The mechanism of ferrous iron binding by suspensions of Bifidobacterium bifidum var. pennsylvanicus

Bifidobacterium bifidum var. pennsylvanicus acquires ferrous iron at a reasonable rate when this bacterium — in the resting state — is incubated with ferrous iron in either a modified Hanks' solution, ot Norris medium in which the bacteria had previously grown. Ferrous iron binding was substantially lower in unused Norris medium containing human milk whey, because it contained iron binding inhibitors that were degraded during bacterial growth. The inhibitors were identified as human milk whey components and bovine casein digest. The affinity of bacteria for ferrous iron was dependent on the iron concentration of the media. In modified Hanks' solution, the affinity of cells for iron at low iron concentrations was some 250-fold greater than that at high iron concentrations. Ferrous iron binding did not take place it a carbon source was omitted from the media or if the cells had previously been heated at 80°C for 20 min. Surface binding of ferrous iron to the bacterial membrane was minimal. Trypsin and microbial proteinase treatment of bacteria did not inhibit iron uptake, making an iron-specific receptor on bifidobacterial cell membranes unlikely. Iron binding by bifidobacteria is an energy-requiring process dependent upon the operation of a heat-sensitive enzyme machinery, and it does not involve an extracellular iron carrier.     

Growth ofNeisseria gonorrhoeae in media deficient in iron without detection of siderophores

A disseminating strain ofNeisseria gonorrhoeae was grown in a liquid, defined medium containing different levels of a strong chelator of ferric iron, ethylenediamine-di (o-hydroxyphenyl acetic acid) (EDDA). The inhibition of growth produced by EDDA was relieved by addition of iron. Filtrates from cultures grown in the media deficient in iron were examined for presence of siderophores, microbial chelators of iron, by colorimetric tests for phenolates and hydroxamates and by two biological assays in which the filtrates were tested for ability to stimulate growth of small inocula and for ability to prevent bactericidal effects of normal human serum. By use of these methods, none of the filtrates displayed evidence of gonococcal siderophores.     

In vitro micropropagation of Primula scotica: a rare Scottish plant

A simple micropropagation method is reported for Primula scotica, a rare plant, endemic to the North of Scotland. The technique involves the clonal proliferation of seed-derived plantlets on either hormone free tissue culture medium or on medium containing benzyl amino purine and indole acetic acid. Average multiplication rates of 4–6 were obtained for plants grown on plant growth regulator supplemented medium. The micropropagation method was applied to four different clones of P. scotica and clonal differences were observed in relation to media type. Some plants from certain clones did display hyperhydricity, however, this was circumvented by using frequent sub-culturing intervals and transferring the plants to hormone-free medium. Plantlets rooted on both media types and displayed normal, true-to-type rosette morphology. The cultures did not callus and development proceeded via shoot and root production only. The in vitro-grown plants could be transferred to ex vitro conditions and a range of growth substrates were assessed for their efficacy in supporting ex vitro growth, with a view to developing longer-term strategies for the transfer and reintroduction of micropropagated P. scotica plants into natural habitats. The simple method described in this paper may offer the potential of being applied to other endangered Primula spp.     

The effect of iron ethylenediaminetetraacetic acid on the growth and metabolism of tomato plants in water culture

Summary Tomato plants were grown in nutrient culture either continuously or discontinuously treated with FeSO₄ or with Fe-EDTA. FeSO₄ rapidly gave a precipitate of Fe₂O₃.nH₂O but remained, under the conditions of the experiments, available to the plants. Fe absorbed from Fe-EDTA in the nutrient medium was less effective in inducing tomato plant growth than iron supplied as ferrous sulphate. This cannot be explained in terms of luxury consumption or by phosphate-induced internal iron immobilization.Plants grown continuously with Fe-EDTA as an iron source showed consistently higher peroxidase and catalase activities. Chlorotic plants treated for a few days with FeSO₄ exhibited mostly equal or higher enzyme activities than those treated with Fe-EDTA. The ratio of green and yellow chloroplast pigments was constant under all the tested levels of iron nutrition.Although there were significant differences in iron utilization, enzyme activities, and some symptoms of slightly modified growth, the main conclusion of this investigation is that, under these strictly comparable conditions, plant metabolism has not been modified considerably by the use of the synthetic iron chelates.     

