The interactive effects of temperature and silicon limitation on the psychrophilic ice diatomPseudonitszchia seriata

Pseudonitzschia seriata, isolated from sea ice in the Canadian Arctic, was grown in silicon-limited batch and semi-continuous culture to determine the effects of temperature on its nutrient utilization. Resource-saturated growth rate (μ_max) increased significantly with temperature from -1.5 to 6°C with a Q₁₀ of 1.63. The efficiency of the algae in using exogenous silicic acid for growth had no significant monotonic relationship with temperature but was significantly (P<0.05) greater in cultures at >0°C than in those at lower temperatures. Silicic acid uptake kinetics did not differ between high and low temperatures. Silicon cell quotas were significantly higher at temperatures <0°C, indicating greater silicon demand at low temperatures.P. seriata should not suffer silicon limitation in its natural ice habitat based on the observed kinetics, but its behaviour provides some support for the suggestion that temperatures <0°C are associated with diminished efficiency of nutrient utilization by cold ocean microalgae.     

DIATOM MINERALIZATION OF SILICIC ACID. VI. THE EFFECTS OF MICROTUBULE INHIBITORS ON SILICIC ACID METABOLISM IN NAVICULA SAPROPHILA1

The role of microtubules in silicon metabolism leading to valve formation was investigated in the pennate diatom Navicula saprophila Lange-Bertalot & Bonik. By using synchronized cells blocked after mitosis and cytokinesis but prior to cell wall formation, effects due to inhibition of mitosis were eliminated. Cells were treated with three anti-microtubule drugs to assess the role of microtubules. Chemical analogs to two of the drugs provided controls for inhibition not related to microtubule disruption. Although all three anti-microtubule drugs reduced cell separation at high concentrations (1 × 10^?3 M), podophyllotoxin was the only drug which reduced cell separation at concentrations lower than 1 × 10^?5 M. None of the drugs at any concentration tested affected cell viability. There was no differential inhibitory effect between the active and inactive drugs on silicic acid transport, total uptake, incorporation, or pool formation. There was no qualitative difference between silica incorporated in treated and untreated cells. A colchicine binding component was isolated from N. saprophila. The characteristics of colchicine binding suggest this component may be tubulin. Microtubules do not appear to be involved in any of the steps of silicon metabolism leading to valve formation and yet they have profound influence on the symmetry and pattern of the mineralized product, the siliceous valve.     

A stable isotope trace method to measure silicic acid uptake by marine phytoplankton.

A tracer method is described that uses the stable isotope ³⁰Si to measure rates of silicic acid uptake by diatom cultures and natural populations of marine phytoplankton. The method involves (i) incubation of organisms requiring silicic acid for growth in the presence of ³⁰Si-labeled silicic acid, (ii) collection of the resulting particulate silicon, (iii) conversion of the particulate silicon to BaSiF₆, (iv) determination of the ³⁰Si content of BaSiF₆ by solid sample mass spectrometry, and (v) calculation of the uptake rate from the ³⁰Si enrichment of the particulate matter during the incubation. The maximum overall error in the uptake rate measurement is ±10%.     

Carbon quality and soil microbial property control the latitudinal pattern in temperature sensitivity of soil microbial respiration across Chinese forest ecosystems

Understanding the temperature sensitivity (Q₁₀) of soil organic C (SOC) decomposition is critical to quantifying the climate–carbon cycle feedback and predicting the response of ecosystems to climate change. However, the driving factors of the spatial variation in Q₁₀ at a continental scale are fully unidentified. In this study, we conducted a novel incubation experiment with periodically varying temperature based on the mean annual temperature of the soil origin sites. A total of 140 soil samples were collected from 22 sites along a 3,800 km long north–south transect of forests in China, and the Q₁₀ of soil microbial respiration and corresponding environmental variables were measured. Results showed that changes in the Q₁₀ values were nonlinear with latitude, particularly showing low Q₁₀ values in subtropical forests and high Q₁₀ values in temperate forests. The soil C:N ratio was positively related to the Q₁₀ values, and coniferous forest soils with low SOC quality had higher Q₁₀ values than broadleaved forest soils with high SOC quality, which supported the “C quality temperature” hypothesis. Out of the spatial variations in Q₁₀ across all ecosystems, gram‐negative bacteria exhibited the most importance in regulating the variation in Q₁₀ and contributed 25.1%, followed by the C:N ratio (C quality), fungi, and the fungi:bacteria ratio. However, the dominant factors that regulate the regional variations in Q₁₀ differed among the tropical, subtropical, and temperate forest ecosystems. Overall, our findings highlight the importance of C quality and microbial controls over Q₁₀ value in China's forest ecosystems. Meanwhile, C dynamics in temperate forests under a global warming scenario can be robustly predicted through the incorporation of substrate quality and microbial property into models.     

