The occurrence and nature of ornithine carbamoyltransferase in senescing apple leaf tissue

Ornithine carbamoyltransferase (EC 2.1.3.3) activity was detected in apple (Pyrus malus L.) leaf tissue from early June to November. Total activity remained relatively constant at 4.1 μmoles citrulline produced per hour per 10 cm² until mid-October when it sharply doubled. Following the first frost of the autumn, the enzyme lost about 80% of its former activity. The enzyme from apple leaf exhibited two pH optima, one at pH 8.6 and the other at pH 7.8, indicating the presence of isozymes or two forms of the enzyme. At pH 8.6, a partially-purified enzyme preparation had binding contrasts for its substrates of 6 mm for carbamyl-phosphate and 4.8 mm for ornithine. At pH 7.8, the Km for carbamyl-phosphate was 1.9 mm and the Km for ornithine was 1.22 mm.     

The appearance of chlorophyll derivatives in senescing tissue

Chlorophylls a-1 and b′, which are breakdown products of chlorophylls a and b respectively, were found in senescing leaves of Phaseolus vulgaris and Hordeum vulgare following excision from the plant. Chlorophyll a-1 was not detected in healthy plants, in senescing attached leaves or in chlorophyll-proteins isolated from senescent tissue. Chlorophyll a-1 formation in excised leaves increased with time for up to 10 days as chlorophyll a levels fell.     

Lipid composition and molecular organization in plasma membrane-enriched fractions from senescing cotyledons

Plasma membrane fractions isolated from cotyledons of Phaseolus vulgaris L. cv. Kinghorn at various stages of senescence showed no significant change in fatty acid saturation with advancing senescence. However, the steroliphospholipid ratio increased by about 400% as senescence intensified. The lipid phase transition temperature of the membranes, which was measured by wide-angle x-ray diffraction, also rose from a point well below the growing temperature for young tissue to about 50°C for membrane from extensively senescent 9-day-old tissue. This means that by day 4 of germination there was a mixture of liquid-crystalline and gel phase phospholipid in the membrane matrices. Crystallinity attributable to sterol-sterol interaction was also apparent in the diffraction patterns for senescent membranes. The co-existence of gel and liquid-crystalline phase phospholipid in the aging membranes as well as the crystalline sterol aggregates presumably render the storage cells of cotyledons leaky and may thus facilitate the translocation of hydrolyzed food reserves into the vascular network.     

Lipoxygenase-mediated production of superoxide anion in senescing plant tissue

Lipoxygenase activity and superoxide (O^.?₂) production by microsomal membranes and cytosol from bean cotyledons increased in parallel as senescence progressed. Superoxide production was heat denaturable and dependent on the availability of linoleate, the substrate for lipoxygenase. The specific inhibitor of lipoxygenase, U28938, caused a parallel reduction in enzyme activity and the formation of O^?₂. These observations demonstrate that lipoxygenase activity mediates the formation of superoxide anion, and support the contention that membrane senescence is attributable to a sequence of reactions in which lipasederived fatty acids are utilized by lipoxygenase to generate O^?₂ and hydroperoxides.     

Protoheme turnover and chlorophyll synthesis in greening barley tissue

Studies in which ¹⁴C-labeled precursors were fed to etiolated barley leaves (Hordeum vulgare L. var. Proctor) yielded chlorophyll and protoheme having similar specific radioactivities. These findings indicate: (a) there appears to be a rapid turnover of protoheme in the absence of net synthesis; (b) both pigments probably originate from a single 5-aminolevulinic acid pool; (c) the efficient utilization of glutamate-1-¹⁴C and the relatively poor utilization of glycine-2-¹⁴C suggest that 5-aminolevulinic acid is probably synthesized by a pathway other than 5-aminolevulinic acid synthetase (succinyl CoA-glycine succinyltransferase) in agreement with previously published work; (d) protoheme turnover appears to be faster under conditions which allow for rapid chlorophyll accumulation; (e) difference spectra indicate that mitochondrial cytochromes make a relatively minor contribution to the total heme in barley leaves. These findings are discussed in the light of current knowledge about tetrapyrrole regulation in photosynthetic organisms.     

