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Ground-based studies and space experiment with potato leaf explants.

This article details the extensive preflight research required to make a plant experiment conform to the constraints imposed by the spaceflight system. Potato explants, each consisting of a leaf, an axillary bud, and small stem section, were flown on USML-2 in the ASTROCULTURE (TM) flight hardware to study tuber formation from the axillary bud during the 16 days of flight. To obtain acceptable explant materials: 1) parent plants had to be grown under reduced light (150 micromoles m-2 s-1 PPF) to ensure uniform bud and tuber development, 2) leaves had to be trimmed to fit the small size of the flight growth chamber, and 3) only young, fully expanded leaves from plants 5-7 weeks old could be used. After six scrubs, the experiment was flown successfully October 20 to November 5 and produced tubers and accumulated starch similar to that produced on ground controls.

Culture Techniques↗

Nitrate concentration effects on NO3-N uptake and reduction, growth, and fruit yield in strawberry.

Strawberries (Fragaria xananassa Duch. 'Osogrande') were grown hydroponically with three NO3-N concentrations (3.75, 7.5, or 15.0 mM) to determine effects of varying concentration on NO3-N uptake and reduction rates, and to relate these processes to growth and fruit yield. Plants were grown for 32 weeks, and NO3-N uptake and nitrate reductase (NR) activities in roots and shoots were measured during vegetative and reproductive growth. In general, NO3-N uptake rates increased as NO3-N concentration in the hydroponics system increased. Tissue NO3- concentration also increased as external NO3-N concentration increased, reflecting the differences in uptake rates. There was no effect of external NO3-N concentration on NR activities in leaves or roots during either stage of development. Leaf NR activity averaged approximately 360 nmol NO2 formed/g fresh weight (FW)/h over both developmental stages, while NR activity in roots was much lower, averaging approximately 115 nmol NO2 formed/g FW/h. Vegetative organ FW, dry weight (DW), and total fruit yield were unaffected by NO3-N concentration. These data suggest that the inability of strawberry to increase growth and fruit yield in response to increasing NO3-N concentrations is not due to limitations in NO3-N uptake rates, but rather to limitations in NO3- reduction and/or assimilation in both roots and leaves.

Culture Media↗

Discontinuous pore fluid distribution under microgravity--KC-135 flight investigations.

Designing a reliable plant growth system for crop production in space requires the understanding of pore fluid distribution in porous media under microgravity. The objective of this experimental investigation, which was conducted aboard NASA KC-135 reduced gravity flight, is to study possible particle separation and the distribution of discontinuous wetting fluid in porous media under microgravity. KC-135 aircraft provided gravity conditions of 1, 1.8, and 10(-2) g. Glass beads of a known size distribution were used as porous media; and Hexadecane, a petroleum compound immiscible with and lighter than water, was used as wetting fluid at residual saturation. Nitrogen freezer was used to solidify the discontinuous Hexadecane ganglia in glass beads to preserve the ganglia size changes during different gravity conditions, so that the blob-size distributions (BSDs) could be measured after flight. It was concluded from this study that microgravity has little effect on the size distribution of pore fluid blobs corresponding to residual saturation of wetting fluids in porous media. The blobs showed no noticeable breakup or coalescence during microgravity. However, based on the increase in bulk volume of samples due to particle separation under microgravity, groups of particles, within which pore fluid blobs were encapsulated, appeared to have rearranged themselves under microgravity.

Alkanes↗

Spectral changes in metal halide and high-pressure sodium lamps equipped with electronic dimming.

Electronic dimming of high-intensity discharge lamps offers control of photosynthetic photon flux (PPF) but is often characterized as causing significant spectral changes. Growth chambers with 400-W metal halide (MH) and high-pressure sodium (HPS) lamps were equipped with a dimmer system using silicon-controlled rectifiers (SCR) as high-speed switches. Phase control operation turned the line power off for some period of the alternating current cycle. At full power, the electrical input to HPS and MH lamps was 480 W (root mean squared) and could be decreased to 267 W and 428 W, respectively, before the arc was extinguished. Concomitant with this decrease in input power, PPF decreased by 60% in HPS and 50% in MH. The HPS lamp has characteristic spectral peaks at 589 and 595 nm. As power to the HPS lamps was decreased, the 589-nm peak remained constant while the 595-nm peak decreased, equaling the 589-nm peak at 345-W input, and the 589-nm peak was almost absent at 270-W input. The MH lamp has a broader spectral output but also has a peak at 589 nm and another smaller peak at 545 nm. As input power to the MH lamps decreased, the peak at 589 diminished to equal the 545-nm peak. As input power approached 428 W, the 589-nm peak shifted to 570 nm. While the spectrum changed as input power was decreased in the MH and HPS lamps, the phytochrome equilibrium ratio (Pfr : Ptot) remains unchanged for both lamp types.

