Tissue specific reactions of sorghum roots to the mycoherbicide Fusarium oxysporum f. sp. strigae versus the pathogenic F. proliferatum

Fusarium oxysporum f. sp. strigae (Foxy 2) is a mycoherbicide against Striga hermonthica. To ensure the safe use of this biocontrol agent, and as part of the risk assessment, this study was aimed at providing cytological evidence that Foxy 2 does not possess pathogenic behavior towards the non-target host sorghum. Therefore, we compared the infection processes and sorghum root tissue reactions towards the pathogenic F. proliferatum to that of Foxy 2 using light- and transmission electron microscopy. Given that during the growth process, hyphae could get into the central cylinder, tissue specific reactions of sorghum to Foxy 2 were also investigated by wounding the roots (exposing the vascular system), and testing for proliferation of hyphae within the vessels. Results showed that 2 weeks after sowing, F. proliferatum had invaded and destroyed all cell types including the central cylinder while Foxy 2 hyphae were located around the outer endodermal layer and were not able to penetrate the latter. There was an increase in blue autoflourescence in the central cylinder and especially the endodermis, probably due to increased phenolics in Foxy 2 infected roots which was not the case for F. proliferatum. This might contribute to the inability of Foxy 2 to penetrate the endodermis. Transmission electron microscopy showed extensive degradation of endodermis and vessel walls into thin translucent layers by F. proliferatum but not by Foxy 2. In the mechanically wounded and infected roots, Foxy 2 could invade the central cylinder close to the wound but was not identified a few millimeters away from the wound. This implies that it was not able to grow within or destroy the central cylinder even when already present within it; probably due to the observed increased phenolics. Thus, exposure of the vascular system did not serve as a route for the invasion of Foxy 2 which therefore could not further cause tracheomycosis. Therefore, Foxy 2 could be seen as a fungus well suited for biocontrol.     

Which Morphological Characteristics Are Most Influenced by the Host Matrix in Downy Mildews? A Case Study in Pseudoperonospora cubensis

Before the advent of molecular phylogenetics, species concepts in the downy mildews, an economically important group of obligate biotrophic oomycete pathogens, have mostly been based upon host range and morphology. While molecular phylogenetic studies have confirmed a narrow host range for many downy mildew species, others, like Pseudoperonospora cubensis affect even different genera. Although often morphological differences were found for new, phylogenetically distinct species, uncertainty prevails regarding their host ranges, especially regarding related plants that have been reported as downy mildew hosts, but were not included in the phylogenetic studies. In these cases, the basis for deciding if the divergence in some morphological characters can be deemed sufficient for designation as separate species is uncertain, as observed morphological divergence could be due to different host matrices colonised. The broad host range of P. cubensis (ca. 60 host species) renders this pathogen an ideal model organism for the investigation of morphological variations in relation to the host matrix and to evaluate which characteristics are best indicators for conspecificity or distinctiveness. On the basis of twelve morphological characterisitcs and a set of twelve cucurbits from five different Cucurbitaceae tribes, including the two species, Cyclanthera pedata and Thladiantha dubia, hitherto not reported as hosts of P. cubensis, a significant influence of the host matrix on pathogen morphology was found. Given the high intraspecific variation of some characteristics, also their plasticity has to be taken into account. The implications for morphological species determination and the confidence limits of morphological characteristics are discussed. For species delimitations in Pseudoperonospora it is shown that the ratio of the height of the first ramification to the sporangiophore length, ratio of the longer to the shorter ultimate branchlet, and especially the length and width of sporangia, as well as, with some reservations, their ratio, are the most suitable characteristics for species delimitation.