• Journal of Microbiology and Biotechnology
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• v.18 no.2
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• pp.234-241
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• 2008

Agrobacterum tumefaciens-mediated transformation (ATMT) is becoming an effective system as an insertional mutagenesis tool in filamentous fungi. We developed and optimized ATMT for two Colletotrichum species, C. falcatum and C. acutatum, which are the causal agents of sugarcane red rot and pepper anthracnose, respectively. A. tumefaciens strain SK1044, carrying a hygromycin phosphotransferase gene (hph) and a green fluorescent protein (GFP) gene, was used to transform the conidia of these two Colletotrichum species. Transformation efficiency was correlated with co-cultivation time and bacterial cell concentration and was higher in C. falcatum than in C. acutatum. Southern blot analysis indicated that about 65% of the transformants had a single copy of the T-DNA in both C. falcatum and C. acutatum and that T-DNA integrated randomly in both fungal genomes. T-DNA insertions were identified in transformants through thermal asymmetrical interlaced PCR (TAIL-PCR) followed by sequencing. Our results suggested that ATMT can be used as a molecular tool to identify and characterize pathogenicity-related genes in these two economically important Colletotrichum species.

• Mycobiology
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• v.32 no.4
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• pp.149-154
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• 2004

Toxin produced by Colletotrichum falcatum Went, the incitant of red rot of sugarcane was isolated, purified and assayed to determine host specificity and identify its chemical nature. The toxin was found to be not host specific as it inhibited germination of various seeds(gram, greengram, blackgram, pea, cowpea, rice and sugarcane) as well as different seedlings viz. tomato, coriander, pea and rice. The toxin consists of two distinct fraction-one fraction having $R_f$, value at 0.36 producing identical red rot lesion when inoculated at leaf midrib of sugarcane, and the other having $R_f$, value at 0.72 not showing any red rot lesion. Chromatogram of high performance liquid chromatography(HPLC) of the red rot lesion causing fraction showed a sharp peak at 1.62 min of retention time(RT), and spectral analysis indicated the presence of following chemical $CH_3$ - groups-C-H, C=O, C-N, $-CH_3,\;-CH_2$ -CH and molecular mass of the compound was 203. - ($M^+,\;C_{11}H_{11}N_2O_2$).

• Journal of Microbiology and Biotechnology
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• v.19 no.12
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• pp.1688-1694
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• 2009

A novel strain of fluorescent pseudomonad (PB-St2) was isolated from surface-sterilized stems of sugarcane grown in Pakistan. The bacterium was identified as Pseudomonas aurantiaca on the basis of 16S rRNA gene sequence analysis and results from physiological and biochemical characteristics carried out with API50 CH and QTS 24 bacterial identification kits. Assays using substrate-specific media for enzymes revealed lipase and protease activities but cellulase, chitinase, or pectinase were not detected. The bacterium was unable to solubilize phosphate or produce indole acetic acid. However, it did produce HCN, siderophores, and homoserine lactones. In dual culture assays on agar, the bacterium showed antifungal activity against an important pathogen of sugarcane in Pakistan, namely Colletotrichum falcatum, as well as for pathogenic isolates of Fusarium oxysporium and F. lateritium but not against F. solani. The antifungal metabolites were identified using thin-layer chromatography, UV spectra, and MALDI-TOFF spectra and shown to be phenazine-1-carboxylic acid (PCA), 2-hydroxyphenazine (2-OH-PHZ), and N-hexanoyl homoserine lactone (HHL) (assessed using only TLC data). The capacity of this bacterium to produce HCN and 2-OH-PHZ, as well as to inhibit the growth of C. falcatum, has not been previously reported.

• Mycobiology
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• v.35 no.3
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• pp.157-158
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• 2007

The alkaloid fuyuziphine was isolated from the whole plant of Fumaria indica. It had inhibitive effect against spore germination of some plant pathogenic fungi (Collectotrichum sp., C. gloeosporioides, C. falcatum, Curvularia maculans, C. lunata, Erysiphe cichoracearum, Helminthosporium pennisetti, Oidium erysiphoides, Ustilago cynodontis, Alternaria chieranthi, A. mel-ongenae, A. brassicicola and A. solam). Curvularia lunata, Oidium erysiphoides, Alternaria brassicicola and A. solani did not germinate at 750 and 1000 ppm and Colletotrichum gloeosporioides, C. falcatum, Curvularia maculans were inhibited at 1000 ppm for 24 hr incubation. Germination of most fungi was significantly inhibited at $100{\sim}750ppm$.

• Journal of Microbiology and Biotechnology
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• v.16 no.2
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• pp.184-192
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• 2006

Six-hundred bacterial strains from human milk and milk from Sahiwal cows, Holstein Friesian cows, and buffaloes were screened for their ability to suppress phytopathogenic fungi under in vitro conditions. A consortium of 3 strains, viz., Bacillus lentimorbus B-30486 (B-30486), B. subtilis B-30487 (B-30487), and B. lentimorbus B-30488 (B-30488), isolated from Sahiwal cow milk resulted in better biological control and plant-growth promotion than single-strain treatments. For commercial-scale production of a bioinoculant, the solid-state fermentation of sugarcane agro-industrial residues, i.e., molasses, press mud, and spent wash, using the consortium of B-30486, B-30487, and B-30488, resulted in a value-added product, useful for enhancing plant growth. The application of the consortium to sugarcane fields infested with Fusarium moniliforme and Colletotrichum falcatum resulted in a reduction of mortality and significantly higher (P=0.05) plant height, number of tillers, and cane girth when compared with the control. Furthermore, under field conditions, the treatment of sugarcane with the consortium resulted in significantly (P=0.05) greater plant growth compared with nonbacterized plants. Accordingly, this is the first report on the effective use of bacteria isolated from milk for biological control and enhancing plant growth under field conditions. Furthormore, a solid-state fermentation technology was developed that facilitates the economic utilization of agro-industrial residues for environmental conservation and improving plant and soil health.