Stimulation of root and somatic embryo production in Euonymus europaeus L. by an inhibitor of polyamine biosynthesis

In vitro formation of roots and somatic embryos is obtained from cotyledon explants of a Spindle tree (Euonymus europaeus L.) cultured on two different media: a medium inducing callus formation and the production of roots, and a medium inducing callus formation, root and somatic embryo production. We studied the effects of -difluoromethylornithine (DFMO), a specific, irreversible inhibitor of ornithine decarboxylase (ODC) on root and somatic embryo production, growth and titers of putrescine in Euonymus explants and explant-derived calli. Early changes in putrescine levels were detected in both cultures before the visible emergence of roots or somatic embryos. DFMO rapidly inhibited putrescine accumulation and growth in non-embryogenic calli and highly stimulated rooting activity. DFMO partially inhibited putrescine accumulation in embryogenic calli. This inhibition had no effects on callus growth but significantly reduced the time of emergence of roots and highly stimulated somatic embryo production. The relationship among putrescine, putrescine metabolism, growth, root and somatic embryo formation is discussed.     

Biochemical correlates of selection for weight-for-age in chickens: twenty-fold higher muscle ornithine decarboxylase levels in modern broilers

Summary Little is known about the biochemical correlates of selection for growth in farm or laboratory animals, or the identity of the gene products affected or produced by trait-genes. Modern broiler chickens have about 8-fold greater breast muscle mass than layer chickens at 7 weeks of age and over 2-fold greater breast muscle mass than their 1972 counterparts. This increase in muscle mass is associated with over 20-fold higher levels of ornithine decarboxylase (ODC) in broiler chickens at 1 week of age as compared with layer strain chickens; there is a comparable increase in a relaxed-selection strain of broilers. The increase in ODC levels is larger than the differences in muscle or body weight between broilers and layers at 7 weeks of age, occurs at an age when there is no difference in weights between the strains and precedes the major growth spurt. Increases in ODC levels and hence polyamine synthesis have been associated with, and usually precede, rapid growth and cell proliferation in a wide range of cell types and organisms in response to many different stimuli. Therefore, the correlation of ODC levels with genetic differences in muscle growth make it worth investigating the control of ODC gene expression in these strains.     

Regulation of N-Methyl-d-aspartate receptor-mediated calcium transport and norepinephrine release in rat hippocampus synaptosomes by polyamines

The role of polyamines (PA) synthesis in NMDA receptor-mediated⁴⁵Ca²⁺ fluxes and norepinephrine release was studied in rat hippocampal synaptosomes. NMDA (50M) caused a sharp (>2-fold) transient increase in PA synthesis regulating enzyme, ornithine decarboxylase (ODC) activity with concomitant elevation in PA levels in the order putrescine>spermidine>spermine. ODC inhibitor, -difluoromethylornithine (DFMO), and NMDA antagonist, 2-amino-5-phosphonovaleric acid (D-AP5), both blocked increases in ODC activity and PA levels. Activation of NMDA receptors induced a sharp (3 to 4-fold) and quick (15 seconds) increase in⁴⁵Ca²⁺ uptake by synaptosomes within 15 seconds of exposure at 37°C. The efflux of⁴⁵Ca²⁺ and³H-norepinephrine (NE) release at 22°C from pre-loaded synaptosomes was also significantly (2 to 4-fold) enhanced by NMDA within 15 seconds. These NMDA receptor-mediated effects on calcium fluxes and NE release were blocked by NMDA receptor-antagonists (DAP-5 and MK-801) and PA synthesis inhibitor, DFMO and the DFMO inhibition nullified by exogenous putrescine. These observations establish that ODC/PA cascade play an important role in transduction of excitatory amino acid mediated signals at NMDA receptors.Special issue dedicated to Dr. Sidney Ochs.     

Polyamine metabolism during seedling development in rice

The main free amines identified during growth and development of rice seedlings were agmatine, putrescine, spermidine, diaminopropane and tyramine. Amine composition differed according to tissue and stages of development. Conjugated amines were only found in roots. We present evidence that arginine decarboxylase (ADC) regulates putrescine during the development of rice seedlings. When ADC action was blocked by DFMA (-DL-difluoromethylarginine, a specific irreversible inhibitor of ADC), polyamine titers and seedling development were diminished; when agmatine or putrescine was added, normal polyamine titers and growth were restored. The effects of DFMA were concentration dependent. DFMO (-DL-difluoromethylornithine, a specific irreversible inhibitor of ornithine decarboxylase or ODC) promoted growth and development at concentrations below 2 mM. This effect was probably related to its unexplained, but consistently observed slight enhancement of rice ADC. When the increase in the concentration of spermidine was prevented by CHA (cyclohexylammonium sulfate), the number of roots increased and the increase in length of leaves and roots was strongly inhibited. The addition of exogenous spermidine at the time of treatment with CHA reversed the inhibition by CHA.Abbreviations ADC arginine decarboxylase - ODC ornithine decarboxylase - DFMA -DL-difluoromethylarginine - DFMO -DL-difluoromethylornithine - CHA cyclohexylammonium sulfate     

