Rhizomatous grass weeds and Ophiobolus graminis Sacc

An account is given of the infection of grass weeds by Ophiobolus granwm in a wheat crop affected by the disease. It was shown that the fungus was carried by the rhizomatous grasses, Agropyron repens and Agrostis spp. and also by Holcus lanatus. Examination of these showed the importance of Agrostis spp. in carrying over the mycelium under field conditions.     

Occurrence of Ophiobolus graminis var. Avenae on wheat crops in the field

Ophiobolus graminis var. Avenae has been found on oat crops in ten counties in Scotland. It has also been shown to attack wheat crops in the same districts.     

Control of survival of Ophiobolus graminis between consecutive crops of winter wheat

Under a glasshouse crop of red clover, Ophiobolus graminis (Sacc.) Sacc. in artificially colonized straws survived for longer on the soil surface than when buried, and for much longer when suspended above the soil; survival in buried straws was somewhat more prolonged in fallow soil than under clover. In field experiments with consecutive crops of winter wheat, under-sowing with red clover was not effective in reducing the incidence of take-all, possibly because of above-ground survival of the pathogen in unploughed straw. Early rotavation, however, significantly reduced disease incidence, probably because of enhanced microbial activity and competition for nitrogen in the well-aerated compost of soil and stubble.     

Effects of nitrogen and glucose on saprophytic survival of Ophiobolus graminis in buried straw

Calcium nitrate prolonged the saprophytic survival of Ophiobolus graminis (Sacc.) Sacc. in artificially colonized straws buried in soil in the laboratory, whether supplied to the soil (at 12·5 or 100 mg nitrogen/100 g soil) or to the straws before colonization (at 0·5 or 1·0 g nitrogen/100 g straw). Glucose (at 2·5 g/100 g soil, and at 10 g/100 g straw) shortened survival. When straws colonized in the presence of 0·5 g nitrogen/100 g straw were buried in soil supplied with 14·1 mg nitrogen/100 g soil, the level of soluble soil nitrogen reached equilibrium at 2–4mg/100g soil; this allowed rapid straw decomposition and, although the added soil nitrogen prolonged survival in straws that remained undecomposed, it also accelerated substrate exhaustion. Addition of 100 mg nitrogen/100 g soil was supra-optimal for survival: although some nitrogen was necessary for maximum survival, the equilibrium concentration of soluble nitrogen (24–56 mg/100 g soil) was high enough in this case to have an inhibitory effect in addition.     

Methoxyhydroquinone, a Growth Inhibitor of Ophiobolus graminis, in Leaves of Oat Seedlings

A methanol extract of leaves of oat seedlings grown in sand cultures in the dark contained a compound which inhibited the growth of Ophiobolus graminis. The inhibitory factor was isolated and proved to be present in the plant as methoxyhydroquinone glucoside. The glucoside was readily hydrolysed to the corresponding aglucone. The methoxyhydroquinone, or possibly its oxydation product, methoxy-P-benzoquinone, was inhibitory to both Ophiobolus graminis var. graminis and Ophiobolus graminis var. avenae, whereas Fusarmm oxysporum var. lycopcrsici was not affected. Synthetic methoxyhydroquinone at 80 mg/l gave a 100% inhibition of Ophiobolus graminis var. graminis. After being exposed to 80 mg/l of the inhibitor for 24 h the mycelium was unable to initiate growth when transferred to a fresh nutrient solution. Only extracts from young leaves showed inhibitory activity, extracts from mature leaves giving no inhibition. The hydroquinone, or its glucoside, was not detected in roots of young seedlings, where avenacin was the only antifungal compound present.     

A scanning electron microscope study of interactions between micro-organisms andGaeumannomyces graminis (Syn.Ophiobolus graminis) on wheat roots

Roots of wheat grown in unsterilized sand inoculated withGaeumannomyces graminis (Sacc.) von Arx and Olivier were examined by scanning electron microscopy. Healthy roots had a mucilaginous covering and were sparsely colonized by bacteria, but asG. graminis colonized the roots the mucilage disappeared and the numbers of bacteria on the surface increased. Lysis of the hyphae occurred, apparently caused by bacteria that colonized the hyphae. Inoculation of wheat in axenic culture with a strain ofPseudomonas fluorescens that was antagonistic toG. graminis in agar gave some protection against the pathogen; lysis of hyphae was observed where protection occurred.     