全球陆地生态系统光合作用与呼吸作用的温度敏感性

基于全球647套通量数据,定量分析了全球尺度下生态系统光合作用和呼吸作用的温度敏感性(Q10)随纬度、气候和植被的分布规律。结果表明:在全球尺度下,光合作用和呼吸过程的温度敏感性(Q10,G和Q10,R)都随纬度的升高而增加,其中Q10,G和Q10,R的均值分别为3.99±0.21和2.28±0.074。除热带多树草原、常绿落叶林外,Q10,G均大于Q10,R值。不同植被类型的温度敏感性存在显著性差异,表现为:针叶林阔叶林;落叶林常绿林,其中生态系统的季节性变异是造成差异的主要原因。当植被类型和纬度区域共同影响Q10值时,植被类型对Q10值的总变异贡献更大。气候类型对Q10,G和Q10,R都有显著影响。在气候带上,干旱带的Q10,G最小,而冷温带的Q10,G最高。不同气候类型下(除温带草原气候外)的Q10,G都大于Q10,R。在极端条件下,温度可能不在是主导因素,而水分对温度敏感性的影响不可忽略,今后的研究需要更多的关注生态系统温度敏感性对水分变化的响应。     

Reduction of metabolism during hibernation and daily torpor in mammals and birds: temperature effect or physiological inhibition?

Summary The present study addresses the controversy of whether the reduction in energy metabolism during torpor in endotherms is strictly a physical effect of temperature (Q₁₀) or whether it involves an additional metabolic inhibition. Basal metabolic rates (BMR; measured as oxygen consumption, ), metabolic rates during torpor, and the corresponding body temperatures (T _b) in 68 mammalian and avian species were assembled from the literature (n=58) or determined in the present study (n=10). The Q₁₀ for change in between normothermia and torpor decreased from a mean of 4.1 to 2.8 with decreasingT _b from 30 to <10°C in hibernators (species that show prolonged torpor). In daily heterotherms (species that show shallow, daily torpor) the Q₁₀ remained at a constant value of 2.2 asT _b decreased. In hibernators with aT _b<10°C, the Q₁₀ was inversely related to body mass. The increase of mass-specific metabolic rate with decreasing body mass, observed during normothermia (BMR), was not observed during torpor in hibernators and the slope relating metabolic rate and mass was almost zero. In daily heterotherms, which had a smaller Q₁₀ than the hibernators, no inverse relationship between the Q₁₀ and body mass was observed, and consequently the metabolic rate during torpor at the sameT _b was greater than that of hibernators. These findings show that the reduction in metabolism during torpor of daily heterotherms and large hibernators can be explained largely by temperature effects, whereas a metabolic inhibition in addition to temperature effects may be used by small hibernators to reduce energy expenditure during torpor.Abbreviation BMR basal metabolic rate     

Adaptation of soil microbial growth to temperature: Using a tropical elevation gradient to predict future changes

Terrestrial biogeochemical feedbacks to the climate are strongly modulated by the temperature response of soil microorganisms. Tropical forests, in particular, exert a major influence on global climate because they are the most productive terrestrial ecosystem. We used an elevation gradient across tropical forest in the Andes (a gradient of 20°C mean annual temperature, MAT), to test whether soil bacterial and fungal community growth responses are adapted to long‐term temperature differences. We evaluated the temperature dependency of soil bacterial and fungal growth using the leucine‐ and acetate‐incorporation methods, respectively, and determined indices for the temperature response of growth: Q₁₀ (temperature sensitivity over a given 10oC range) and T_min (the minimum temperature for growth). For both bacterial and fungal communities, increased MAT (decreased elevation) resulted in increases in Q₁₀ and T_min of growth. Across a MAT range from 6°C to 26°C, the Q₁₀ and T_min varied for bacterial growth (Q_10–20 = 2.4 to 3.5; T_min = ?8°C to ?1.5°C) and fungal growth (Q_10–20 = 2.6 to 3.6; T_min = ?6°C to ?1°C). Thus, bacteria and fungi did not differ significantly in their growth temperature responses with changes in MAT. Our findings indicate that across natural temperature gradients, each increase in MAT by 1°C results in increases in T_min of microbial growth by approximately 0.3°C and Q_10–20 by 0.05, consistent with long‐term temperature adaptation of soil microbial communities. A 2°C warming would increase microbial activity across a MAT gradient of 6°C to 26°C by 28% to 15%, respectively, and temperature adaptation of microbial communities would further increase activity by 1.2% to 0.3%. The impact of warming on microbial activity, and the related impact on soil carbon cycling, is thus greater in regions with lower MAT. These results can be used to predict future changes in the temperature response of microbial activity over different levels of warming and over large temperature ranges, extending to tropical regions.     