Scaled-up proliferation and regeneration of Nerine in liquid cultures Part II. Ontogeny of somatic embryos and bulblet regeneration

The ontogeny of somatic embryos was followed in liquid cultured Nerine tissue. Proliferating, nodular meristematic clusters were maintained in bubble bioreactors in a medium supplemented with 0.25 M 1-naphthalene acetic acid (NAA), 10 M 6-benzyladenine (BA) and 8.7 M Paclobutrazol (PAC). Regeneration of plantlets from this tissue was limited. Omission of PAC from the medium induced proembryogenic clusters. Embryo development and maturation were enhanced in flask cultures by substituting N⁶-(isopentenyl) adenine (2iP) for BA and elevating the sucrose concentration in the medium to 6%. High rates of embryo germination occurred in a growth regulator-free, low (3%) sucrose medium. Bulblet-bearing plantlets developed on agar-solidified, auxin-supplemented media. Recurrent embryogenesis occurred in long term growth regulator-free, or high sucrose media. The potential of using the somatic embryogenesis pathway for micropropagation of Nerines is described.     

No role of the 5′ untranslated region of ornithine decarboxylase mRNA in the feedback control of the enzyme

The polyamines are ubiquitous in nature and appear to fulfil several important functions, mostly related to growth, in the cell. The first, and often rate-limiting, step in the biosynthesis of the polyamines is catalysed by ornithine decarboxylase (ODC), which is subject to a variety of control mechanisms. The polyamines exert a strong feedback regulation of the expression - as well as the degradation of the enzyme. The regulation of ODC expression appears to occur at the translational level. The ODC mRNA contains a long GC-rich 5 untranslated region (UTR), which has been demonstrated to hamper the translation of the mRNA. However, it has not yet been conclusively established whether this part of the mRNA fulfils any function in relation to the polyamine-mediated control of ODC synthesis. In the present study, we have used stable transgenic CHO cells, expressing either full-length ODC mRNA or 5 UTR-truncated ODC mRNA, to elucidate the role, if any, of the 5 UTR in the translational regulation of the enzyme by polyamines. No differences in regulatory properties were observed between the cells expressing the full-length ODC mRNA and those expressing the ODC mRNA devoid of most the 5 UTR. The cell lines down-regulated ODC (synthesis as well as activity) to the same extent upon exposure to an excess of polyamines, demonstrating that the feedback control of ODC mRNA translation occurs by a mechanism independent of the major part of the 5 UTR of the ODC mRNA.     

Carbohydrate status in roots of two soybean cultivars: A possible parameter to explain different efficiencies concerning phosphate uptake

The carbohydrate levels in the roots of two soybean cultivars (Century, Woodworth) were determined during the early vegetative growth stage. Based on Michaelis-Menten kinetics Century was characterized as being more efficient at phosphate uptake when compared to Woodworth. The experiments were carried out at normal nutrition (+P treatment) as well as under a condition of moderate phosphate stress (–P treatment). The stress situation was created by withdrawal of phosphate from the nutrient solution for five or nine days. Compared to the +P-treatment the deprivation of phosphate led to a reduction of more than 50% in the phosphate content of the plant tissue.The carbohydrates in the roots exhibited differences between both cultivars with Century generally having higher sucrose levels. During the morning the +P treatment of Century exhibited 10 mg g^-1 and in the afternoon 16 mg g^-1 sucrose (plant dry weight basis). This compares to 8 mg g^-1 sucrose (morning) and 13 mg g^-1 sucrose (afternoon) for Woodworth. Phosphate stress increased the levels of hexoses (glucose, fructose) in both varieties significantly. The sucrose levels remained higher for the Century variety even during phosphate stress. We conclude that the shoot of the Century variety supplies its root with an increased amount of energy in form of carbohydrates compared to the Woodworth variety. This may enable the plant to take up phosphate more efficiently. It is also remarkable that the Century variety reduces the pH of the nutrient medium more rapidly. This active proton deposition may create a high membrane potential which is known to be necessary for phosphate uptake.     

