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In planta detection of Macrophomina phaseolina from jute (Corchorus olitorius) by a sodium acetate-based direct PCR method

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Title In planta detection of Macrophomina phaseolina from jute (Corchorus olitorius) by a sodium acetate-based direct PCR method
Not Available
 
Creator Chinmay Biswas , Piyali Dey , Kunal Mandal , Subrata Satpathy & P. G. Karmakar
 
Subject s ITS primer. dPCR . Sequencing
 
Description Not Available
Macrophomina phaseolina (Tassi) Goid. is
the most important pathogen of jute and primarily
causes seedling blight, leaf spot and stem rot. The pathogen was detected from field samples by a simple method of direct PCR (dPCR) which obviates the steps of
DNA extraction. The leaf bits were treated with a lysis
buffer at 65°C for 25 min, whereas the stem pieces were
initially incubated at 65°C for 5 min followed by incubation at 60°C for 25 min and the lysate was used as
PCR template. Based on the type of tissue, the composition and concentration of lysis buffer systems were
optimized. For leaf samples the optimized buffer system
composed of 20 mmol l
-1 tris (hydroxymethyl
aminomethane (Tris)-Cl (pH 8.0)), 1.5 mmol l
-1 ethylene
diamine tetra acetate (EDTA) (pH 8.0), 1.4 mol l
-1 sodium acetate and 200 μg/ml proteinase K. Further, 3%
PVP (w/v) and β-mercaptoethanol (1% w/v) were added
into the buffer. In case of stem samples, PVP was not
applied and higher concentrations were used for other
components. M. phaseolina could be detected from both
leaf and stem samples generating amplicon of 350 bp.
This is the first report of detecting M. phaseolina by a
direct PCR method without DNA extraction.
Not Available
 
Date 2020-08-06T08:39:10Z
2020-08-06T08:39:10Z
2014-05-16
 
Type Research Paper
 
Identifier Not Available
Not Available
http://krishi.icar.gov.in/jspui/handle/123456789/39000
 
Language English
 
Relation Not Available;
 
Publisher Springer