Growth of swimming muscles and its metabolic cost in larvae of whitefish at different temperatures

The temperature relationship of routine metabolic rate (R_r) of non-feeding, non-growing Coregonus lavaretus larvae between 2 and 15°C is characterized by Q₁₀-values ranging from l.8-2.45. The rate of growth, based on weight determinations, of first-feeding larvae amounted to 3.5, 7.6 and 9.4% day-¹ at 5, 10 and 12°C respectively, from which Q₁₀-values between 4.0 and 4.8 can be calculated. The rate of increase of muscle mass between 5 and 10°C, based on the determination of the cross-sectional area of inner muscle fibres, resulted in a Q₁₀-value of 4.5. Water temperature influenced the pattern of growth of the inner muscle fibres. At hatching, after 360 day degrees, total muscle mass of larvae reared at 4 and 8°C was independent of temperature, but at 4°C the rate of mass increase owed more to hyperplasia (increase in fibre number) than to hypertrophy (increase in fibre mass), whereas at 8°C the opposite was the case. The calculation of power budgets (including the metabolic cost of growth) of first-feeding larvae yielded net conversion efficiencies (K₂) increasing with temperature from 46.3% at 5°C to 54.7% at 12°C. Comparing our data with literature data two general conclusions can be drawn. (1) In first-feeding larvae the net, but not the gross, conversion efficiency of food energy increases with temperature. This is due to net energy input being characterized by a much higher Q₁₀-value than energy expenditures. (2) In embryos of freshwater fish so far investigated hyperplasia plays a greater role in the increase of fibre mass than hypertrophy at the lower temperature, whereas in embryos of marine fish hyperplasia prevails at the higher temperature. It is suggested that this discrepancy correlates with the high concentration of free amino acids in the eggs of marine species which provide an additional, easily available, source of metabolic energy absent in freshwater species.     

THE TOCOPHEROL,VITAMIN K,AND RELATED ISOPRENOID QUINONE COMPOSITION OF A UNICELLULAR RED ALGA (PORPHYRIDIUM CRUENTUM)1

The total lipids of axenically cultivated cells of Porphyridium cruentum were extracted with aqueous methanol-chloroform mixture and fractionated into neutral and polar lipids by silicic acid column chromatography. Thin-layer and reversed-phase paper chromatographic analyses of the neutral lipid fractions revealed the occurrence of plastoquinones (PQ) A and C, vitamin K₁ (K), ubiquinone-10 (Q₁₀), α-tocopherol (α-T), and α-tocopherolquinone (α-TQ) in the photoautotrophically cultured alga, and the same quinones but no tocopherol in the alga grown photoheterotrophically on glycerol. The plastoquinone A and vitamin K₁ were isolated, identified, and estimated by spectroscopic methods. The results indicated the following decreasing order of concentrations: autotrophic culture, PQ A > K > Q₁₀ > PQ C, α-TQ, α-T; heterotrophic culture, PQ A > Q₁₀ > K > PQ C, α-TQ. Except for the absence of plastoquinone B, the overall quinonoid composition was in general agreement with those previously reported for multicellular members of Rhodophyta, but the concentration level in total lipid was markedly lower.     

