Effect of naphthalene and aqueous crude-oil extracts on the green flagellate Chlamydomonas angulosa. VI. Phosphate uptake and retention

1985 ◽  
Vol 63 (4) ◽  
pp. 829-833 ◽  
Author(s):  
T. C. Hutchinson ◽  
C. Soto ◽  
J. A. Hellebust

Chlamydomonas angulosa when grown in normal Bold's basal medium (BBM) (1.7 mM Pi) has a very high cell P content (120 fg-at. P∙cell−1) and low C: P atomic ratio (35: 1) and is only able to show net Pi uptake in media with Pi concentrations higher than 0.1 mM when grown in the light. The presence of light enhances net phosphate uptake or decreases net phosphate loss under all external Pi concentrations. Cells transferred to low-Pi media will, however, grow rapidly while reducing their cell P to almost 1/10 of the initial level, indicating that most of the cell P in cells grown in high-P media represents reserve P, probably in the form of polyphosphates. The presence of naphthalene and crude-oil components in the culture media decreases net uptake of Pi at high external Pi concentrations and increases the rate of Pi loss at low external concentrations. Naphthalene present at 100% saturation level in BBM causes rapid loss of a large fraction of cell P under all conditions tested. The phosphorus lost appears as reactive phosphorus in the medium, suggesting that cell polyphosphates are rapidly converted to Pi, which then leaks out of the cells in response to the presence of saturating naphthalene concentrations in the medium. BBM with 50% naphthalene saturation causes much less loss of cell P under most of the incubation conditions, and net uptake can take place at the highest external Pi concentrations tested in the light. Media saturated with crude oil are even less effective than 50% naphthalene saturated media in causing P efflux, or in reducing net Pi uptake, although at low external P concentrations, uptake of P is decreased compared with the control.


1985 ◽  
Vol 63 (4) ◽  
pp. 834-840 ◽  
Author(s):  
J. A. Hellebust ◽  
C. Soto ◽  
T. C. Hutchinson

Chlamydomonas angulosa grows equally well on nitrate and ammonium as sources of nitrogen. The presence of ammonium decreases nitrate uptake by less than 10% in short-term experiments. The presence of nitrate has no significant effect on short-term uptake of the ammonium analogue methylamine. Cells grown in nitrate media possess considerable methylamine uptake capacity during early exponential growth. This uptake capacity falls rapidly as the cells enter the declining growth phase. When cells are transferred to nitrogen-deficient media, the uptake capacity for methylamine increases threefold to fourfold in 24 h. The half-saturation constants (Km) for nitrate and methylamine uptake of this alga are 0.4 mM and 90 μM, respectively. When C. angulosa cells are transferred from control Bolds basal medium (BBM) to 50% naphthalene saturated or aqueous crude oil saturated media, the initial uptake rate for nitrate increases by a factor of two or decreases by a factor of one-third, respectively, as compared with that of cells transferred to control BBM. However, cells incubated in closed incubation systems with naphthalene or aqueous crude oil saturated media for 3 days lose nitrate when resuspended in control media. Cells transferred to media containing naphthalene up to 20% saturation show no immediate decrease in methylamine transport, while higher naphthalene concentrations cause an immediate decrease in transport activity. However, cells incubated in 50% naphthalene saturated media in a closed system for 2–4 h actually show increased methylamine transport activity when the incubation system is opened to allow escape of the hydrocarbon. Prolonged exposure to 50% naphthalene saturated media, however, causes progressive loss of transport activity.



1975 ◽  
Vol 53 (2) ◽  
pp. 118-126 ◽  
Author(s):  
C. Soto ◽  
J. A. Hellebust ◽  
T. C. Hutchinson

