Assessment of Arsenite,Arsenate, and Chromate Phytotoxicity Based on the Activity of Seed Germination and Growth (Root & Shoot) of Various Plant Seeds

Effects of different concentrations of arsenite, arsenate, and chromate on seed germination, root length (RL), and shoot length (SL) in four seed types, chosen from preliminary tests with eight seed types, were investigated to assess the toxicity of the tested metals. The sensitivities of the four different seed types toward germination, relative RL (RRL), and relative SL (RSL) varied with each metal. In a comparison, the germination of the seeds was more sensitive to the tested metals than the other chosen endpoints (RL and SL). Arsenite was generally more restrictive to all the endpoints (germination, root, and shoot growth) than arsenate and chromate. Lactuca (garden lettuce) was also generally more sensitive to the tested metals than the other seed types. The correlation between RRL and RSL varied depending on the seed type and metal tested. However, significant correlations (r² > 0.85) of these were observed with Lactuca seeds, which appeared to be an optimal plant with respect to the tolerance of the tested metals. The differences in the toxicities of metals toward different plant species should be taken into account in the bioassessment of metals-contaminated sites. Thus, this study encourages the need to combine the three endpoints of various seeds in the evaluation of toxicities of metals.     

A Novel ELISA Format for the Rapid and Sensitive Detection of Staphylococcal Enterotoxin A

Staphylococcal food poisoning is one of the leading causes of bacterial food poisoning each year. Detection kits for staphylococcal enterotoxins are commercially available and the assays can require from one and a half to twenty-four hours to complete with detection limits ranging from 0.5 to 2 ng enterotoxin per gram of food. We have successfully demonstrated a microsphere-packed capillary (MPC) ELISA for the detection of staphylococcal enterotoxin A (SEA) and have compared it to two commercially available kits. The MPC assay detected a lower amount of SEA in ham, chicken, cheese, and bean sprouts than either of the two commercially available kits. In addition, the novel MPC assay was completed in less than ten minutes, as compared to three and twenty-four hours for the two commercially available kits. This research also demonstrated that the MPC ELISA can contain integrated positive and negative controls and has the potential to simultaneously detect and identify multiple enterotoxins.     

Varying the ratio of 15N-labelled ammonium and nitrate-N supplied to creeping bent: effects on nitrogen absorption and assimilation, and plant growth

The relative rates of ammonium and nitrate-N uptake and assimilation by creeping bent ( Agrostis stolonifera ), were investigated for plants grown in soil and supplied with three different ratios of ammonium and nitrate-N. Following two preliminary defoliations, plants were supplied with the equivalent of 150 kg N ha⁻¹, given as ¹⁵N-(differentially) labelled NH₄⁺ and NO₃⁻-N in three different ratios (20:80, 50:50 and 80:20), followed by sequential destructive harvests of shoots and roots at four points during a 35-d regrowth period. Maximum use of labelled nitrogen and 'exhaustion' of soil mineral nitrogen reserves occurred much earlier when plants were supplied with half or more of their nitrogen as ammonium, than occurred when they were supplied predominately with nitrate-N. The lack of consistency in the patterns of ammonium and nitrate-N absorption, however, implied that the plants had no specific preference for either nitrogen form. Supplying plants with different combinations of ammonium and nitrate produced distinctive differences in plant morphology. In the high nitrate treatment, plants preferentially partitioned resources into shoot and stolon formation, whereas in the high ammonium treatment, resources were preferentially partitioned into root production. These changes in plant morphology might be adaptations to aid species survival in environments associated with a predominance of either nitrogen form.     

Determination of Monosaccharides,Disaccharides, and Oligosaccharides in the Same Plant Sample by High-Performance Liquid Chromatography

A method for determining low-molecular carbohydrates based on the use of HPLC with refractometric detection was modified. In contrast to previous methods, the determination of the qualitative and quantitative composition of sugars in plant extracts was performed without their preliminary separation into monosaccharides, disaccharides, and oligosaccharides. The exclusion of an additional separation stage made it possible to reduce twofold the time of analysis of a single sample, to save expensive materials necessary for carbohydrate HPLC, and to increase the useful life of the analytical column.     

