An improved method of DNA isolation suitable for PCR-based detection of begomoviruses from jute and other mucilaginous plants.
A relatively quick and inexpensive modified cetyl trimethylammonium bromide method for extraction of DNA from leaf materials containing large quantities of mucilage is described. The modification including use of more volume of extraction buffer and dissolving crude nucleic acid pellet in 1 M NaCl, reduced markedly the viscosity of the mucilage and thus in the final purification step yielded a larger quantity of mucilage-free DNA suitable for subsequent PCR-based detection of begomoviruses. The method was standardized with jute samples with yellow mosaic disease and validated with different other mucilaginous-hosts with low titre of begomoviruses. DNA isolated using this method showed consistency in yield and compatibility with PCR for detection of begomoviruses from different mucilaginous plant species. The method was compared for efficacy with other reported methods and it was found to be superior over the existing methods described for isolation of DNA from mucilaginous hosts. Thus the method described could be used on a wider scale for reliable and consistent detection of begomoviruses from mucilaginous hosts for characterization and variability study.
Ghosh
Raju
R
Plant Virus Laboratory and Biotechnology Unit, Division of Crop Protection, Central Research Institute for Jute and Allied Fibres, Barrackpore, Kolkata 700120, West Bengal, India.
Paul
Sujay
S
Ghosh
Subrata Kumar
SK
Roy
Anirban
A
D053839:Begomovirus / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D000077286:Cetrimonium; D002593:Cetrimonium Compounds / Q000737:chemistry; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D010935:Plant Diseases / Q000821:virology*; D018515:Plant Leaves / Q000737:chemistry / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods; D011933:Reagent Kits, Diagnostic
D002593:Cetrimonium Compounds; D004279:DNA, Viral; D011933:Reagent Kits, Diagnostic; D000077286:Cetrimonium