Comparisons have been made of the effects of crude oil extracts and naphthalene on the photosynthesis of the alga Chlamydomonas angulosa. The addition of the hydrocarbon naphthalene to C. angulosa cultures causes an immediate and almost complete loss of photosynthetic capacity. When cells that have been incubated in closed systems containing naphthalene-saturated media are transferred to open systems which allow loss by volatilization of naphthalene, the rate of recovery of photosynthesis is inversely proportional to the length of incubation in the closed system with naphthalene. Incubation of cells in open systems with media containing aqueous crude oil extracts has no significant effect on their photosynthetic capacity as compared with that of cells incubated in control media. However, the photosynthetic capacity is decreased when cells are incubated in closed systems with media containing aqueous crude oil extracts. This is followed by a gradual recovery with time of incubation after treatment in the closed systems.Experiments using labelled naphthalene indicate that C. angulosa cells accumulate naphthalene from the medium in closed systems for up to 7 days. When such cells are transferred to fresh media not containing naphthalene, there is an almost immediate loss of the aromatic hydrocarbon from the cells. However, when cells are retained in their original medium after opening the culture system to allow naphthalene to escape, the decrease in naphthalene content per cell appears dependent upon resumption of cell division and occurs in a stepwise manner, which suggests cell retention rather than an actual loss of naphthalene to the medium.



1981 ◽  
Vol 59 (5) ◽  
pp. 742-749 ◽  
Author(s):  
T. C. Hutchinson ◽  
J. A. Hellebust ◽  
C. Soto

The effect of exposure to naphthalene and to aqueous extracts of crude oil on contents of manganese and potassium in cells of Chlamydomonas angulosa has been measured simultaneously by neutron activation analysis. Decreases of both manganese and potassium from treated cells commenced within a very short time of treatment initiation, and may be due to hydrocarbon-induced membrane damage. In control experiments, manganese uptake into cells was found to be light dependent, and potassium uptake was even more so. In cells treated with a 100% saturated naphthalene solution in Bold's basal medium (BBM), little cellular manganese remained after 6 h of exposure and less than 2% was left within 24 h. Potassium loss was even more rapid and complete. Rates of loss for both elements in a 50% saturated naphthalene solution in BBM and in an aqueous crude oil extract were slower but showed a similar pattern. For potassium in both light or dark exposures, an initial increase in cellular content occurred. Sodium acetate was added to assess heterotrophic growth, and the effects of this addition on the loss of the two elements are discussed. Over a short time, it appeared to act as an alternative to light energy in restoring membrane function.



1982 ◽  
Vol 60 (4) ◽  
pp. 358-363
Author(s):  
A. Thuillier ◽  
P. Neumann

Ceratocystis coerulescens, C. fimbriata, C. ips, and C. minor were tested for production of sexual fruiting bodies, and C. penicillata and C. piceae for asexual fruiting bodies. Ceratocystis fimbriata produced perithecia easily on standard culture media, but there were marked differences between the two strains tested (503, 560). Strain 503 had a good production of fruiting bodies on malt agar (M) and a basal nutrient solution (N). Strain 560 fared better than 503 on Leonian agar (L), but did not fructify on M and N. Supplementing media with various wood extracts produced better results. M + maple sapwood extracts and L + poplar sapwood extracts gave the best results with strain 503, and L + pine sapwood extracts was the best with strain 560.Production of coremia was also influenced by the basal medium and the kind of extracts added as supplements. Fir and maple extracts stimulated the production of fruiting bodies, whereas pine and poplar extracts had no or very little stimulating effects. In every other species tested, the production of fruiting bodies was none or very irregular. [Journal translation]



Genetika ◽  
2006 ◽  
Vol 38 (2) ◽  
pp. 129-136
Author(s):  
Velichka Rodeva ◽  
Stanislava Grozeva ◽  
Velichka Todorova

Callusogenesis and regeneration ability of cotyledon and hypocotyl explants from three Bulgarian pepper varieties in MS basal medium supplemented with l-3mg/l BAP. l.0mg/1 IAA and 0.5mg/l GA3 was studied. In the different variants of culture medium was registered high level of callusogenesis and organogenesis in both type of explants from the all varieties. The highest percentage of plant-regenerants is established in cotyledon explants (from 3.3 to 18.3) in variant 3 of the culture medium containing 3mg/l BA. In the process of micropropagation by stem explants of the same studied pepper varieties the addition of the vitamins C. B12. Casein hydrolysate and Ferulic acid had a stimulation effect on the plant growth in height and rooting. In result of anther cultivation from three pepper varieties and four breeding lines the highest percentage of embryo structure formation was registered in varieties Albena and Strjama (12.0 and 13.8 respectively). The Bulgarian peppers are recalcitrant and their in vitro answer is different depending from the explants type, genotype and the culture media composition.



