Mycosporine-like amino acids (MAAs) profile of a rice-field cyanobacterium Anabaena doliolum as influenced by PAR and UVR

The mycosporine-like amino acid (MAA) profile of a rice-field cyanobacterium, Anabaena doliolum, was studied under PAR and PAR + UVR conditions. The high-performance liquid chromatographic analysis of water-soluble compounds reveals the biosynthesis of three MAAs, mycosporine-glycine (lambda (max) = 310 nm), porphyra-334 (lambda (max) = 334 nm) and shinorine (lambda (max) = 334 nm), with retention times of 4.1, 3.5 and 2.3 min, respectively. This is the first report for the occurrence of mycosporine-glycine and porphyra-334 in addition to shinorine in Anabaena strains studied so far. The results indicate that mycosporine-glycine (monosubstituted) acts as a precursor for the biosynthesis of the bisubstituted MAAs shinorine and porphyra-334. Mycosporine-glycine was under constitutive control while porphyra-334 and shinorine were induced by UV-B radiation, indicating the involvement of UV-regulated enzymes in the biotransformation of MAAs. It seems that A. doliolum is able to protect its cell machinery from UVR by synthesizing a complex set of MAAs and thus is able to survive successfully during the summer in its natural brightly lit habitats.     

Characterization of salinity-tolerant mutant of Anabaena doliolum exhibiting multiple stress tolerance

Results show that an isolated mutant of the cyanobacterium Anabaena doliolum is a fast-growing strain. It exhibits approximately twofold higher NaCl tolerance than the wild type. It also reveals cross-resistance against the herbicide 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), drug bacitracin, and LiCl. Further, an improved LiCl tolerance property of both the mutant and wild-type strains at high concentration of NaCl (40 mM) may be interpreted in terms of competitive inhibition of the Li⁺ uptake by Na⁺ ions, whereas bacitracin resistance in these organisms is described to be the result of an alteration in the drug transporting channels of membrane. The multiple stress tolerance property of the A. doliolum may be attributed to altered membrane characteristics in the mutant strain, leading to reduced intake of such toxicants. Received: 6 November 2001 / Accepted: 14 December 2001     

Isolation and characterization of salinity resistant mutant of a nitrogen-fixing cyanobacterium, Anabaena doliolum

D.V. SINGH. A.K. TRIPATHI AND H.D. KUMAR. 1991. Sodium chloride, up to 20 mmol/l concentration, had a positive effect on acetylene reducing activity and photosynthetic oxygen evolution of a paddy field cyanobacterium, Anabaena doliolum. Beyond 20 mmol/l level of salinity adverse effects appeared. A mutant resistant to 200 mmol/l NaCl was isolated by nitrosoguanidine mutagenesis. The mutant, NaCl-R₂₀₀, showed about 20–25% more nitrogenase activity and photosynthetic oxygen evolution than the parent. Better capacity of nitrogen fixation and photosynthesis possibly could help the mutant in synthesis of osmotic stabilizer to resist the salinity stress.     

Cobalt-induced inhibition of growth in cyanobacteria Anabaena doliolum and Anacystis nidulans: interaction with sulphur containing amino acids

Cobalt, above 1 microM concentration was growth inhibitory for both A. doliolum and A. nidulans. Its toxicity was mitigated by sulphur containing amino acids (cystine and cysteine), however, methionine could not mitigate the cobalt toxicity at all.     

Effects of amino acids on methionine-sulfoximine-induced heterocyst formation in Anabaena

Heterocyst development in ammonia-grown cultures of Anabaena variabilis and Anabaena 7120 was fully induced by the amino-acid analog methionine sulfoximine (MSO) at concentrations of 0.5–1.0 M. Glutamine, glutamate, aspartate, and alanine at 0.5 mM blocked the induction of heterocysts by MSO in A. variabilis. With Anabaena 7120, glutamine and glutamate were fully effective and alanine partially effective in preventing MSO-induced heterocyst formation. In MSO-treated algae, glutamine synthetase activity was reduced to less than 15% of control values within 4–6 h. Inactivation of the enzyme was prevented by all four amino acids tested.     

