Biological and chemical detection of fumonisins produced on agar medium by Fusarium verticillioides isolates collected from corn in Sohag, Egypt.

Fusarium verticillioides (Sacc.) Nirenberg is among the most common Fusarium species corn pathogens worldwide, and has been recognized as a fumonisin B1 (FB1) and fumonisin B2 (FB2) producer. In the present work, extracts of 58 F. verticillioides isolates from corn samples collected from Sohag Governorate, Egypt, were tested for their biotoxicity and production of fumonisin toxins. Forty-four Fusarium verticillioides isolates out of 58 tested produced FB1 or FB1 and FB2 (15 and 29 isolates, respectively) on potato-sucrose agar medium, detected by TLC, whereas the other 14 isolates did not produce fumonisin toxins. HPLC crude extract analysis confirmed the results from TLC plates. Brine shrimp larvae as well as the Gram-negative bacteria Pseudomonas aeuroginosa showed low bio-sensitivity towards the F. verticillioides crude extract toxicity, whereas the Gram-positive bacteria Bacillus cereus and Bacillus subtilis, especially B. subtilis, showed higher sensitivity towards the tested Fusarium crude extracts. These results enabled us to bio-evaluate and chemically detect fumonisin mycotoxins using a simple agar medium technique.

Increased Salinity Tolerance of Cowpea Plants by Dual Inoculation of an Arbuscular Mycorrhizal Fungus Glomus clarum and a Nitrogen-fixer Azospirillum brasilense.

Pot greenhouse experiments were carried out to attempt to increase the salinity tolerance of one of the most popular legume of the world; cowpea; by using dual inoculation of an Am fungus Glomus clarum and a nitrogen-fixer Azospirillum brasilense. The effect of these beneficial microbes, as single- or dual inoculation-treatments, was assessed in sterilized loamy sand soil at five NaCl levels (0.0~7.2 ds/m) in irrigating water. The results of this study revealed that percentage of mycorrhizal infection, plant height, dry weight, nodule number, protein content, nitrogenase and phosphatase activities, as well as nutrient elements N, P, K, Ca, Mg were significantly decreased by increasing salinity level in non-mycorrhized plants in absence of NFB. Plants inoculated with NFB showed higher nodule numbers, protein content, nitrogen concentration and nitrogenase activities than those of non-inoculated at all salinity levels. Mycorrhized plants exhibited better improvement in all measurements than that of non-mycorrhized ones at all salinity levels, especially, in the presence of NFB. The concentration of Na(+) was significantly accumulated in cowpea plants by rising salinity except in shoots of mycorrhizal plants which had K(+)/Na(+) ratios higher than other treatments. This study indicated that dual inoculation with Am fungi and N-fixer Azospirillum can support both needs for N and P, excess of NaCl and will be useful in terms of soil recovery in saline area.

A biological assay for the detection of Myrothecium spp. produced macrocyclic trichothecenes.

A rapid, inexpensive bioassay to detect Myrothecium spp.-produced macrocyclic trichothecenes was developed. Media containing Myrothecium isolates were inoculated with Chlorella vulgaris, Ustilago maydis and Trichoderma viride. Based on width of the inhibition zone, isolates could be classified as highly toxigenic, non-toxigenic and intermediate. Whereas, C. vulgaris and U. maydis showed significant differences in their response to toxigenic and non-toxigenic isolates, T. viride did not. Production of roridins and verrucarins by the toxigenic isolates (by bioassay) was confirmed by thin layer chromatography and high performance liquid chromatography. This bioassay system, combined with confirmation chemical analyses, increases our ability to detect toxigenic fungal isolates.

Macrocyclic trichothecenes produced by Stachybotrys isolated from Egypt and eastern Europe.

Twenty seven isolates of Stachybotrys chartarum, S. albipes, S. kampalensis and S. microspora from Egypt and Eastern Europe were tested for production of macrocyclic trichothecenes. Twenty of the 27 isolates, grown on rice seeds, were toxic to brine shrimp larvae. Based on TLC and HPLC analyses, 5 macrocyclic trichothecenes (verrucarin J, roridin E, satratoxins F, G & H) as well as trichoverrols were identified. When grown in liquid culture on rice extract medium, only 3 isolates were toxic and produced verrucarin J, roridin E and satratoxins G & H. Extracts from mycelial mats were more toxic than culture filterates of two isolates grown on rice extract and both contained the same macrocyclic trichothecenes (285.5 mg/4 L), in addition to trichoverrols A & B (31 mg/4 L) found in mycelial mats only. When grown on 3% sucrose Czapek's medium supplemented with peptone and yeast extract (still cultures), all isolates were non-toxic to brine shrimp and no trichothecenes could be detected in the extracts.

The mystery of trichothecene antibiotics in Baccharis species.

The Brazilian higher plant Baccharis coridifolia has been shown to synthesize de novo a series of highly toxic macrocyclic trichothecene antibiotics heretofore found to be produced only by fungi. These compounds are produced only by female plants that have undergone pollination. Neither the male nor female plant is sensitive to the toxic effects of trichothecenes, whereas North American Baccharis species are. The macrocyclic trichothecenes found in B. coridifolia are the same as those produced by Myrothecium fungi, and it is suggested that the plant has acquired the toxin-producing genes from this fungus.