Glucose metabolism in detached leaves of tomato plants grown with ammonium and nitrate as nitrogen sources

The metabolism of exogenous glucose-¹⁴C in the light and the dark was studied in the detached leaves of tomato plants grown with ammonium nitrogen and nitrate nitrogen. In the light, ¹⁴CO₂, release and incorporation of glucose into insoluble materials were hardly affected at all by the nitrogen sources. Among the soluble labelled amino acids, serine had the greatest amount of label in the ammonium-plants while aspartate had the greatest amount in the nitrate-plants. This aspartate was synthesized from C₃-compounds by carboxylation. During dark-light transition, the change in the composition of soluble amino acids was more rapid in the ammonium-plants than in the nitrate-plants. In the dark, ¹⁴CO₂-release, which was ten times as much as in the light, was larger in the ammonium-plants than in the nitrate-plants; but the synthesis of high molecular compounds from glucose in the ammonium-plants was about half that in the nitrate-plants. So, it is considered that respiration operates sufficiently in the ammonium-plants. The effects of DCMU and a 100% O₂ atmosphere on glucose metabolism in both groups of plants were studied and the respiration of leaves in the light was discussed.     

Proteomic analysis of shade-avoidance response in tomato leaves

The aim of this project was to investigate the molecular mechanisms of shade-avoidance response in tomato ( Solanum lycopersicum) plants. Plants were grown in direct sunlight in ambient temperature and in an adjacent environment under shade cloth. Leaves were harvested, and protein expression differences were investigated using two-dimensional differential in-gel electrophoresis and nanoflow high-performance liquid chromatography-tandem mass spectrometry. Striking differences in plant physiology and protein expression were observed. Plants grown in the shade grew very tall but bore almost no fruit and displayed a dramatic reduction in the accumulation of Rubisco and a number of other metabolic enzymes. We have identified, quantified, and classified 59 protein features found to be up- or down-regulated as part of a shade-avoidance response in S. lycopersicum and correlated these with phenotypic data. A large group of proteins related to metabolism and respiration were greatly reduced in accumulation in shade-grown plants, and there was also evidence of significant proteolysis occurring. Four stress-related proteins appear to be constitutively expressed as a result of heat acclimation, while three distinct stress-related proteins appear to accumulate as part of the shade-avoidance response. The identification and functional classification of all 59 differentially accumulating proteins is presented and discussed.     

The role of antioxidative metabolism of tomato leaves in long‐term salt‐stress response

The antioxidative protection system as adaptation strategy to high soil salinity in the leaves of two tomato (Lycopersicon esculentum Mill.) hybrids (Buran F1 and Berberana F1) was investigated. Changes in the activity of superoxide‐dismutase (SOD, EC 1.15.1.1), peroxidase (POD, EC 1.11.1.7), ascorbate peroxidase (APX, EC 1.11.1.11), as well as total and oxidized ascorbate concentrations (AA and DHA) in the plant leaves subjected to three salinity levels (EC 3.80 dS m⁻¹, 6.95 dS m⁻¹, and 9.12 dS m⁻¹) relative to non‐saline control were analyzed during the fruiting phase. The obtained results clearly indicate a relation between SOD activity and AA concentration in the antioxidative protection without any peroxidase‐related H₂O₂ detoxification. Increased SOD activity accompanied by high AA concentration was noticed at all salinity levels, but the response of hybrids was specific for the particular salt concentration. The first salinity level (EC 3.80 dS m⁻¹) induced the highest level of AA in the Buran F1 (70%), while in Berberana F1 hybrid leaves the highest AA concentration (64%) was noticed at the third salinity level (9.12 dS m⁻¹). All salinity levels caused a decline in POD and APX activities in both hybrids. The possibility of a predominant role of ascorbate and SOD in the antioxidative protection of mature tomato leaves under long‐term salt stress is discussed.     

Changes in proline-14C metabolism in barley seedlings germinating at low temperature

Changes in the metabolic pattern of proline-¹⁴C were examined in barley seedlings germinated at the low temperature of 2°C (LT) and compared with those germinated at 25°C (HT). In the LT shoots. proline-¹⁴C incorporation was higher in the cationic fraction and lower in the acid-neutral fraction than that of HT, respectively. More proline-¹⁴C of LT was converted to other amino acids, especially to acidic amino acids in free amino acid state, than was the HT proline-¹⁴C which had a comparatively wide distribution.

