Quantitation of DNA of Propidium Iodide-stained Nuclei from Ipomoea Species and Sweetpotato Using DNA Flow Cytometry
Sweetpotato, Ipomoea batatas is in the morning glory family, Convolvulaceae, genus Ipomoea, group Batatas. It has many wild Ipomoea relatives that serve as a reservoir of many needed pest and stress-resistance genes. A major barrier to introgression of useful genes is the ploidy gap—sweetpotato is a hexaploid and wild Ipomoeas are diploids and tetraploids. The wild species can be successfully crossed using 2n pollen or by first increasing ploidy by colchicine treatment. The ploidy of such hybrid offpsring can be determined by DNA flow cytometry. My objective was to develop a technique to determine DNA content in Ipomoea and values for DNA content for the major Ipomoea species using the EPIC flow cytometer with a UV detector. Nuclei were extracted and pretreated with cellulase and pectolyase before staining with propidium iodide (PI). A highly linear relationship was found between the DNA content determined by DNA flow cytometry and the ploidy of the closest sweetpotato relatives as determined by chromosome counts. These species were diploid I. trifida, tetraploid I. batatas, and hexaploid I. batatas. DNA content was most similar among other diploid Ipomoea species in the group Batatas and was significantly different in other Ipomoeas not in group Batatas.