Qualitative and quantitative changes in nitrogenous compounds in senescing leaf and bark tissues of the apple

Quantitative and qualitative changes in proteins and ethanol-soluble nitrogen were followed in senescing leaf and bark tissues of ‘Golden Delicious’ apple trees (Malus domestica Borkh.). While senescing leaves lost 46% of their proteins, total bark protein increased 240% during senescence. However, the protein nitrogen gain in bark tissue was about the same as the protein nitrogen loss in leaf tissue per unit fresh weight of tissues. The pattern of bark protein accumulation appears to be gradual from early August to November and sequential from lower to higher molecular weight species of proteins. The final electrophoretic profile of total bark proteins was established at the later stages of senescence. By late November, 89% of the nitrogen in the bark tissue was found in proteins with 11% in the ethanol-soluble fractions. The total protein content of dormant bark tissue was 3.5% per gram dry tissue. Fractionation of the total bark proteins by DEAE-cellulose chromatography indicated that the final upsurge of bark proteins observed in November was associated primarily with one group of proteins (Peak III).     

Studies on senescing tobacco leaf disks with special reference to peroxidase

Summary Senescing tobacco leaf disks were treated with hydroxyproline (hypro). Chlorophyll breakdown and -amino-nitrogen increase were partially inhibited, but the decrease in soluble protein was stimulated. The normal increase in absolute acid-phosphatase activity was reversed, but the increase in peroxidase activity occurred although after a lag phase. Kinetin inhibition of senescence was reversed by hypro and kinetin actually reinforced the effects of hypro. Proline only partially overcame the effects of hypro plus kinetin. In the presence of kinetin, proline considerably stimulated the increase in peroxidase activity.Discussed are the way in which hypro may be acting, and the possibility of peroxidase being involved in proline hydroxylation.     

Photosynthesis, leaf resistances, and ribulose-1,5-bisphosphate carboxylase degradation in senescing barley leaves

The relationship between loss of ribulose-1,5-bisphosphate carboxylase (RuBPCase) and the decline in photosynthesis during the senescence of barley primary leaves was assessed. Loss of RuBPCase accounted for about 85% of the decrease in soluble protein. RuBPCase was highly correlated with in vitro RuBPCase activity (r = 0.95) and gross photosynthesis (r = 0.96). However, the rate of photosynthesis per milligram RuBPCase increased during the early stages of leaf senescence. The concentration of nonreducing sugars was negatively correlated (1% level) with photosynthesis. Free α-amino N, in contrast to nonreducing sugars, declined markedly during senescence. A decrease in chlorophyll and an increase in in vitro protease activity was observed, but these changes did not appear to be closely related to the decline in photosynthesis and RuBPCase. Mesophyll resistance increased at the same rate that photosynthesis and RuBPCase declined. Stomatal resistance increased more rapidly than mesophyll resistance and accounted for about 24% of the total increase in resistance to CO₂ diffusion. The concentration of CO₂ in the intercellular air spaces decreased during the last stage of senescence. Although loss of RuBPCase probably is the primary event responsible for the decline in photosynthesis during leaf senescence, other factors such as in vivo regulation and stomatal aperture must also be considered.     

Metabolic regulation in the senescing tobacco leaf: I. Changes in pattern of p incorporation into leaf disc metabolites

Changes in phosphate metabolism were explored in discs from tobacco (Nicotiana tabacum) leaves of three contrasting types: green leaves which were fully expanded and attached to the plant, leaves which had yellowed following excision and dark starvation, and leaves which had yellowed while attached to the plant. 2,4-Dinitrophenol at 10⁻⁵m stimulated the respiration rate of discs from green and yellow-detached leaves only slightly, but markedly stimulated that of discs from yellow-attached leaves. Following a 10-minute uptake period the incorporation of ³²P-orthophosphate into phosphate esters and lipids of discs from yellow-detached leaves was resistant to 2,4-dinitrophenol, whereas in discs from green and yellow-attached leaves it was inhibited by 2,4-dinitrophenol. Incorporation into a salt-soluble fraction containing unidentified nucleotide material showed converse behavior in that it was stimulated by 2,4-dinitrophenol in discs from green and yellow-attached leaves; in discs from yellow-detached leaves it was resistant to 2,4-dinitrophenol. In discs from yellow-detached and yellow-attached leaves there was a shift in the labeling pattern of phosphate esters toward increased label in hexose phosphates at the expense of adenine nucleotides, 3-phosphoglycerate, and phosphoenolpyruvate. It is concluded that incorporation into phosphate esters in discs from yellow-detached leaves is by substrate level phosphorylation coupled to enhanced aerobic glycolysis. In discs from yellow-attached leaves, on the other hand, incorporation depends on oxidation phosphorylation, and it is suggested that the shift in labeling pattern is caused by senescence-induced changes in activity of glycolytic enzymes.     

