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

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.

1970 ◽  
Vol 48 (10) ◽  
pp. 1091-1095 ◽  
Author(s):  
W. K. Kim

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.


1970 ◽  
Vol 48 (9) ◽  
pp. 1617-1623
Author(s):  
A. O. Jackson ◽  
D. J. Samborski ◽  
R. Rohringer ◽  
W. K. Kim

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.


1968 ◽  
Vol 46 (12) ◽  
pp. 1533-1536 ◽  
Author(s):  
A. J. Roos ◽  
A. M. Spronk ◽  
E. A. Cossins

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.


1970 ◽  
Vol 16 (10) ◽  
pp. 947-951 ◽  
Author(s):  
A. L. Branen ◽  
T. W. Keenan

Diacetyl reductase (diacetyl:reduced nicotinamide adenine dinucleotide (NADH) oxidoreductase, EC. 1.1.1.5) has been isolated from Lactobacillus casei. Cell sonication, ammonium sulfate fractionation, Sephadex gel filtration, DEAE-cellulose chromatography, and alumina gel adsorption were used to obtain the partially purified enzyme. Both NADH oxidase and diacetyl reductase activity were associated with the same fraction at all stages in purification. Growth in media containing added pyruvate resulted in a 10-fold increase in the NADH oxidase activity and a 3-fold increase in the diacetyl reductase activity of crude cell extracts on a protein basis. Purified preparations showed maximal reductase and oxidase activities at pH 4.5 and 5.0, respectively. Lineweaver–Burke plots yielded intersecting lines when NADH and diacetyl concentrations were varied, suggesting a flavin-linked reaction. The absorption spectrum of the purified preparation was characteristic of that of a flavoprotein. The product of the reduction of diacetyl was identified as acetoin. Acetoin and methylene blue were inactive as acceptors.


1972 ◽  
Vol 43 (6) ◽  
pp. 799-813 ◽  
Author(s):  
R. Corrocher ◽  
B. K. Bhuyan ◽  
A. V. Hoffbrand

1. The composition of guinea-pig liver folates and the biochemical route of formation of liver folates from injected tritium-labelled pteroylglutamic acid (folic acid) have been studied. 2. Endogenous folate was measured by microbiological assay with Lactobacillus casei and Streptococcus faecalis, with and without deconjugation of whole liver pteroylpolyglutamates (conjugated folates). Individual folate compounds were identified by microbiological assay after fractionation of liver folates by DEAE cellulose ion-exchange column chromatography. 3. Liver folate in the guinea-pig consists of about 84–87% reduced pteroylpolyglutamates with more than three glutamate moieties/molecule, about 12–15% reduced pteroyltriglutamates, about 1% reduced pteroyldiglutamates and only traces of reduced pteroylmonoglutamates. 4. About 53% of the liver folate consists of methylated derivatives. 5. Injected pteroylglutamic acid was first rapidly reduced and formylated or methylated. Glutamate moieties were then added, probably singly, to form di-, tri- and poly-glutamates. This was a relatively slow process with a hold-up at the triglutamate stage. 6. The proportion of the labelled pteroylglutamic acid in the polyglutamate form approximated to the proportion of endogenous folates in this form after 3–4 days. 7. The amount of radioactive folate in the liver increased progressively from 1 to 84 h after injection of a standard amount of radioactive pteroylglutamic acid.


Author(s):  
Dan VODNAR ◽  
Floricuța RANGA ◽  
Oana POP ◽  
Carmen SOCACIU

Tea is rich in polyphenols and phenolics compounds that have been widely reported to have beneficial health effects. The aim of this study was to investigate the ability of catechins, major phenols in tea, to act as growth factors for Lactobacillus casei during lactic acid fermentation. Major catechins presented in tea extracts are gallocatechin, catechin, epigalocatechin, galocatechingalat, epicatechingalat, catechingalat. Different concentrations of green tea and black tea extracts have been added to MRS model media. Growth and viability of Lactobacillus casei was positively affected by the addition of green tea and black tea extracts. This indicates that green tea and black tea addition exert protective effect on Lactobacillus casei growth in MRS media, increasing the viability and biomass production, probably by acting as metabolic enhancer. This results indicated the posibility of using green tea and black tea as metabolic enhancers for the growth of Lactobacillus casei. 


2019 ◽  
Author(s):  
Rui Shi ◽  
Dilip R. Panthee

AbstractBacterial spot (BS) is one of the most devastating foliar bacterial diseases of tomato caused by multiple species of Xanthomonas. We performed the RNA-Seq analysis of three tomato lines with different level of resistance to Xanthomonas perforans race T4 to study the differentially expressed genes (DEGs) and transcript-based sequence variations.Analysis between inoculated and control samples revealed that resistant line PI 270443 had more DEGs (834), followed by susceptible line NC 714 (373), and intermediate line NC 1CELBR (154). Gene functional analysis based on Gene Ontology (GO) terms revealed that more GO terms (51) were enriched for up-regulated DEGs in the resistant line PI 270443, and more down-regulated DEGs (67) were enriched in the susceptible line NC 714. The specific analysis for DEGs in biotic stress pathway using MapMan software showed more up-regulated biotic stress pathway DEGs (67) for PI 270443 compared to more down-regulated DEGs (125) for susceptible NC 714 line. One interesting feature was that resistant PI 270443 has three up-regulated DEGs for PR-protein, and susceptible line NC 714 has one down-regulated R gene, which is disease-related.Analysis of sequence variations called from RNA-Seq reads against the reference genome of susceptible Heinz 1706 showed that chr11 which has multiple reported resistance QTLs to BS race T4 is identical between two resistant lines, PI 270443 and NC 1CELBR, suggesting that these two lines share the same resistance QTLs on this chromosome. Several loci for PR-resistance proteins with sequence variation between the resistant and susceptible tomato lines were identified near the known Rx4 resistance gene on chr11. These findings may be useful for further molecular breeding of tomato.


1982 ◽  
Vol 47 (1) ◽  
pp. 349-359 ◽  
Author(s):  
Ludmila Vašáková ◽  
Miroslav Štefl

Three glutamate kinases (GK 1, GK 2 and GK 3) have been found in the leaves of winter wheat (Triticum aestivum L). They were separated by salting out with ammonium sulphate and by chromatography on DEAE-cellulose in a concentration gradient of KCl and on Sephadex G-100. GK 1 belongs to the biosynthesis of L-proline, which inhibits it according to the principle of feedback, GK 2 to the biosynthesis of glutamine; the function of GK 3 has not been found. The partically purified oligomer GK 1 had a molecular weight of 254 000 and dissociated to subunits of molecular weights 84 000 and 42 000. In a K+-phosphate buffer, 50 mmol/l, it was active in a pH range of 6.3 to 8.0, with an optimum of 7.2. An amine buffer inhibited it completely. The reaction required Mg2+ ions as activators and L-glutamate and ATP as substrates. ATP in a concentration above 70 mmol/l inhibited it. The steady-state kinetics at the optimum pH have shown that GK 1 produces a complex co-operativity of the substrates and belongs to the slowly dissociating hysteretic enzymes of the type Np P.


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