Influence of Zn and Pb on Rhizopogon roseolus mycelium - energy dispersion spectroscopy and cytochemical investigation

Mycelium isolated from fruitbodies of <em>Rhizopogon roseolus</em>, collected from calamine wastes in Poland, was cultivated on agar media supplemented with Zn or Pb salts. The stimulation of exudate production by the aerial mycelium and the mycelium growing on the surface of the media, accompanied by the change of mycelium pigmentation, was found as a result of Zn application. The presence of Pb resulted mainly in the stimulation of crystalloid production on the surface of mycelium, in direct contact with the medium. Exudate droplets formed on the surface of mycelium cultivated on media with and without the Zn addition, were investigated by means of cytochemical tests (PATAg and Gomori-Swift reaction). In the control media most droplets gave a diffused, positive reaction to both tests. In media supplemented with Zn salts, besides the droplet-like material described in the control also another kind of exudate was observed. It was characterized by the collar showing apositive Gomori-Swift reaction, while the rest of the exudate had an oily appearance and gave a faint or no reaction to both tests. Comparative research by means of scanning electron microscopy accompanied by energy dispersion spectroscopy, was carried out showing the differences in exudate and in mycelia composition as a result of the Zn and Pb presence in the medium.

Structural and cytochemical investigation of the leaf of silverleaf nightshade (Solanum elaeagnifolium), a drought-resistant alien weed of the Greek flora

Light and scanning electron microscopy as well as histochemical reactions were employed to investigate the leaf structure and secretory activity of an adventive perennial pest that is rapidly dispersing in the Mediterranean region. Interesting structural characteristics, not common among the well-adapted native Mediterranean phanerophytes, were observed. Although ‘unconventional’, these structural characteristics produce an ecologically equivalent, xeromorphic leaf on a plant that proves to be competitive and very efficient under the Mediterranean climatic conditions.

Cytochemical Investigation of the Antagonistic Interaction Between a Microsphaeropsis sp. (Isolate P130A) and Venturia inaequalis

In an attempt to better understand the mode of action of the antagonistic fungus Microsphaeropsis sp., the interaction between this fungus and Venturia inaequalis was studied, using both light and electron microscopy. Cytological observations indicated that the antagonistic interaction between the two fungi likely involves a sequence of events, including (i) attachment and local penetration of Microsphaeropsis sp. into V. inaequalis hyphae; (ii) induction of host structural response at sites of potential antagonist entry; (iii) alteration of host cytoplasm; and (iv) active multiplication of antagonistic cells in pathogen hyphae, leading to host cell breakdown and release of the antagonist. The interaction was investigated further by gold cytochemistry. The use of gold-complexed β-1,4-exoglucanase and wheat germ agglutinin/ovomucoid-gold complex to localize cellulosic β-1,4-glucans and chitin monomers, respectively, resulted in regular labeling of V. inaequalis cell walls. This finding supports other studies refuting the classification of ascomycetes as only a glucan-chitin group. At an advanced state of parasitism, the labeling pattern of cellulose and chitin, which clearly showed that the level of integrity of these compounds was affected, suggested the production of cellulolytic and chitinolytic enzymes by Microsphaeropsis sp. Wall appositions formed in V. inaequalis in response to the antagonist's attack contained both cellulose and chitin. However, penetration of this newly formed material frequently succeeded. This study provides the first detailed picture of the cytological events associated with mycoparasitism in V. inaequalis.