Mechanistic insights into the dual inhibition strategy for checking Leishmaniasis

Leishmaniasis ^{1 1}These authors contributed equally.Communicated by Ramaswamy H. SarmaCommunicated by Ramaswamy H. Sarma is an endemic disease mainly caused by the protozoan Leishmania donovani (Ld). Polyamines have been identified as essential organic compounds for the growth and survival of Ld. These are synthesized in Ld by polyamine synthesis pathway comprising of many enzymes such as ornithine decarboxylase (ODC), spermidine synthase (SS), and S-adenosylmethionine decarboxylase. Inhibition of these enzymes in Ld offers a viable prospect to check its growth and development. In the present work, we used computational approaches to search natural inhibitors against ODC and SS enzymes. We predicted three-dimensional structures of ODC and SS using comparative modeling and molecular dynamics (MD) simulations. Thousands of natural compounds were virtually screened against target proteins using high throughput approach. MD simulations were then performed to examine molecular interactions between the screened compounds and functional residues of the active sites of the enzymes. Herein, we report two natural compounds of dual inhibitory nature active against the two crucial enzymes of polyamine pathway of Ld. These dual inhibitors have the potential to evolve as lead molecules in the development of antileishmanial drugs.     

Involvement of Polyamines in the Action of Transforming Growth Factor β and Interleukin-1 on Cultured Chick Embryo Fibroblasts

In this study we have examined the relationship between growth factor-induced proliferation and ODC/polyamine levels. TGFβ promotes cell growth and enhances [³H]-thymidine incorporation in chick embryo fibroblasts maintained in a serum-depleted medium. The action on DNA synthesis declines in the second day of treatment. IL-1 does not affect proliferation or [³H]-thymidine incorporation either when it is added alone or in combination with TGFβ. The response of the cells to TGFβ is associated with a significant stimulation of ODC activity and Put, Spd levels together with an enhancement of the Spd/polyamines ratio. IL-1, which does not act on cell proliferation, fails to activate ODC and to increase polyamine levels, thus indicating that the ODC/polyamine system is most likely to be an important link in the chain of events that leads to growth factor-induced proliferation. © 1997 John Wiley & Sons, Ltd.     

Polyamine metabolism and in vitro cell multiplication and differentiation in leaf explants of Chrysanthemum morifolium Ramat

Arginine decarboxylase (ADC), ornithine decarboxylase (ODC), diamine oxydase (DAO) free amine and conjugated amine titers were estimated in leaf explants of Chrysanthemum morifolium Ramat. var. Spinder cultivated in vitro in relation to hormone treatment. Addition of benzyladenine (BA) to a basal medium caused the formation of buds on the explants. BA plus 2,4 dichlorophenoxyacetic acid (2,4 D) caused callus formation and proliferation. Formation of roots was obtained by addition of indolylacetic acid (IAA). Arginine decarboxylase (ADC) ornithine decarboxylase (ODC) and diamine oxidase (DAO) activities increased during the first days of culture when cell multiplication was rapid, followed by a sharp decline as the rate of cell division decreased and differentiation took place. DAO activities increased rapidly in proliferating and growing organs and decreased during maturity. This increase was concomitant with ADC and ODC activities and polyamine content (free and conjugated polyamines). The biosynthesis and oxidation of polyamines which occurred simultaneously in physiological states of intense metabolism such as cell division or organ formation were directly correlated. In callus cultures DAO activity was blocked throughout development and regulated neither the cellular levels of polyamines nor polyamine conjugates. Levels of polyamine conjugates were high in callus cultures throughout development. In foliar explants cultivated on a medium promoting callus, inhibition of ODC activity by DFMO (-DL-difluoromethylornithine, a specific enzyme-activated ODC inhibitor) resulting in an amide deficiency facilated the expression of differentiated cell function; substantial activation of DAO was observed until the emergence of the buds. On a medium promoting bud formation, -OH ethylhydrazine (DAO inhibitor) promoted callus formation without differentiation. In this system DAO activity was blocked and there were high levels of polyamines, especially polyamine conjugates, throughout the culture period. The relationship among free and conjugated polyamines related biosynthetic enzyme activities, DAO activities, cell division and organ formation is discussed.Abbreviations ADC = arginine decarboxylase - ODC = ornithine decarboxylase - DOA = diamine oxidase - DFMA = -DL-difluoromethylarginine - DFMO = -DL-difluoromethylornithine - Put = putrescine     