Effect of iron on adherence and cytotoxicity of Entamoeba histolytica to CHO cell monolayers

Iron is an essential element for almost all living organisms. The possible role of iron for growth, adherence and cytotoxicity of Entamoeba histolytica was evaluated in this study. The absence of iron from TYI-S-33 medium stopped amebic growth in vitro. However, iron concentrations in the culture media of 21.4-285.6 microM did not affect the growth of the amebae. Although growth was not retarded at these concentrations, the adhesive abilities of E. histolytica and their cytotoxicities to CHO cell monolayer were correlated with iron concentration. Amebic adhesion to CHO cell monolayers was significantly reduced by low-iron (24.6 +/- 2.1%) compared with 62.7 +/- 2.8 and 63.1 +/- 1.4% of amebae grown in a normal-iron and high-iron media, respectively. E. histolytica cultured in the normal- and high-iron media destroyed 69.1 +/- 4.3% and 72.6 +/- 5.7% of cultured CHO cell monolayers, but amebae grown in the low-iron medium showed a significantly reduced level of cytotoxicity to CHO cells (2.8 +/- 0.2%). Addition of divalent cations other than iron to amebic trophozoites grown in the low-iron medium failed to restore levels of the cytotoxicity. However, when E. histolytica grown in low-iron medium were transferred to normal-iron medium, the amebae showed completely restored cytotoxicity within 7 days. The result suggests that iron is an important factor in the adherence and cytotoxicity of E. histolytica to CHO cell monolayer.     

Effects of various iron supply on oxidative stress development and ferritin formation in the common ice plants

Mesembryanthemum crystallinum L. plants were grown from seeds in perlite. At the age of 4 weeks (juvenile plants) or 6 weeks (adult plants), they were transferred on nutrient media with different Fe³⁺ content brought in as Fe₂(SO₄)₃—EDTA complex (pH 6.0): control, iron deficit, and iron “excess”. Adult plants grown in media differing in iron content were subjected to salinity (300 mM NaCl) during the last 8 days of growth. Biochemical analyses were performed after plant fixation in liquid nitrogen; simultaneously, the samples for electron microscopy were taken. Different content of available Fe³⁺ in medium, especially under salinity conditions, changed sharply the content of chlorophyll and proline, the rate of lipid peroxidation, the level of H₂O₂, the activities of antioxidant enzymes in the leaves and roots, the number and sizes of plastoglobules, and ferritin formation in plastids. Joint action of salinity and iron deficit enhanced oxidative stress development, whereas iron excess hampered oxidative reaction development, reduced the rate of lipid peroxidation, and increased the chlorophyll content. At iron excess, plastoglobule lysis in plastids did not occur, their number and sizes increased, and ferritin deposits appeared, whereas the latter were absent at iron deficit.     

Effect of various exogenous and endogenous factors on microspore embryogenesis in Indian mustard (<Emphasis Type="Italic">Brassica juncea</Emphasis> (L.) Czern and Coss)

Summary The influence of donor plant growth environment, microspore development stage, culture media and incubation conditions on microspore embryogenesis was studied in three Indian B. juncea varieties. The donor plants were grown under varying environments: field conditions, controlled conditions, or a combination of the two. The correlation analysis between the bud size and microspore development stage revealed that the bud size is an accurate marker for donor plants grown under controlled conditions, however, the same does not hold true for the field-grown plants. The buds containing late uninucleate microspores collected from plants grown under normal field conditions up to bolting stage and then transferred to controlled environment were observed to be most responsive with genotypic variability ranging from 10 to 35 embryos per Petri dish, irrespective of the other factors. NLN medium containing 13% sucrose was found to be most suitable for induction of embryogenesis The fortification of this medium with activated charcoal, polyvinylpyrrolidone, colchicine, or growth regulators (6-benzylaminopurine and 1-naphthaleneacetic acid) was observed to be antagonistic for microspore embryogenesis, while silver nitrate (10 μM) had a significant synergistic effect. A post-culture high-temperature incubation of microspores at 32.5±1°C for 10–15 d was found most suitable for high-frequency production of microspore embryos. The highest frequency of microspore embryogenesis (78 embryos per Petri dish) was observed from the late uninucleate microspores (contained in bud sizes 3.1–3.5 nm irrespective of genotype) cultured on NLN medium containing 13% sucrose and silver nitrate (10 μM), and incubated at 32.5°C for 10–15 d.     