In the LT protein fractions. more proline-¹⁴C was incorporated into the cytoplasmic protein than into the cell wall protein. On the contrary, the radioactivity of the lIT cell wall increased more distinctively. proline-¹⁴C in the two proteins of LT was a little less converted to other amino acids than in those of HT. A little higher radioactivity was found in the aspartate and glutamate of the LT protein hydrolysates. The hydroxyproline which is closely related with proline had a little lower level of radioactivity in the LT cell wall.     

Absorption, disposition, metabolism, and excretion of [3-(14)C]caffeic acid in rats

Male Sprague-Dawley rats ingested 140 × 10(6) dpm of [3-(14)C]trans-caffeic acid, and over the ensuing 72 h period, body tissues, plasma, urine, and feces were collected and the overall levels of radioactivity determined. Where sufficient radioactivity had accumulated, samples were analyzed by HPLC with online radioactivity and tandem mass spectrometric detection. Nine labeled compounds were identified, the substrate and its cis isomer, 3'-O- and 4'-O-sulfates and glucuronides of caffeic acid, 4'-O-sulfates and glucuronides of ferulic acid, and isoferulic acid-4'-O-sulfate. Four unidentified metabolites were also detected. After passing down the gastrointestinal tract, the majority of the radiolabeled metabolites were excreted in urine with minimal accumulation in plasma. Only relatively small amounts of an unidentified (14)C-labeled metabolite were expelled in feces. There was little or no accumulation of radioactivity in body tissues, including the brain. The overall recovery of radioactivity 72 h after ingestion of [3-(14)C]caffeic acid was ～80% of intake.     

Mineral deficiencies effect on resistance-related enzymes activities in tomato leaves

Abstract

Crop yield and quality are limited by different biotic and abiotic factors. This study focused on the ability of phenylalanine ammonia-lyase, ascorbate peroxidase and polyphenol oxidase to impart resistance to mineral deficiencies. Tomato plants (Lycopersicon esculentum L.) were grown for 48, 72 and 96?hr with different specific deficiencies of minerals, including Mg²⁺, Ca²⁺, Fe²⁺, PO₄^2?, K⁺, NO₃^? and SO₄^2?, and then the soluble protein content and enzymatic activities (phenylalanine ammonia-lyase, polyphenol oxidase and ascorbate peroxidase) were analyzed. The impact of the mineral deficiencies varied in the tomato leaves. Mineral deficiencies that caused early significant protein accumulation were detected in tomato leaves grown in hydroponic cultures deficient in Ca²⁺, PO₄^2? and NO₃^? after 48, 72 and 96?hr of incubation, whereas the soluble protein level was drastically reduced in tomato leaves grown in cultures deficient in Mg²⁺, Fe²⁺, K⁺ and SO₄^2? that reveal remarkable time-dependent increase in resistance-related enzymes' activities. The resistancerelated enzymes' activities were not correlated with the soluble protein levels. These findings suggest that these activities depend on the amount of foliar nutrients, particularly Mg²⁺, Fe²⁺, K⁺ and SO₄^2?. These nutrients act synergistically on the phenolic compound biosynthesis and degradation-dependent enzymes, and this synergistic process could be considered as a mechanism of acclimation for plants against specific mineral deficiencies.     

Ammonium concentration in solution affects chlorophyll meter readings in tomato leaves

Abstract

Combinations of NH₄‐N:NO₃‐N usually result in higher tomato (Lycopersicon esculentum Mill.) yields than when either form of nitrogen (N) was used alone. Leaf chlorophyll content is closely related to leaf N content, but the effect of the NH₄‐N:NO₃‐N ratio on leaf greenness was not clear. The objective of this study was to determine the influence of NH₄‐N:NO₃‐N ratios on chlorophyll meter (SPAD) readings, and evaluate the meter as a N status estimator and tomato yield predictor in greenhouse production systems. Fruit yield and SPAD readings increased as the amount of NH₄‐N in solution increased up to 25%, while higher ratios of NH₄‐N resulted in a decline in both. The N concentration in tomato leaves increased as concentration of NH₄‐N in solution increased. Fruit yield increased as chlorophyll readings increased. SPAD readings, total N in leaves, fresh weight of shoots, and fruit yield all showed a quadratic response to NH₄‐N, reaching a peak at 25 or 50% of N as NH₄‐N. SPAD readings taken at the vegetative and flowering stages of growth had the highest correlation (r²=0.54) with N concentration in leaves, but this could not be used as a reliable estimate of N status and fruit yield. Lack of correspondence between high N concentration values and fruit yield indicated a detrimental effect of NH₄‐N on chlorophyll molecules or chloroplast structure. The SPAD readings, however, may be used to determine the optimum NH₄‐N concentration in solution to maximize fruit yield.     

