Stereospecificity and electrogenicity of amino acid transport in Riccia fluitans

In the aquatic liverwort Riccia fluitans, membrane depolarization (_m), change in membrane conductance (g_m), and current-voltage (I-V) characteristics in the presence of different amino acids as well as the uptake of ¹⁴C-labeled amino acids were measured. L-isomers of the tested amino acids generate larger electrical effects (_m, g_m) than D-isomers, and the I-V characteristics show that the positive electrical inward-current of 20 mA m^-2 generated by 0.5 mM D-serine is only about 50% of the current generated by adding 0.5 mM L-serine. Whereas - and -amino acids rapidly depolarize the membrane to the same extend, with -aminobutyric acid (-AB) and dipeptides no significant electrical effects have been measured. The uptake kinetics of ¹⁴C-labeled amino acids display three components: (I) A saturable high-affinity component with K_s-values of 48 M D-alanine, 12 M -aminoisobutyric acid (AIB), 9 M L-alanine, 8 M L-proline, and 6 M L-serine, respectively; (2) an apparently linear low-affinity component, and (3) an also linear but unspecific component at concentrations >20 times the given K_s-value. Uptake of ¹⁴C-labeled AIB can be inhibited competitively by all tested neutral amino acids, the L-isomers being more effective than the D-isomers, as well as by ammonium or methylamine. Vice versa, AIB competitively inhibits uptake of L-serine and L-alanine. It is concluded that an uncharged stereospecific carrier for the investigated amino acids exists in the plasmalemma of Riccia fluitans. Accumulation ratios of about 50 suggest secondary active transport driven by a transmembrane electro-chemical gradient (mainly _m) which is generated by the electrogenic proton pump. It is suggested that this carrier binds to the amino group forming either a charged binary complex with positively charged amines (Felle 1980), or an uncharged complex with -AB or dipeptides, whereas electrogenic transport of - and -amino acids is mediated by a ternary carrier complex, probably charged by a proton.Symbols and Abbreviations _m membrane potential (mV) - E_co equilibrium potential (mV) of the transport system - g_m membrane (slope) conductance (Sm^-2) - g_m change in g_m - I-V curve current-voltage curve - AIB -aminoisobutytric acid - -AB -aminobutyric acid     

Improvement of growth and camptothecin yield by altering nitrogen source supply in cell suspension cultures of Camptotheca acuminata

Nitrate at 70m gave the highest biomass of Camptotheca acuminata in suspension culture in MS medium, but a NH₄ ⁺/NO₃ ^– molar ratio of 5:1 (giving a total of 40 m N) gave the maximum camptothecin yield. A two-stage flask culture system was established to improve culture efficiency; cell dry weight, camptothecin content and yield was increased by 30%, 280% and 340%, respectively when compared with those of control, reaching up to 36g l^–1, 0.36mgg^–1, and 12.8mgl^–1, respectively.     

Mikrospektralphotometrische Untersuchungen zum Speichermechanismus der kationischen Farbstoffe Viktoriablau B und 4R durch die lebende und tote Pflanzenzelle

Zusammenfassung Die Abhängigkeit der Absorptionsmaxima wässeriger Farbstoff-Lösungen konstanten pH-Wertes von der Farbstoffkonzentration deutet darauf hin, daß die angewandten Farbstoffe Assoziat bilden.Bei Ausschüttelungsversuchen in organischen Lösungsmitteln ergeben Viktoriablau B und 4R je nach der Polarität des Solvens unterschiedliche Absorptionskurven.Natriumnucleinat bedingt eine positive Metachromasie; die jeweilige Lage des Maximums wird von der Natriumnucleinatkonzentration bestimmt.Rutin bedingt positive und negative Metachromasie.Plasma und Zellkern der Oberepidermis von Allium cepa-Schuppenblättern zeigen nach Vitalfärbung mit Viktoriablau B und 4R übereinstimmend ein Absorptionsmaximum bei 625–630 nm für Viktoriablau B und bei 600–605 nm für Viktoriablau 4R. Die mit Viktoria-blau B vital gefärbten vollen" Zellsäften der Unterepidermis zeigen ein Absorptionsspektrum mit einem Maximum bei 630 nm.Aus der Lage der Absorptionsmaxima und dem Verlauf der Absorptionskurven kann geschlossen werden, daß die Färbung des lebenden Plasmas und Zellkernes auf eine Anreicherung der Farbstoffteilchen vorwiegend in frei vorliegenden polaren Lipiden vom Lezithintyp und zum Teil in freien Fettsäuren beruht, während es sich bei der Färbung des fixierten Plasmas und Zellkernes um eine Bindung der benutzten Farbstoffe an Nucleinsäuren handelt. Die Speicherung der vollen" Zellsäfte beruht auf einer salzartigen Komplexbildung zwischen Farbstoffteilchen und zelleigenen Flavonolen.