Dose-Response Relationship, Radiation↗

Photomorphogenesis, photosynthesis, and seed yield of wheat plants grown under red light-emitting diodes (LEDs) with and without supplemental blue lighting.

Red light-emitting diodes (LEDs) are a potential light source for growing plants in spaceflight systems because of their safety, small mass and volume, wavelength specificity, and longevity. Despite these attractive features, red LEDs must satisfy requirements for plant photosynthesis and photomorphogenesis for successful growth and seed yield. To determine the influence of gallium aluminium arsenide (GaAlAs) red LEDs on wheat photomorphogenesis, photosynthesis, and seed yield, wheat (Triticum aestivum L., cv. 'USU-Super Dwarf') plants were grown under red LEDs and compared to plants grown under daylight fluorescent (white) lamps and red LEDs supplemented with either 1% or 10% blue light from blue fluorescent (BF) lamps. Compared to white light-grown plants, wheat grown under red LEDs alone demonstrated less main culm development during vegetative growth through preanthesis, while showing a longer flag leaf at 40 DAP and greater main culm length at final harvest (70 DAP). As supplemental BF light was increased with red LEDs, shoot dry matter and net leaf photosynthesis rate increased. At final harvest, wheat grown under red LEDs alone displayed fewer subtillers and a lower seed yield compared to plants grown under white light. Wheat grown under red LEDs+10% BF light had comparable shoot dry matter accumulation and seed yield relative to wheat grown under white light. These results indicate that wheat can complete its life cycle under red LEDs alone, but larger plants and greater amounts of seed are produced in the presence of red LEDs supplemented with a quantity of blue light.

Biomass↗

Response of the water status of soybean to changes in soil water potentials controlled by the water pressure in microporous tubes.

Water transport through a microporous tube-soil-plant system was investigated by measuring the response of soil and plant water status to step change reductions in the water pressure within the tubes. Soybeans were germinated and grown in a porous ceramic 'soil' at a porous tube water pressure of -0.5 kpa for 28 d. During this time, the soil matric potential was nearly in equilibrium with tube water pressure. Water pressure in the porous tubes was then reduced to either -1.0, -1.5 or -2.0 kPa. Sap flow rates, leaf conductance and soil, root and leaf water potentials were measured before and after this change. A reduction in porous tube water pressure from -0.5 to -1.0 or -1.5 kPa did not result in any significant change in soil or plant water status. A reduction in porous tube water pressure to -2.0 kPa resulted in significant reductions in sap flow, leaf conductance, and soil, root and leaf water potentials. Hydraulic conductance, calculated as the transpiration rate/delta psi between two points in the water transport pathway, was used to analyse water transport through the tube-soil-plant continuum. At porous tube water pressures of -0.5 to-1.5 kPa soil moisture was readily available and hydraulic conductance of the plant limited water transport. At -2.0 kPa, hydraulic conductance of the bulk soil was the dominant factor in water movement.

Biological Transport↗

Effects of CO2 on stomatal conductance: do stomata open at very high CO2 concentrations?