Influences of soil volume and an elevated CO2 level on growth and CO2 exchange for the Crassulacean acid metabolism plant Opuntia ficus-indica

Effects of the current (38 Pa) and an elevated (74 Pa) CO₂ partial pressure on root and shoot areas, biomass accumulation and daily net CO₂ exchange were determined for Opuntia ficus-indica (L.) Miller, a highly productive Crassulacean acid metabolism species cultivated worldwide. Plants were grown in environmentally controlled rooms for 18 weeks in pots of three soil volumes (2 600, 6 500 and 26 000 cm³), the smallest of which was intended to restrict root growth. For plants in the medium-sized soil volume, basal cladodes tended to be thicker and areas of main and lateral roots tended to be greater as the CO₂ level was doubled. Daughter cladodes tended to be initiated sooner at the current compared with the elevated CO₂ level but total areas were similar by 10 weeks. At 10 weeks, daily net CO₂ uptake for the three soil volumes averaged 24% higher for plants growing under elevated compared with current CO₂ levels, but at 18 weeks only 3% enhancement in uptake occurred. Dry weight gain was enhanced 24% by elevated CO₂ during the first 10 weeks but only 8% over 18 weeks. Increasing the soil volume 10-fold led to a greater stimulation of daily net CO₂ uptake and biomass production than did doubling the CO₂ level. At 18 weeks, root biomass doubled and shoot biomass nearly doubled as the soil volume was increased 10-fold; the effects of soil volume tended to be greater for elevated CO₂. The amount of cladode nitrogen per unit dry weight decreased as the CO₂ level was raised and increased as soil volume increased, the latter suggesting that the effects of soil volume could be due to nitrogen limitations.     

Environmental influences on growth and defence responses of the invasive shrub,Lonicera maackii,to simulated and real herbivory in the juvenile stage

Background and Aims

Tolerance and defence against herbivory are among the many mechanisms attributed to the success of invasive plants in their novel ranges. Because tolerance and defence against herbivory differ with the ontogeny of a plant, the effects of herbivore damage on plant fitness vary with ontogenetic stage and are compounded throughout a plant''s lifetime. Environmental stresses such as light and nutrient limitations can further influence the response of the plant. Much is known about the response of plants in the seedling and reproductive adult stages, but less attention has been given to the pre-reproductive juvenile stage.

Methods

Juvenile plants of the North American invasive Lonicera maackii were exposed to simulated herbivory under high and low light and nitrogen availability and growth, allocation patterns and foliar defensive chemistry were measured. In a second experiment, complete nutrient availability and damage type (generalist caterpillar or simulated) were manipulated.

Key Results

Juvenile plants receiving 50 % defoliation had lower total biomass and a higher root^:^shoot ratio than controls for all treatment combinations except low nitrogen/low light. Low light and defoliation increased root^:^shoot ratio. Light, fertilization and defoliation had little impact on foliar defensive chemistry. In the second experiment, there was a reduction in total biomass when caterpillar damage was applied. The root^:^shoot ratio increased under low soil fertility and was not affected by defoliation. Stem-diameter growth rates and specific leaf area did not vary by damage type or fertilization. Foliar protein increased through time, and more strongly in defoliated plants than in controls, while peroxidase activity and total flavonoids decreased with time. Overall, resource limitations were more influential than damage in the growth of juvenile L. maackii plants.

Conclusions

The findings illustrate that even when resources are limited, the tolerance and defence against herbivory of a woody invasive plant in the juvenile stage may contribute to the establishment and persistence of some species in a variety of habitats.     

The effects of CO2, temperature and their interaction on the growth and yield of carrot (Daucus carota L.)

