Spring Mobilization of Protein Nitrogen in Apple Bark

Mobilization of protein nitrogen in the spring was studied in bark of stems of unringed and double stem-ringed apple rootstocks M.7 given different nitrogen treatments. A ready protein hydrolysis occurred; the proteins contributed the greater part of the storage nitrogen exported to the growing parts. Protein hydrolysis was little affected by the supply of newly absorbed nitrogen. Movement of nitrogen out of the bark between the rings could not be demonstrated. Protein breakdown in the isolated bark sections was slightly reduced. Arginine was the predominant amino acid in the proteins of the trees with a high level of storage nitrogen but was not conspicuous in the low-nitrogen trees. The protein composition changed little during hydrolysis. Only the share of arginine in the high-nitrogen trees dropped appreciably. It is suggested that the high-nitrogen trees possess a special storage protein characterized by a high arginine content. Analysis of the nitrogen fraction of isolated bark sections showed that the composition of the soluble nitrogen was characterized by a high level of asparagine and especially of arginine, and was quite different from the composition of the proteins. The data suggest that the asparagine in particular originated largely from transformation of the various amino acids set free during protein hydrolysis.     

苹果胚性细胞原生质体培养获得再生新梢

苹果原生质体再生愈伤组织已有报道,但再生植株的报道仅有一例。以往的工作,多采用茎、叶、叶愈伤组织或悬浮培养物分高原生质体,培养成愈伤组织后不易发生器官分化。本文首次报道从新红星苹果胚珠愈伤组织建立的胚性悬浮细胞系分离原生质体,经培养获得无根绿苗的结果。 1.材料及悬浮细胞系的建立试材为中国农科院果树所果园内14年生新红星苹     

苹果树新梢生长动态的数学模型

研究苹果树新梢生长动态,建立了一类反映植物生长动态的数学模型．所建模型不仅包含了经典的植物生理模型如 Ｌｏｇｉｓｔｉｃ方程等,而且能很好地表达不同树势的红富士新梢生长动态．     

In vitro Shoot and Root Formation in the Apple Cultivar Akero

Regulation of in vitro shoot and root formation and the histologicalorigin of newly formed shoots was studied in the apple cultivarÅkerö. Both composition of mineral elements and benzylaminopurine(BAP) concentration affected shoot multiplication. Similar numbersof shoots were obtained with Lepoivre and MS medium after twosucceeding subcultures, but Murashige and Skoog medium was preferabledue to production of longer shoots. The optimum BAP concentrationwax around 8.8 µM. Higher concentrations caused vitrifiedshoots. The rooting ability increased with numbers of subcultures.Also the concentration of indol-3-yl butyric acid (IBA) affectedrooting. A strong interaction between numbers of subculturesand IBA concentration was obtained. After insufficient numbersof subcultures, when shoots were still difficult to root, increasingIBA concentration exerted little effect on rooting. When shootshad reached an ‘easy-to-root condition’ root initiationdepended on IBA concentration, showing an optimum at 2.5 µM.Supraoptimal IBA concentrations delayed root initiation. Dark treatment of shoots during the root-initiation phase increasedrooting ability. The most positive effect was obtained at suboptimalIBA concentrations. Anatomical studies revealed both axillaryand adventitious shoots. Two kinds of adventitious structureswere demonstrated. Malus domestica Borkh. apple cultivar Åkerö, in vitro propagation, anatomy, origin of shoots     

Copper Excess Impairs Mobilization of Storage Proteins in Bean Cotyledons

Germination represents a limiting stage of plant life cycle. One of the underlying metabolic activities following imbibition of seed is the reserve mobilization. Seeds of bean (Phaseolus vulgaris L. var. soisson nain hatif) were germinated by soaking in distilled water or 200?μM CuCl(2). Storage proteins breakdown and amino acids freeing from reserve tissues were investigated. Compared to the control, Cu caused a reduction in germination rate, embryo growth, and in mobilization of cotyledonary biomass. The failure in albumin and globulin hydrolysis after the exposure to the pollutant was argued by (1) higher contents of remaining proteins than control ones, (2) persistence of some polypeptide bands resolved by polyacrylamide gel electrophoresis of albumin and globulin-rich fractions, and (3) decrease in the availability of amino acids. Nitrogen starvation in embryonic axis should be associated with the Cu-imposed delay in growth.     

The Structure of an Arabinogalactan Isolated from Bamboo Shoot

It was found that azide bound to alcohol oxidase non-covalently and caused a color change from yellow to red. Alcohol oxidase was purified with a high yield from methanol-limited chemostat-grown cells of Candida boidinii S2 as an enzyme-azide complex by a simple procedure. That is, a cell-free extract was prepared from cells treated with a cationic detergent, Cation M2, and alcohol oxidase amounted to more than 80% of the soluble protein. Inactivation of the enzyme by H₂O₂ and aldehydes was decreased in the complex. The results were discussed as to the production of various aldehydes with the yeast cells.     