2020 ◽  
Vol 21 (11) ◽  
Author(s):  
Yupi ISNAINI ◽  
Titien Ngatinem Praptosuwiryo

Abstract. Isnaini Y, Praptosuwiryo TNg. 2020. In vitro spore germination and early gametophyte development of Cibotium barometz (L.) J. Sm. in different media. Biodiversitas 21: 5373-5381. Cibotium barometz (L.) J. Sm. is known as the golden chicken fern and included in Appendix II of CITES. It is an important export commodity for traditional and modern medicine. Globally, populations of this species are under significant pressure due to overexploitation in the wild. In vitro culture is one of the technologies used for ex-situ propagation and conservation of rare and endangered ferns and lycophytes. This study’s objectives were: (i) to observe in vitro spore germination and early gametophyte development of C. barometz, and (ii) to determine the best culture medium for rapid spore germination and early development of the gametophytes. The sterilized spores were sown in half-strength Murashige & Skoog (½MS) basal medium supplemented with combinations of 6-Benzylaminopurine (BAP) and α-Naphthalene acetic acid (NAA). A factorial combination of four BAP concentrations (0, 2, 4, and 6 mg L-1) with four concentrations of NAA (0; 0.01; 0.03 and 0.05 mg L-1) created 16 treatments replicated in a Completely Randomized Design. Spore germination of C. barometz was observed to be Vittaria-type, and its prothallial development was Drynaria-type. Spore germination started 7-14 days after sowing. Young heart-shape gametophytes consisting of 110-240 cells were formed in 45-61 days after sowing. The two best spore culture media for rapid spore germination and development of C. barometz gametophytes were ½ MS with or without 2 mg L-1 BAP.



Author(s):  
Odilon D. Kaboré ◽  
Sylvain Godreuil ◽  
Michel Drancourt

Traditionally recognized as environmental bacteria, Planctomycetes have just been linked recently to human pathology as opportunistic pathogens, arousing a great interest for clinical microbiologists. However, the lack of appropriate culture media limits our future investigations as no Planctomycetes have ever been isolated from patients’ specimens despite several attempts. Several Planctomycetes have no cultivable members and are only recognized by 16S rRNA gene sequence detection and analysis. The cultured representatives are slow-growing fastidious bacteria and mostly difficult to culture on synthetic media. Accordingly, the provision of environmental and nutritional conditions like those existing in the natural habitat where yet uncultured/refractory bacteria can be detected might be an option for their potential isolation. Hence, we systematically reviewed the various natural habitats of Planctomycetes, to review their nutritional requirements, the physicochemical characteristics of their natural ecological niches, current methods of cultivation of the Planctomycetes and gaps, from a perspective of collecting data in order to optimize conditions and the protocols of cultivation of these fastidious bacteria. Planctomycetes are widespread in freshwater, seawater, and terrestrial environments, essentially associated to particles or organisms like macroalgae, marine sponges, and lichens, depending on the species and metabolizable polysaccharides by their sulfatases. Most Planctomycetes grow in nutrient-poor oligotrophic environments with pH ranging from 3.4 to 11, but a few strains can also grow in quite nutrient rich media like M600/M14. Also, a seasonality variation of abundance is observed, and bloom occurs in summer-early autumn, correlating with the strong growth of algae in the marine environments. Most Planctomycetes are mesophilic, but with a few Planctomycetes being thermophilic (50°C to 60°C). Commonly added nutrients are N-acetyl-glucosamine, yeast-extracts, peptone, and some oligo and macro-elements. A biphasic host-associated extract (macroalgae, sponge extract) conjugated with a diluted basal medium should provide favorable results for the success of isolation in pure culture.



