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Title: | In planta detection of Macrophomina phaseolina from jute (Corchorus olitorius) by a sodium acetate-based direct PCR method. |
Other Titles: | Not Available |
Authors: | Biswas, C., Dey, P., Mandal, K., Satpathy, S. and Karmakar, P.G. |
ICAR Data Use Licennce: | http://krishi.icar.gov.in/PDF/ICAR_Data_Use_Licence.pdf |
Author's Affiliated institute: | ICAR::Central Research Institute for Jute and Allied Fibres |
Published/ Complete Date: | 2014 |
Project Code: | Not Available |
Keywords: | ITS primer dPCR Sequencing |
Publisher: | Not Available |
Citation: | Not Available |
Series/Report no.: | Not Available; |
Abstract/Description: | 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. |
Description: | Not Available |
ISSN: | Not Available |
Type(s) of content: | Research Paper |
Sponsors: | Not Available |
Language: | English |
Name of Journal: | Phytoparasitica |
NAAS Rating: | 7.14 |
Volume No.: | 42(5) |
Page Number: | 673-676 |
Name of the Division/Regional Station: | Not Available |
Source, DOI or any other URL: | Not Available |
URI: | http://krishi.icar.gov.in/jspui/handle/123456789/28177 |
Appears in Collections: | CS-CRIJAF-Publication |
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