The use of histological techniques, combined with the use of dyes, facilitates the visualisation of fungi present in tissues, allowing the details of plant–pathogen interactions in many different pathosystems to be revealed. | Turkish Journal of Botany Research Note Turk J Bot (2013) 37: 784-787 © TÜBİTAK doi: New staining technique for fungal-infected plant tissues João Paulo Rodrigues MARQUES, Marli Kasue Misaki SOARES, Beatriz APPEZZATO-DA-GLORIA* Department of Biological Sciences, Luiz de Queiroz College of Agriculture, University of São Paulo, Piracicaba, Brazil Received: Accepted: Published Online: Printed: Abstract: The use of histological techniques, combined with the use of dyes, facilitates the visualisation of fungi present in tissues, allowing the details of plant–pathogen interactions in many different pathosystems to be revealed. The purpose of this study is to present a new double-staining method that uses cotton blue and safranin to identify fungal structures in plant tissues. With this staining method, the fungal hyphae stain blue and plant cell walls stain orange. The new method was tested on 2 pathosystems. The technique distinguishes between fungal and plant cell walls, giving it great practical utility. This is a staining technique that can be used routinely in laboratories and facilitates the creation of permanent slides. Key words: Citrus black spot, cotton blue, postbloom fruit drop, safranin, staining 1. Introduction Fungal diseases are responsible for structural and physiological disorders in a large number of crops, which leads to decreased productivity and economic losses for producers. Structural studies using histological techniques on the mode of penetration and colonisation of plant tissues by fungi indicate in detail the interactions of the pathogen with the plant tissue. These studies reveal important aspects for the understanding of the monocycle of diseases. Techniques in plant anatomy that are designed to observe and characterise the behaviour of pathogens on their hosts are commonly used. These techniques are used in conjunction with a wide .
