Folate derivatives in ungerminated and germinated uredospores of wheat stem rust

1970 ◽  
Vol 48 (9) ◽  
pp. 1617-1623
Author(s):  
A. O. Jackson ◽  
D. J. Samborski ◽  
R. Rohringer ◽  
W. K. Kim
Keyword(s):  
Growth Factors ◽  
Glutamic Acid ◽  
Detailed Analysis ◽  
Stem Rust ◽  
Lactobacillus Casei ◽  
Deae Cellulose ◽  
Wheat Stem Rust ◽  
Streptococcus Faecalis ◽  
Peak Fraction ◽  
Methyl Tetrahydrofolate

Extracts of ungerminated uredospores and of uredospores germinated for 6 h and 12 h were assayed for folates with Lactobacillus casei, Streptococcus faecalis, and Pediococcus cerevisiae. These organisms did not respond to the extracts without conjugase treatment, indicating that most of the folates were present in conjugated form with more than three glutamic acid moieties per molecule. During the 6-h and 12-h germination periods the content of growth factors for L. casei declined to 70.9% and 46.0% of initial levels. In this same period, the content of growth factors for S. faecalis declined to 54.5% and 15.2% of initial levels, indicating an increase in the proportion of methylated folates during uredospore germination.This trend was confirmed in a detailed analysis of folate components after fractionation of the extracts on DEAE-cellulose columns. The folate profiles consisted of five peaks. Two peak fractions present in profiles from ungerminated spores contained mostly formylated folates and were greatly reduced or absent in profiles from germinated spores. A peak fraction composed of a 5-methyl-tetrahydrofolate conjugate was not observed in profiles of ungerminated spores but was predominant in those from spores germinated for 12 h.

Effect of excision and benzimidazole treatment on folate content of wheat leaves and wheat leaf chloroplasts

1970 ◽  
Vol 48 (10) ◽  
pp. 1091-1095 ◽  
Author(s):  
W. K. Kim
Keyword(s):  
Lactobacillus Casei ◽  
Deae Cellulose ◽  
Streptococcus Faecalis ◽  
Crude Extracts ◽  
Wheat Leaf ◽  
Wheat Leaves ◽  
Peak Fraction

Crude extracts were prepared from freshly harvested wheat leaves, and from leaves floated on water or on solutions of benzimidazole. Extracts were also prepared from chloroplasts isolated with non-aqueous solvents from similar leaves. All extracts were fractionated on DEAE-cellulose columns. Folate content was determined with Lactobacillus casei, Streptococcus faecalis, and Pediococcus cerevisiae as assay organisms.Total folates increased in leaves after excision whether benzimidazole was present or not. The increase was mainly due to an increase of conjugates of 5-methyltetrahydrofolate. Similar folate drifts were observed in chloroplasts isolated from these samples. Peak fraction analyses indicated that methylated folate conjugates were converted to 5-methyltetrahydrofolate, and formylated folates disappeared in leaves floated on water. Benzimidazole treatment arrested the degradation of conjugated folates, except that of a formylated folate found only in chloroplasts.

Folate derivatives in healthy and rust-infected primary leaves of wheat

1969 ◽  
Vol 47 (12) ◽  
pp. 1161-1169 ◽  
Author(s):  
R. Rohringer ◽  
W. K. Kim ◽  
D. J. Samborski
Keyword(s):  
Growth Factors ◽  
Lactobacillus Casei ◽  
Deae Cellulose ◽  
Resistant Line ◽  
Streptococcus Faecalis ◽  
Susceptible Line ◽  
Crude Extracts ◽  
Wheat Leaves ◽  
Chicken Pancreas

Six days after inoculation, crude extracts were prepared from healthy and rust-infected wheat leaves of a susceptible line and from healthy and rust-infected wheat leaves of a resistant line. Extracts from rust-infected leaves of both lines contained 50% more folates, active as growth factors for Lactobacillus casei, than did those from healthy leaves. Rust-infected leaves did not differ from healthy leaves in their content of folates active as growth factors for Streptococcus faecalis and Pediococcus cerevisiae.All extracts were fractionated on DEAE-cellulose columns. The eluate fractions were treated with chicken pancreas conjugase and assayed with L. casei. All extracts yielded similar folate profiles consisting of five major and three minor peaks. Conjugase treatment and differential assay of the peak fractions with L. casei, S. faecalis, and P. cerevisiae indicated that the eluates contained folates methylated at N-5 and folates formylated at N-10 in various states of oxidation and conjugation. The eluates also contained a compound believed to be 5-formyltetrahydropteroylglutamate, small amounts of pteroylglutamate, traces of tetrahydropteroylglutamate, and several unidentified folates.The increase of folate levels in rust-infected leaves was due almost entirely to increases of 5-methyl-tetrahydropteroylgiutamate and its conjugates. The folate composition of resistant-reacting leaves did not differ appreciably from that of susceptible-reacting leaves.Radioactivity was not incorporated into either 5-methyltetrahydropteroyltriglutamate or into methionine when 5-[methyl-14C]-tetrahydropteroylglutamate was fed to wheat leaves.