PHYSIOLOGICAL RESPONSES OF ECKLONIA RADIATA (LAMINARIALES) TO A LATITUDINAL GRADIENT IN OCEAN TEMPERATURE1

We tested the ability of sporophytes of a small kelp, Ecklonia radiata (C. Agardh) J. Agardh, to adjust their photosynthesis, respiration, and cellular processes to increasingly warm ocean climates along a latitudinal gradient in ocean temperature (～4°C). Tissue concentrations of pigment and nutrients decreased with increasing ocean temperature. Concurrently, a number of gradual changes in the metabolic balance of E. radiata took place along the latitudinal gradient. Warm‐acclimatized kelps had 50% lower photosynthetic rates and 90% lower respiration rates at the optimum temperature than did cool‐acclimatized kelps. A reduction in temperature sensitivity was also observed as a reduction in Q₁₀‐values from cool‐ to warm‐acclimatized kelps for gross photosynthesis (Q₁₀: 3.35 to 1.45) and respiration (Q₁₀: 3.82 to 1.65). Respiration rates were more sensitive to increasing experimental temperatures (10% higher Q₁₀‐values) than photosynthesis and had a higher optimum temperature, irrespective of sampling location. To maintain a positive carbon balance, E. radiata increased the critical light demand (E_c) exponentially with increasing experimental temperature. The temperature dependency of E_c was, however, weakened with increasing ocean temperature, such that the critical light demand was relaxed in kelp acclimated to higher ocean temperatures. Nevertheless, calculations of critical depth limits suggested that direct effects of future temperature increases are unlikely to be as strong as effects of reduced water clarity, another globally increasing problem in coastal areas.     

Ventilation and control of acid-base status during temperature acclimation in the crab,Cancer magister

Summary Frequencies of scaphognathite (ventilatory,f _sc) and heart (f _h) pumping, oxygen consumption ( ), and hemolymph oxygen, carbon dioxide and pH levels were measured in adult Dungeness crabs (Cancer magister) during 7–10 day periods of exposure to 7, 12, and 17°C seawater. Ventilation volume ( ) was calculated for individual animals fromf _sc and a previously determined relationship between stroke volume and animal mass. increases (Q₁₀=2.3) with temperature were associated with larger increases inf _sc (Q₁₀=3.3) and (Q₁₀=3.5) and smaller increases inf _h (Q₁₀=1.5). The incidence of unilateral scaphognathite pumping and pausing decreased as temperature rose.Postbranchial oxygen tension was maintained in vivo but hemolymph oxygen content decreased both in vivo and in vitro as temperature rose. Postbranchial carbon dioxide tension did not change significantly but relative alkalinity was maintained as temperature rose by loss of hemolymph bicarbonate. The effects of increased ventilation volume and potential mechanisms of bicarbonate regulation are discussed.The responses of the essentially subtidalCancer magister are compared with those of subtidal, intertidal and terrestrial crabs demonstrating that the concepts of acid-base regulation developed for water and air breathing vertebrates are also applicable to water and air breathing crabs, and that intertidal crabs may exhibit transitional states.This work was supported by Grant No. A.5762 National Research Council of Canada     

SYNCHRONIZATION OF MITOCHONDRIAL DNA SYNTHESIS IN CHINESE HAMSTER CELLS (LINE CHO) DEPRIVED OF ISOLEUCINE

Mitochondrial DNA (mit-DNA) synthesis was compared in suspension cultures of Chinese hamster cells (line CHO) whose cell cycle events had been synchronized by isoleucine deprivation or mitotic selection. At hourly intervals during cell cycle progression, synchronized cells were exposed to tritiated thymidine ([³H]TdR), homogenized, and nuclei and mitochondria isolated by differential centrifugation. Mit-DNA and nuclear DNA were isolated and incorporation of radioisotope measured as counts per minute ([³H]TdR) per microgram DNA. Mit-DNA synthesis in cells synchronized by mitotic selection began after 4 h and continued for approximately 9 h. This time-course pattern resembled that of nuclear DNA synthesis. In contrast, mit-DNA synthesis in cells synchronized by isoleucine deprivation did not begin until 9–12 h after addition of isoleucine and virtually all [³H]TdR was incorporated during a 3-h interval. We have concluded from these results that mit-DNA synthesis is inhibited in CHO cells which are arrested in G₁ because of isoleucine deprivation and that addition of isoleucine stimulates synchronous synthesis of mit-DNA. We believe this method of synchronizing mit-DNA synthesis may be of value in studies of factors which regulate synthesis of mit-DNA.     

