Epicuticular Wax of the First Leaves of Two Barley Cultivars Studied by Cryo Scanning Electron Microscopy

Chemical fixation and critical point drying of plant tissue prior to scanning electron microscopy often degrades the outermost layer of the specimen, i.e. the epicuticular wax. Knowledge of the unaltered wax, including its morphology, is important because the wax constitutes the interface between the plant and its surroundings. Having worked previously with barley leaves (anatomy, infection biology of pathogens), we found it of interest to examine and describe the wax morphology on barley leaves and to determine whether or not there were differences between cultivars. Hence, the morphology of the epicuticular wax on the first leaf of two spring barley cultivars has been studied by scanning electron microscopy following cryofixation.
The wax on both cultivars consists chiefly of scales with a small proportion of rods and granules. There is little variation in wax morphology over each leaf side, and only minor differences between the abaxial and adaxial sides of the same leaf. Distinct differences are, however, observed between the two cultivars, especially regarding the thickness and the shape of the wax scales. The wax bodies cover the entire surface of a leaf except for parts of trichomes and guard cells, and some scattered wax free areas.
The morphology and distribution of wax are discussed in relation to studies made on barley leaves using different techniques. Also, the implications of wax morphology for disease resistance are briefly discussed.     

Extracellular Matrix, Esterase and the Phytotoxin Prehelminthosporol in Infection of Barley Leaves by Bipolaris sorokiniana

Light microscopy and cryo-scanning electron microscopy showed that hyphae of Bipolaris sorokiniana adhered to the wax surfaces of barley leaves by means of an extensive extracellular matrix (ECM). Prehelminthosporol, the major non-host specific phytotoxin formed by B. sorokiniana was immunolocalized in large amounts in the ECM surrounding the hyphae. Similarly, esterase activity involved in degradation of the cuticular wax surface was found in the ECM. Therefore, it appears that the ECM is not only important for adhesion of the fungus to its host, but also functions as a sink of phytotoxins and lytic enzymes important for infection of the host plant.