Calcium-phosphate mediated DNA transfer into HEK-293 cells in suspension: control of physicochemical parameters allows transfection in stirred media. Transfection and protein expression in mammalian cells

This is the first report of transient transfection of suspended cells with purified plasmid DNA in bioreactors or spinner flasks. DNA/calcium phosphate complexes were pumped or injected directly into stirred cultures of the immortalized human embryo kidney cell line 293 (HEK-293) which had been adapted to growth in suspension. We identified culture conditions suitable for this approach and modified the protocol for the generation of precipitate complexes, based on our earlier work. In order to stabilize the DNA-vehicle-complex in the culture medium, we identified pH ranges and ion-concentrations which prevent dissolution or aggregation of the precipitate particles. Such conditions maintained suspended fine particles in spinners or bioreactors for up to 6 hr. During that period, cells and precipitate complexes interacted sufficiently to allow DNA transfer and subsequent expression of recombinant protein. In a simple 5 day batch process, with a starting density of 0.3 × 10⁶ cells mL^-1, about 0.5 mg L^-1 of a recombinant tissue plasminogen activator variant was observed.     

Reversal of the growth inhibitory effect of α-difluoromethylornithine by putrescine but not by other divalent cations

Summary Treatment with -difluoromethylornithine (DFMO), an enzyme-activated irreversible inhibitor of ornithine decarboxylase (ODC), depletes the putrescine and spermidine content, and reduces the growth rate of Ehrlich ascites tumor cells.The addition of putrescine, which is the immediate precursor of spermidine, promptly replenished the intracellular putrescine and spermidine pools and completely reversed the antiproliferative effect of DFMO. A sequential accumulation of spermine, spermidine and putrescine was observed.1,3-diaminopropane, a lower homolog of putrescine, did not reverse the antiproliferative effect of DFMO, despite its structural similarity and identical positive charge. By inhibiting remaining ODC activity, resistant to 5 mM DFMO, and possibly by inhibiting spermine synthase activity, 1,3-diaminopropane produced a further decrease in total polyamine content by reducing the spermine content.Mg²⁺, which can replace putrescine in many in vitro reactions, completely lacked the capacity to reverse the antiproliferative effect of putrescine and spermidine deficiency.Abbreviations DFMO -difluoromethylornithine - ODC ornithine decarbxylase     

Different effects of two brassinosteroids on growth,auxin and cytokinin content in tobacco callus tissue

Two synthetic brassinosteroids, 24-epibrassinolide (24-epiBR) and 2,3, 17-trihydroxy-5-androstan-6-one (THA-BR), exhibit different effects on growth of tobacco callus tissue. When added to a culture medium containing growth-limiting amounts of auxin, 24-epiBR reduced and THA-BR increased the fresh weight yield of tissue up to 53% and 207%, respectively, after 6 weeks of cultivation. The stimulatory and inhibitory effects of the two brassinosteroids on tissue growth occurred over a broad range of concentrations without a pronounced maximum corresponding to the yes or no type of response. Different effects of 24-epiBR and THA-BR on tissue growth were inversely proportional to the content of endogenous cytokinins. Maximum contents of predominant cytokinins N⁶-(²-isopentenyl)adenine (iP) and trans-zeatin (Z) in tissues supplied with 24-epiBR in growth-inhibiting concentrations were up to 3.7 fold and 3.4 fold higher, respectively, as compared to tissues grown on media containing growth-stimulating concentrations of THA-BR. Stimulation of tissue growth by THA-BR correlated with content of endogenous IAA and an inverse correlation was found between the content of endogenous IAA and cytokinins in tissues supplied with 24-epiBR. THA-BR exhibited weak cytokinin-like activity in a bioassay based on stimulation of growth of lateral buds of pea while 24-epiBR was inactive. Results indicate that the qualitatively different effects of the two brassinosteroids on growth of tobacco tissue may reflect their different influence on content of endogenous cytokinin.Abbreviations BR(s) brassinosteroid(s) - 24-epiBR 24-epibrassinolide - THA-BR 2,33, 17-trihydroxy-5-androstan-6-one - CK(s) cytokinin(s) - iP N⁶-(²-isopentenyl)adenine - [9R]iP N⁶-(²-isopentenyl)adenosine - Z trans-zeatin - [9R]Z ribosyl-trans-zeatin - ABA abscisic acid - IAA indole-3-acetic acid - NAA naphtalene-1-acetic acid - DEAE cellulose diethylaminoethyl cellulose - HPLC high performance liquid chromatography - ELISA enzyme linked immuno-sorbent assay     