Naphthoquinone contents of in vitro cultured plants and cell suspensions of Dionaea muscipula and Drosera species

In vitro cultured carnivorous plants were grown on a hormone-free medium. They produced the following naphthoquinones: Dionaea muscipula (plumbagin: 5.3%), Drosera rotundifolia (7-methyljuglone: 0.6%), D. binata (plumbagin: 1.4%), and D. capensis (7-methyljuglone: 0.5%). A red, slow-growing suspension culture of D. muscipula was maintained in a modified McCowns Woody Plant (McC) medium and produced plumbagin (2.59%) after 30 days growth. A suspension culture of D. rotundifolia grew slowly as multicoloured small aggregates only in a modified Murashige and Skoog (MS) medium. No quantifiable amounts of naphthoquinones were produced. Several cell lines of D. capensis were developed. Green aggregates grown in a modified MS medium contained 7-methyljuglone (0.33%) and differentiated into plants when placed onto hormone-free medium. Pink cultures grown in modified McC medium contained 7-methyljuglone (1.24%), while dark red cultures produced ca. 1% in both modified McC and MS media. Though the latter medium was significantly better with regard to biomass production, cells excreted a mucin when cultured in both media (0.21 g dry mucin/g dry cells in McC) and (0.16 g dry mucin/g dry cells in MS). Effects of the presence or absence of light during the growth period of 30 days showed that there was no effect on biomass and only slight effects on mucin production and naphthoquinone contents.     

Utilization of cathodically-produced hydrogen from mild steel byDesulfovibrio species with different types of hydrogenases

Summary Desulfovibrio (D.) vulgaris Hildenborough with a highly active Fe-containing periplasmic hydrogenase,D. salexigens British Guiana with a Fe–Ni–Se periplasmic hydrogenase, andD. multispirans with a Fe–Ni cytoplasmic hydrogenase utilized cathodically-produced hydrogen from mild steel as the only energy source for activity and growth. Changes on the mild steel surface occurred during growth of these bacteria. The concentration of iron sulfide, a corrosion product of mild steel, increased over time, andDesulfovibrio species had an active hydrogenase when they were grown in lactate/sulfate media. This hydrogenase may be any of the three types found in the genus,Desulfovibrio. The concentration of iron in the media affected the production and activity of the Fe-hydrogenase fromD. vulgaris Hildenborough. With an iron-limited medium, the specific activity and the total amount of the periplasmic hydrogenase was less than found with a non-iron limited media.     

Influence of growth medium on antifungal activity of neem oil (<Emphasis Type="Italic">Azadirachta indica</Emphasis>) against <Emphasis Type="Italic">Lagenidium giganteum</Emphasis> and <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis>

The inhibition of mycelial growth of Lagenidium giganteum by neem oil was lower than that of Metarhizium anisopliae in PYG and Emerson’s YpSs agar media. However, neem oil did not inhibit the mycelial growth of L. giganteum in sunflower seed extract agar medium, but did it inhibit the mycelial growth of M. anisopliae. The minimum inhibitory concentration of neem oil for L. giganteum was higher than that for M. anisopliae. The minimum fungicidal concentration of neem oil in PYG medium was lower than in YpSs for both fungi. The spores of L. giganteum grown in SFE medium could be used with neem oil for vector control.     

Jasmonate effects on in vitro tuberization and tuber bulking in two potato cultivars (Solanum tuberosum L.) under different media and photoperiod conditions

Summary Jasmonic acid (JA) effects on in vitro tuberization of potato nodal explants cvs. Sangre and Russet Burbank were tested under liquid and solid media conditions and 0,8, and 16h photoperiod. Explants taken from stock plants grown on 2.5μM JA-supplemented medium tuberized first, particularly in darkness. The most pronounced benefits of the JA pretreatment were recorded under 16h photoperiod, which is known to inhibit tuberization. Cultivar Sangre benefited from the JA preconditioning of stock plants more than Russet Burbank. Russet Burbank required the JA supplement in tuberization media to reach the same degree of stimulation. Overall, microtubers produced either from JA preconditioned stock plants or on the JA-containing tuberization media were more uniform and larger than from other treatments. Eight hours photoperiod was by far the best treatment for the production of high-quality uniform microtubers. JA conditioning of stock plants prior to taking explants for tuberization is being proposed as a treatment enhancing the quality of microtubers.     

