Detection of Chrysanthemum Yellows Mycoplasmalike Organism by Dot Hybridization and Southern Blot Analysis

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Abstract
Chrysanthemum yellosw (CY) mycoplasmalike organism (MLO) was detected in plants of Catharanthus roseus (periwinkle), Chrysanthemum frutescens, and Chrysanthemum carinatum by dot hybridizations of biotinylated cloned DNA probes to nucleic acid extracts from infected plants. Probes containing DNA fragments from an eastern U.S. aster yellows (AY) MLO or from a Connecticut periwinkle little leaf (0-1) MLO hybridized with nucleic acid extracted from CY-diseased but not healthy plants. Of five probes used in Southern blot analyses, one probe containing extrachromosomal DNA from O-1 MLO hybridized with extrachromosomal but not chromosomal DNA of CY MLO. Three probes containing chromosomal DNA of AY MLO and one containing chromosomal DNA of O-1 MLO hybridized with chromosomal DNA of CY MLO. The nucleotide sequence homologies between the probes and nucleic acid of CY mLO are evidence of genetic relatedness of a European MLO and American MLOs and provide diagnostic tools for CY MLO detection.