Potato and wheat plants were grown for 50 d at 400, 1000 and 10000 micromoles mol-1 carbon dioxide (CO2). and sweetpotato and soybean were grown at 1000 micromoles mol-1 CO2 in controlled environment chambers to study stomatal conductance and plant water use. Lighting was provided with fluorescent lamps as a 12 h photoperiod with 300 micromoles m-2 s-1 PAR. Mid-day stomatal conductances for potato were greatest at 400 and 10000 micromoles mol-1 and least at 1000 micromoles mol-1 CO2. Mid-day conductances for wheat were greatest at 400 micromoles mol-1 and least at 1000 and 10000 micromoles mol-1 CO2. Mid-dark period conductances for potato were significantly greater at 10000 micromoles mol-1 than at 400 or 1000 micromoles mol-1, whereas dark conductance for wheat was similar in all CO2 treatments. Temporarily changing the CO2 concentration from the native 1000 micromoles mol-1 to 400 micromoles mol-1 increased mid-day conductance for all species, while temporarily changing from 1000 to 10000 micromoles mol-1 also increased conductance for potato and sweetpotato. Temporarily changing the dark period CO2 from 1000 to 10000 micromoles mol-1 increased conductance for potato, soybean and sweetpotato. In all cases, the stomatal responses were reversible, i.e. conductances returned to original rates following temporary changes in CO2 concentration. Canopy water use for potato was greatest at 10000, intermediate at 400, and least at 1000 micromoles mol-1 CO2, whereas canopy water use for wheat was greatest at 400 and similar at 1000 and 10000 micromoles mol-1 CO2. Elevated CO2 treatments (i.e. 1000 and 10000 micromoles mol-1) resulted in increased plant biomass for both wheat and potato relative to 400 micromoles mol-1, and no injurious effects were apparent from the 10000 micromoles mol-1 treatment. Results indicate that super-elevated CO2 (i.e. 10000 micromoles mol-1) can increase stomatal conductance in some species, particularly during the dark period, resulting in increased water use and decreased water use efficiency.

Biomass↗

Culturability as an indicator of succession in microbial communities.

Successional theory predicts that opportunistic species with high investment of energy in reproduction and wide niche width will be replaced by equilibrium species with relatively higher investment of energy in maintenance and narrower niche width as communities develop. Since the ability to rapidly grow into a detectable colony on nonselective agar medium could be considered as characteristic of opportunistic types of bacteria, the percentage of culturable cells may be an indicator of successional state in microbial communities. The ratios of culturable cells (colony forming units on R2A agar) to total cells (acridine orange direct microscopic counts) and culturable cells to active cells (reduction of 5-cyano-2,3-ditolyl tetrazolium chloride) were measured over time in two types of laboratory microcosms (the rhizosphere of hydroponically grown wheat and aerobic, continuously stirred tank reactors containing plant biomass) to determine the effectiveness of culturabilty as an index of successional state. The culturable cell:total cell ratio in the rhizosphere decreased from approximately 0.25 to less than 0.05 during the first 30-50 days of plant growth, and from 0.65 to 0.14 during the first 7 days of operation of the bioreactor. The culturable cell:active cell ratio followed similar trends, but the values were consistently greater than the culturable cell:total cell ratio, and even exceeded I in early samples. Follow-up studies used a cultivation-independent method, terminal restriction fragment length polymorphisms (TRFLP) from whole community DNA, to assess community structure. The number of TRFLP peaks increased with time, while the number of culturable types did not, indicating that the general decrease in culturability is associated with a shift in community structure. The ratio of respired to assimilated C-14-labeled amino acids increased with the age of rhizosphere communities, supporting the hypothesis that a shift in resource allocation from growth to maintenance occurs with time. Results from this work indicate that the percentage of culturable cells may be a useful method for assessing the successional state of microbial communities.

Amino Acids↗

Distribution of microbial communities associated with the dominant high marsh plants and sediments of the United States East Coast.

Microbial communities in the sediment and associated with the dominant type of standing dead plant were collected from the high marsh zones of 10 sites along the eastern coast of the United States from Maine to Florida. Microbial community composition was examined using T-RFLP, and bacterial and fungal abundance was determined microscopically. Within the sediment, community composition was strongly correlated with latitude, indicating that biogeographical factors are important determinants of sediment community composition, whereas abundance was positively and strongly correlated with sediment organic matter content. A strong biogeographical effect was observed for both bacterial and fungal abundance on standing dead plants, but there was no clear relationship between community composition and latitude. Microbial community composition was more similar among plants of the same type (i.e., related plant species) suggesting that plant type (i.e., substrate quality) is primarily responsible for the determining community composition on standing dead plants.

Bacteria↗

Dissolution kinetics of a lunar glass simulant at 25 degrees C: the effect of pH and organic acids.

