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Research Article
 

Effect of Pseudomonas fluorescens for the Management of Insecticide Resistant Helicoverpa armigera Hubner (Lepidoptera: Noctuidae)



P. Duraimurugan and A. Regupathy
 
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ABSTRACT

In the present study, the talc-based formulation of Pseudomonas fluorescens strain (Pf1) was tested against H. armigera in cotton, okra and pigeonpea. The susceptibility of H. armigera fed on P. fluorescens treated and untreated plants was bioassayed against cypermethrin on cotton bolls, okra fruits and pigeonpea pods. The susceptibility of third instar larvae of H. armigera to cypermethrin, fed on P. fluorescens treated plants did not differ significantly with the untreated plants. However, there was variation in the protein banding pattern among the P. fluorescens treated and untreated plants with or without infestation of H. armigera. Protein bands of molecular weight 83 and 40 kDa in cotton, 130 and 52 kDa in okra were observed in the Pseudomonas treated plants with infestation of H. armigera.

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  How to cite this article:

P. Duraimurugan and A. Regupathy , 2005. Effect of Pseudomonas fluorescens for the Management of Insecticide Resistant Helicoverpa armigera Hubner (Lepidoptera: Noctuidae). Asian Journal of Plant Sciences, 4: 445-448.

DOI: 10.3923/ajps.2005.445.448

URL: https://scialert.net/abstract/?doi=ajps.2005.445.448

INTRODUCTION

Helicoverpa armigera Hubner has become India’s number one agricultural pest[1]. In India, it causes severe damage to a variety of crops like cotton, pigeonpea, okra, chickpea, tomato and sunflower. The damage by H. armigera is estimated at more than Rs 2000 crores ($ 450 m) nationally with 15% decline in the cotton yield[2]. More than 75% of the insecticides used in cotton are being targeted towards H. armigera[3]. Of which, synthetic pyrethroids constitute 50-70%[4]. This high selection pressure led to the development of resistance in H. armigera. The utilization of plant’s own defense mechanism is the subject of current interest in the management of pests and diseases. Induced protection of plants against various pests and pathogen by biotic and abiotic inducers has been reported in many crops[5]. Of these, the induced protection by selected strains of non-pathogenic, root-colonizing Plant Growth Promoting Rhizobacteria (PGPR) has been shown to be capable of inducing pest and disease resistance in addition to promoting plant growth. This phenomenon is commonly referred to as rhizobacteria mediated Induced Systemic Resistance (ISR). However, the reports on the use of PGPR for induced resistance against arthropod pests are limited. In the present study, a talc-based powder formulation was tested to assess the susceptibility of H. armigera to synthetic pyrethroids after feeding on inducing agent P. fluorescens treated plants.

MATERIALS AND METHODS

Susceptibility of H. armigera to synthetic pyrethroids after feeding on P. fluorescens treated plant sources: To assess the PGPR mediated induced systemic resistance against the susceptibility of H. armigera to synthetic pyrethroids after feeding on inducing agent treated plants, a pot culture study was undertaken by using completely randomized design with four replications. The talc-based formulation of PGPR, Pseudomonas fluorescens (Strain Pf1) containing 2.5 to 3.0x108 cfu g-1 obtained from the Department of Plant Pathology, Centre for Plant Protection Studies, Tamil Nadu Agricultural University, Coimbatore, India was applied as seed treatment, soil application and foliar spray. The dry talc powder was mixed with the seeds of cotton, okra and pigeonpea at the rate of 20 g kg-1 of seeds along with rice gruel at the rate of 50 mL kg-1 of seeds[6].

Five gram of talc-based formulation per pot was added 30 days after planting[7]. The talc based product was dissolved in water (20 g L-1) and allowed to settle for 1 h, filtered through muslin cloth and the filtrate was sprayed 30 days after planting[8]. Untreated checks without bacterial treatment were also maintained. Foliar spray with water 30 days after planting was carried out in the untreated checks.

The different treatments on cotton, pigeonpea and okra are T1-cotton treated with P. fluorescens, T2-cotton untreated, T3-okra treated with P. fluorescens, T4-okra untreated, T5-pigeonpea treated with P. fluorescens, T6-pigeonpea untreated. H. armigera, first instar larvae of H. armigera were released on to the bolls of cotton, pods of pigeonpea and fruits of okra and the introduced part of the plants were covered with polyethylene cover and allowed to feed. The larvae grown upto early third instar were bioassayed for susceptibility to cypermethrin by following the bouquet bioassay/foliar residue/terminal bud bioassay method[9]. The LC25 and LC50 values of cypermethrin on cotton bolls, okra fruits and pigeonpea pods were determined through probit analysis[10].

Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE) analysis of crude protein of Pseudomonas treated plants against H. armigera: The total protein was assessed at molecular level for the systemic leaves of P. fluorescens induced cotton, okra and pigeonpea plants with and without infestation of H. armigera and it was compared with the untreated plants. The leaf (with and without feeding of H. armigera) tissues were collected and immediately homogenized. One gram of powdered leaf samples was homogenized with 1 mL of 0.1 M sodium phosphate buffer (pH 7.0) under 4°C. The homogenate was centrifuged for 20 min at 10000 rpm. The supernatant was used for the SDS-PAGE[11].

RESULTS AND DISCUSSION

Susceptibility of H. armigera to synthetic pyrethroid after feeding on P. fluorescens treated plant sources: There was no significant difference in the susceptibility of H. armigera fed on P. fluorescens treated and untreated host plants as the LC50 values were comparable (Table 1). The LC50 value of cypermethrin for the P. fluorescens induced plants was 4.50, 4.07 and 5.28 μg/larva, while the normal plants registered a LC50 of 4.75, 3.90 and 4.95 μg/larva on cotton bolls, okra fruits and pigeonpea pods, respectively.

In addition to direct antagonism and plant growth promotion, certain isolates of fluorescent pseudomonads, interestingly bring about induction of systemic resistance against infection by herbivores[5,12]. Delivery of P. fluorescens through seed, soil, root or foliage leads to the reduction in the incidence of pests[6,7]. But in the present investigation there was no significant difference in the susceptibility of H. armigera which fed on P. fluorescens treated and untreated plants as the LC50 values for cypermethrin were comparable (Table 1). This is contrary to the reports of Murugan[6], who observed that the lower level LC50 value of the quinalphos, chlorpyriphos and profenophos to H. armigera on the jasmonic acid and P. fluorescens induced tomato plants compared to normal plants. Unlike the pathogens, P. fluorescens did not kill the insects through the host plant. Its application brings some physiological changes in host plants that prevent the insects from feeding[13]. This may be the reason, why Pseudomonas induced plant infested with H. armigera did not vary with the concentration mortality response with the normal plants.

SDS-PAGE analysis of crude protein of Pseudomonas treated plants against H. armigera: It was found that there was variation in the protein banding pattern among the Pseudomonas treated and untreated plants with or without infestation (Fig. 1). The intensity of bands were very high in H. armigera infested Pseudomonas treated plants, where as it was less in case of Pseudomonas untreated plants without infestation. Pseudomonas induced proteins with molecular weight of 83 and 40 kDa against H. armigera in cotton. In okra, it was noticed that protein with low relative mobility of 130 and 52 kDa with high intensity was induced. In pigeonpea, there was no induction of protein. However, 10 kDa protein was observed in Pseudomonas treated and untreated plants only after H. armigera infestation (Fig. 1).

Table 1: Susceptibility of H. armigera larvae to cypermethrin after feeding on P. fluorescens treated and untreated plants
Image for - Effect of Pseudomonas fluorescens for the Management of Insecticide
Resistant Helicoverpa armigera Hubner (Lepidoptera: Noctuidae)
LL-Lower Limit; UL -Upper Limit

Image for - Effect of Pseudomonas fluorescens for the Management of Insecticide
Resistant Helicoverpa armigera Hubner (Lepidoptera: Noctuidae)
M:Medium range protein marker
1:Untreated plant without infestation of H. armigera
2:Untreated plant with infestation of H. armigera
3:P. fluorescens pretreated plant without infestation of H. armigera
4:P. fluorescens pretreated plant with infestation of H. armigera
Fig. 1:SDS-PAGE analysis of proteins induced by H. armigera in P. fluorescens pretreated cotton, okra and pigeonpea

The crude protein profile of P. fluorescens treated plants after infestation with H. armigera showed variation in the protein banding pattern compared to untreated plants with or without infestation. Radjacommare et al.[7] reported that the Cnaphalocrocis medinalis infested Pseudomonas treated rice plants induced proteins with molecular weight of 41 kDa compared to the untreated plants. This is in agreement with the present investigation. However the induced proteins observed in the present study were not sufficient to make susceptibility in H. armigera against pyrethroids.

ACKNOWLEDGMENTS

The financial support from the Common Fund for Commodities (CFC) Europe, International Cotton Advisory Committee (ICAC) USA and Natural Resource Institute (NRI), UK is acknowledged.

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