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Department of Agricultural Biotechnology, Seoul National University, Seoul 151-921, Korea(Received on September 1, 2009; Accepted on February 4, 2010)Staining profiles and bacterial morphology were com-pared in Xanthomonas citri pv. citri by a transmissionelectron microscopy. Four types of negative stainingregimes were employed depending on culture mediaand heavy metal stains. The bacterial cells grown on LBagar media often appeared clustered on the support-ing film. Meanwhile, individual bacterial cells could bereadily found on the preparations from LB brothmedia. Typical rod-shaped cells (ca. 1 µm in length) andtheir flagella were observed in either 2% uranyl acetate(UA) or 2% neutralized potassium phosphotungstate(PTA) staining. The UA-stained bacteria often showedrelatively intact cell morphology and rather positivelystained cells with a thin electron-dense stain depth aroundbacteria. The PTA-stained bacteria were characterizedby the wrinkled cell surface where the stain was entra-pped in grooves. In addition, distinct electron-dense staindepth was evident around the PTA-stained preparations.Numerous fimbriae could be mostly observed from thePTA-stained preparations of the two culture media, butnot from the UA-stained preparations. Keywords :citrus canker, fimbriae, flagella, negative staining,Xanthomonas citri pv. citri. Negative staining is a simple and rapid method in trans-mission electron microscopy to observe the morphologyand structure of particulate specimens such as bacteria,viruses, cell components, and isolated macromolecules(Hayat, 2000). The staining is usually achieved by heavymetal stain solutions which occupy hydrated regions aroundand within the objects, and form a cast upon drying onto athin supporting film (Hoppert and Holzenburg, 1998).Ideally, negative stains should not bind to the objects whichare represented by stain-excluding areas, that is, theyremain electron-translucent in transmission electron micro-scopy (Hoppert and Holzenburg, 1998). However, a certaindegree of interaction between charged groups of the speci-men and staining ions may occur and produce unwantedpositive staining (Hayat, 2000).The two commonly used heavy metal stains for negativestaining include uranyl acetate (UA; C

Different Profiles of the Negatively Stained Citrus Canker Bacterium Xanthomonas citri pv. citri Depending on Culture Media and Heavy Metal Stains