Experimental evidence for the attenuating effect of SOM protection on temperature sensitivity of SOM decomposition

The ability to predict C cycle responses to temperature changes depends on the accurate representation of temperature sensitivity (Q₁₀) of soil organic matter (SOM) decomposition in C models for different C pools and soil depths. Theoretically, Q₁₀ of SOM decomposition is determined by SOM quality and availability (referred to here as SOM protection). Here, we focus on the role of SOM protection in attenuating the intrinsic, SOM quality dependent Q₁₀. To assess the separate effects of SOM quality and protection, we incubated topsoil and subsoil samples characterized by differences in SOM protection under optimum moisture conditions at 25 °C and 35 °C. Although lower SOM quality in the subsoil should lead to a higher Q₁₀ according to kinetic theory, we observed a much lower overall temperature response in subsoil compared with the topsoil. Q₁₀ values determined for respired SOM fractions of decreasing lability within the topsoil increased from 1.9 for the most labile to 3.8 for the least labile respired SOM, whereas corresponding Q₁₀ values for the subsoil did not show this trend (Q₁₀ between 1.4 and 0.9). These results indicate the existence of a limiting factor that attenuates the intrinsic effect of SOM quality on Q₁₀ in the subsoil. A parallel incubation experiment of ¹³C‐labeled plant material added to top‐ and subsoil showed that decomposition of an unprotected C substrate of equal quality responds similarly to temperature changes in top‐ and subsoil. This further confirms that the attenuating effect on Q₁₀ in the subsoil originates from SOM protection rather than from microbial properties or other nutrient limitations. In conclusion, we found experimental evidence that SOM protection can attenuate the intrinsic Q₁₀ of SOM decomposition.     

Changes in the temperature sensitivity of SOM decomposition with grassland succession: implications for soil C sequestration

Understanding the temperature sensitivity (Q₁₀) of soil organic matter (SOM) decomposition is important for predicting soil carbon (C) sequestration in terrestrial ecosystems under warming scenarios. Whether Q₁₀ varies predictably with ecosystem succession and the ways in which the stoichiometry of input SOM influences Q₁₀ remain largely unknown. We investigate these issues using a grassland succession series from free‐grazing to 31‐year grazing‐exclusion grasslands in Inner Mongolia, and an incubation experiment performed at six temperatures (0, 5, 10, 15, 20, and 25°C) and with four substrates: control (CK), glucose (GLU), mixed grass leaf (GRA), and Medicago falcata leaf (MED). The results showed that basal soil respiration (20°C) and microbial biomass C (MBC) logarithmically decreased with grassland succession. Q₁₀ decreased logarithmically from 1.43 in free‐grazing grasslands to 1.22 in 31‐year grazing‐exclusion grasslands. Q₁₀ increased significantly with the addition of substrates, and the Q₁₀ levels increased with increase in N:C ratios of substrate. Moreover, accumulated C mineralization was controlled by the N:C ratio of newly input SOM and by incubation temperature. Changes in Q₁₀ with grassland ecosystem succession are controlled by the stoichiometry of newly input SOM, MBC, and SOM quality, and the combined effects of which could partially explain the mechanisms underlying soil C sequestration in the long‐term grazing‐exclusion grasslands in Inner Mongolia, China. The findings highlight the effect of substrate stoichiometry on Q₁₀ which requires further study.     

Rate of RNA synthesis as a function of temperature in Chinese hamster lung fibroblasts (V-79)

The rates of intracellular RNA synthesis at various temperatures between 33 and 41 °C were determined in Chinese hamster lung fibroblasts by measuring average amounts of [³H]uridine incorporated per cell per unit of time. The energy of activation and Q₂₀ for intracellular RNA synthesis were calculated from the slopes of the relative rates of RNA synthesis in hamster fibroblasts vs time, plotted on Arrhenius coordinates. The incorporation of uridine into RNA is characterized by an energy of activation of 19 200 calories/mole and a Q₁₀ of 2.71. The absolute rates of RNA synthesis were determined at various temperatures, with values ranging from 1.55 to 0.60 × 10⁻¹⁵ g RNA/min/cell at 41 to 33 °C, respectively.     

COPPER TOXICITY TO SKELETONEMA COSTATUM (BACILLARIOPHYCEAE)1,2

Copper toxicity to Skeletonema costatum (Grev.) Cleve has been studied in batch cultures of chemically defined culture media. The alga is relatively insensitive to cupric ion activity, demonstrating no effect on growth up to (Cu²⁺) = 10^?8.5 M. Cultures inoculated from stationary phase stocks exhibit a prolongation of the lag phase with increasing copper concentrations near and above the point of precipitation of the copper. The toxicity of copper is a function of the silicic acid concentration in the medium. This effect is observed in a range of Si(OH)₄ concentrations (10^?5 M to 10^?4 M) above known values for the saturation of silicon uptake kinetics, thus suggesting an influence of copper on silicate metabolism.     