Response of Anabaena species to different nitrogen sources

Nitrogenase activity, ammonia excretion and glutamine synthetase (GS) activity were examined in five strains of Anabaena (A. anomala ARM 314, A. fertilissima ARM 742, A. variabilis ARM 310, A. oryzae ARM 313 and A. oryzae ARM 570) in the presence of 2.5 mM NO3-N (KNO3), 2.5 mM NH-4-N [(NH4)2SO4] and diatomic nitrogen (N2). Ammonium-N was more inhibitory to nitrogenase activity as compared to NO3-N in all the strains. Maximum GS activity was exhibited in NO3-N medium, irrespective of the cyanobacterial strains studied. Uninduced release of ammonia was observed in all the species, with A. oryzae ARM 313 and Anabaena variabilis ARM 310 exhibiting maximum excretion of 0.25-0.31 and 0.27-1.23 mu moles NH4 mg Chl(-1) respectively on the 15th day of incubation. The glutamine synthetase activity of A. oryzae ARM 313 was relatively very high as compared to Anabaena variabilis ARM 310. There was no nitrate reductase activity in any of the Anabaena sp. grown on NH3-N or N2-N on the 15th day of incubation.     

Regulation of urea uptake by ammonia in the cyanobacterium Anabaena doliolum

Abstract The urea uptake system was studied with regard to its repression and derepression in the cyanobacterium, Anabaena doliolum . The uptake of urea was energy-dependent and was repressed in ammonia grown cells. Repression of the urea uptake by ammonium did not require ammonium assimilation or de novo protein synthesis, suggesting that ammonium itself was the repressor signal. The derepression of the urea uptake system, however, required de novo protein synthesis and glutamine synthetase activity.     

Some Observations on the Pattern of Sporulation in a Blue-green Alga, Anabaena doliolum

Sporulation in Anabaena doliolum begins in the middle of thetwo heterocysts and proceeds towards the heterocystous ends.Two inorganic nitrogen sources—potassium nitrate and ammoniumchloride inhibit sporulation, whereas glucose promotes it. Duringsporulation, the reductive ability of the heterocyst graduallydiminishes. It is concluded that spore differentiation in this alga is controlledby critical levels of nitrogen and of sugar in the cell. Thecritical levels are probably regulated by the heterocyst.     

Cadmium mitigates ultraviolet-B stress in Anabaena doliolum: Enzymatic and non-enzymatic antioxidants

Impact of ultraviolet-B (UV-B) and Cd, applied individually and in combination, measured in terms of oxygen-evolution, chlorophyll (Chl) and protein contents, lipid peroxidation, and enzymatic and non-enzymatic antioxidants of Anabaena doliolum, revealed a greater oxidative damage induced by UV-B than by Cd. While superoxide dismutase (SOD) showed a greater stimulation by UV-B than Cd, the activities of catalase (CAT) and glutathione reductase (GR) declined after UV-B treatment. Cd treatment, however, enhanced the activities of ascorbate peroxidase (APX) and GR. CAT activity increased at low but decreased at high dose of Cd. Increase in carotenoid (Car) content in UV-B treated cells suggested a shielding effect of Car against UV-B stress. A 15-and 10-fold rise in α-tocopherol (α-TOC) content at high dose of Cd and/or UV-B offered testimony to the antioxidant role of α-TOC.     

The regulation of aromatic amino acid biosynthesis in amino acid liberating mutant strains of Anabaena variabilis

Mutant strains of Anabaena variabilis which are resistant to the tryptophan analogue, 6-fluorotryptophan, liberated a wide range of amino acids although none liberated tryptophan in detectable quantities. Four strains (FT-7, FT-8, FT-9, FT-10) produced predominantly alanine together with small amounts of phenylalamine and tyrosine, strain FT-2 liberated mainly phenylalanine and tyrosine and strain FT-6 liberated mainly glutamate, NH ₄ ⁺ and several unidentified ninhydrin-positive compounds. Two forms of 3-deoxy-D-arbinoheptulosonate 7-phosphate (DAHP) synthase were identified in the parent strain, a tyrosine-sensitive form and a phenylalanine-sensitive form. In strains FT-2 and FT-6 the phenylalanine-sensitive enzyme was not detected and in strain FT-7 it was apparently deregulated with respect to inhibition by phenylalanine. No deregulation of anthranilate synthase was observed but mutant strains were found to have higher specific activities of this enzyme than the parent strain.Abbreviations chla chlorophyll a - 6-FT 6-fluorotryptophan - DAHP 3-deoxy-D-arabinoheptulosonate 7-phosphate - PEP phosphoenolpyruvate     

Stereoselective routes towards the synthesis of unusual bis(amino acids) and cyclic amino acids

Stereoselective syntheses are described of bridged bis(glycines) as conformationally constrained substitutes for cystine, and of cyclic -amino acids where the -carbon of the amino acid is part of a five-, six- or seven-membered ring which may hold a hydroxy group as a threonine analogue.     

