens .

The bacterial suspension was prepared in sterile water and 1 ml was added to the petriplates. To this King’s

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fluorescens .

Treatment

Diameter of zone of inhibition (cm) 1.10

T1 - Econeem plus [Azadirachtin 1%] (0.2%) T2 - Neem oil (3%)

0.00

T3 - Azadin plus [Azadirachtin 0.03%] (0.5%)

0.00

T4 - Control

0.00

water and 1 ml was added to the petriplates. To this King’s B medium was added and mixed well to get seeded medium. Appropriate concentration of neem formulations were made in sterile water. Sterile filter paper discs of 10mm diameter were soaked in the solution. After 30 minutes the filter paper discs were taken out aseptically, dried and placed to the center of the bacteria seeded King’s B medium in petri plates. The filter paper discs soaked in sterile water served as control. The plates were kept for incubation at 28° C for 3 days. Observations on the zone of inhibition were recorded. In vitro evaluation of fungicides on Pseudomonas fluorescens An in vitro study was conducted to test the effect of fungicides on Pseudomonas fluorescens by inhibition zone technique. Design : Completely Randomised Design (CRD)

Plate 1a. Effect ofEconeem plus (Azadirachtin 1%) at 0.2% on P. fluorescens

Plate 1b. Effect of neem oil (3%) on P. fluorescens

Replications : 3 Treatments : T1 - Carbendazim 50% WP (0.1%) T2 - Isoprothiolane 40% EC (0.15%) T3 - Kitazin 48% EC (0.1%) T4 - Hexaconazole 5% EC (0.2%) T5 - Mancozeb 75% WP (0.25%) T6 - Copper Oxy Chloride 50% WP (0.3%) T7 - Carbendazim12% + Mancozeb 63% WP (0.15%)

Plate 1c. Effect of Azadin plus (Azadirachtin 0.03%) at 0.5% on P.fluorescens

T8 - Wettable Sulphur (0.5%) T9 - Bordeaux mixture (1%) T10 - Control

well and allowed to boil. Then other chemicals were added and heated until all dissolve. 200 ml of the media was added to 250 ml conical flasks, plugged with cotton plugs, and sterilized in autoclave at 15lb pressure and 121°C for 20 minutes. In vitro evaluation of neem formulations on Pseudomonas fluorescens : An in vitro study was conducted to test the effect of neem formulations on Pseudomonas fluorescens by inhibition zone technique. Design : Completely Randomised Design (CRD) Replications : 5 Treatments : T1 - Econeem plus [Azadirachtin 1%] (0.2%) T2 - Neem oil (3%) T3 - Azadin plus [Azadirachtin 0.03%] (0.5%)

The bacterial suspension was prepared in sterile water and 1 ml was added to the petriplates. To this King’s B medium was added and mixed well to get seeded medium. Appropriate concentrations of fungicides were made in sterile water. Sterile filter paper discs of 10mm diameter were soaked in the solution. After 30 minutes the filter paper discs were taken out aseptically, dried and placed to the center of the bacteria seeded King’s B medium in petri plates.The filter paper discs soaked in sterile water served as control. The plates were kept for incubation at 28° C for 3 days. Observations on the zone of inhibition were recorded. In vitro evaluation of insecticides on Pseudomonas fluorescens : An in vitro study was conducted to test the effect of insecticides on Pseudomonas fluorescens by inhibition zone technique. Design : Completely Randomised Design (CRD)

T4 - Control

Study of effect of neem formulations, fungicides and insectcides on growth

143

Table-2 : Effect of fungicides on Pseudomonas fluorescens Treatment

T1 - Carbendazim (0.1%)

Diameter of zone of inhibition (cm) 0.00c

T2 - Isoprothiolane (0.15%)

0.00c

T3 - Kitazin (0.1%) T4 - Hexaconazole (0.2%)

0.00c

T5 - Mancozeb (0.25%)

0.00c 0.00c

T6 - Copper Oxy Chloride (0.3%)

1.83a

T7 - Carbendazim 12% + Mancozeb 63% (0.15%)

0.00c

T8 - Wettable Sulphur (0.5%)

1.33b

T9 - Bordeaux Mixture (1%)

0.00c

T10 = Control CD (0.05)

Plate 2f. Effect of hexaconazole (0.2%) on P. fluorescens

0.00c 0.09329 Plate 2g. Effect of mancozeb (0.25%) on P. fluorescens


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Plate 2h. Effect of carbendazim12% + mancozeb 63% (0.15%) on P.fluorescens

Plate 2a. Effect of Copper oxy chloride (0.3%) on P. fluorescens Plate 2i. Effect of bordeaux mixture (1%) on P.fluorescens

Replications : 3 Treatments : T1 - Quinalphos 25% EC (0.15%) T2 - Phosalone 35% EC(0.2%) T3 - Dimethoate 30% EC (0.05%) Plate 2b. Effect of wettable sulphur (0.5%) on P.fluorescens

T4 - Chlorpyriphos 20% EC (0.02%) T5 - Acephate 75% SP (0.16%) T6 - Imidacloprid 30.5% (0.05%) T7 - Flubendiamide 39.35% (0.01%) T8 - Control The bacterial suspension was prepared in sterile

Plate 2e. Effect of kitazin (0.1%) on P. fluorescens


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