DEVELOPMENT OF SCAR MARKER FOR IDENTIFICATION OF CHAETOMIUM GLOBOSUM - A POTENTIAL BIOCONTROL AGENT

摘要

In this study we report the application of sequence-characterized amplified region (SCAR) marker for the detection of Chuetomium globosum. DNA was extracted from pure cultures of fifteen Chaetomium globosum isolates (Cg1-Cg15) and one isolates each of C. reflexum (Cr) and C perlucidum (Cp). 12 URP primers were used for DNA amplification of all Chuetomium isolates mentioned as above. URP-2R produced a unique DNA band of 1.9 kb in all the isolates of C. globosum but not in C. reflexum and C. perlucidum. This band was eluted and ligated into pGEMT vector. Transformed colonies (White) were used for sequencing and four primer pairs were designed. PCR was performed using all the four synthesized Primers. Amplification with primer Cg 5P (forward primer) 5"- CAC CAA TCG CAC ACT TTG ACC-3" and Cg2 (2) (reverse primer) 5"- ACT GAT CGC ACA CTC CAC CTCT -3" produced a unique DNA band of 1.9 kb in all the isolates of Chuetomium glohosum but it was absent in C.perlucidum, C.reflexum, C.cupreum, Ccochlioides as well as in other fungal genera Aspergillus flavus, Bipoluris sorokiniana and Fusarium moniliformue. Diagnostic PCR was performed using the primer pair IV. The results showed that this SCAR marker can clearly distinguish Chaelomium glohosum at inter specific level as well as inter generic level. Our data provided the foundation for a precise and rapid PCR-based diagnostic system for Chuetomium glohosum. a biocontrol agent at its site of application.