兰州市典型绿化树种叶性状间的权衡关系

植物叶性状的权衡关系反映了植物对环境的高度适应性及其在复杂生境下的自我调控能力,了解半干旱区城市绿化树种叶性状间的权衡关系,比较不同生长型(针叶和阔叶)树种异速生长的差异有助于进一步认识植物的进化机制。以兰州市典型绿化树种(13种针叶树和47种阔叶树)为对象,测量部分叶性状,采用标准化主轴估计和系统独立比较分析的方法比较不同性状间的权衡关系。结果表明:不同生长型树种叶面积、叶体积与叶片干重之间均为等速生长关系,针叶和阔叶树种间并无显著差异;叶厚与叶面积间为"此消彼长"的权衡关系,但并不显著;阔叶叶长与叶宽间异速生长斜率为0.764。兰州市典型绿化树种叶性状间的权衡可能是长期适应黄土高原特殊生境的自适应过程,也说明部分性状间存在协变关系,阔叶绿化树种的权衡表明其符合"快速投资-收益"特征。     

Proteomic analysis of leaf peroxisomal proteins in greening cotyledons of Arabidopsis thaliana

Leaf peroxisomes are present in greening cotyledons and contain enzymes of the glycolate pathway that functions in photorespiration. However, only a few leaf peroxisomal proteins, that is hydroxypyruvate reductase (HPR), glycolate oxidase (GO) and alanine:glyoxylate aminotransferase 1 (AGT1), have been characterized, and other functions in leaf peroxisomes have not been solved. To better understand the functions of leaf peroxisomes, we established a method to isolate leaf peroxisomes of greening cotyledons. We analyzed 53 proteins by MALDI-TOF MS and then identified 29 proteins. Among them, five proteins are related to the glycolate pathway, four proteins function in scavenging of hydrogen peroxide and additionally 20 novel leaf peroxisomal proteins were identified. In particular, protein kinases and protein phosphatase were first identified as peroxisomal proteins suggesting that protein phosphorylation is one of the regulatory mechanisms in leaf peroxisomes. Novel leaf peroxisomal proteins contained five PTS1-like proteins that have sequences where one amino acid is substituted with another one in PTS1 sequences. The PTS1 motif was suggested to have novel PTS1 sequences.     

Water balance in leaf tissue

Samples of the leaf tissue (14cm²) were placed in a plexiglass chamber which consisted of three parts. Water absorbed by the leaf tissue on one side of the sample was transported through the middle part of the sample to the opposite side and was transpirated there. The intensity of transpiration the intensity of water absorption and water saturation deficit (w.s.d.) were determined simultaneously in this tissue by gravimetry. Water balance was studied either in saturated samples of leaf tissue or in tissue where w.s.d. (10%, 20%, 30%, 40%) was established in advance. Although conditions for water absorption in leaf segments were optimal, w.s.d. originated in the saturated leaf tissue under all given external conditions (evaporation from 41.7 to 17.8 mg cm^?2 h^?1). W.s.d. which was established in advance for the most part increased during the experiment and reached even high values (more than 60%). the equilibration was reached only under conditions of low evaporation and initial w.s.d. higher than 20% in young leaves and higher than 30% in adult leaves. A positive correlation between the ratio of the intensity of water absorption to the intensity of transpiration and w.s.d. was found only under conditions of lower evaporation (17.8 and 23.2mg cm^?2h^?1). The maximal values of w.s.d. were limited in this way. Water balance was studied: 1. in leaf tissue of upper, middle and lower leaves of fodder cabbage, 2. in leaf tissue of middle leaves of young and adult plants of fodder cabbage, 3. in leaf tissue of dicots (fodder cabbage) with different vessel orientation in respect to water transport, 4. in leaf tissue of monocots (banana-tree) with water transport upright to the vessel orientation. Considerable change of water balance was observed when the water transport was prolonged by two incisions in the middle part of the sample. Results of all these experiments revealed the possibility of water stress origin even in leaf tissue sufficiently supplied with water.     

Lipid biosynthesis by isolated plastids from greening pea, Pisum sativum

Isolated etioplasts from 8-day-old dark-grown pea seedlings incorporated [1-(14)C]acetate into lipid at a relatively low rate. Plastids from seedlings that had been illuminated for at least 2 hr showed an enhanced incorporation provided the plastids were illuminated during incubation with the labeled acetate. Dark incubation or the addition of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) decreased the acetate-incorporating activity of the developing chloroplasts to the level observed with etioplasts. Light had a marked effect on the type of fatty acid into which acetate was incorporated by the developing chloroplasts. Unsaturated fatty acids (mostly oleic acid) accounted for 60-80% of the incorporated label if the plastids were illuminated, but in the dark or in the presence of DCMU the unsaturated acids accounted for only 0-15% of the label incorporated into lipid. The effect of ATP on incorporation was dependent on the maturity of the chloroplasts; mature pea chloroplasts were inhibited by ATP, whereas in developing plastids there was a slight stimulation by ATP. Inhibition of acetate incorporation into lipid by DCMU appears to be due to inhibition of noncyclic phosphorylation. Incorporation was restored by reduced 2,3,5,6-tetramethylphenylenediamine, which restored phosphorylation, but not by reduced N,N,N',N'-tetramethylphenylenediamine.     