Chlamydomonas reinhardtii: duration of its cell cycle and phases at growth rates affected by temperature

Synchronized cultures of the green alga Chlamydomonas reinhardtii were grown photoautotrophically under a wide range of environmental conditions including temperature (15–37°C), different mean light intensities (132, 150, 264 μmol m⁻² s⁻¹), different illumination regimes (continuous illumination or alternation of light/dark periods of different durations), and culture methods (batch or continuous culture regimes). These variable experimental approaches were chosen in order to assess the role of temperature in the timing of cell division, the length of the cell cycle and its pre- and post-commitment phases. Analysis of the effect of temperature, from 15 to 37°C, on synchronized cultures showed that the length of the cell cycle varied markedly from times as short as 14 h to as long as 36 h. We have shown that the length of the cell cycle was proportional to growth rate under any given combination of growth conditions. These findings were supported by the determination of the temperature coefficient (Q ₁₀), whose values were above the level expected for temperature-compensated processes. The data presented here show that cell cycle duration in C. reinhardtii is a function of growth rate and is not controlled by a temperature independent endogenous timer or oscillator, including a circadian one.     

Modelling respiration of vegetation: evidence for a general temperature‐dependent Q10

Temperature responses of rates of respiratory CO₂ efflux from plants, soils, and ecosystems are frequently modelled using exponential functions with a constant Q₁₀ near 2.0 (fractional change in rate with a 10 °C increase in temperature). However, we present evidence that Q₁₀ declines with short‐term increases in temperature in a predictable manner across diverse plant taxa. Thus, models using a constant Q₁₀ are biased, and use of a temperature‐corrected Q₁₀ may improve the accuracy of modelled respiratory CO₂ efflux in plants and ecosystems in response to temperature and predicted global climate changes.     

Body temperature and metabolic rate during natural hypothermia in endotherms

During daily torpor and hibernation metabolic rate is reduced to a fraction of the euthermic metabolic rate. This reduction is commonly explained by temperature effects on biochemical reactions, as described by Q ₁₀ effects or Arrhenius plots. This study shows that the degree of metabolic suppression during hypothermia can alternatively be explained by active downregulation of metabolic rate and thermoregulatory control of heat production. Heat regulation is fully adequate to predict changes in metabolic rate, and Q ₁₀ effects are not required to explain the reduction of energy requirements during hibernation and torpor.Abbreviations BMR basal metabolic rate - BW body weight - C thermal conductance - C_HL thermal conductance as derived from HL - C_HP thermal conductance as derived from HP - HL heat loss - HP heat production - MR metabolic rate - RQ respiratory quotient - T_a ambient temperature - T_b body temperature     

Ultrastructural evidence for uptake of silicon-containing silicic acid analogs byUrtica pilulifera and incorporation into cell wall silica

Summary In natural environments the stinging nettle plant,Urtica pilulifera, bears stinging cells in which electron dense silica deposits occupy a significant volume of the cell wall. Plants were grown in hydroponic solutions with and without supplements of silicic acid, the chemical form of silicon available to biological systems to determine if this plant and the stinging cells will grow normally under conditions of silicon starvation. In separate experiments, several analogs of silicic acid were added as supplements to the hydroponic solution to determine whether silicic acid binding sites had detectably different specificities for the different molecular structures of the analogs. The analogs [(R-)_nSi(-OH)_m] have the following structures (R, n, m): (1)-H, 1, 3; (2)-CH₃, 1, 3; (3)-CH₃, 2, 2; (4)-CH₃, 3, 1; (5)-CH₂CH₃, 1, 3; and (6)-C₆H₅, 1, 3. Electron microscopy was used as an assay for the uptake and incorporation of analogs into an electron dense silica-like product in the stinging cell wall. The results indicate that cell wall silica production occurred only when the analog contained at least three hydroxyl groups. The morphology and ontogeny of the plant was normal except for: 1, the appearance of green spots on the leaves when the analog contained two or more hydroxyl groups, and 2, total blockage of flowering by the two methyl derivative of silicic acid, (CH₃)₂Si(OH)₂.