Thermal and spectrophotometric analysis of some fungal melanins and soil humic compounds

Seven types of fungal melanins were compared with humic compounds from chernozem and podzol soils and with commercial humic acid. The results indicate some similarities but also distinct differences between the melanins and humic substances in spectral characteristics in the UV and visible regions and in their resistance to thermal degradation.     

The distribution of nitrogen in some highly organic tropical volcanic soils

The qualitative and quantitative distribution of N-compounds in 10 tropical soils, and in a number of humic materials extracted from representative samples thereof, was determined after 6 N HCl hydrolysis.Eighty to 98% of the total N in the soils and humic materials was hydrolysable by 6n HCl. Slightly less than one half the hydrolysable N in the soils and humic fractions consisted of amino acids. Well-drained soils and fulvic acids extracted from them contained unusually high concentrations of the acidic amino acids, aspartic and glutamic acids. Between 80 and 95% of the amino acids in the soils was accounted for in the humic materials + NaOH-insoluble organic residues. NH⁺₄-N released by acid hydrolysis was generally higher for the soil samples than for the humic materials. Amino sugar-N constituted relatively small proportions of the total N in the soils and humic fractions.Our data suggest that large quantities of amorphous allophanic materials coupled with relatively high enzymic activity are responsible for the observed accumulation of acidic amino acids in the well-drained tropical volcanic soils.     

Volatilization of sulfur from unamended and sulfate-treated soils

Volatilization of sulfur from unamended and sulfate-treated soils was studied by sensitive gas chromatographic techniques using a flame photometric detector fitted with a sulfur filter. The soils employed were surface samples of 25 Iowa soils selected to obtain a wide range in properties. No release of volatile sulfur compounds was detected when 11 of these soils were incubated under aerobic or waterlogged conditions before or after treatment with sulfate (400 μg sulfate S/g soil). Fourteen soils released volatile sulfur compounds when incubated under waterlogged conditions before and after addition of sulfate, but only 4 of these soils released volatile sulfur compounds when incubated under aerobic conditions. Where volatilization of sulfur was observed, the volatile sulfur detected was identified as dimethyl sulfide or as dimethyl sulfide associated with smaller amounts of carbonyl sulfide, carbon disulfide, methyl mercaptan, and (or) dimethyl disulfide. No trace of hydrogen sulfide was detected. Where release of volatile sulfur was observed, the amount of sulfur volatilized at 30°C in 60 days under aerobic or waterlogged conditions was very small and did not account for more than 0–05% of the sulfur in the unamended or sulfate-treated soils studied. It is concluded that gaseous loss of sulfur from unamended or sulfate-treated soils is insignificant under conditions likely to be encountered in the field.     

Quantitation of o- and p-sulfamoylbenzoic acids in commerical saccharin by high-performance liquid chromatography.

Quantitation of o- and p-sulfamoylbenzoic acid residues in saccharin and its sodium salt is achieved by a method comprising methanolic extraction and high-performance ion exchange chromatography. A commercially available anion exchange column was employed with an aqueous buffered (pH 9.2) mobile phase. As little as 80 ppm of the ortho-isomer and 25 ppm of the para-isomer can be accurately determined. The levels of detectability (2 times noise) are estimated as 8 ppm (0.16 mug on column) and 2.5 ppm (0.05 mug on column), respectively. Recoveries from saccharin ranged from 92.7 to 96.5% (ortho) and from 92.2 to 103.3% (para). Recoveries from the sodium salt ranged from 93.1 to 104.4% (ortho) and from 93.5 to 97.8% (para). Of 9 other potential saccharin impurities tested separately, only one was found to interfere slightly in the chromatographic part of the procedure.     

Comparison of dieldrin, lindane, and DDT extractions from serum, and gas-liquid chromatography using glass capillary columns.

