Resolution of aflatoxins B1, B2, G1, and G2 by high-pressure liquid chromatography.

Aflatoxins were completely resolved as sharp peaks in the order BU-B2-G1-G2 by high-pressure liquid chromatography on a small particle (10 mum) porous silica gel column in 7-13 min (B1 through G2) by a water-saturated chloroform-cyclohexane-acetonitrile elution solvent (25+7.5+1.0), with detection by ultraviolet absorbance at 360 nm. The relationship between peak height and amount injected was linear over a 5-400 ng range for each aflatoxin. Both retention times and peak heights were highly reproducible, multiple injections of mixed standards giving coefficients of variation of 1.0-1.4% (retention time) and 1.6-2.8% (peak height) for the 4 aflatoxins. Detection was highly sensitive, with mean peak height, mm/ng, of 7.1 (B1), 6.4 (B2), 4.5 (G1), and 4.1(G2), allowing detection of 1-2 ng of each aflatoxin.     

Gas-liquid chromatographic analysis of ethephon and fenoprop residues in apples and their decline before and after harvest.

Ethephon (2-chloroethylphosphonic acid) and fenoprop (2-(2,4,5-trichlorophenoxy) propionic acid) may be determined in the same apple sample. After extraction with methanol, 2 separate methylation procedures were required to quantitatively convert each compound. Ethephon was esterified with diazomethane and analyzed by a flame photometric detector in the P-mode. Fenoprop was esterified with boron trifluoride/methanol and analyzed by electron capture gas chromatography. Average recoveries were about 95% at 0.05 ppm for both compounds. The limit of detection was 0.05 ppm for ethephon and 0.01 ppm for fenoprop in a 1 g sample. The persistence of both compounds before and after harvest was studied. Ethephon and fenoprop were applied simultaneously to apple trees at the recommended concentrations of 300 and 20 ppm, respectively. Ethephon residues averaged 1.6, 0.75, and 0.4 ppm at 2 hr, 10 days, and after washing at 13 days, respectively. The corresponding fenoprop residues were 0.70, 0.025, and 0.024 ppm.     

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.     

Multiple forms of acetyl-naphthyl-esterase activity in soil organic matter

Acetyl-naphthyl-esterase activity has been identified and characterized in organic matter extracted from an A₁ horizon of an Alpine podzol. The temperature optimum of the esterase is about 75° C and its activity rises with increasing pH, without reaching a maximum value in the tested range. The Michaelis constant has been determined as Km = 2.950 mM. Pronase does not disrupt esterase activity. Electrofocusing in acrylamide gel shows several peaks of enzyme activity, which correspond with humic isoelectric bands. The location of acetyl-esterase activity in organic matter is discussed.     

Studies on soil fumigation—III: Effects on enzyme activities,bacterial numbers and extractable ninhydrin reactive compounds

Fumigation of a field soil with chloropicrin and methyl bromide, either singly or in combination, differentially decreased soil enzyme activities and viable bacterial numbers and increased the amounts of ninhydrin reactive compounds extractable with acidified Tris buffer. Chloropicrin treatment was more effective than methyl bromide.The rates of hydrolysis of both an amide and a peptide derivative were decreased by chloropicrin treatment and remained relatively low despite changes in activities over 325 days. By contrast, caseinase activity initially was decreased by both chloropicrin and methyl bromide fumigation, but activities of the fumigated soils recovered to exceed those of untreated soils. Thereafter, caseinase activities of fumigated and untreated soils exhibited relatively large fluctuations, which were partly associated with seasonal drying of the soils in the field.Chloropicrin but not methyl bromide fumigation markedly depressed the viable bacterial populations, which subsequently increased to be above those of the untreated soils. There was no consistent relationship between the release of ninhydrin reactive compounds following fumigation and changes in bacterial numbers or changes in enzyme activity. Autolytic reactions are probably important in the early stages of amino-nitrogen release in fumigated soils. Net gains in caseinase activity may be partly due to the production de novo of extracellular proteases by microorganisms or to the release of intracellular proteases from killed cells.