![]() At least five individual bacterial cells were focused on in order to detect any differences in shape and cell wall intactness. In order to study the effect of nanocopper on bacterial cell, the nanocopper-treated (0.2 ppm) strain ITCC BD0003 of Xap was viewed on a scanning electron microscopic (SEM) under low vacuum state after 48 h of incubation. Scanning electron microscopic analysis of nanocopper treated Xap In the quantitative assay, YP broth was amended with nanocopper as described earlier and, after incubation at 48 h, the CFU count (CFU ml −1) was compared with the control (YP without nanocopper but inoculated with Xap). The observation was recorded as positive for suppression of growth and negative for appearance of bacterial growth. ![]() A YP plate not amended with nanocopper but streaked with Xap was kept as a control. For the qualitative assay, the YP agar amended with different doses of nanocopper was streaked with Xap strain ITCC BD0003 and kept for incubation at 28☌ for 96 h. The in vitro growth inhibition activity of nanocopper doses was analyzed qualitatively (in solid YP agar media) and quantitatively (in YP liquid broth). Nanocopper formulation was added to the growth medium and adjusted to a final concentration of 20, 16, 8, 4, 2, 0.5, 0.4, and 0.2 ppm. The nanocopper formulation used for this study was produced by Micro Chemicals (Mandsaur, Madhya Pradesh, India). In vitro efficacy of nanocopper against Xap Well separated fluidal colonies of the strain were selected for further studies after growth at 48 h at 28☌ at 200 rpm in YP broth. The bacterial cells were revived from stored culture by streaking on a yeast peptone (YP) (yeast extract 5 g, peptone 10 g, agar 20 g and water 1 L, pH 7.0 ) agar plate and incubating at 28☌ for 72–96 h. punicae (Xap) strain ITCC BD0003, isolated from pomegranate leaves in Delhi and stored as single colonies in 50% glycerol at −70☌, was used in this study. Keeping in view this objective, based on our expertise on other bacterial pathogens (bacterial blight of rice and bean), in the present study we investigated the efficacy and possible mode of action of nano-based formulation of copper (nanocopper) against pomegranate blight bacterium. Therefore, the develop of environment-friendly strategies for the disease management is necessary. In the OHM, the antibiotic streptocycline is used in a dose (>500 ppm) more than twice higher in comparison with the optimal dose (100–200 ppm). Presently, orchard health management (OHM) strategies were found to be effective to certain extent. Treatments by chemicals had limited success against the disease (Kumar et al. Several management options have been investigated, involving the application of antibiotics, chemicals and other cultural practices. Up to now, very little information is available on the disease management aspects. 2010) and Pakistan (Akhtar and Bhatti 1992). 1965) until in recent reports from South Africa (Petersen et al. The bacterial blight of pomegranate is known to be confined mainly in India (Chand and Kishun 1991 Hingorani and Mehta 1952 Hingorani and Singh 1959 Patel et al. The disease was of minor importance until the recent appearance of the Xap in epidemic form in many pomegranate growing states of India, including Karnataka and Maharashtra. punicae (Xap) strongly damaged the pomegranate production, inducing large economical losses to the Indian growers (Mondal and Sharma 2009). However, bacterial blight caused by Xanthomonas axonopodis pv. Recently, improved varieties, such as Bhagva and Ganesh, have been introduced in Maharashtra state, where they cover more than 90 per cent of cultivated areas (Mondal and Singh 2009). Pomegranate ( Punica granatum), being a favorite table fruit of tropical and sub-tropical regions of the world, has emerged as commercial fruit in many Indian states including Maharashtra, Andhra Pradesh, Uttar Pradesh, Gujarat, Rajasthan, Karnataka and Tamil Nadu (Mondal and Mani 2009).
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