Title
Detection of sweetpotato chlorotic stunt crinivirus (SPCSV) by non-radioactive nucleic acid spot hybridization (NASH) technique
Date Issued
01 January 2002
Access level
metadata only access
Resource Type
conference paper
Publisher(s)
International Society for Horticultural Science
Abstract
Sweetpotato viral disease (SPVD) caused by the synergistic interaction of sweetpotato feathery mottle potyvirus (SPFMV) and sweetpotato chlorotic stunt crinivirus (SPCSV) is difficult to recognize in infected plants of most cultivars. At present, specific monoclonal and polyclonal antibodies are available to detect both viral components of the disease. However, low concentration of SPCSV in plant tissues makes difficult its serological detection. A non-radioactive specific probe was developed to detect SPCSV using the published sequence of its coat protein gene. The non-radioactive probe was labeled by a random priming labeling technique with a fluoresceinated-nucleotide and assayed by Nucleic Acid Spot Hybridization (NASH). Different extraction buffers and modifications in the hybridization steps were assayed. Samples from field-grown sweetpotato showing typical SPVDsymptoms and samples without symptoms were collected and assayed by NCMELISA and NASH techniques. Comparative results showed that the non-radioactive NASH technique is slightly better than NCM-ELISA when samples are extracted with buffer 5X SSC and hybridized at 70°C for 4 h, using the hybridization buffer used routinely at CIP for Nick Translation NASH technique. This technique represents an important sensitive tool for the detection of SPCSV in "seed" multiplication approaches and also in programs devoted to development of SPVDresistant genotypes.
Start page
129
End page
133
Volume
583
Language
English
OCDE Knowledge area
Horticultura, Viticultura Agricultura
Scopus EID
2-s2.0-78249270492
ISSN of the container
05677572
ISBN of the container
9789066059856
Conference
Acta Horticulturae
Sources of information: Directorio de Producción Científica Scopus