1982 ◽  
Vol 60 (8) ◽  
pp. 1495-1502 ◽  
Author(s):  
J. A. Hellebust ◽  
C. Soto ◽  
T. C. Hutchinson

Chlamydomonas angulosa shows a relatively weak capacity for heterotrophic growth on acetate in the dark. The cell doubling time on acetate in the dark is about 62 h as compared with about 17 h for growth in the absence of acetate under optimal light conditions. The cell's capacity for acetate uptake and its isocitrate lyase activity are also relatively low. No [14C]glucose is taken up by the cells, in agreement with the inability of C. angulosa to grow on glucose in the dark or of glucose to stimulate growth under light-limiting conditions. The Km and Vmax for acetate uptake for cells cultured on 12 h light: 12 h dark cycle, at the end of a 12-h dark period, are 0.05 mM and 1.3 fmol cell−1 min−1, respectively. The uptake capacity increases strongly upon dark incubation, while the presence of light during short uptake experiments enhances the uptake rate. There is only a moderate 2.5 times increase in isocitrate lyase activity upon incubation of light-grown cells in the dark with acetate.Cells incubated in 100% naphthalene-saturated media undergo a rapid and complete loss of acetate uptake capacity. Media, 50% saturated with naphthalene or 100% saturated with aqueous crude oil extracts, cause an initial stimulation of acetate uptake. Media, 50% saturated with aqueous crude oil extracts cause a much longer (at least 7 days) stimulation of acetate uptake capacity. Similarly, 50% saturated-naphthalene media cause an increase in isocitrate lyase activity on a per cell basis over control cells. While 50% naphthalene-saturated media permit almost no cell division in closed acetate-containing cultures in the dark, very dramatic increases in cell size occur over long time periods. Moderate concentrations of crude oil components dissolved in algal culture media, thus, permit significant rates of acetate uptake and assimilation but inhibit cell division. It is possible that the observed stimulation of acetate uptake by moderate concentrations of crude oil components is due to the following reasons: (i) hydrocarbon-induced permeabilization of the cell membrane and (ii) an increased availability of energy from photosynthetic light reactions owing to decreased CO2 photoassimilation.



HortScience ◽  
1994 ◽  
Vol 29 (6) ◽  
pp. 683-685 ◽  
Author(s):  
Hazel Y. Wetzstein ◽  
Choongsik Kim ◽  
Harry E. Sommer

Effects of autoclaving volume, gelling agent (Bactoagar, Gel-gro, Phytagar), and basal salts [Murashige and Skoog (MS); Woody Plant Medium (WPM); Gamborg B5 (GB)] on gel strength and pH of tissue culture media were tested. Gel strength was significantly affected by gelling agent and basal medium. MS media were generally softer than comparable WPM or GB media. As the vessel volume during autoclaving decreased, gel strength significantly decreased with Phytagar and Bactoagar gelling agents; Gel-gro had greater gel strength at the intermediate volume of medium autoclave. In all cases, autoclaving resulted in a pH decrease of 0.2 to 0.5 pH units. Lower pH values were associated with softer gels. The type of gelling agent did not greatly affect the postautoclave pH; mean values among gelling agents were within 0.05 pH units. Postautoclave pH of MS medium was lower than that of WPM or GB. This study verifies the need to observe uniform sterilization protocols to maintain consistency in the chemical and physical properties of media.



HortScience ◽  
2012 ◽  
Vol 47 (10) ◽  
pp. 1495-1498 ◽  
Author(s):  
Rebecca C.-C. Hsu ◽  
Yung-I Lee

The histological and histochemical changes in developing seeds of Cypripedium debile Rchb. f., a native slipper orchid species with horticultural potential, were investigated. The effects of timing for seed collection, culture media, and cultural conditions were also examined. The optimum germination percentage occurred when mature seeds were collected and sowed on 1/4 Murashige and Skoog basal medium. Besides, the liquid culture promoted germination of mature seeds. This finding is contrary to most other Cypripedium species, which are relatively easy to germinate with immature seeds. Moreover, two notable cytological changes of C. debile were observed. First, the suspensor cell protruded beyond the micropyle opening of the inner seedcoat, making the inner seedcoat not substantial. Second, Nile red staining indicated that the deposition of cuticular material on the seedcoat was fragmentary. It is proposed that the less hydrophobic nature of the seedcoat makes mature seeds of C. debile easier to obtain water and nutrients for germination.



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