The Structures of Water-soluble Polysaccharides Isolated from Bamboo Shoot

The structures of two polysaccharides reported in the previous paper were studied by means of methylation analysis and the Smith degradation. As a result, it was concluded that the water-soluble xylan consisted essentially of a (l→4)-linked β-d-xylopyranosyl chain and contained l-arabinofuranosyl residues linked through the C–l as terminal side units. Unambiguous information concerning the residues of d-glactose and d-glucuronic acid as the constituents of the xylan has not been obtained. For the arabinogalactan, evidence was obtained for an interesting structure having a backbone chain of (l→3)-linked β-d-galactopyranosyl residues to which the terminal arabinose residues were attached at the C–6 position as the most prevalent side chains.     

Growth in Axenic Culture of Isolated Shoot Apices of Eichhornia crassipes

Apical meristem culture of Eichhornia crassipes has shown that for successful regeneration, the excised meristem dome must be associated with at least the youngest leaf primordium as part of the explant and a culture medium containing coconut milk (10 %, v/v), IAA (0.1 mg/l) and kinetin (1 mg/l) as growth supplements with 2 % sucrose as carbon source.     

Mobilization of Nitrogen in Fruiting Plants of a Cultivar of Cowpea

Patterns of flow of nitrogen were constructed for the post-anthesisdevelopment of symbiotically-dependent cowpea (Vigna unguiculataWalp. cv. Vita 3-Rhizobium CB756). Nitrogen fixed after floweringcontributed 40% of the fruits' total intake of N, mobilizationof N fixed before flowering the remaining 60%. Leaflets, nodulatedroot, stem plus petioles, and peduncles contributed mobilizedN in the approximate proportions 5: 2: 1: 1 respectively. Eachfruit drew on all available current sources of N, but N fromleaves was distributed preferentially to closest fruit(s), andlower fruits monopolized the N exported from nodulated rootsduring late fruiting. Rates of nitrogen fixation declined parallel with decreasingnet photosynthesis of shoots. Leaflets at upper reproductivenodes mobilized 70–77% of their N and declined steeplyin net photosynthesis rate per unit chlorophyll or per unitribulose-l, 5-bisphosphate carboxylase (RuBPCase)² before abscisingduring mid- to late fruiting, whereas leaflets at lower vegetativenodes (1–3) mostly failed to abscise, lost 44–57%of their N and maintained photosynthetic activity throughoutfruiting. Peptide hydrolase activity was examined in extracts of leaflets,roots and nodules, by autodigestion of extracts, or in assaysusing bovine haemoglobin and purified RuBPCase isolated fromcowpea as substrates. Hydrolase activities during fruiting werebroadly related to N loss from plant organs, but asynchronyin peaks of activity against different protein substrates indicateddistinct groups of hydrolases within single organs. Hydrolaseactivity of leaflet extracts against RuBPCase was highly andpositively correlated with in vivo rates of loss of RuBPCasefrom the same leaflets, and preferential degradation of thisprotein occurred during leaflet senescence. Key words: Nitrogen, Mobilization: Cowpea     

14C-Studies on Apple Trees III. The Influence of Season on Storage and Mobilization of Labelled Compounds

The present paper reports an attempt to elucidate the storage and mobilization processes in 1-year-old apple rootstocks by studying the ¹⁴C content of different parts of the tree following application of ¹⁴CO₂. The translocation of the ¹⁴C taken up from the leaves to other parts proceeds at the highest rate during the first few days after the application of ¹⁴C. The distribution in shoots, trunk and roots after application of ¹⁴C during May, July, August, and in part of September depends in particular upon the intensity of growth in the various parts. From the lime of the application of ¹⁴C until leaf-fall, 40–50 per cent of the ¹⁴C initially absorbed disappears from the tree. After exposure to ¹⁴C during October, and in part of September, a relatively large part of the ¹⁴C applied goes to the root. In this case there is a considerable reduction in the ¹⁴C-conlent from leaf-fall to the following spring after leafing, especially in the root, although relatively speaking reduction is also considerable in the bark of the parts above ground, and it is most pronounced in the methanol (80 %) soluble fraction. The reduction takes place primarily during spring, and comprises, after application during October, for the whole tree 20–25 per cent of the ¹⁴C initially absorbed. Only 13–17 per cent of this amount was recovered in the newly developed shoots and leaves in the following June.     