Stands of carrot (Daucus carota L.) were grown in the field within polyethylene-covered tunnels at a range of soil temperatures (from a mean of 7·5°C to 10·9°C) at either 348 (SE = 4·7) or 551 (SE = 7·7) μmol mol⁻¹ CO₂. The effect of increased atmospheric CO₂ concentration on root yield was greater than that on total biomass. At the last harvest (137d from sowing), total biomass was 16% (95% CI = 6%, 27%) greater at 551 than at 348 μmol mol⁻¹ CO₂, and 37% (95% CI = 30%, 44%) greater as a result of a 1°C rise in soil temperature. Enrichment with CO₂ or a 1°C rise in soil temperature increased root yield by 31% (95% CI = 19%, 45%) and 34% (95% CI = 27%, 42%), respectively, at this harvest. No effect on total biomass or root yield of an interaction between temperature and atmospheric CO₂ concentration at 137 DAS was detected. When compared at a given leaf number (seven leaves), CO₂ enrichment increased total biomass by 25% and root yields by 80%, but no effect of differences in temperature on plant weights was found. Thus, increases in total biomass and root yield observed in the warmer crops were a result of the effects of temperature on the timing of crop growth and development. Partitioning to the storage roots during early root expansion was greater at 551 than at 348 μmol mol⁻¹ CO₂. The root to total weight ratio was unaffected by differences in temperature at 551 μmol mol⁻¹CO₂, but was reduced by cooler temperatures at 348 μmol mol⁻¹ CO₂. At a given thermal time from sowing, CO₂ enrichment increased the leaf area per plant, particularly during early root growth, primarily as a result of an increase in the rate of leaf area expansion, and not an increase in leaf number.     

Heat Shock Increases the Tolerance of Plants to UV-B Radiation: 1. Growth,Development, and Water Supply to Tissues

The effects of heat shock (HS); UV-B irradiation; and the consecutive action of these factors on the growth, development, and water supply of seven-day-old melon (Melo sativusSager., cv. Torpeda) seedlings were investigated. Depending on the HS severity, we observed growth stimulation (after treatment at 45°C for 1 h), growth retardation (after treatment at 45°C for 2 h or at 48°C for 1 h), or complete growth inhibition and cell death (after treatment at 45°C for 3 h or at 55°C for 1 h). UV-B irradiation (18.3 kJ/(m²h)), depending on its duration, stimulated (5–10 min), retarded (60 min), or resulted in complete growth inhibition and plant death (90 min). HS treatment (at 45°C for 1 h) prior to UV-B irradiation (for 1 h) favorably affected both the growth and water balance of seedlings. Apparently, the HS pretreatment increases the tolerance of seedlings to high doses of UV-B radiation.     

NaCl胁迫对红砂萌发的影响及萌发期耐盐性评价

探究盐胁迫对红砂种子萌发及萌发后生长和生理生化指标的影响,为解析红砂萌发期耐盐机制及后续研究提供数据支撑。以盐生植物红砂种子为试验材料,NaCl溶液模拟盐胁迫,研究不同浓度NaCl(0、50、100、150、200、300和400 mmol/L)处理下,红砂种子萌发及萌发后根长、株高、生理生化指标的变化趋势。结果显示:(1)与对照相比,50 mmol/L处理的红砂萌发率升高,植株根长、株高显著增加;随着处理浓度的升高,红砂的萌发时间推迟,萌发率减少,盐害率增加,根长、株高下降显著。当盐浓度增加到400 mmol/L时,红砂不能萌发,但仍保持萌发活力,400 mmol/L胁迫35 d的未萌发种子经复萌处理后,相对萌发率为105.43%。(2)随着NaCl浓度的增加,红砂中的丙二醛(MDA)、脯氨酸(Pro)含量显著增加,过氧化物酶(POD)、超氧化物歧化酶(SOD)活性呈现不同程度的增加。(3)分别以相对萌发率、植株全长下降到对照的50%为标准,确定红砂的萌发耐盐阈值为273 mmol/L,最适盐浓度为45.78 mmol/L;萌发后生长耐盐阈值为388.19 mmol/L,生长最适盐浓度为50.59 mmol/L。红砂可在胁迫环境下长期保持萌发活力,待萌发条件适宜时,可快速集中大量萌发;红砂生长阈值浓度大于萌发阈值浓度,说明萌发期是其在盐胁迫下完成生活史的关键时期;随着盐浓度的增加,红砂的生长受到不同程度的抑制,植株在一定胁迫程度内可通过提高POD、SOD活性清除MDA,增加Pro含量从而减少胁迫危害,以维持正常的生理代谢。     