A non-antibiotic selection system uses the phosphomannose-isomerase (PMI) gene forAgrobacterium-mediated transformation of Chinese cabbage

To establish a non-antibiotic selection system that utilizes the phosphomannose-isomerase (PMI) gene for Chinese cabbage transformation, we first determined the optimum mannose concentration for selecting transformed cells. Hypocotyl and cotyledon expiants that were grown on media containing more than 5 g L^-1 mannose did not induce green calli but, rather became chlorotic and withered before dying. In contrast, media containing 20 g L^-1 sucrose plus 5 g L^-1 mannose proved suitable for selection. We then used this particular level of mannose to transform hypocotyl tissues. Within 6 weeks, shoots were regenerated from some of the calli; subsequently, these plants were transplanted to pots and grown in the greenhouse. A 514-bp PCR fragment was obtained from most transformants but not from the non-transformed plants. Southern blot analysis also revealed the expectedPMI gene in those PCR-confirmed transgenic plants. RT-PCR of total RNA was performed to confirmPMI expression. We have now demonstrated that this gene does not inhibit the growth of transgenic plants, and that this selection system can be applied to Chinese cabbage transformation.     

Growth and chlorophyll production in plant callus tissues grown in vitro

Summary Growth, nutrition and chlorophyll development were studied in chlorophyllous callus tissues isolated from the following edible angiospermous plants: carrot root, crown gall of tomato, endive embryo, leaf petiole and stem of lettuce, leaf petiole of parsley, pea stem and rose stem. Growth patterns of these tissues in vitro were sigmoid. Synthetic media produced less growth, in terms of fresh weight increase, than media containing coconut milk, a highly complex and little understood natural substance. Murashige and Skoog's synthetic medium proved useful for satisfactory growth and chlorophyll production in a number of tissues. Its usefulness was further increased by additional amounts of copper sulphate, potassium nitrate and monobasic ammonium phosphate. Increased levels of iron and magnesium inhibited growth. Incorporation of yeast extract in the tobacco-high-salts-medium produced the highest amount of growth and chlorophyll formation in endive tissue. Presence of exogenous sucrose was essential for the continued good growth of the above callus tissues in vitro. Highest amount of growth took place either in white light or in the dark. Different tissues had different responses to high or low intensities of light. Endive and carrot tissues produced in vitro were palatable to human taste. Endive tissue was particularly good as it also differentiated many small rosettes of leaves, shoots and had a mild aromatic flavor typical of the endive plants grown in nature.     

A theoretical explanation of the Piper-Steenbjerg effect

The relation between plant yield and plant nutrient concentration is sometimes found to be negative, a phenomenon called the Piper-Steenbjerg (PS) effect. A model was used to examine the underlying causes of the PS effect, and the conditions under which it is most likely to occur. The model uses the nutrient productivity concept for plant growth and a nutrient uptake equation in which root growth rate and external nutrient concentration determine the uptake rate. The study suggests that the PS effect occurs when the fast growth of plants grown in an initially higher nutrient medium eventually leads to a more rapid depletion of external nutrients than the slow growth of plants grown in an initially lower nutrient medium. The fast growth of plants combined with a rapid decrease of nutrient uptake leads to a fall in plant nutrient concentration. When these large plants with very low nutrient concentrations are compared with the smaller, slow-growing plants, a PS effect may be found depending on the time at which the plants are harvested, and on the range of initial values of the external nutrient content. When it occurs, the effect is greatest when the depletion volume per unit new root (V_d) is lowest, and when the mobility of nutrients in the medium is highest (α=1). The results are sufficiently general to apply to a variety of nutrients, plant species and growth media.     

Accumulation of periplasmic enterobactin impairs the growth and morphology of Escherichia coli tolC mutants

TolC is the outer membrane component of tripartite efflux pumps, which expel proteins, toxins and antimicrobial agents from Gram‐negative bacteria. Escherichia coli tolC mutants grow well and are slightly elongated in rich media but grow less well than wild‐type cells in minimal media. These phenotypes have no physiological explanation as yet. Here, we find that tolC mutants have highly aberrant shapes when grown in M9‐glucose medium but that adding iron restores wild‐type morphology. When starved for iron, E. coli tolC mutants synthesize but cannot secrete the siderophore enterobactin, which collects in the periplasm. tolC mutants unable to synthesize enterobactin display no growth or morphological defects, and adding exogenous enterobactin recreates these aberrations, implicating this compound as the causative agent. Cells unable to import enterobactin across the outer membrane grow normally, whereas cells that import enterobactin only to the periplasm become morphologically aberrant. Thus, tolC mutants grown in low iron conditions accumulate periplasmic enterobactin, which impairs bacterial morphology, possibly by sequestering iron and inhibiting an iron‐dependent reaction involved in cell division or peptidoglycan synthesis. The results also highlight the need to supply sufficient iron when studying TolC‐directed export or efflux, to eliminate extraneous physiological effects.     