小金海棠和山定子幼苗根自由空间铁累积量和活化量

不同基因型苹果幼苗根系自由空间铁累积量和活化利用能力不同。在缺铁胁迫条件下,抗缺铁的苹果基因型小金海棠幼苗与对缺铁敏感的山定于幼苗相比,根系自由空间铁累积量大,且它对此铁库的活化利用能力强。此外,供给铁源不同,在植物根中形成的自由空间铁库大小就不同;不同植物基因型对此铁库的活化利用能力也不同,因此,根自由空间铁库的大小及植物对该铁库中铁的活化能力大小可作为不同基因型苹果铁营养效率的筛选指标。     

The Effect of the Autumn Senescence of Leaves on the Internal Cycling of Nitrogen for the Spring Growth of Apple Trees

The internal cycling of nitrogen (N) has been studied in applerootstocks grown in sand culture and subjected to a constantN supply, or defoliation, or withholding the N supply in theautumn in order to manipulate the amount of N stored over thewinter. The trees subsequently received either no N or 8–0mol N m^–3 (labelled with ¹⁵N to 498 atom%) with the irrigationthe following spring in order to determine the effect of thecurrent N supply on the remobilization of N for leaf growth. Provision of an autumnal N supply delayed leaf senescence andreduced the amount of N withdrawn from leaves from 156 mg Nplant^–1 to 91 mg N plant^–1. Loss of protein ribulose1,5-bisphosphate carboxylase/oxygenase (RUBISCO) accounted for83–87% of the soluble protein N lost during leaf senescence,there being a preferential loss of RUBISCO compared with othersoluble leaf proteins. Remobilization of N from perennial woody tissues (stems androots) in the spring was used predominantly for leaf growth.The amount of N remobilized depended upon the size of the Nstore, but was unaffected by the current N supply, demonstratingthat fertilization of trees does not alter the efficiency withwhich they cycle N. Degradation of RUBISCO in the autumn accountedfor between 32% and 48% of the N subsequently remobilized forleaf growth the following spring, suggesting that RUBISCO hasa role as a summer store for N. Key words: Malus domestica, Borkh, nitrogen, senescence, ribulose 1, 5-bisphosphate carboxylase, oxygenase, storage, remobilization     

Trypanosoma cruzi Intracellular Amastigotes Isolated by Nitrogen Decompression Are Capable of Endocytosis and Cargo Storage in Reservosomes

Epimastigote forms of Trypanosoma cruzi (the etiologic agent of Chagas disease) internalize and store extracellular macromolecules in lysosome-related organelles (LROs) called reservosomes, which are positive for the cysteine protease cruzipain. Despite the importance of endocytosis for cell proliferation, macromolecule internalization remains poorly understood in the most clinically relevant proliferative form, the intracellular amastigotes found in mammalian hosts. The main obstacle was the lack of a simple method to isolate viable intracellular amastigotes from host cells. In this work we describe the fast and efficient isolation of viable intracellular amastigotes by nitrogen decompression (cavitation), which allowed the analysis of amastigote endocytosis, with direct visualization of internalized cargo inside the cells. The method routinely yielded 5x10⁷ amastigotes—with typical shape and positive for the amastigote marker Ssp4—from 5x10⁶ infected Vero cells (48h post-infection). We could visualize the endocytosis of fluorescently-labeled transferrin and albumin by isolated intracellular amastigotes using immunofluorescence microscopy; however, only transferrin endocytosis was detected by flow cytometry (and was also analyzed by western blotting), suggesting that amastigotes internalized relatively low levels of albumin. Transferrin binding to the surface of amastigotes (at 4°C) and its uptake (at 37°C) were confirmed by binding dissociation assays using acetic acid. Importantly, both transferrin and albumin co-localized with cruzipain in amastigote LROs. Our data show that isolated T. cruzi intracellular amastigotes actively ingest macromolecules from the environment and store them in cruzipain-positive LROs functionally related to epimastigote reservosomes.     

Long-Term Nitrogen Storage and Soil Nitrogen Availability in Post-Fire Lodgepole Pine Ecosystems

