title: Rapid protocol for visualization of rust fungi structures using fluorochrome Uvitex 2B.
abstract: BACKGROUND
  Histological examination using fluorochromes is one of the standard methods for observation of microorganisms in tissues and other compartments. In the study of fungi, especially those that cannot be cultured in axenic media such as biotrophic fungi, histological examination of processes associated with the fungal growth, differentiation, infection and other cellular functions can lead to the better understanding of host-parasite interactions. Fluorescence microscopy coupled with Fluorochrome Uvitex 2B have been extensively utilized to study rust fungi structures and host-pathogen interactions. In this study, we report development of a rapid staining protocol of the rust fungus Puccinia triticina using fluorochrome Uvitex 2B. The newly developed rapid procedure was compared with a standard staining technique to observe in planta fungal infection structures development during the wheat-Puccinia triticina interaction.
  
  
  RESULTS
  While significantly reducing the time for staining, the rapid protocol described here was equally efficient or better compared to standard procedure in detecting fungal infection structures using Uvitex 2B. In the rapid staining procedure, pre-heating of the stain increased efficiency to detect all the infection structures including haustoria with highly reduced background noise from plant tissue.
  
  
  CONCLUSIONS
  This staining process described here is simple and quick. It can be completed in 4 h, which is of 6 times faster than the standard Uvitex 2B staining procedure.
authors.affiliation: Department of Plant Pathology, North Dakota State University, Dept. 7660, PO Box 6050, Fargo, ND 58108-6050 USA.
authors.forename: Sheshanka
authors.initials: S
authors.lastname: Dugyala
authors.affiliation: Department of Animal Sciences, North Dakota State University, Dept. #7630, PO Box 6050, Fargo, ND 58108-6050 USA.
authors.forename: Pawel
authors.initials: P
authors.lastname: Borowicz
authors.affiliation: Department of Plant Pathology, North Dakota State University, Dept. 7660, PO Box 6050, Fargo, ND 58108-6050 USA.
authors.forename: Maricelis
authors.initials: M
authors.lastname: Acevedo
keywords: Fluorescence microscopy; Fungus; Histology; Host-pathogen interaction; Infection structures; Leaf rust; Puccinia triticina; Staining; Uvitex 2B; Wheat