Utilization of phosphorus by pasture plants supplied with myo-inositol hexaphosphate is enhanced by the presence of soil micro-organisms

A range of pasture grass (Danthonia richardsonii and Phalaris aquatica) and legume (Medicago polymorpha, M. sativa, Trifolium repens and T. subterraneum) species showed limited capacity to obtain phosphorus (P) from inositol hexaphosphate (IHP), when grown in either sterile agar (pH 5.0 or 5.5) or sand-vermiculite media (pH 5.0). The total P content of shoots from IHP-supplied plants grown in agar was between 20% and 34% of that for seedlings supplied with an equivalent amount of P as inorganic phosphate (P_i), while in sand-vermiculite, the total P content of IHP-grown plants was between 5 and 10% of control plants. The poor ability of plants to utilize P from IHP resulted in significantly lower tissue P concentrations and, in general, reduced plant dry weight accumulation. In contrast, the P nutrition of plants supplied with IHP was significantly improved by inoculating media with either a cultured population of total soil micro-organisms or with a specific isolate of Pseudomonas sp., selected for its ability to release phosphate from IHP (strain CCAR59; Richardson and Hadobas, 1997 Can. J. Micro. 43, 509-516). In agar and sand-vermiculite media, respectively, the P content of IHP-grown plants increased with inoculation by up to 3.9- and 6.8-fold, such that the dry weight and P content of the plant material were equivalent to those observed for control plants supplied with P_i. However, the response to inoculation was dependent on the growth medium and the source of micro-organisms used. In sand-vermiculite, the cultured population of soil micro-organisms was effective when IHP was supplied at an equivalent level of P_i required for maximum plant growth. By comparison, inoculation of plants with the Pseudomonas strain was only effective at very high levels of IHP supply (×36), whereas in agar a response to inoculation occurred at all levels of IHP. The ability of pasture plants to acquire P from phytate was, therefore, influenced by the availability of IHP substrate, which was further affected by the presence of soil micro-organisms. Our results show that in addition to having an effect on the sorption characteristics of the growth media, soil micro-organisms also provided a source of phytase for the dephosphorylation of phytate for subsequent utilization of P_i by plants.     

Growth and solute accumulation in 3-week-old seedlings of Agropyron elongatiun stressed with sodium and potassium salts

Agropyron elongatum [Host. (Beauv.)] [^cv. Arizona Glendale, was grown in liquid medium salinized with either NaCl, KCI, or a 50:50 mixture of these two salts at osmotic potentials ranging from 0 to –1.6 MPa. The amount of growth in 21 days was measured, and extracts were made of the shoots at this time. The extracts were assayed for low-molecular-weight organic compounds (glucose, fructose, sucrose, be-taine, proline) and inorganic solutes (Na⁺, K⁺, Cl^?, P.). The purpose was to determine if there was any correlation between the harmful effect of salinity on growth and the concentrations of solutes in tissues. Growth inhibition of A. elongatum was roughly proportional to the osmotic potential of the growth medium and was independent of the ionic composition of the salinizing salts. Total monovalent cation (the sum of Na⁺ and K⁺) concentrations and the ratio of these two cations in leaves were mainly a function of the ionic compostion of the salt in growth media, and, to a lesser degree, of osmotic potentials. F At an osmotic potential of –0.2 MPa, total monovalent cation in leaves was the same as in non-stressed plants. However, if the salinizing salt contained NaCl, there was an increase in foliar Na⁺ with a balancing decrease in K⁺. At stress levels between –0.4 and –1,6 MPa, and, if the media were salinized with either 100% NaCl or a 50:50 mixture of NaCl and KCI, total monovalent cation concentrations remained constant at a value that was twice that in non-stressed plants. Although total monovalent cation concentrations were equal in plants grown under these two salinity conditions, the K⁺/Na⁺ ratios shifted from a value of 1:2 in plants grown in 100% NaCl to 3:1 in plants subjected to the 50:50 mixture. If 100% KCI was used to salinize media, total monovalent cation was 80% of its concentration in NaCl-treated plants in the range of –0.4 to -1.2 MPa. At –1.6 MPa due to 100% KCI, total monovalent cation was double that in plants subjected to -0.4 MPa. In the range of osmotic potentials from–0.2 to –1.2 MPa, the chloride:cation ratio was 1:2. At –1.6 MPa the ratio changed to 3:4. Proline started accumulating in leaves of A. elongatum when the tissue concentration of total monovalent cation exceeded 200 μ (g fresh weight)^?1. Above this threshold value of total monovalent cation, the proline concentration of leaves was 6% of the amount of total monovalent cation that exceeded 200 umol (g fresh weight)¹.     