THE METABOLISM OF PROPIONATE BY WHEAT STEM RUST UREDOSPORES

1963 ◽  
Vol 41 (3) ◽  
pp. 737-743 ◽  
Author(s):  
H. Reisener ◽  
A. J. Finlayson ◽  
W. B. McConnell ◽  
G. A. Ledingham
Keyword(s):  
Carbon Dioxide ◽  
Amino Acids ◽  
Glutamic Acid ◽  
Stem Rust ◽  
Water Extract ◽  
Hot Water ◽  
Insoluble Residue ◽  
Soluble Carbohydrates ◽  
Wheat Stem Rust ◽  
Carbon 14

When uredospores of wheat stem rust were shaken for 3 hours with phosphate buffer (pH 6.2) containing propionate-1-C14, -2-C14, or -3-C14, about 55% of the carbon-14 was removed from the solution. With propionate-1-C14, most of the carbon-14 taken up was released as carbon dioxide-C14, whereas about 20% and 31% of propionate carbon 2 and carbon 3, respectively, was incorporated into the spores. The specific activity of a fraction consisting of the free amino acids of a hot-alcohol and hot-water extract of the spores increased markedly with increase in the position number of propionate in which the carbon-14 was located. A similar relation was observed for other fractions such as soluble carbohydrates, ether-soluble material, organic acids, and insoluble residue from spores. The most active amino acids isolated were glutamic acid, γ-aminobutyric acid, and alanine. Partial degradations showed that with propionate-2-C14 the carboxyl groups of glutamic acid were especially radioactive, whereas with propionate-3-C14 the internal carbons were most radioactive.It is concluded that propionate metabolism in the rust spores involved conversion of carbon 1 to carbon dioxide, and utilization of carbons 2 and 3 as acetate with carbon 2 behaving as the carboxyl carbon.

UTILIZATION OF ADDED SUBSTRATES BY UREDOSPORES OF WHEAT STEM RUST

1956 ◽  
Vol 2 (6) ◽  
pp. 559-563 ◽  
Author(s):  
P. Shu ◽  
A. C. Neish ◽  
G. A. Ledingham
Keyword(s):  
Carbon Dioxide ◽  
Amino Acids ◽  
Fatty Acids ◽  
Glutamic Acid ◽  
Metabolic Activity ◽  
Stem Rust ◽  
Volatile Fatty Acids ◽  
The Other ◽  
Wheat Stem Rust ◽  
Better Than

Uredospores of wheat stem rust utilized a number of externally-supplied, labelled carbohydrates, amino acids, and volatile fatty acids. The carbon of these substrates appeared in the spore materials and in the carbon dioxide. This metabolic activity, though very weak, is definite. D-Mannose, D-mannitol, D-glucose, sucrose, and D-fructose were utilized better than the other carbohydrates. Glutamic acid gave the highest yield of carbon dioxide while the basic amino acids, L-arginine and L-lysine, were more efficiently incorporated into the spore material.

THE METABOLISM OF PROPIONATE BY WHEAT STEM RUST UREDOSPORES

1963 ◽  
Vol 41 (1) ◽  
pp. 737-743 ◽  
Author(s):  
H. Reisener ◽  
A. J. Finlayson ◽  
W. B. McConnell ◽  
G. A. Ledingham
Keyword(s):  
Carbon Dioxide ◽  
Amino Acids ◽  
Glutamic Acid ◽  
Stem Rust ◽  
Water Extract ◽  
Hot Water ◽  
Insoluble Residue ◽  
Soluble Carbohydrates ◽  
Wheat Stem Rust ◽  
Carbon 14

When uredospores of wheat stem rust were shaken for 3 hours with phosphate buffer (pH 6.2) containing propionate-1-C14, -2-C14, or -3-C14, about 55% of the carbon-14 was removed from the solution. With propionate-1-C14, most of the carbon-14 taken up was released as carbon dioxide-C14, whereas about 20% and 31% of propionate carbon 2 and carbon 3, respectively, was incorporated into the spores. The specific activity of a fraction consisting of the free amino acids of a hot-alcohol and hot-water extract of the spores increased markedly with increase in the position number of propionate in which the carbon-14 was located. A similar relation was observed for other fractions such as soluble carbohydrates, ether-soluble material, organic acids, and insoluble residue from spores. The most active amino acids isolated were glutamic acid, γ-aminobutyric acid, and alanine. Partial degradations showed that with propionate-2-C14 the carboxyl groups of glutamic acid were especially radioactive, whereas with propionate-3-C14 the internal carbons were most radioactive.It is concluded that propionate metabolism in the rust spores involved conversion of carbon 1 to carbon dioxide, and utilization of carbons 2 and 3 as acetate with carbon 2 behaving as the carboxyl carbon.