二氧化硫衍生物对玉米种子萌发、幼苗生长和细胞分裂的影响

研究3种二氧化硫衍生物硫酸钠、亚硫酸钠和亚硫酸氢钠对玉米种子萌发、幼苗生长和细胞分裂的影响。结果表明,一定浓度(0.1 mmol·L^-1)的硫酸盐和亚硫酸盐处理液能促进玉米种子萌发和幼苗生长,但浓度超过2.5 mmol·L^-1时能抑制种子萌发和幼苗生长。较高浓度的3种盐溶液对根尖分生区细胞的分裂具有抑制作用,并导致其中的部分细胞异常,出现细胞核固缩,甚至产生质壁分离。3种二氧化硫衍生物的毒性排序为：亚硫酸氢钠＞亚硫酸钠＞硫酸钠。     

Deep simple epicotyl morphophysiological dormancy in seeds of two Viburnum species, with special reference to shoot growth and development inside the seed

Background and Aims

In seeds with deep simple epicotyl morphophysiological dormancy, warm and cold stratification are required to break dormancy of the radicle and shoot, respectively. Although the shoot remains inside the seed all winter, little is known about its growth and morphological development prior to emergence in spring. The aims of the present study were to determine the temperature requirements for radicle and shoot emergence in seeds of Viburnum betulifolium and V. parvifolium and to monitor growth of the epicotyl, plumule and cotyledons in root-emerged seeds.

Methods

Fresh and pre-treated seeds of V. betulifolium and V. parvifolium were incubated under various temperature regimes and monitored for radicle and shoot emergence. Growth of the epicotyl and cotyledons at different stages was observed with dissecting and scanning electron microscopes.

Key Results

The optimum temperature for radicle emergence of seeds of both species, either kept continuously at a single regime or exposed to a sequence of regimes, was 20/10 °C. GA₃ had no effect on radicle emergence. Cold stratification (5 °C) was required for shoot emergence. The shoot apical meristem in fresh seeds did not form a bulge until the embryo had grown to the critical length for radicle emergence. After radicle emergence, the epicotyl–plumule and cotyledons grew slowly at 5 and 20/10 °C, and the first pair of true leaves was initiated. However, the shoot emerged only from seeds that received cold stratification.

Conclusions

Seeds of V. betulifolium and V. parvifolium have deep simple epicotyl morphophysiological dormancy, C_1bB (root)–C₃ (epicotyl). Warm stratification was required to break the first part of physiological dormancy (PD), thereby allowing embryo growth and subsequently radicle emergence. Although cold stratification was not required for differentiation of the epicotyl–plumule, it was required to break the second part of PD, thereby allowing the shoot to emerge in spring.     

A Rapid and Simple Method for the Determination of Picogram Levels of 3-Methoxytyramine in Brain Tissue Using Liquid Chromatography with Electrochemical Detection

Abstract: A rapid and simple technique using solvent extraction, ion-pairing extraction, and high pressure liquid chromatography with electrochemical detection has been developed for the determination of 3-methoxytyramine in striata of rats killed by microwave irradiation. The method is specific and reproducible (coefficient of variation among replications, ±4%); recovery of authentic 3-methoxytyramine added to the samples is 45–50%. 3-Methoxytyramine levels found with this technique in rat striata were 15 ± 1.7 ng/g. The method has a sensitivity of about 0.2 pmol per brain sample. Monoamine oxidase inhibition with pargyline increased 3-methoxytyramine levels in rat striata, while catechol- O -methyltransferase inhibition with 3',4'-dihydroxy-2 methylpropiophenone completely depleted 3-methoxytyramine. The effects of nomifensine, quipazine, caroxazone, piribedil, and D-amphetamine were also examined. The 3-methoxytyramine concentrations in the brains of animals killed by decapitation or by microwave irradiation were compared.     

