Differentiation of Bulgarian Isolates from Cucumber Mosaic Virus by Serological Methods

Abstract

Cucumber mosaic virus (CMV) is of great importance to the Bulgarian economy and hence a detailed knowledge of its diversity under local geographic and climatic conditions is required. An extended study was carried out on CMV strains the currently occur in Bulgaria. Fifty‐one isolates and strains found in different regions and various crops were biologically characterized and serologically differentiated into subgroups I and II using different variants of enzyme‐linked immunosorbent assay (ELISA) [double antibody sandwich (DAS)‐, antigen‐coated plate (ACP)‐, triple antibody sandwich (TAS)‐ with poly and monoclonal antibodies] and immunodiffusion tests. The ELISA modifications with monoclonal antibodies individually (ACP) or in combination with polyclonal antibodies (TAS‐ELISA) are suitable for mass screening of CMV isolates. The hyperimmune sera against strains from CMV subgroups I and II were very efficient for use in isolate differentiation via gel double immunodiffusion. The results obtained correlated with the polymerase chain reaction and restriction fragment length polymorphism data reported by other authors. The majority of the isolates belonged to subgroup I, whereas 10, mainly from tomato and pepper, belonged to subgroup II. Most of the subgroup II isolates came from the north of Bulgaria. The results of the present study will help to clarify the virus epidemiology and to develop specific control measures.

Molecular Techniques for Detection of Microbial Pathogens

“…Plate-trapped antigen (PTA)-ELISA was used to screen the hybridoma cells producing monoclonal antibodies against Sugarcane yellow leaf virus (ScYLV) and for routine detection of the virus in infected plant tissues (Korimbocus et al 2002). PTA-ELISA was employed to detect Cucumber mosaic virus (CMV) in various crops and different regions of Bulgaria using individual MABs (Hristova et al 2002). By using the MAB 2B5 in indirect antigen-coated plate-ELISA, Sugarcane mosaic virus was detected in maize leaf samples showing dwarf mosaic symptoms in China (Jiang et al 2003).…”

“…Plate-trapped antibody (PTA)-ELISA technique has been shown to be sufficient in detecting some plant viruses in infected tissues. By using specific antisera Sugarcane yellow leaf virus (SCYLV) (Korimbocus et al 2002) Tomato yellow leaf curl virus (TYLCV), Tomato mosaic virus (ToMoV) (Abouzid et al 2002), Cucumber mosaic virus (Hristova et al 2002) and Sugarcane mosaic virus (Jiang et al 2003) have been detected reliably. Antigen-coated plate (ACP)-ELISA format was employed to detect Maize streak virus (MSV) and Maize mottle chlorotic stunt virus (MMCSV) in leaf samples from germplasm in Nigeria.…”

“…Likewise, triple antibody-sandwich (TAS)-ELISA also has been reported to be useful in detecting virus-infected plants. The viruses that could be detected successfully by TAS-ELISA are CMV (Hristova et al 2002) Faba bean necrotic yellows virus (FBNYV), Potato virus Y (Ounouna et al 2002) and CMV (Yu et al 2005).…”