The dissolution kinetics of a simulated lunar glass were examined at pH 3, 5, and 7. Additionally, the pH 7 experiments were conducted in the presence of citric and oxalic acid at concentrations of 2 and 20 mM. The organic acids were buffered at pH 7 to examine the effect of each molecule in their dissociated form. At pH 3, 5, and 7, the dissolution of the synthetic lunar glass was observed to proceed via a two-stage process. The first stage involved the parabolic release of Ca, Mg, Al, and Fe, and the linear release of Si. Dissolution was incongruent, creating a leached layer rich in Si and Ti which was verified by transmission electron microscopy (TEM). During the second stage the release of Ca, Mg, Al, and Fe was linear. A coupled diffusion/surface dissolution model was proposed for dissolution of the simulated lunar glass at pH 3, 5, and 7. During the first stage the initial release of mobile cations (i.e., Ca, Mg, Al, Fe) was limited by diffusion through the surface leached layer of the glass (parabolic release), while Si release was controlled by the hydrolysis of the Si-O-Al bonds at the glass surface (linear release). As dissolution continued, the mobile cations diffused from greater depths within the glass surface. A steady-state was then reached where the diffusion rate across the increased path lengths equalled the Si release rate from the surface. In the presence of the organic acids, the dissolution of the synthetic lunar glass proceeded by a one stage process. The release of Ca, Mg, Al, and Fe followed a parabolic relationship, while the release of Si was linear. The relative reactivity of the organic acids used in the experiments was citrate > oxalate. A thinner leached layer rich in Si/Ti, as compared to the pH experiments, was observed using TEM. Rate data suggest that the chemisorption of the organic anion to the surface silanol groups was responsible for enhanced dissolution in the presence of the organic acids. It is proposed that the increased rate of Si release is responsible for the one stage parabolic release of mobile cations and the relatively thin leached layer compared to experiments at pH 3 and 5.

Agriculture↗

Simultaneous quantification of poly-dispersed anionic, amphoteric and nonionic surfactants in simulated wastewater samples using C18 high-performance liquid chromatography-quadrupole ion-trap mass spectrometry.

This paper describes the development of a guantitative method for direct and simultaneous determination of three frequently encountered surfactants, amphoteric (cocoamphoacetate, CAA), anionic (sodium laureth sulfate, SLES), and nonionic (alcohol ethoxylate, AE) using a reversed-phase C18 HPLC coupled with an ESI ion-trap mass spectrometer (MS). Chemical composition, ionization characteristics and fragmentation pathways of the surfactants are presented. Positive ESI was effective for all three surfactants in agueous methanol buffered with ammonium acetate. The method enables rapid determinations in small sample volumes containing inorganic salts (up to 3.5 g L(-1)) and multiple classes of surfactants with high specificity by applying surfactant specific tandem mass spectrometric strategies. It has dynamic linear ranges of 2-60, 1.5-40, 0.8-56 mg L(-1) with R2 egual or greater than 0.999, 0.98 and 0.999 (10 microL injection) for CAA, SLES, and AE, respectively.

Anions↗

Microbial structural diversity estimated by dilution-extinction of phenotypic traits and T-RFLP analysis along a land-use intensification gradient.

The present work tested whether the relationship between functional traits and inoculum density reflected structural diversity in bacterial communities from a land-use intensification gradient applying a mathematical model. Terminal restriction fragment length polymorphism (T-RFLP) analysis was also performed to provide an independent assessment of species richness. Successive 10-fold dilutions of a soil suspension were inoculated onto Biolog GN(R) microplates. Soil bacterial density was determined by total cell and plate counts. The relationship between phenotypic traits and inoculum density fit the model, allowing the estimation of maximal phenotypic potential (Rmax) and inoculum density (KI) at which Rmax will be half-reduced. Though Rmax decreased with time elapsed since clearing of native vegetation, KI remained high in two of the disturbed sites. The genetic pool of bacterial community did not experience a significant reduction, but the active fraction responding in the Biolog assay was adversely affected, suggesting a reduction in the functional potential.

Bacteria↗

Relative effectiveness of kinetic analysis vs single point readings for classifying environmental samples based on community-level physiological profiles (CLPP).

