This study shows that the superoxide radical (O 2˙" ), a direct indicator of oxidative stress, is involved in the differentiation of the phytopathogenic filamentous fungi Rhizoctonia solani, Sclerotinia sclerotiorum, Sclerotium rolfsii and Sclerotinia minor, shown by using superoxide dismutase (SOD) mimetics to decrease their sclerotial differentiation. The production rate of O 2˙" and SOD levels in these fungi, as expected, were significantly lowered by the SOD mimetics, with concomitant decrease of the indirect indicator of oxidative stress, lipid peroxidation.
INTRODUCTIONSclerotial differentiation is a primitive form of differentiation exhibited by certain phytopathogenic filamentous fungi. It is expressed by four main types of sclerotia (compact bodies of aggregated hyphae), which are loose, terminal, lateral-chained and lateral-simple. These types are represented mainly, but not solely, by Rhizoctonia solani, Sclerotinia sclerotiorum, Sclerotium rolfsii and Sclerotinia minor, respectively (Chet et al., 1967(Chet et al., , 1969Chet & Henis, 1975;Le Tourneau, 1979;Willetts, 1971Willetts, , 1978Willetts, , 1997Willetts & Wong, 1980). These fungi are of great agricultural interest because they cause stem-rot diseases in a wide variety of economically important agricultural crops. Therefore, elucidating the mechanisms of sclerotium biogenesis of these fungi may lead to the development of non-toxic ways to combat plant diseases, which could be used as alternatives to the traditional toxic fungicides. Moreover, understanding sclerotial differentiation is very important because it will help to understand more complex forms of differentiation in other organisms as well as whether their mechanisms share common biochemical pathways.It has been proposed that sclerotial differentiation is related to high oxidative stress (Georgiou, 1997), based on the finding that sclerotium biogenesis in S. rolfsii was accompanied by the accumulation of high levels of lipid peroxidation products. This finding has been supported by other studies that developed into a theory of oxidative stress as an inducer of sclerotial metamorphosis (Georgiou et al., 2006). Specifically, hydroxyl radical scavengers, vitamin C and b-carotene (both also acting as endogenous fungal antioxidants), decreased sclerotial differentiation in S. rolfsii, S. minor, S. sclerotiorum and R. solani. In more recent studies, we demonstrated a strong relationship between the thiol redox state of these fungi and their sclerotial differentiation (Patsoukis & Georgiou, 2007a, b, 2008a. Specifically, in R. solani, S. rolfsii and S. minor, there was a decrease in the total amount of reduced thiols during transformation of the undifferentiated mycelium into sclerotium, which, for S. sclerotiorum was accompanied by an increase in the total oxidized components. Moreover, the antioxidant thiol redox state modulator N-acetylcysteine inhibited sclerotial differentiation in S. rolfsii, R. solani and S. sclerotiorum.Until now, our studies have associated only indirect indicators ...