Exogenous Calcium Enhances the Formation of Vegetative Buds,Flowers and Roots in Tobacco Pith Explants Cultured in the Absence of Exogenous Hormones

Pith explants excised from the apical stem internodes of vegetative, flowering, and fruiting tobacco plants were cultured on hormone-free medium in the presence or absence of CaCl₂ (3 mM). The aim was to determine the role of exogenous calcium (Ca²⁺), applied at the concentration normally present in the Murashige and Skoog (1962) medium, in organ formation obtainable in the absence of the exogenous hormonal input. Exclusive formation of vegetative buds was obtained from explants excised from vegetative plants (pure vegetative programme); vegetative buds and flowers (and occasionally roots) on the same sample were obtained from explants from flowering plants (mixed flowering programme); whereas roots, very occasionally associated with vegetative buds and flowers on the same sample, were obtained from explants from fruiting plants (mixed rooting programme). Histological analysis showed that the organs always exhibited indirect regeneration. Exogenous Ca²⁺ promoted the formation of meristemoids and the first phases of their growth into organs, but did not change the realization of the organogenic programme and did not affect callogenesis. Instead, the influence of exogenous Ca²⁺ changed with the programme, when considering the last phases of organ growth (i.e., macroscopic development and elongation), and the appearance of morphological anomalies in the organs.     

Predicting the impact of increasing carbon dioxide concentration and temperature on seed germination and seedling establishment of African grasses in Brazilian Cerrado

Global climate changes and biological invasions are environmental disturbances that may interact synergistically, causing loss of biodiversity. As the early stages of development are the most sensitive and easily affected by these constraints, this study investigated the effects of increased carbon dioxide (CO₂) and temperature, as forecasted for 2100, on seed germination and early development of three species of invasive African grasses that have gradually replaced landscapes of the Brazilian Cerrado biome. It was observed that these parameters affected percentage and rate of germination in Urochloa brizantha, rate of germination and mean germination time in Urochloa decumbens and accelerated autotrophy acquisition in U. brizantha, U. decumbens and Megathyrsus maximus. Regarding root elongation, all species showed changes in total length, absolute and relative growth rate, but at different stages of development or time intervals, with increased temperature being more significant than increased CO₂, probably due to seed reserves still being the main carbon sources at this stage. Taken together, the results indicate that the effects of CO₂ and increased temperature are species specific and highlight the greatest potential of U. brizantha to germinate, and of U. decumbens for seedling establishment under these environmental changes.     

A Method for the Rapid Determination of Formic and Acetic Acids in Tobacco

The yeast SPF1 gene encodes a novel P-type ATPase, the substrate of which specificity has not been identified. It is required for sensitivity to SMKT, a killer toxin produced by the halotolerant yeast Pichia farinosa. To investigate the function of Spf1p, Asp487, the putative phosphorylation site of Spf1p, was replaced by Asn. Expression of the altered SPF1, with Asp487 replaced by Asn, did not suppress the SMKT-resistant phenotype of spf1 mutants, suggesting that the catalytic activity of this ATPase is required for acquisition of sensitivity to SMKT. Subcellular fractionation experiments indicated that the fractionation pattern of Spf1p was similar to that of an early Golgi protein, Och1p. Cells lacking Spf1p had an abnormal fractionation pattern of Sec12p. The spf1 disruptant also showed increased expression of Kar2p and sensitivity to tunicamycin. The glycosylation-defective phenotype and possible role of Spf1p in the secretory pathway are discussed.     

Rootless Aquatic Plant Aldrovanda Vesiculosa: Physiological Polarity, Mineral Nutrition, and Importance of Carnivory

Various ecophysiological investigations are presented in Aldrovanda vesiculosa, a rootless aquatic carnivorous plant. A distinct polarity of N, P, and Ca tissue content per dry mass (DM) unit was found along Aldrovanda shoots. Due to effective re-utilization, relatively small proportions of N (10 – 13 %) and P (33 – 43 %) are probably lost with senescent leaf whorls, while there is complete loss of all Ca, K, and Mg. The total content of starch and free sugars was 26 – 47 % DM along adult shoots, with the maximum in the 7^th – 10^th whorls. About 30 % of the total maximum sugar content was probably lost with dead whorls. The plant was found to take up 5 – 7 times more NH₄ ⁺ to NO₃ ^– from a mineral medium. Under nearly-natural conditions in an outdoor cultivation container, catching of prey led to significantly more rapid growth than in unfed plants. DM of the fed controls was 48 % higher than in the unfed plants. The controls produced 0.69 branches per plant, while the unfed plants did not produced any. However, the N and P content per DM unit increased by 6 – 25 % in the apices and the first 6 whorls in the unfed variant, as compared to the fed controls. It may be suggested that carnivory is very important for Aldrovanda.