Please use this identifier to cite or link to this item: http://ir.nbu.ac.in/handle/123456789/4459
Title: Serological and molecular detection of Macrophomina phaseolina, causing root rot of Citrus reticulata
Other Titles: NBU Journal of Plant Sciences, Vol.6, No.1 (March 2012) p 77-86
Authors: Chakraborty, B.N.
Chakraborty, U.
Rai, K.
Sunar, K.
Dey, P.L.
Keywords: Citrus reticulata
Macrophomina phaseolina
PTA-ELISA
DIBA
Immunofluorescence
rDNA
ITS region
Issue Date: Mar-2012
Publisher: University of North Bengal
Abstract: Polyclonal antibodies (PAbs) were raised against mycelial antigens of Macrophomina phaseolina a causal organism of root rot disease of mandarin plants. IgG was purified and further packaged into immunological formats such as immuno diffusion, Plate trapped antigen (PTA)-ELISA, dot immunobinding assay, Western blot analysis and indirect immunofluorescence for quick and accurate detection of pathogen from soil. Indirect staining of mycelia and sclerotia of M. phaseolina with homologous PAb and labeling with goat antirabbit IgG conjugated with FITC developed strong fluorescence in young hyphal tips and sclerotia of M. phaseolina. Genomic DNA prepared from mycelia of M. phaseolina was purified and PCR amplification of 18S rDNA was done using ITS region specific primer pair. The amplified DNA was sequenced and aligned against ex-type strain sequences from NCBI GenBank using BLAST and phylogenetic analysis was obtained using MEGA4 software. Amplification of ITSI region of the rDNA can be considered as a rapid technique for identifying pathogens successfully in all cases.
URI: http://ir.nbu.ac.in/handle/123456789/4459
ISSN: 0974-6927
Appears in Collections:NBU Journal of Plant Sciences, Vol.6, No.1 (March 2012)

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