Long-term, landscape patterns in inorganic nitrogen (N) availability and N stocks following infrequent, stand-replacing fire are unknown but are important for interpreting the effect of disturbances on ecosystem function. Here, we present results from a replicated chronosequence study in the Greater Yellowstone Ecosystem (Wyoming, USA) directed at measuring inorganic N availability (ion-exchange resin bags) and ecosystem N pools among 77 lodgepole pine stands that varied in age and density. Inorganic N availability ranged from 0.07 to 3.20 μN bag⁻¹ d⁻¹ and nitrate (NO₃⁻) was, on average, 65% of total resin-sorbed N. Total ecosystem N stocks (live + detrital + soil) averaged 109.9 ± 3.0 g N m⁻² (range = 63.7–185.8 g N m⁻²). Live N was 14%, detrital N was 29%, and soil N was 57% of total stocks. Soil NO₃⁻, total ecosystem N, live N, and detrital N generally increased with stand age, but soil N stocks decreased. Models (AIC_c) to predict soil N availability and N stocks included soil P, soil Ca, bulk density, and pH in addition to age (adj R ² ranged from 0.18 to 0.53) and density was included only for live N stocks. Patterns of N stocks and N availability with density were strongest for young stands (<20 years) regenerating from extensive fire in 1988; for example, litterfall N stocks increased with density (adj R ² = 0.86, P < 0.001) but inorganic N availability declined (adj R ² = 0.47, P < 0.003). Across the complex Yellowstone landscape, we conclude that N stocks and N availability are best predicted by a combination of local soil characteristics in addition to factors that vary at landscape scales (stand density and age). Overall, total ecosystem N stocks were recovered quickly following stand-replacing fire, suggesting that moderate increases in fire frequency will not affect long-term landscape N storage in Greater Yellowstone. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Author contributions   EAHS, MGT, and MGR conceived the study; DMK performed field research; EAHS and DMK oversaw laboratory analyses and analyzed data; EAHS wrote the paper.     

Storage of Conidia of Penicillium chrysogenum in Liquid Nitrogen

Conidiated slope cultures of derivative of Penicillium chrysogenum Wis 54-1255 were stored at -196 or +4 C for a period of 3.5 years. After this time, the viability fell to 68% in the former case and to 4% in the latter. At the end of the experiment, 65 single conidial isolates from each series were tested for penicillin yield. Among those from conidia stored at -196 C, the spread of penicillin yields did not differ markedly from that of 65 single conidial isolates made as controls prior to storage. However, 18% of those from conidia stored at +4 C formed a subpopulation with substantially lower penicillin titers than those of control isolates. Storage at -196 C may reduce or prevent a possible source of penicillin yield decay, namely, the selection of spontaneous mutants of low titer present in small numbers in the original culture and selected, as viability decreased, by virtue of their increased longevity relative to that of the parental culture.     

Characterization of the Major Protease Involved in the Soybean beta-Conglycinin Storage Protein Mobilization

Protease C1, the protease responsible for the initial degradation of the α′ and α subunits of the soybean β-conglycinin storage protein (Glycine max [L.] Merrill), has been purified. The enzyme was found by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to have a molecular weight of 70,000 and a pH optimum of 3.5 to 4.5. Susceptibility to protease inhibitors indicates that protease C1 is a serine protease. Study of the proteolytic intermediates generated suggests that the cleavage of the α′ and α subunits of β-conglycinin by protease C1 results in intermediates that are 1 or 2 kilodaltons smaller than the native α′ and α subunits. Following that, a succession of intermediates exhibiting molecular masses of 70.0 and 58.0 kilodaltons, then 63.0, 61.0, 55.0, and 53.5 kilodaltons, are observed. A 50.0- and a 48.0- kilodalton intermediate are the final products of protease C1 action. Comparison of these intermediates with the prominent anti-β-conglycinin cross-reacting bands that increase during the first few days of germination and early growth show that protease C1 plays an important physiological role, but not an exclusive one, in the living plant.     

Occurrence of a Temperature-induced Phase Transition in Mitochondria Isolated from Apple Fruit

Mitochondria were isolated from fruit of six cultivars of apples differing in susceptibility to the physiological disorder, low temperature breakdown. The state 3 rate of succinate-dependent oxygen uptake and the motion of a spin label were measured at from 0 to 25 C. Arrhenius plots of the data showed that the apparent energy of activation of both respiration and motion of the spin label increased abruptly at low temperatures indicative of a temperature-induced phase change in the membrane lipids. The changes were detected with mitochondria from all of the cultivars, but the temperature at which the changes occurred did not correlate with the susceptibility of the cultivars to low temperature breakdown.     

Long-Term Storage of Infective Microsporidian Spores in Liquid Nitrogen

Thirty-one species of microsporidia, isolated from insects and stored in liquid nitrogen for up to 25 yr, were infectious when removed from liquid nitrogen. The natural hosts of all of these microsporidia were terrestrial insects, representing six different insect orders: Coleoptera, Diptera, Hemiptera, Hymenoptera, Lepidoptera, and Orthoptera. All microsporidia from terrestrial insects that were tested survived storage in liquid nitrogen, while Nosema algerae , a microsporidium from aquatic mosquito hosts did not survive freezing in liquid nitrogen. A Nosema species from the alfalfa weevil, Hypera postica , lost some infectivity in a water storage medium after 25 yr in liquid nitrogen. Liquid nitrogen storage of microsporidian spores in 50% and 100% glycerol media reduced loss of infectivity and is recommended for extended storage of microsporidia from terrestrial insect hosts.