Effects of light and kinetin on amaranthin synthesis induced by cAMP

The effects of cyclic 3′,5′-adenosine monophosphate (cAMP) on amaranthin synthesis in the dark, or in the presence of kinetin or light were investigated in isolated cotyledons of Amaranthus tricolor and A. caudatus. The results suggest that sites or modes of action of cAMP and kinetin are not separated and differ from those of light and that the nucleotide cannot be considered a messenger involved in amaranthin formation stimulated by kinetin or by light.     

Effects of G protein and cGMP on phytochrome-mediated amaranthin synthesis in Amaranthus caudatus seedlings

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Characterization of dopa betaine,tyrosine betaine and N-dimethyltyrosine from Lobaria laetevirens

DOPA betaine, tyrosine betaine and N-dimethyltyrosine were isolated from the lichen Lobaria laetevirens. Their structures were determined by ra     

Photocontrol of betaxanthin synthesis in Celosia plumosa seedlings

Light stimulates the betaxanthin accumulation in Celosia plumosa. The induction is partially reversed by far-red and inhibited by actinomycin D, puromycin, salicylaldoxime and 2,4-dinitrophenol, while 3-(3,4-dichlorophenyl)-1,1-dimethylurea has an inhibitory effect only when photosynthesis is operative. In darkness betaxanthins synthesis is promoted by kinetin.     

Effect of rare earth element europium on amaranthin synthesis in Amarathus caudatus seedlings

Considering the resemblances between Eu³⁺ and Ca²⁺ in their atomic radius and structures of the valence electron, the effects of Eu³⁺ on amaramthin synthesis in Amarathus caudatus seedling were studied. Eu³⁺ had both promoting and inhibiting effects on amaramthin synthesis. The optimum promoting concentration and half inhibiting concentration of Eu³⁺ to synthesis of amaranthin were 0.4 mmol/L and 2.5 mmol/L, respectively. In the dark, A₂₃₁₈₇ (ions carrier) could carry Eu³⁺ into cells through the Ca²⁺ channel. When Ca²⁺ was chelated with EGTA, the synthesis of amaranthin could be partly retrieved by Eu³⁺. Eu³⁺ treatment could also activate Ca²⁺-ATPase on plasma membrane. Moreover, the sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns of total proteins from the plants treated by Eu³⁺ and Ca²⁺ were similar but slightly different in the contents. It suggested that the effects of Eu³⁺ and Ca²⁺ on amaranthin synthesis were similar. After being treated by Eu³⁺ or Ca²⁺, the outside Ca²⁺ could enter into cells to promote synthesis of amaranthin. The results above indicated that Eu³⁺ might replace Ca²⁺ in the calcium/calmidulin-dependent phytochrome signal transduction system and play important roles in plant development by promoting calcium transportation across plasma membrane.     

Effect of white and far-red light on betalain formation

Light-induced betalain synthesis has been studied in three species of Amaranthaceae. The results obtained suggest that the response to short-term irradiation with white light is mediated by phytochrome, while the effects of prolonged illumination are controlled by the photosynthetic system. Betalain accumulation stimulated by prolonged far-red irradiation is controlled chiefly by the photosynthetic system, the participation of phytochrome being of minor importance.     

The effects of hormones and inhibitors on amaranthin synthesis in seedlings of Amaranthus tricolor

Summary Both gibberellic acid and abscisic acid inhibit the light induced synthesis of amaranthin in Amaranthus tricolor seedlings. The auxin, indolyl-3-acetic acid has no effect. The protein/RNA inhibitors, cycloheximide and 8-azaguanine, also reduced the levels of amaranthin produced.     