THE UTILIZATION OF AMMONIUM CHLORIDE-15N BY UREDOSPORES OF WHEAT STEM RUST

1966 ◽  
Vol 44 (11) ◽  
pp. 1511-1518 ◽  
Author(s):  
W. B. McConnell ◽  
E. W. Underhill
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Glutamic Acid ◽  
Ammonium Chloride ◽  
Stem Rust ◽  
Organic Nitrogen ◽  
Ammonia Nitrogen ◽  
Ammonium Nitrogen ◽  
Wheat Stem Rust ◽  
Carbon 14

When uredospores of wheat stem rust, Puccinia graminis van tritici (race 15B), were incubated with a 3 mM solution of ammonium chloride-15N, a significant amount of nitrogen 15 was converted into organic nitrogen. Most of this organic nitrogen 15 was found in the ethanol and water extracts, with lesser amounts in the buffer and in extracted spores.Amino acids extracted from the spores all contained excess nitrogen 15. Nitrogen 15 from the inorganic source was diluted by factors of 1.7 and 2.7 in free aspartic and glutamic acids respectively; these amino acids were the most heavily labeled with the isotope. Proline was the most weakly labeled amino acid, the nitrogen 15 being diluted by a factor of 102. Good incorporation of nitrogen 15 into glutamic acid compared to simultaneous poor incorporation into the biochemically related amino acid, proline, parallels previous observations made during carbon 14 experiments with rust uredospores.Fourteen "bound" amino acids were isolated after acid hydrolysis of extracted spores. All contained nitrogen 15, the dilution of the added ammonia nitrogen ranging from 96 for glutamic acid to 7660 for proline.The results are taken as evidence that uredospores of wheat stem rust can incorporate ammonium nitrogen into free amino acids and into proteins.

5-Methyltetrahydrofolic acid and other folate derivatives in germinating pea seedlings

1968 ◽  
Vol 46 (12) ◽  
pp. 1533-1536 ◽  
Author(s):  
A. J. Roos ◽  
A. M. Spronk ◽  
E. A. Cossins
Keyword(s):  
Folic Acid ◽  
Lactobacillus Casei ◽  
Deae Cellulose ◽  
Methyl Derivative ◽  
Streptococcus Faecalis ◽  
Feeding Experiments ◽  
Pea Seedlings ◽  
Young Leaves

The folate derivatives present in germinating pea seedlings were isolated by chromatography on DEAE-cellulose and assayed with Lactobacillus casei, Streptococcus faecalis, and Pediococcus cerevisiae. The major folate derivative in the cotyledons, developing embryos, and young leaves was identified as 5-methyltetrahydrofolate. Smaller amounts of 10-formyltetrahydrofolate were also present in these tissues. Synthesis of these derivatives in the cotyledons was inhibited by aminopterin. Feeding experiments showed that the 5-methyl derivative was rapidly synthesized from folic acid-2-14C.

THE METABOLISM OF VALERATE-3-C14 AND -5-C14 BY WHEAT STEM RUST UREDOSPORES

1964 ◽  
Vol 42 (3) ◽  
pp. 327-332 ◽  
Author(s):  
H. Reisener ◽  
A. J. Finlayson ◽  
W. B. McConnell
Keyword(s):  
Carbon Dioxide ◽  
Amino Acids ◽  
Glutamic Acid ◽  
Stem Rust ◽  
Phosphate Buffer ◽  
Free Amino Acids ◽  
Free Amino ◽  
Methyl Groups ◽  
Wheat Stem Rust ◽  
Carbon 14

Uredospores of wheat stem rust took up about 90% of the carbon-14 present either as valerate-3-C14 or as valerate-5-C14 in M/30 phosphate buffer pH 6.2 in 3 hours. The initial valerate concentration was 0.017 mM and spores were supplied at the rate of 250 mg/30 ml of buffer. Carbon 3 of the valerate was largely respired as carbon dioxide but carbon 5 was extensively incorporated into spore components. Free amino acids contained about 40% of the radioactivity in the spores labelled with valerate-5-C14 and glutamic acid was highly labelled. Carbon 1 contained 8.1% and carbon 5, 3.8% of the carbon-14 in this glutamic acid and thus internal carbons contained 88%. The results with valerate-3-C14 and with valerate-5-C14 compare well with those of experiments done earlier with propionate-1-C14 and propionate-3-C14 respectively. It is concluded that propionate is formed from carbons 3, 4, and 5 of valerate, and thus that carbon 3 is converted to carbon dioxide, and carbons 4 and 5 to the carboxyl and methyl groups respectively of acetate.

Sign in / Sign up

Export Citation Format

Share Document