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Department of Biotechnology
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Division of Entomology and Nematology
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    ABSTRACT: To understand the factors that contribute to the variations in colony forming units (CFU) in different bacteria during spread-plating. Employing a mix culture of vegetative cells of ten organisms varying in cell characteristics (Gram-reaction, cell-shape and cell-size), spread-plating to the extent of just drying the agar surface (50-60 s) was tested in comparison with the alternate spotting-and-tilt-spreading (SATS) approach where 100 μl inoculum was distributed by mere tilting of plate after spotting as 20-25 micro-drops. The former imparted a significant reduction in CFU by 20% over the spreader-independent SATS approach. Extending the testing to single organisms, Gram-negative proteobacteria with relatively larger cells (Escherichia, Enterobacter, Agrobacterium, Ralstonia, Pantoea, Pseudomonas and Sphingomonas spp.) showed significant CFU reduction with spread-plating except for slow-growing Methylobacterium sp., while those with small rods (Xenophilus sp.) and cocci (Acinetobacter sp.) were less affected. Among Gram-positive non-spore formers, Staphylococcus epidermidis showed significant CFU reduction while S. haemolyticus and actinobacteria (Microbacterium, Cellulosimicrobium and Brachybacterium spp.) with small rods / cocci were unaffected. Vegetative cells of Bacillus pumilus and B. subtilis were generally unaffected while others with larger rods (B. thuringiensis, Brevibacillus, Lysinibacillus and Paenibacillus spp.) were significantly affected. A simulated plating study coupled with Live-Dead bacterial staining endorsed the chances of cell disruption with spreader impaction in afflicted organisms. Significant reduction in CFU could occur during spread-plating due to physical impaction injury to bacterial cells depending on the spreader usage and the variable effects on different organisms are determined by Gram-reaction, cell-size and cell-shape. The inoculum spreader could impart physical disruption of vegetative cells against a hard surface. Possibility of CFU reduction in sensitive organisms and the skewed selection of hardier organisms during spread-plating, and the recommendation of SATS as an easier and safer alternative for CFU enumerations. This article is protected by copyright. All rights reserved.
    Journal of Applied Microbiology 12/2013; DOI:10.1111/jam.12412
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    ABSTRACT: Wider germplasm diversity and transferability of the antioxidant parameters is the primary requirement for the development of high antioxidant tomato cultivars. The present study was conducted to screen the tomato genotypes including hybrids, varieties, cherry tomatoes, wild species, elite germplasm lines, inter-specific hybrids and backcross populations for antioxidant activity and other quality parameters to select high antioxidant lines with good total soluble solids (TSS) for further usage in crop improvement programmes. Wild species and inter-specific hybrids between LA-1777 (Solanum habrochaites) and an elite genotype 15SBSB recorded very high antioxidant capacity (FRAP), DPPH radical scavenging ability, higher phenols and flavonoids. Inter-specific hybrids also recorded very high total soluble solids (TSS). Significantly higher total carotenoids, lycopene and vitamin C were observed in IIHR-249-1 with moderately higher TSS. Cherry tomato lines IIHR-2866, 2865 and 2864 recorded 4 to 5 times more β-carotene than the commercial hybrids/varieties. Tomato line IIHR-249-1 can be used for improving the antioxidant capacity, total carotenoids and lycopene in tomato breeding programmes. Cherry tomato lines IIHR-2866, 2865 and 2864 can be used for improving the β-carotene content. LA-1777 and inter-specific hybrids could be used for developing tomato lines rich in antioxidants as well as TSS.
    Journal of the Science of Food and Agriculture 08/2013; 94(5). DOI:10.1002/jsfa.6359
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    ABSTRACT: Rapid, precise and timely identification of invasive pest insects such as aphids is important and a challenge worldwide due to their complex life cycles, parthenogenetic reproduction, sex and colour morphs. In this respect, DNA barcoding employing a 658 bp fragment of 5' region of the mitochondrial cytochrome oxidase I (CO-I) gene is an effective tool in addressing the above. In the present study, we employed CO-I for discriminating 142 individuals representing 32 species of aphids from India. Sequence analyses revealed that the intraspecific and interspecific distances ranged from zero to 3.8% and 2.31 to 18.9%, respectively. In addition, the study also showed for the first time the prevalence of three cryptic species, namely Brevicoryne brassicae (Linnaeus), Hyperomyzus carduellinus (Theobald) and Brachycaudus helichrysi (Kaltenbach) from India. Our work has clearly demonstrated that DNA barcoding is an efficient and accurate method for identification of aphid species (including cryptic species), an approach that potentially could play an important role in formulating viable pest management strategies, more especially biocontrol.
    Bulletin of entomological research 05/2013; DOI:10.1017/S0007485313000278


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    Bangalore, India
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National meet on Betelvine - Farmers, Traders and Researchers Interface, IIHR Bangalore; 02/2014

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