Grain yield and incidence of take-all (Ophiobolus graminis Sacc.) in wheat grown in different crop sequences

The yield of wheat and the incidence of take-all were measured in crops grown in six different 4-year sequences, repeated in 3 successive years. The first crop of winter wheat grown after oats or beans yielded 13–23 cwt/acre (1632–2887 kg/ha) more grain than wheat after wheat or barley. Spring wheat after oats yielded 2–5 cwt/acre (250–625 kg/ha) more than spring wheat after wheat. The smaller yields of wheat after wheat or barley were caused mostly by greater prevalence of take-all. Regression analysis indicates that each 1 % increase in straws with take-all decreased yield of winter wheat by 0·6%. Take-all was more prevalent in the second and third successive wheat crops after oats than in the fourth crop.     

Structural Basis for Substrate Specificity of Mammalian Neuraminidases

The removal of sialic acid (Sia) residues from glycoconjugates in vertebrates is mediated by a family of neuraminidases (sialidases) consisting of Neu1, Neu2, Neu3 and Neu4 enzymes. The enzymes play distinct physiological roles, but their ability to discriminate between the types of linkages connecting Sia and adjacent residues and between the identity and arrangement of the underlying sugars has never been systematically studied. Here we analyzed the specificity of neuraminidases by studying the kinetics of hydrolysis of BODIPY-labeled substrates containing common mammalian sialylated oligosaccharides: 3′Sia-LacNAc, 3′SiaLac, SiaLe_x, SiaLe_a, SiaLe_c, 6′SiaLac, and 6′SiaLacNAc. We found significant differences in substrate specificity of the enzymes towards the substrates containing α2,6-linked Sia, which were readily cleaved by Neu3 and Neu1 but not by Neu4 and Neu2. The presence of a branching 2-Fuc inhibited Neu2 and Neu4, but had almost no effect on Neu1 or Neu3. The nature of the sugar residue at the reducing end, either glucose (Glc) or N-acetyl-D-glucosamine (GlcNAc) had only a minor effect on all neuraminidases, whereas core structure (1,3 or 1,4 bond between D-galactose (Gal) and GlcNAc) was found to be important for Neu4 strongly preferring β3 (core 1) to β4 (core 2) isomer. Neu3 and Neu4 were in general more active than Neu1 and Neu2, likely due to their preference for hydrophobic substrates. Neu2 and Neu3 were examined by molecular dynamics to identify favorable substrate orientations in the binding sites and interpret the differences in their specificities. Finally, using knockout mouse models, we confirmed that the substrate specificities observed in vitro were recapitulated in enzymes found in mouse brain tissues. Our data for the first time provide evidence for the characteristic substrate preferences of neuraminidases and their ability to discriminate between distinct sialoside targets.     

Structural Basis for Substrate Specificity in Human Monomeric Carbonyl Reductases

Carbonyl reduction constitutes a phase I reaction for many xenobiotics and is carried out in mammals mainly by members of two protein families, namely aldo-keto reductases and short-chain dehydrogenases/reductases. In addition to their capacity to reduce xenobiotics, several of the enzymes act on endogenous compounds such as steroids or eicosanoids. One of the major carbonyl reducing enzymes found in humans is carbonyl reductase 1 (CBR1) with a very broad substrate spectrum. A paralog, carbonyl reductase 3 (CBR3) has about 70% sequence identity and has not been sufficiently characterized to date. Screening of a focused xenobiotic compound library revealed that CBR3 has narrower substrate specificity and acts on several orthoquinones, as well as isatin or the anticancer drug oracin. To further investigate structure-activity relationships between these enzymes we crystallized CBR3, performed substrate docking, site-directed mutagenesis and compared its kinetic features to CBR1. Despite high sequence similarities, the active sites differ in shape and surface properties. The data reveal that the differences in substrate specificity are largely due to a short segment of a substrate binding loop comprising critical residues Trp229/Pro230, Ala235/Asp236 as well as part of the active site formed by Met141/Gln142 in CBR1 and CBR3, respectively. The data suggest a minor role in xenobiotic metabolism for CBR3.

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Effect of Ophiobolus graminis infection on the assimilation and distribution of 14C in wheat

The uptake of ¹⁴C and movement of ¹⁴C-labelled assimilates in wheat plants inoculated with Ophiobolus graminis was examined following exposure of the second youngest leaf to ¹⁴CO₂. Autoradiographs of plants with infected seminal roots showed that assimilates were not translocated past the sites of root infection but accumulated in the undamaged portions of infected root systems, in particular the developing crown roots. There was no evidence that assimilates accumulated in the vicinity of O. graminis lesions. The net assimilation of ¹⁴CO₂ by wheat plants over a 5 h feeding period was not significantly affected by O. graminis infection. However, infection reduced the amount of ¹⁴C lost through respiration. Infection delayed the transfer of labelled assimilates from the fed leaf to the remainder of the plant but increased the proportion translocated to the roots. The latter effect was not apparent when infected plants were continuously irrigated during, and for 20 h following, the feeding period.