The relative effectiveness of average-well-color-development-normalized single-point absorbance readings (AWCD) vs the kinetic parameters mu(m), lambda, A, and integral (AREA) of the modified Gompertz equation fit to the color development curve resulting from reduction of a redox sensitive dye from microbial respiration of 95 separate sole carbon sources in microplate wells was compared for a dilution series of rhizosphere samples from hydroponically grown wheat and potato ranging in inoculum densities of 1 x 10(4)-4 x 10(6) cells ml-1. Patterns generated with each parameter were analyzed using principal component analysis (PCA) and discriminant function analysis (DFA) to test relative resolving power. Samples of equivalent cell density (undiluted samples) were correctly classified by rhizosphere type for all parameters based on DFA analysis of the first five PC scores. Analysis of undiluted and 1:4 diluted samples resulted in misclassification of at least two of the wheat samples for all parameters except the AWCD normalized (0.50 abs. units) data, and analysis of undiluted, 1:4, and 1:16 diluted samples resulted in misclassification for all parameter types. Ordination of samples along the first principal component (PC) was correlated to inoculum density in analyses performed on all of the kinetic parameters, but no such influence was seen for AWCD-derived results. The carbon sources responsible for classification differed among the variable types with the exception of AREA and A, which were strongly correlated. These results indicate that the use of kinetic parameters for pattern analysis in CLPP may provide some additional information, but only if the influence of inoculum density is carefully considered.

Carbon↗

Effect of elevated carbon dioxide on nutritional quality of tomato.

Tomato (Lycopersicon esculentum Mill.) cvs. Red Robin (RR) and Reimann Philipp (RP) were grown hydroponically for 105 d with a 12 h photoperiod, 26 degrees C/22 degrees C thermoperiod, and 500 micromol m-2 s-1 PPF at either 400, 1200, 5000, or 10,000 micromol mol-1 (0.04, 0.12, 0.50, 1.00 kPa) CO2. Harvested fruits were analyzed for proximate composition, total dietary fiber, nitrate, and elemental composition. No trends were apparent with regard to CO2 effects on proximate composition, with fruit from all treatments and both cultivars averaging 18.9% protein, 3.6% fat, 10.2% ash, and 67.2% carbohydrate. In comparison, average values for field-grown fruit are 16.6% protein, 3.8% fat, 8.1% ash, and 71.5% carbohydrate (Duke and Atchely, 1986). Total dietary fiber was highest at 10,000 micromol mol-1 (28.4% and 22.6% for RR and RP) and lowest at 1000 micromol mol-1 (18.2% and 15.9% for RR and RP), but showed no overall trend in response to CO2. Nitrate values ranged from 0.19% to 0.35% and showed no trend with regard to CO2. K, Mg, and P concentrations showed no trend in response to CO2, but Ca levels increased from 198 and 956 ppm in RR and RP at 400 micromol mol-1, to 2537 and 2825 ppm at 10,000 micromol mol-1. This increase in Ca caused an increase in fruit Ca/P ratios from 0.07 and 0.37 for RR and RP at 400 micromol mol-1 to 0.99 and 1.23 for RR and RP at 10,000 micromol mol-1, suggesting that more dietary Ca should be available from high CO2-grown fruit.

Calcium↗

Hydroponic potato production on nutrients derived from anaerobically-processed potato plant residues.

Bioregenerative methods are being developed for recycling plant minerals from harvested inedible biomass as part of NASA's Advanced Life Support (ALS) research. Anaerobic processing produces secondary metabolites, a food source for yeast production, while providing a source of water soluble nutrients for plant growth. Since NH4-N is the nitrogen product, processing the effluent through a nitrification reactor was used to convert this to NO3-N, a more acceptable form for plants. Potato (Solanum tuberosum L.) cv. Norland plants were used to test the effects of anaerobically-produced effluent after processing through a yeast reactor or nitrification reactor. These treatments were compared to a mixed-N treatment (75:25, NO3:NH4) or a NO3-N control, both containing only reagent-grade salts. Plant growth and tuber yields were greatest in the NO3-N control and yeast reactor effluent treatments, which is noteworthy, considering the yeast reactor treatment had high organic loading in the nutrient solution and concomitant microbial activity.

Anaerobiosis↗

Atmospheric leakage and method for measurement of gas exchange rates of a crop stand at reduced pressure.