The effect of polyamine biosynthesis inhibition on growth and differentiation of the phytopathogenic fungus Sclerotinia sclerotiorum

We studied the effects of several polyamine biosynthesis inhibitors on growth, differentiation, free polyamine levels and in vivo and in vitro activity of polyamine biosynthesis enzymes in Sclerotinia sclerotiorum. -Difluoromethylornithine (DFMO) and -difluoromethylarginine (DFMA) were potent inhibitors of mycelial growth. The effect of DFMO was due to inhibition of ornithine decarboxylase (ODC). No evidence for the existence of an arginine decarboxylase (ADC) pathway was found. The effect of DFMA was partly due to inhibition of ODC, presumably after its conversion into DFMO by mycelial arginase, as suggested by the high activity of this enzyme detected both in intact mycelium and mycelial extracts. In addition, toxic effects of DFMA on cellular processes other than polyamine metabolism might have occurred. Cyclohexylamine (CHA) slightly inhibited mycelial growth and caused an important decrease of free spermidine associated with a drastic increase of free putrescine concentration. Methylglyoxal bis-[guanyl hydrazone] (MGBG) had no effect on mycelial growth. Excepting MGBG, all the inhibitors strongly decreased sclerotial formation. Results demonstrate that sclerotial development is much more sensitive to polyamine biosynthesis inhibition than mycelial growth. Our results suggest that mycelial growth can be supported either by spermidine or putrescine, while spermidine (or the putrescine/spermidine ratio) is important for sclerotial formation to occur. Ascospore germination was completely insensitive to the inhibitors.     

Cationic responses of organs and haemolymph of Biomphalaria pfeifferi (Krauss), Biomphalaria glabrata (Say) and Helisoma trivolvis (Say) (Gastropoda: Planorbirdae) to cationic alterations of the medium

The cationic responses of haemolymph, mantle collar, headfoot, gut + digestive diverticulum and ovotestis + albumin gland of three planorbids to cationic changes in the media are explored. Body organs exhibited cationic homeostasis, although attained with difficulty under very low Ca : Mg and Ca : Na ratios. Haemolymph imbalances were obtained at very low calcium concentrations and at very low Ca : Mg and Ca : Na ratios in the medium; this may be linked to competition for Ca²⁺ uptake sites in the epithelium by Mg²⁺ and possibly Na⁺ ions.Fecundity and shell growth in relation to calcium concentrations and to Ca : Mg and Ca : Na ratios are examined.Normal internal cationic levels, under optimal conditions, were obtained for each species. Significant interspecific differences for haemolymph sodium were found; magnesium levels were slightly higher in Biomphalaria spp. than in Helisoma trivolvis; potassium levels were all similar. Amounts of tissue calcium decreased as follows: Mantle collar head-foot ovotestis + albumin gland-gut + diverticulum. Tissue magnesium levels in the gut were low. H. trivolvis had highest tissue calcium and sodium; B. glabrata had highest tissue potassium.     

Changes in nutrient composition and pH of the culture medium during in vitro shoot proliferation of crabapple and pear

Immature soybean (Glycine max (L.) Merr) embryos, or cotyledons isolated from them, were cultured on modified MS medium containing B5 vitamins and NAA (50 M) to induce somatic embryogenesis. The effects of media variables, dissection treatments and light conditions were investigated in this system. The efficiency of embryogenesis increased as sugar concentration decreased from 12 to 1.5%; sucrose and glucose were similarly effective as carbon sources. In an examination of the effects of medium pH, values between pH 5.0 and 7.0 gave similar embryogenesis efficiencies, but the frequency of normal embryos was greater in media with low pH values. In buffered medium (10 mM MES), a pH of 5.0 was inhibitory to embryogenesis, and most normal embryos were produced at pH 5.5. Under various dissection treatments, embryogenesis efficiency and root and callus production were increased by tissue damage. Somatic embryogenesis was observed both in darkness and in light, although embryo development was impaired under high light (80 E m^-2 s^-1) conditions. The ability of somatic embryos to germinate was closely correlated with embryo normality, and was influenced little by the hormone content of germination media. Of various media tested for their ability to support the growth of germinated embryos, a medium based on hydroponic nutrient salts, supplemented with yeast extract, and gelled with Difco-Bacto agar gave the best plantlet growth.Abbreviations E m^-2 s^-1 microEinsteins per square meter per second - NAA -napthalene acetic acid - N50 MS salts with B5 vitamins and 50 M NAA (Napthalene acetic acid) - MES 2(n-morpholino) ethanesulphuric acid - BAP benzylamino purine - IBA indole butyric acid This paper (No. 86-3-97) is published with the approval of the director of the Kentucky Agricultural Experiment Station.     

