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ABSTRACT: We propose a simple technique for bacterial and yeast cfu estimations from diverse samples with no prior idea of viable counts, designated as single-plate-serial-dilution-spotting (SP-SDS) with the prime recommendation of sample anchoring (100 stocks). For pure cultures, serial dilutions were prepared from 0.1 OD (100) stock and 20µl aliquots of six dilutions (101 to 106) were applied as 10-15 micro-drops in six sectors over agar-gelled medium in 9-cm plates. For liquid samples 100 to 105 dilutions, and for colloidal suspensions and solid samples (10% w/v), 101 to 106 dilutions were used. Following incubation, at least one dilution level yielded 6-60 cfu per sector comparable to the standard method involving 100µl samples. Tested on diverse bacteria, composite samples and Saccharomyces cerevisiae, SP-SDS offered wider applicability over alternative methods like drop-plating and track-dilution for cfu estimation, single colony isolation and culture purity testing, particularly suiting low resource settings.
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ABSTRACT: The study was taken up to assess if the media constituents played any role in governing the variable colony characteristics or pathogenicity of the bacterial wilt pathogen, Ralstonia solanacearum cultured on the widely employed Kelman medium. The effects due to the constituents 2,3,5-triphenyl tetrazolium chloride (TTC), peptone, casein hydrolysate and glucose on colony characteristics were investigated using -80°C stored culture of strain 'NH-Av01' (race 1, biovar 3) isolated from tomato. Comparing the pigment inducing TTC from two brands, its source or mode of storage/incorporation did not impart any significant effects. The source of peptone, on the other hand, displayed striking effects on the extent of colony growth, fluidity and red pigmentation depending on type, brand or batch / lot of manufacture as documented with 20 different formulations. Significant differences in the pathogenicity of isolate derived from different peptone sources in seedling-challenge assay on tomato were observed. The observations on peptone effects were endorsed with four other isolates belonging to distinct geographic locations, crops (eggplant, chilli, ginger) or races (race 1 or 4). The peptone source did not influence the pathogen-responses in biovar tests but notably altered the pattern of lawn formation and inhibition zone development during antagonistic assays. Casein hydrolysate displayed some variable effects while glucose source had no effect. This study brings to light the significant modifying effects by the peptone-constituent in Kelman medium on the physiology of R. solanacearum and the virulence of isolate and the need to consider the source of media components during culture maintenance, host-pathogen interaction studies or microbe-microbe interaction investigations.
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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.
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