14-3-3 protein down-regulates key enzyme activities of nitrate and carbohydrate metabolism in potato plants

The 14-3-3 protein is one of the best candidates for coordinating all plant metabolic pathways. To verify this suggestion transgenic potato plants with repression of one (J4 and J5 plants), two (G1 plants), and six (G3 plants) constitutive 14-3-3 protein isoforms as well as plants overexpressing the 14-3-3 protein were studied. Reduction in the 14-3-3 protein level in the J4 and J5 transformants, the G1 transformants, and the G3 transformants was close to 29, 41.5, 38, and 55%, respectively. In the case of the 14-3-3 overexpressing plants (J2), a 30% increase in protein content was detected. Changes in nitrate reductase (NR), sucrose phosphate synthase (SPS), and starch synthase (SS) activities in the transgenic plants perfectly reflect the overall 14-3-3 protein level. The highest increase in enzyme activities was observed for the G3 plants and the lowest for the J4 transformants. The same was detected for the measured metabolites. The highest increase in the protein, starch, and sucrose levels was detected in the tubers from the G3 transgenic plants. Because there was almost no change in the isoform ratio in the transgenic plants when compared to the control, it is suggested that it is the overall content of the 14-3-3 protein, rather than the content of particular isoforms, which plays a crucial role in the regulation of enzyme activities and thus in metabolite synthesis. The properties of the 14-3-3 overexpressing plants are very similar to those of the control ones, suggesting that the protein is in excess in the nontransformants and a further increase in its content is not recognized by cell metabolism. A considerable influence of the 14-3-3 protein level on potato plant metabolism was demonstrated. This effect was observed in key metabolic enzyme activities and metabolite content as well. A high variability between mean values, representing individual transgenes, with respect to nitrate reductase, sucrose phosphate synthase, and starch synthase activities in the examined genotypes was noted. These changes were closely correlated with metabolite levels, among them protein, starch, reducing sugars, and sucrose. The results obtained for the five types of transgenic potato plants in comparison with the control were statistically assessed using discriminate function and cluster analyses.     

[噻唑基-2-14C]-毒氟磷在产蛋鸡体内的分布

为阐明毒氟磷在动物体内的分布代谢特征,并深入认识毒氟磷的安全性和膳食风险,本研究选择白羽产蛋鸡为试验对象,[噻唑基-2-¹⁴C]-毒氟磷为同位素示踪剂,研究了毒氟磷在产蛋鸡体内的排泄分布特征。结果表明,毒氟磷在产蛋鸡体内排泄水平高,首次给药24 h后即排泄出当日给药量的82.04%,连续给药7 d后的累计排泄率为82.24%。毒氟磷在组织中的总残留仅占引入量的3.81%,其中胃中残留量占比相对最高,占引入量的2.14%,而肺、肾、脂肪、胰腺中的放射性残留量均不超过引入量的0.01%,膳食评估结果表明在上述内脏组织中的毒氟磷残留无膳食风险。蛋、肌肉、心、脑、脾、卵巢等组织中未检测到放射性残留。本研究为科学评价毒氟磷在家禽中的安全性提供了试验依据。     