Effect of continuous far red on betaxanthin and betacyanin synthesis

Seedlings of Celosia plumosa under prolonged irradiation with far red light synthesize chlorophyll α and betaxanthin. Levulinic acid and 2,4-dinitrophenol, inhibitors of chlorophyll synthesis and cyclic photophosphorylation respectively, reduce betaxanthin synthesis. Pigment formation is also inhibited by actinomycin-D and puromycin, but is unaffected by 3-(3,4-dichlorophenyl)-1,1-dimethylurea, an inhibitor of noncyclic photophosphorylation. These findings are evidence of the involvement of photosynthesis through cyclic photophosphorylation, in the far red HER associated with betaxanthin synthesis. Under continuous far red seedlings of Amaranthus tricolor synthesize only chlorophyll α. Lack of betacyanin formation is ascribed to the inactive status of the genes involved in the pigment synthesis.     

The Effect of Phenylalanine on DOPA Synthesis in PC12 Cells

DOPA synthesis from phenylalanine was studied in PC12 cells incubated with m-hydroxybenzylhydrazine, to inhibit aromatic L-amino acid decarboxylase. DOPA synthesis rose with increasing concentrations of either phenylalanine or tyrosine; maximal rates (~55 pmol/min/mg protein for tyrosine; ~40 pmol/min/mg protein for phenylalanine) occurred at a medium concentration of ~10 M for either amino acid. The K_m for either amino acid was about 1 M (medium concentration). At tyrosine concentrations above 30 M, DOPA synthesis declined; inhibition was observed at higher concentrations for phenylalanine (300 M). These effects were most notable in the presence of 56 mM potassium. Measurements of intracellular phenylalanine and tyrosine suggested the K_m for either amino acid is 20–30 M; maximal synthesis occurred at 120–140 M. In the presence of both phenylalanine and tyrosine, DOPA synthesis was inhibited by phenylalanine only at a high medium concentration (1000 M), regardless of medium tyrosine concentration. The inhibition of DOPA synthesis by high medium tyrosine concentrations was antagonized by high medium phenylalanine concentrations (100, 1000 M). Together, the findings indicate that for PC12 cells, phenylalanine can be a significant substrate for tyrosine hydroxylase, is a relatively weak inhibitor of the enzyme, and at high concentrations can antagonize substrate inhibition by tyrosine.     

The pathway of betalain biosynthesis: Effect of cytokinin on enzymic oxidation and hydroxylation of tyrosine in Amaranthus tricolor seedlings

The effect of exogenously added tyrosine or l -3-(3,4-dihydroxyphenyl) alanine on the accumulation of betacyanin in response to cytokinin in Amaranthus tricolor (L.) var. tricolor half-seedlings depends on the age of the seedlings and the treatment of the seedlings prior to induction of pigment by benzyladenine. Neither extracted polyphenol oxidase, peroxidase or tyrosine hydroxylase activity, nor in vivo tyrosine hydroxylation is increased in response to exposure of seedlings to cytokinin. However a small percentage of the polyphenol oxidase activated or unmasked by Triton X-100 treatment of membrane fractions is increased after cytokinin treatment of half-seedlings for 22 h. It is concluded that cytokinin control may be on a multi-enzyme membrane-located complex involving part of the polyphenol oxidase activity of the tissue (possibly an isoenzyme), and that the majority of the polyphenol oxidase activity in Amaranthus is a constitutive membrane enzyme which is not involved in betacyanin synthesis. Although cytokinins do not affect in vivo tyrosine hydroxylation, this activity follows closely the accumulation of betacyanin which is first detectable about 6.5 h after the application of cytokinin. Only a very low level of in vivo hydroxylation can be demonstrated in half-seedlings treated for 6 h either with or without cytokinin but it begins to increase shortly after this time. A large increase in this activity by 16 h (independent of cytokinin) can be almost completely (79%) prevented by chloramphenicol (300 μM) although the drug increases accumulation of betacyanin. At this concentration about 30% of the protein synthesis in inhibited. In vitro tyrosine hydroxylation is, however, not reduced in half-seedlings treated with chloramphenicol during 16 h induction nor is extractable polyphenol oxidase reduced. It is concluded that chloramphenicol is inhibiting the synthesis of some protein essential for in vivo hydroxylation other than the activity measured during in vitro hydroxylation and that the inhibition of synthesis of 79% in vivo hydroxylation still leaves enough activity for maximum betacyanin synthesis.     

