title: A multiplex nested PCR assay for simultaneous detection of Corchorus golden mosaic virus and a phytoplasma in white jute (Corchorus capsularis L.).
abstract: A multiplex nested PCR assay was developed by optimizing reaction components and reaction cycling parameters for simultaneous detection of Corchorus golden mosaic virus (CoGMV) and a phytoplasma (Group 16Sr V-C) causing little leaf and bunchy top in white jute (Corchorus capsularis). Three sets of specific primers viz. a CoGMV specific (DNA-A region) primer, a 16S rDNA universal primer pair P1/P7 and nested primer pair R16F2n/R2 for phytoplasmas were used. The concentrations of the PCR components such as primers, MgCl2 , Taq DNA polymerase, dNTPs and PCR conditions including annealing temperature and amplification cycles were examined and optimized. Expected fragments of 1 kb (CoGMV), 674 bp (phytoplasma) and 370 bp (nested R16F2n/R2) were successfully amplified by this multiplex nested PCR system ensuring simultaneous, sensitive and specific detection of the phytoplasma and the virus. The multiplex nested PCR provides a sensitive, rapid and low-cost method for simultaneous detection of jute little leaf phytoplasma and CoGMV. Based on BLASTn analyses, the phytoplasma was found to belong to the Group 16Sr V-C.
authors: Crop Protection Division, Central Research Institute for Jute and Allied Fibres (CRIJAF), Barrackpore, Kolkata, West Bengal, India. chinmaybiswas@rediffmail.com
authors: C
authors: C
authors: Biswas
authors: P
authors: P
authors: Dey
authors: S
authors: S
authors: Satpathy
mesh_terms: D053839:Begomovirus / Q000235:genetics / Q000302:isolation & purification*; D032381:Corchorus / Q000235:genetics / Q000382:microbiology* / Q000821:virology; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D009029:Mosaic Viruses / Q000235:genetics / Q000302:isolation & purification; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D010802:Phylogeny; D045582:Phytoplasma / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D010935:Plant Diseases / Q000382:microbiology* / Q000821:virology*; D018515:Plant Leaves / Q000382:microbiology / Q000821:virology; D012336:RNA, Ribosomal, 16S / Q000235:genetics
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D004279:DNA, Viral; D012336:RNA, Ribosomal, 16S