Benzyladenine induction of buds and somatic embryogenesis in Picea omorika (Pančić) Purk.

Somatic embryogenesis and adventitious bud formation, initiated from shoot explants of Picea omorika is described. Benzyladenine (BA) as the only growth regulator, added to modified Von Arnold and Eriksson medium, induced formation of both adventitious buds and embryogenic tissue. Optimal BA concentration for bud induction was 4.5 M and further bud development and plantlet formation was achieved on growth regulator-free medium. The embryogenic tissue formation was induced when the explants were first grown on the medium with high BA content (22.5 M) and then transferred to medium without growth regulators. Subsequent proliferation of embryogenic tissue was accomplished by subculturing on medium containing 9 M 2,4-dichlorophenoxyacetic acid and 4.5 M BA, and further embryo development was achieved on medium with 12 M abscisic acid. Embryos cultured on growth regulator-free medium formed roots and rooted plantlets were successfully established in soil in the greenhouse.     

Zinc-phosphorus interactions in two cultivars of tomato (Lycopersicon esculentum L.) grown in chelator-buffered nutrient solutions

Zinc-phosphorus interactions have been frequently studied using a diverse number of crop species, but attainment of reproducible Zn deficiencies, especially severe ones, has been hampered by the use of conventional hydroponic solutions wherein contaminating levels of Zn are often near-adequate for normal growth. We utilized novel, chelator-buffered nutrient solutions for precise imposition of Zn deficiencies. Tomato (Lycopersicon esculentum L. cv. Jackpot or Celebrity) seedlings were grown for 15 to 18 d in nutrient solutions containing 200, 600, or 1200 M P, and 0 to 91 M total Zn. Computed free Zn²⁺ activities were buffered at 10^-10.3 M by inclusion of a 100-M excess (above the sum of the micronutrient metal concentrations) of the chelator DTPA. At total added Zn=0, acute Zn deficiency resulted in zero growth after seedling transfer, and plant death prior to termination. Free Zn²⁺ activities 10^-10.6 M resulted in Zn deficiencies ranging from mild to severe, but activities 10^-11.2 were required to cause hyperaccumulation of shoot P to potentially toxic levels. Despite severe Zn deficiency (i.e. ca. 20% of control growth), tissue Zn levels were usually much higher than the widely reported critical value of 20 mg kg^-1, which may be an artifact of the selection of DTPA for buffering free Zn²⁺. Across Zn treatments, increasing solution P depressed growth slightly, especially in Celebrity, but corresponding increases in tissue P (indicative of enhanced P toxicity) or decreases in tissue Zn (P-induced Zn deficiency) were not observed. The depressive effect of P was also not explained by reductions in the water-soluble Zn fraction. Within 40 h, restoration of Zn supply did not ameliorate high leakage rates (as measured by K⁺ efflux) of Zn-deficient roots. Similarly, transfer of Zn-sufficient plants to deficient solutions did not induce leakiness within 40 h. Foliar sprays of ZnSO₄ almost completely corrected both Zn deficiency and membrane leakiness of plants grown in low-Zn solutions. Hence, maintenance of root membrane integrity appears to depend on the overall Zn nutritional status of the plant, and not on the presence of certain free Zn²⁺ levels in the root apoplasm.     

Changes in polyamine synthesis and concentrations during chick embryo development

We have measured the activities of the two rate controlling enzymes in polyamine synthesis, L-ornithine decarboxylase (ODC) and S-adenosyl-L-methionine decarboxylase (SAMDC), and the concentrations of the polyamines, putrescine, spermidine and spermine, in the developing chick embryo from laying to hatching. The embryo exhibited major peaks in the ODC and SAMDC activities as well as in the concentrations of all three polyamines at 15 h (gastrulation), 23-30 h (early organogenesis), days 4-5 (mid-organogenesis), and days 12-17 (organ growth and maturation). In the 4 and a half-day-old embryo, ODC activity and polyamine concentrations were about twice as high in the head region as compared to the trunk region. In the 14-day-old embryo, the highest ODC and SAMDC activities were found in lung, intestine and kidney, and there was a positive correlation between the enzyme activities and the growth rates of most organs/tissues.