Effect of the phosphonic analogue of tyrosine on tyrosine iodination

Summary Depositing ofdl-1-amino-2-(p-hydroxyphenyl)-ethylphosphonic acid (Tyr-P) on the chicken embryo induced a dose dependent decrease of the iodine uptake by the embryonic thyroid. Tyr-P interfered on iodination of tyrosine when tested with hog thyroid peroxidase (TPO) and with bovine lactoperoxidase (LPO); the analogue was recognized by the two enzymes but its affinity for TPO and LPO was respectively 3 and 7 fold higher compared with that of the natural substrate, suggesting that Tyr-P may act as an iodine trap.     

Action of the phosphonic analogue of tyrosine and of other tyrosine derivatives on rat liver tyrosine aminotransferase

Summary Tyrosine transamination has been investigatedin vitro with a preparation of rat liver tyrosine aminotransferase in the presence of several structural derivatives of the substrate, including the phosphonic analogue. The transamination by tyrosine aminotransferase (TAT) needs the presence in the substrate molecule of free amino and carboxylic groups, a three-carbon aliphatic chain, a para-phenolic hydroxylic function and al-configuration. Some tyrosine analogues can markedly disturb the Tyr-TAT association: the chief structural modifications are (i) the removal of the free amine function in a compound still possessing a para-hydroxylic and a carboxylic group, (ii) the change of the carboxylic function by another acidic group, especially a phosphonic one, (iii) a disubstitution in positions 3 and 5. In every situation, the presence of a parahydroxylic group is compulsory to observe an inhibitory effect.     

Biosynthesis of 3,4-dihydroxyphenylalanine in Vicia faba

Radioactive shikimic acid and l-tyrosine were shown to be efficient precursors of 3,4-dihydroxyphenylalanine (DOPA) in Vicia faba. [1-¹⁴C]Acetate and l[U-¹⁴C]phenylalanine were not incorporated into tyrosine or DOPA. Thus the synthesis of DOPA occurs via the shikimic acid pathway and tyrosine or a very closely related metabolise. Phenolase was present in etiolated plants in much larger quantities after a brief light exposure whereas DOPA concentration was relatively constant during all stages of plant growth. Partially purified phenolase did not catalyze the conversion of tyrosine to DOPA and does not appear to have a role in DOPA synthesis.     

Effect of tyrosine administration on dopa accumulation in light- and dark-adapted retinas from normal and diabetic rats

The interaction of tyrosine concentration and lighting on in vivo dihydroxyphenylalanine (dopa) accumulation rate was studied in retinas of normal and diabetic rats. In both groups of rats, dopa accumulation and in vitro hydroxylase activity were higher in retinas exposed to light than in those adapted to darkness. In light-adapted diabetic rats, though, retinal tyrosine level, dopa accumulation, and in vitro tyrosine hydroxylase activity were all below normal. In both normal and diabetic rats exposed to light, tyrosine injection raised retinal tyrosine concentrations and stimulated dopa accumulation. Injection of tyrosine into dark-adapted rats raised retinal tyrosine level but did not enhance dopa accumulation. Together, these results suggest that in vivo retinal amacrine cells will vary their dopa accumulation rate as a function of substrate supply, but only in the light, when tyrosine hydroxylase is activated. They further indicate that dopa accumulation rate remains sensitive to tyrosine supply in the light-activated diabetic retina.     

Effect of environment pH on storage glucan synthesis in three thermophilic algae

Plectonema nostocorum, a thermophilic cyanophyte which lives under alkaline conditions at pHs approaching 13, forms a storage glucan showing a maximum absorption of its iodine complex almost identical with that of another thermophilic cyanophyte, Oscillatoria princeps, which exists at a more neutral pH, and with that of the acidophilic thermophile, Cyanidium caldarium. Gel electrophoretic patterns of the storage glucan-forming isozymes of Plectonema do not differ essentially from those of Oscillatoria. The a₂ phosphorylase isozyme appears to be primer-independent, and resembles the a₂ isozymes of both Oscillatoria and Cyanidium. The isozymes responsible for forming α-1,6-glucosidic branched linkages in Plectonema are of the b.e. type (able to further branch amylopectin), rather than of the Q type (able to branch amylose only to amylopectin).     