Stereoselective routes towards the synthesis of unusual bis(amino acids) and cyclic amino acids

Summary Stereoselective syntheses are described of bridged bis(glycines) as conformationally constrained substitutes for cystine, and of cyclic α-amino acids where the α-carbon of the amino acid is part of a five-, six- or seven-membered ring which may hold a hydroxy group as a threonine analogue.     

Comparative ultrastructure of a filamentous mutant and the wild type ofAgmenellum quadruplicatum

Summary A filamentous variant of the marine coccoid cyanophycean alga,Agmenellum quadruplicatum has been produced after treatment of cells with the chemical mutagen, N-methyl-N-nitro-N-nitrosoguanidine (NTG). The mutation to the filamentous condition is stable and has been compared ultrastructurally with the unicellular wild type. The structural basis for the maintenance of the filamentous state is due to a failure of the homogeneously-structured transverse wall to differentiate into the stratified longitudinal wall. A manifestation of this condition is the continued synthesis of the investment membrane resulting in proliferations at each constriction. The wild type cells are nearly spherical while the filamentous mutant cells are smaller and elongate. An analysis of the structure and morphology of the nucleoplasmic regions is presented for both strains and a correlation between the morphology of these regions and the unicellular and filamentous conditions is made.     

Distribution of amino acids in bran, embryo and milled endosperm and shifts in storage protein subunits of in vitro-selected and lysine-enhanced mutant and wild type rice

Experiments were designed to test whether an enhanced lysine phenotype from in vitro selections was expressed in milled rice, Oryza sativa L. Analysis of the endosperm tissue after removal of the aleurone and outer layers proved the grains maintained enhanced lysine after milling. Mutant endosperm had 15% greater lysine than the control and the enhanced lysine was correlated with reduced glutamic acid in two different mutants. Glutelins separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed increases in the protein quantity of the 33-kDa alpha and 21-kDa beta subunits.     

Protein and free amino acids in a high lysine maize double mutant

A comparative study of free amino acids and protein fractions of normal with a double mutant (su₁ o₂) was made, during endosperm development in segregating ears of a maize synthetic. Zein content showed striking differences in the two genotypes, being 7.7 and 6 times greater in the normal endosperm at 24 and 47 days after pollination respectively. This observed decrease in zein synthesis, coded by sugary-1/opaque-2 genes, causes an accumulation of alanine, glutamic and aspartic acids, glutamine and asparagine in the high lysine endosperm mutant.     

Synthesis and regulation of D-xylanase from Cellulomonas flavigena wild type and a mutant

The xylanolytic system from Cellulomonas flavigena was enhanced by adding cellulose to the growth medium. The Solka floc:xylan (60:40 w/w) mixture induced xylanase synthesis by more than 3-fold over that induced by growing C. flavigena, wild type and its mutant PN-120 on pure xylan. The hydrolysis pattern of sugar cane bagasse and xylan indicated the presence of debranching endo-;-xylanase activity.     

Transport of basic amino acids by the dinitrogen-fixing cyanobacterium Anabaena PCC 7120

Two transport systems for L-arginine were evident in Anabaena sp. strain PCC 7120: a high-affinity one (Km, 1.7 microM) that accumulated arginine within the cells through an energy-requiring process and another one that exhibited low affinity for L-arginine (Km, 0.75 mM) and was unable to accumulate the substrate. Both systems were inhibited by L-canavanine, L-lysine, and L-ornithine. Two systems were also evident for L-lysine uptake (Km, 1.9 and 110 microM, respectively). After selection for resistance to canavanine or hydroxylysine, independent mutants were isolated which were impaired in the high-affinity uptake of arginine and lysine. A common permease appears, therefore, to be involved in the high-affinity transport of these basic amino acids. Both the high- and the low-affinity systems can contribute to the growth of Anabaena sp. on L-arginine. However, arginine did not effectively repress either nitrogenase or nitrate reductase.