Rats were given an oral dose of 14C-labeled chlorinated pesticides to obtain serum containing p,p'-DDT, dieldrin, or lindane. Simple hexane and formic acid-hexane extraction methods, involving pretreatment of the serum with formic acid, were compared by radiometric and by paper chromatographic and gas chromatographic analysis. In vivo binding of chlorinated pesticides to constituents of the serum does not necessarily prohibit their isolation by simple hexane extractions, provided that the extraction is very vigorous and at least 5 min long. Stable emulsions were broken by cooling in liquid nitrogen or Dry Ice-acetone. The hexane extraction method described yields quantitative recovery of the pesticides studied, whereas the formic acid-hexane method is quantitative for p,p'-DDT, 93% for dieldrin, and 89% for lindane. Gas chromatographic comparison of both methods, using human serum, shows that the hexane method extracts 16% more beta-BHC, 7% more dieldrin and HCB, and 4% more p,p'-DDE from serum than does the formic acid-hexane method. The difference for p,p'-DDT is not significant. Gas chromatography with glass capillary columns and an all-glass solids injection system yielded detection limits as low as 15 fg. Data show that the use of an internal standard considerably improves the precision of quantitation.     

Infection of wheat seminal roots by varieties of Phialophora radicicola and Gaeumannomyces graminis

The growth of isolates of Phialophora radicicola var. radicicola, P. radicicola var. graminicola, Gaeumannomyces graminis var. graminis, G. graminis var. tritici and Leptosphaeria narmari was compared on the coleoptiles and roots of wheat seedlings. Fungal growth was measured as the extent and density of dark runner hyphae. All except P. radicicola var. graminicola grew on coleoptiles and all grew on roots although only G. graminis var. tritici extensively colonized the root stele. Growth rate on roots was positively correlated with that on agar, P. radicicola var. graminicola and L. narmari growing at about half the rate of the other fungi; hyphal density was high for P. radicicola var. graminicola but relatively low for the other fungi. For P. radicicola var. radicicola, P. radicicola var. graminicola and G. graminis var. tritici growing from buried inocula, the extent and density of hyphae up roots towards the seed was similar to that down, but G. graminis var. tritici caused chocolate-brown stelar discoloration up roots only.Root invasion by P. radicicola var. radicicola, P. radicicola var. graminicola and G. graminis var. tritici was described from sections. Each gave a different pattern of hyphae and host response within an inoculum layer, and progressive changes occurred away from the inoculum. Studies of the rate of penetration by each fungus and the rate and pattern of death of cortical cells explained the differences between fungi. G. graminis var. tritici penetrated living cells in advance of other soil micro-organisms, and hence by hyaline hyphae inducing much lignituber formation as a host resistance reaction. P. radicicola var. graminicola penetrated only senescent or dead cells in association with other soil microorganisms, and hence by dark hyphae, inducing little lignituber formation. P. radicicola var. radicicola was intermediate in all these respects. The high hyphal density of P. radicicola var. graminicola was due to the colonization of cortical cells and spaces by dark, clearly visible, rather than hyaline hyphae, which are invisible in unstained roots. Cell death in the outer cortex explained the observed progressive restriction of growth by all fungi to the inner cortex with increasing distance from the inoculum. Spread by G. graminis var. tritici up roots was ectotrophic relative to the stele but down roots hyphae spread rapidly within the stele. Stelar reactions suggested as resistance mechanisms occurred up roots only. Their absence down roots is attributed to infection disrupting stelar transport.     

Decomposition of algal cells and components and their stabilization through complexing with model humic acid-type phenolic polymers

Axenic cultures of Anacystis, Microcoleus, Plectonema and Synechococcus isolated from Greenfield sandy loam and of Anabaena flos-aquae, Nostoc muscorum and Chlorella pyrenoidosa from other sources were cultured under light and constant aeration and with [U-¹⁴C]-glucose in the nutrient medium. Whole cells, cell walls, cytoplasm and extracellular polysaccharides of selected species readily decomposed in the soil and after 22 weeks between 61 and 81% of the added C had evolved as CO₂. Complexing of cell wall and cytoplasmic preparations from A. flos-aquae and N. muscorum with model humic acid-type phenolic polymers reduced decomposition of the cell walls by 40% and of the cytoplasm by 70%. Over 50% of the residual 14C activity in the soil amended with whole algal cells remained in the 0.5% NaOH-extracted soil. With exception of Microcoleus sp. more of the residual ¹⁴C from cell walls, cytoplasm and polysaccharide fractions was present in the humic acid or fulvic acid fractions.