Benefits associated with the stalk of Gallionella ferruginea,evaluated by comparison of a stalk-forming and a non-stalk-forming strain and biofilm studies in situ

Factors that regulate and induce stalk formation by the iron-oxidizing and stalk-forming bacterium Gallionella ferruginea were studied in laboratory cultures and in situ. A stalk-forming strain, Sta⁺, and a non-stalk-forming strain, Sta^-, were used for comparative studies of the benefits associated with the stalk. Two different growth media were used: a ferrous sulfide medium (FS-medium), with slow oxidation of iron giving high concentrations of toxic oxygen radicals and a ferrous carbonate medium (FC-medium), with fast iron oxidation giving low concentration of the toxic oxygen radicals. It was found that Sta⁺ cells grown in the FS-medium survived 3 weeks longer than Sta^- cells grown in the FS-medium. When each strain was grown in the FC-medium, the Sta^- cells had an advantage and survived 8 weeks longer than the Sta⁺ cells. No difference in survival was found for Sta⁺ cells grown in FS-medium compared to growth in FC-medium. In laboratory cultures, the average stalk length per cell values were 7–2.5 times higher (92 h and 150–300 h growth, respectively) in a medium with 620 m iron than in a medium with 290 m iron. Gallionella ferruginea Sta⁺ outcompeted Sta^- cells when inoculated as mixed populations in FC-medium. It has previously been suggested that stalk formation in vitro is induced by oxygen. To confirm this observation, biofilm development in natural waters was studied in two wells, one with trace amounts of oxygen (LH) and one without (TH). A dense biofilm developed on surfaces exposed to flowing well LH water, but no biofilm developed in well TH. Stalks were formed in water samples from both wells when allowed to make contact with air. This work demonstrates for the first time that the stalk has a protecting function against the toxic oxygen radicals formed during the chemical iron oxidation. It also shows that it is the oxidation rate of the ferrous iron and not its concentration that is harmful to the cells. The stalk gives G. ferruginea a unique possibility to colonize and survive in habitats with high contents of iron, inaccessible for bacteria without a defense system against the oxidation of iron. Correspondence to: L. Hallbeck     

Growth characteristics and toxin production in batch cultures of Anabaena flos-aquae: effects of culture media and duration

The effects of media and culture duration on growth, macromolecular composition and toxicity of an anatoxin- a-producing freshwater cyanobacterium Anabaena flos-aquae (UTEX 2383) were evaluated. The four media A₃M₇, CB, MA and B-12 influenced growth in terms of cell number, chlorophyll-a content and specific growth rate. A₃M₇ medium supported the best growth. The macromolecular composition of cultured cells, viz. total carbohydrate, protein and lipid content varied with media and culture duration reaching maximum concentration at various growth periods. The differences were significant due to interaction of the culture medium and duration. Toxicity of cells grown in different media was compared by Artemia salina bioassay and mouse units. The cells grown in A₃M₇ medium showed highest toxicity and the optimum culture duration was 5 weeks. In terms of both growth characteristics and toxicity the media can be ranked as A₃M₇, MA, CB and B-12 in decreasing order.     

Iridescent material and the effect of iron on its production byPseudomonas aeruginosa

The nutritional conditions controlling iridescence inPseudomonas aeruginosa were studied using synthetic media solidified with agar. Iron and magnesium were growth-limiting factors in media solidified with dialysed agar. Iridescence only occurred on iron-deficient media and was not suppressed by adding Ca, Cu, Mn and Zn to these media. The ultraviolet absorption spectrum of the iridescent material was almost identical to the spectrum of the pyo I substances which are 2-alkyl-4-quinolinols.The amount of material produced was inversely proportional to the iron content of the medium. Small amounts of material were produced by cells grown at levels of iron optimal for growth. Synthesis of 2-alkyl-4-quinolinol may be a normal metabolic process in the iridescent strains ofPseudomonas aeruginosa. It was enhanced by anthranilic acid and tryptophan; kynurenine and kynurenic acid had no effect. The results can be explained if it is assumed that the activity of iron-requiring enzymes catalizing the breakdown of tryptophan is reduced.Even in the presence of anthranilic acid or tryptophan no material was produced by a non-iridescent strain.