The variable pressure growth chamber (VPGC) was used in a 34-day functional test to grow a wheat crop using reduced pressure (70 kPa) episodes totalling 131 hours. Primary goals of the test were to verify facility and subsystem performance at 70 kPa and to determine responses of a wheat stand to reduced pressure and modified partial pressures of carbon dioxide and oxygen. Operation and maintenance of the chamber at 70 kpa involved continuous evacuation of the chamber atmosphere, leading to CO2 influx and efflux. A model for calculating CO2-exchange rates (net photosynthesis and dark respiration) was developed and tested and involved measurements of chamber leakage to determine appropriate corrections. Measurement of chamber leakage was based on the rate of pressure change over a small pressure increment (70.3 to 72.3 kPa) with the pump disabled. Leakage values were used to correct decreases and increases in chamber CO2 concentration resulting from net photosynthesis (Ps) and dark respiration (DR), respectively. Composite leakage corrections (influx and efflux) at day 7 of the test were 9% and 19% of the changes measured for Ps and DR, respectively. On day 33, composite corrections were only 3% for Ps and 4% for DR. During the test, the chamber became progressively tighter; the leak rate at 70.3 kPa decreasing from 2.36 chamber volumes/day pretest, to 1.71 volumes/day at the beginning of the test, and 1.16 volumes/day at the end of the test. Verification of the short-term leakage tests (rate of pressure rise) were made by testing CO2 leakage with the vacuum pump enabled and disabled. Results demonstrate the suitability of the VPGC or conducting gas exhange measurements of a crop stand at reduced pressure.

Air Conditioning↗

Glycine betaine accumulation, ionic and water relations of red-beet at contrasting levels of sodium supply.

Exposure of plants to sodium (Na) and salinity may increase glycine betaine accumulation in tissues. To study this, red-beet cvs. Scarlet Supreme and Ruby Queen, were grown for 42 days in a growth chamber using a re-circulating nutrient film technique with 0.25 mmol/L K and either 4.75 mmol/L (control) or 54.75 mmol/L (saline) Na (as NaCl). Plants were harvested at weekly intervals and measurements were taken on leaf water relations, leaf photosynthetic rates, chlorophyll fluorescence, chlorophyll levels, glycine betaine levels, and tissue elemental composition. Glycine betaine accumulation increased under salinity and this accumulation correlated with higher tissue levels of Na in both cultivars. Na accounted for 80 to 90% of the total cation uptake under the saline treatment. At final harvest (42 days), K concentrations in laminae ranged from approximately 65-95 micromoles g-1 dry matter (DM), whereas Na in shoot tissue ranged from approximately 3000-4000 micromoles g-1. Leaf sap osmotic potential at full turgor [psi(s100)] increased as lamina Na content increased. Glycine betaine levels of leaf laminae showed a linear relationship with leaf sap [psi(s100)]. Chlorophyll levels, leaf photosynthetic rates, and chlorophyll fluorescence were not affected by Na levels. These results suggest that the metabolic tolerance to high levels of tissue Na in red-beet could be due to its ability to synthesize and regulate glycine betaine production, and to control partitioning of Na and glycine betaine between the vacuole and the cytoplasm.

Beta vulgaris↗

Stomatal conductance of lettuce grown under or exposed to different light qualities.

BACKGROUND AND AIMS: The objective of this research was to examine the effects of differences in light spectrum on the stomatal conductance (Gs) and dry matter production of lettuce plants grown under a day/night cycle with different spectra, and also the effects on Gs of short-term exposure to different spectra. METHODS: Lettuce (Lactuca sativa) plants were grown with 6 h dark and 18 h light under four different spectra, red-blue (RB), red-blue-green (RBG), green (GF) and white (CWF), and Gs and plant growth were measured. KEY RESULTS AND CONCLUSIONS: Conductance of plants grown for 23 d under CWF rose rapidly on illumination to a maximum in the middle of the light period, then decreased again before the dark period when it was minimal. However, the maximum was smaller in plants grown under RB, RGB and GF. This demonstrates that spectral quality during growth affects the diurnal pattern of stomatal conductance. Although Gs was smaller in plants grown under RGB than CWF, dry mass accumulation was greater, suggesting that Gs did not limit carbon assimilation under these spectral conditions. Temporarily changing the spectral quality of the plants grown for 23 d under CWF, affected stomatal responses reversibly, confirming studies on epidermal strips. This study provides new information showing that Gs is responsive to spectral quality during growth and, in the short-term, is not directly coupled to dry matter accumulation.

Culture Media↗