CO2与养分交互作用对番茄幼苗叶片碳、氮积累及碳、氮比动态的影响

营养液栽培条件下,以番茄(品种,合作906)为材料,研究CO2施肥与4种不同养分供应强度的交互作用对番茄幼苗生长及其叶片中的碳、氮浓度与碳、氮比动态变化的影响。结果表明,在不同营养液养分浓度下,CO2施肥能增加番茄幼苗生物量的积累,提高生长速度;增加番茄幼苗叶片中氮、碳积累量与吸收速率;而且对CO2作用效果的响应随营养液养分浓度的提高而增加。在所有处理中碳、氮积累量与吸收速率随生育期的延长呈上升趋势。说明在番茄育苗后期要增加施肥量,而且在CO2施肥的情况下施肥量增加的量要大。CO2施肥对生长在不同营养液中番茄叶片中的碳、氮比在不同生长阶段的影响是不同的,但在同一CO2浓度条件下,番茄幼苗各个取样阶段均表现为碳、氮比随营养液浓度的降低而增加。对番茄幼苗碳、氮积累量、总干生物量与生长时间的关系研究表明,氮积累量、总干生物量与生长时间均符合二次曲线变化。     

Accumulation of copper in soils and leaves of tomato plants in greenhouses in Turkey

Copper (Cu)‐containing fertilizers, fungicides, and bactericides has been used extensively in the greenhouses in the Antalya province in recent decades with the use of such materials tending to increase year by year. The level of DTPA‐extractable Cu was determined in 210 soils (0–20 and 20–40 cm depth) and 105 leaf samples obtained from 105 tomato greenhouses. The DTPA‐extractable Cu contents of the soils taken from the 0–20 depth ranged between 0.76 and 88.03 mg kg^‐1 (mean 7.79 mg kg^‐1). The percentage of soils containing DTPA‐extractable Cu greater than the critical toxicity level (20 mg kg^‐1) was 8.1. The Cu content of tomato leaf samples ranged between 2.4 and 1,490 mg kg^‐1 (mean 166.5 mg kg^‐1). The Cu concentration in leaf samples was very high due to the intensive use of foliar applied Cu‐containing chemicals. As a consequence, 24.8% of the leaf tomato samples analyzed contained over 200 mg Cu kg^‐1, the maximum accepted tolerance level. Therefore, it may be necessary to reduce the use of Cu‐containing pesticides and fertilizers, or at least reduce the amount of Cu‐containing fertilizers being used in those greenhouses where Cu‐containing pesticides have been or are being used.     

Persistence, fate, and metabolism of [(14)C]metalaxyl in typical Indian soils

The biodegradation of ring-labeled [(14)C]metalaxyl in six Indian soils was examined. The total recovery of radioactivity from soil was 100 +/- 6% of the applied radioactivity. Volatile organics and (14)CO(2) were detected at lower levels. This suggests that neither mineralization nor volatilization is a major route of metalaxyl dissipation. The most rapid degradation of metalaxyl was observed in Bannimantap soil, in which the half-life of metalaxyl was 36 days. An inverse relationship was found when half-lives were plotted against microbial biomass and soil clay content. However, soil total organic carbon did not correlate with metalaxyl persistence. Five metabolites detected by thin-layer chromatography were more polar than metalaxyl.     

短期昼间亚高温胁迫对番茄光合作用的影响

为了探讨日光温室春末夏初亚高温环境对番茄光合作用的影响,采用人工气候室模拟环境,在番茄第1花序第1花开花当天,以25℃为对照,研究35℃昼间亚高温处理对番茄光合作用日变化的影响。结果表明：在适宜光照度条件下,3 d以下35℃昼间亚高温处理可提高番茄净光合速率,而6 d以上昼间亚高温处理可明显降低番茄的净光合速率;同时适宜温度（对照,25℃）处理的番茄净光合速率日变化未出现“光合午休”现象;而35℃昼间亚高温处理的番茄净光合速率日变化出现轻度“光合午休”现象;昼间亚高温处理可提高番茄叶片气孔导度、降低气孔限制值、增加开放气孔的比率、提高胞间CO2浓度和蒸腾速率。这些结果说明,在适宜光照条件下,番茄遭受昼间亚高温胁迫的初期可通过调节自身一些相关反应来适应环境,避免光合速率的降低,而当胁迫时间达到6 d以上时则会对光合作用产生不良影响,同时番茄“光合午休”现象不仅与光照度有关,而且与温度有关;昼间亚高温对番茄光合速率的影响与气孔无关,是由非气孔因素限制的。     

Uptake and metabolic fate of [14C]-2,4-dichlorophenol and [14C]-2,4-dichloroaniline in wheat (Triticum aestivum) and soybean (Glycine max)