Phase transformations in a model mesenchymal tissue

Connective tissues, the most abundant tissue type of the mature mammalian body, consist of cells suspended in complex microenvironments known as extracellular matrices (ECMs). In the immature connective tissues (mesenchymes) encountered in developmental biology and tissue engineering applications, the ECMs contain varying amounts of randomly arranged fibers, and the physical state of the ECM changes as the fibers secreted by the cells undergo fibril and fiber assembly and organize into networks. In vitro composites consisting of assembling solutions of type I collagen, containing suspended polystyrene latex beads ( approximately 6 microm in diameter) with collagen-binding surface properties, provide a simplified model for certain physical aspects of developing mesenchymes. In particular, assembly-dependent topological (i.e., connectivity) transitions within the ECM could change a tissue from one in which cell-sized particles (e.g., latex beads or cells) are mechanically unlinked to one in which the particles are part of a mechanical continuum. Any particle-induced alterations in fiber organization would imply that cells could similarly establish physically distinct microdomains within tissues. Here we show that the presence of beads above a critical number density accelerates the sol-gel transition that takes place during the assembly of collagen into a globally interconnected network of fibers. The presence of this suprathreshold number of beads also dramatically changes the viscoelastic properties of the collagen matrix, but only when the initial concentration of soluble collagen is itself above a critical value. Our studies provide a starting point for the analysis of phase transformations of more complex biomaterials including developing and healing tissues as well as tissue substitutes containing living cells.     

Lipid peroxidation and antioxidative defense systems in early leaf growth

Changes in lipid peroxidation activities—superoxide dismutase (SOD), peroxidase (POD), and antioxidation—have been studied in leaves ofCucumis sativus during early growth until full expansion in situ. The content of malonyldialdehyde (MDA) rapidly decreased and then increased gradually. The changes in SOD, POD, and antioxidation activities were similar to each other and inverse to the changes in MDA contents. In the first phase, decrease in MDA is accompanied by increase in the activities of antioxidative defense systems, and in the second, exponential growth phase, increase in MDA content is accompanied by the decrease in defense systems.This study was supported by National Natural Science Foundation of China.     

Lipid peroxidation and antioxidative defense systems in early leaf growth

Changes in lipid peroxidation activities—superoxide dismutase (SOD), peroxidase (POD), and antioxidation—have been studied in leaves ofCucumis sativus during early growth until full expansion in situ. The content of malonyldialdehyde (MDA) rapidly decreased and then increased gradually. The changes in SOD, POD, and antioxidation activities were similar to each other and inverse to the changes in MDA contents. In the first phase, decrease in MDA is accompanied by increase in the activities of antioxidative defense systems, and in the second, exponential growth phase, increase in MDA content is accompanied by the decrease in defense systems.     

城市绿化植物叶片表面特征对滞尘能力的影响

以西安市21种常见绿化植物为对象,采用人工降尘方法测定植物叶片的最大滞尘量,研究植物叶片表面绒毛、润湿性、表面自由能及其分量对滞尘能力的影响.结果表明： 21种供试植物叶片的最大滞尘量在0.8～38.6 g·m^-2,不同树种最大滞尘量差异显著,物种间相差40倍以上.叶片表面绒毛数量及其形态、分布特征对滞尘能力具有重要影响,可能与绒毛和颗粒物间的作用方式有关.除叶片表面着生绒毛的悬铃木、国槐、榆叶梅和毛梾4个物种外,其他植物叶片接触角与最大滞尘量均呈显著负相关.接触角较小、易润湿的植物叶片最大滞尘量在2.0～8.0 g·m^-2,而接触角较大的银杏、三叶草、紫叶小檗和鸡爪槭的最大滞尘量均＜2.0 g·m^-2.叶片表面自由能主要表现分子间色散力的作用,而极性分量对表面自由能的贡献低于20%,可能与叶片表面含有的非极性或弱极性物质有关.最大滞尘量与叶片表面自由能及其色散分量呈显著正相关,而与极性分量的相关关系不显著.