Methyl jasmonate differentially affects tocopherol content and tyrosine amino transferase activity in cultured cells of Amaranthus caudatus and Chenopodium quinoa

Tocopherols are lipid-soluble compounds synthesised exclusively by photosynthetic organisms. In this study, in vitro callus cultures were established from two plants that are naturally rich in tocopherols, Amaranthus caudatus and Chenopodium quinoa, in order to examine whether callus cultures were able to produce these compounds at levels comparable to those observed in planta . In both species, cotyledon explants produced the best callus induction and, once established, callus cultures were grown under two different hormonal treatments to check for effects of growth and to induce chloroplast differentiation in the cells. A rapid differentiation of chloroplasts occurred only in C. quinoa cell aggregates grown in the presence of benzyladenine, leading to the production of a homogeneous green callus. In both species, only α-tocopherol was produced by callus cultures, although levels were much lower than in planta, and the production was not influenced by the hormonal conditions. Interestingly, cell cultures of the two species responded in different ways to methyl jasmonate (MJ). In A. caudatus cultures, treatment with 100 μ m MJ increased the production of α-tocopherol up to fivefold, and the inductive effect was influenced by the hormonal composition of the medium. This increase in α-tocopherol was associated with a proportional increase in tyrosine aminotransferase (TAT) activity, one of the key enzymes involved in tocopherol biosynthesis. By contrast, in C. quinoa cultures, elicitation with MJ did not have any effect, neither on tocopherol production, nor on TAT activity. These results are discussed in relation to chloroplast differentiation and the interplay between jasmonates and phytohormones.     

弗氏柠檬酸细菌酪氨酸酚解酶基因在大肠杆菌中的克隆与表达

以基因组DNA为模板,利用PCR技术从弗氏柠檬酸细菌（Citrobacter freundii)中扩增得到含有酪氨酸酚解酶基因的DNA片段,定向连续到质粒pUC118上,得到重组质粒pTPL,将此重组质粒转化到受体菌E.colXL-1-Blue MRF′中,通过蓝白斑鉴定挑出阳性菌株。从此阳性菌株中提取质粒pTPL并将此质粒转入到E.coliJM109中,用E.coliJM109（pTPL)制备高活性的酪氨酸酚解酶。对质粒稳定性的研究表明,E.coliJM109（pTPL)在无选择压力下37℃连续培养50代以上,质粒丢失率仅有15％,说明质粒基本稳定。     

Effect of divalent metal ions on the synthesis of oligosaccharide side chains of Neurospora crassa glycoproteins

Neurospora crassa membrane preparations incorporated mannose from GDP-mannose-[¹⁴C] in the presence of Mg²⁺ into a polyprenol lipid and side chains of protein acceptor(s), which are labile on hydrolysis in weak base. The addition of Mn²⁺ to the reaction mixtures does not affect mannosyl lipid synthesis but it stimulates the transfer of mannose to larger oligosaccharide chains resistant to β-elimination and the transfer of a second mannosyl unit to form an O-glycosidically linked mannobiosyl side chain. Incubation of particulate preparations with polyprenol-mannose-[¹⁴C] in the presence of Mg²⁺ and Mn²⁺ also results in the transfer of a single mannose to the protein. When non-radioactive GDP-mannose is added to this reaction mixture, however, β-elimination yields mannobiose. The mannobiose is labeled in the reducing sugar only. These results indicate that the first mannose of this mannobiosyl side chain is transferred via a lipid intermediate, but the second mannose is transferred directly from GDP-mannose. In the presence of Mg²⁺ and Mn²⁺, mannose apparently is also transferred from polyprenol-mannose-[¹⁴C] to side chains which are resistant to hydrolysis.