The uptake and metabolism of [14C]-2,4-dichlorophenol (DCP) and [14C]-2,4-dichloroaniline (DCA) were investigated in wheat and soybean. Seeds were exposed to a nutrient solution containing 50 microM of one of two radiolabeled compounds, and plant organs were harvested separately after 18 days of growth. In wheat, uptake of [14C]-2,4-DCP was 16.67 +/- 2.65 and 15.50 +/- 2.60% of [14C]-2,4-DCA. In soybean, uptake of [14C]-2,4-DCP was significantly higher than [14C]-2,4-DCA uptake, 38.39 +/- 2.56 and 18.98 +/- 1.64%, respectively. In the case of [14C]-2,4-DCP, the radioactivity absorbed by both species was found mainly associated with roots, whereas [14C]-2,4-DCA and related metabolites were associated with aerial parts, especially in soybean. In wheat, nonextractable residues represented 7.8 and 8.7% of the applied radioactivity in the case of [14C]-2,4-DCP and [14C]-2,4-DCA, respectively. In soybean, nonextractable residues amounted to 11.8 and 5.8% of the total radioactivity for [14C]-2,4-DCP and [14C]-2,4-DCA, respectively. In wheat, nonextractable residues were nearly equivalent to extractable residues for [14C]-2,4-DCP, whereas they were greater for [14C]-2,4-DCA. In soybean, the amount of extractable residues was significantly greater for both chemicals. However, in both species, nonextractable residues were mainly associated with roots. Isolation of soluble residues was next undertaken using excised shoots (wheat) or excised fully expanded leaves including petioles (soybean). Identification of metabolite structures was made by comparison with authentic standards, by enzymatic hydrolyses, and by electrospray ionization-mass spectrometric analyses. Both plant species shared a common metabolism for [14C]-2,4-DCP and [14C]-2,4-DCA since the malonylated glucoside conjugates were found as the final major metabolites.     

Soil metabolism of a new herbicide, [14C]Pyribenzoxim, under flooded conditions

To elucidate the fate of a new pyrimidinyloxybenzoic herbicide, pyribenzoxim, a soil metabolism study was carried out with [14C]pyribenzoxim applied to a sandy loam soil under flooded conditions. The material balance of applied radioactivity ranged from 96.4 to 104.4% and from 96.1 to 101.9% for nonsterile and sterile soils, respectively. The half-life of [14C]pyribenzoxim was calculated to be approximately 1.3 and 9.4 days for nonsterile and sterile soils, respectively. The metabolites identified during the study were 2,6-bis(4,6-dimethoxypyrimidin-2-yloxy)benzoic acid (M1) and 2-hydroxy-6-(4,6-dimethoxypyrimidin-2-yloxy)benzoic acid (M2), resulting from the cleavage of the ester bond and subsequent hydrolysis. The nonextractable radioactivity levels increased to 37.8% for nonsterile conditions at 50 days after treatment and to 38.2% for sterile conditions at 60 days after treatment. Fractionation of the nonextractable soil residues indicated that bound radioactivity was associated mainly with humin fraction. No significant volatile products or [14C]carbon dioxide was observed during the study. On the basis of these results, pyribenzoxim is considered to undergo rapid degradation in soil by microbial and chemical reactions, mainly hydrolysis, which limits its transfer to and accumulation in lower soil layers and groundwater. Therefore, the possibility of environmental contamination from the use of pyribenzoxim is expected to be very low.     

基于边缘链码信息的番茄苗重叠叶面分割算法

针对番茄穴盘苗自动移栽机在检测幼苗特征参数时,由于叶面重叠导致提取特征参数不准确问题,提出一种基于重叠叶面边缘链码信息逐层分割算法。首先将穴盘苗原图像预处理得到的边缘二值图进行重叠叶面边缘拐点检测和链码统计,然后通过拐点配对进行叶面逐层分割,对真实分割拐点对进行线性插值,最终实现重叠叶面分割。试验结果表明,采用逐层重叠叶面分割算法具有旋转不变性,针对72孔和128孔2种穴盘规格,重叠叶面分割成功率分别为100%和96%,分割平均耗时分别为0.835和0.99 s。此分割算法分割速度快、精确度高,可满足实际工厂化全自动移栽作业要求。