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Microspectrophotometric investigations into the mechanism of accumulation of the cationic dyes Victoria blue B and 4R by the living and dead plant cell

Summary Since maximum absorption of aqueous dye solutions with stable pH is dependent upon the concentration of the dye, it is supposed that the used dyes build associates.In shake out experiments with organic solvents Victoria blue B and 4R show different absorption curves according to the polarity of the solvent.Sodium nucleinate causes positive metachromasy; the respective site of the maximum is determined by the concentration of sodium nucleinate.Rutin causes positive and negative metachromasy.After vital staining with Victoria blue B and 4R both the plasma and the nucleus of cells from the upper epidermis of scale leaves of Allium cepa show an absorption maximum at 625–630 nm for Victoria blue B and 600–605 nm for Victoria blue 4R. After vital staining by Victoria blue B, the full cell saps of cells from the lower epidermis show an absorption spectrum with a maximum at 630 nm.The position of the absorption maxima and the course of the absorption curves lead to the conclusion, that the staining of the living plasma and nucleus is due to an accumulation of the dye particles principally in free polar lipids of the lecithine type and partly in free fatty acids, whereas the staining of the fixed plasma and nucleus is caused by the used dyes bound to nucleic acids. The accumulation of full cell saps is due to a salt-like complex formation between dye particles and flavonoles genuine to the cell.

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Die Absorptionskurven an der lebenden und toten Zelle wurden mit dem UMSP I aufgenommen, das Herrn Prof. Dr. H. Drawert — Institut für Allgemeine Botanik, Hamburg — von der Deutschen Forschungsgemeinschaft zur Verfügung gestellt worden ist. Deswegen spreche ich Herrn Prof. Dr. H. Drawert meinen ergebenen Dank aus. Mein Dank gilt auch Herrn A. Zoumpos für die Umzeichnung der Textabbildungen.     

Growth stimulation ofEuphorbia lathyris L. by GA4+7 and BA

Container-grownEuphorbia lathyris plants were treated with foliar sprays of various combinations of BA and GA₄₊₇ or 0–3600 mg L^–1 Promalin (11 BA + GA₄₊₇) in separate experiments. GA₄₊₇ and Promalin stimulated plants to grow taller. BA and Promalin promoted axillary shoot growth. Multiple applications of Promalin stimulated branching more than single treatments. Dry weight accumulation was stimulated only if the growth regulators were applied to 28–33-cm and not to 56-cm tall plants. Chemical names used: (1, 2, 4a, 4b, 10)-2,4a,7-trihydroxy-1-methyl-8-methylenegibb-3-ene-1,10-dicarboxylic acid 1,4a-lactone (GA₄₊₇),N-(phenylmethyl)-H-purin-6-amine (BA), and Promalin [11 (wt/wt) GA₄₊₇ and BA].The use of the name Promalin or other trade names does not imply endorsement to the exclusion of other products or vendors that may also be suitable.     

Tissue culture ofKarwinskia humboldtiana—a plant producing toxins with antitumoural effects

Isolated embryos ofKarwinskia humboldtiana were cultured in vitro. The growth of embryos and development to plantlets on woody plant medium supplemented with indole-3-acetic acid 6.10^-2 mol l^–1, gibberellic acid (GA₃) 3.10^-2 mol l^–1, and 6-benzylaminopurine (BA) 2 mol l^–1 was obtained. Multiplication of shoots and rooting of excised shoots has been achieved. Callus formation on modified Murashige-Skoog medium supplemented with 1-naphthaleneacetic acid 10 mol l^–1, GA₃ 14 mol l^–1, and kinetin 5 mol l^–1 on hypocotyls, or on root cultures on medium supplemented with 2.4-dichlorophenoxyacetic acid 10 mol l^–1 and BA 10 mol l^–1 was induced.Abbreviations BA 6-benzylaminopurine - 2,4-d 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid - IAA indole-3-acetic acid - NAA 1-naphthaleneacetic acid - TEM transmission electron microscopy     

Secondary structure of the entomocidal toxin from Bacillus thuringiensis subsp. kurstaki HD-73

The secondary structure of the toxin fromBacillus thuringiensis subsp.kurstaki (Btk) HD-73 was estimated by Raman, infrared, and circular dichroism spectroscopy, and by predictive methods. Circular dichroism and infrared spectroscopy gave an estimate of 33–40% -helix, whereas Raman and predictive methods gave approximately 20%. Raman and circular dichroism spectra, as well as predictive methods, indicated that the toxin contains 32–40% -sheet structure, whereas infrared spectroscopy gave a slightly lower estimate. Thus, all of these approaches are in agreement that the native conformation of Btk HD-73 toxin is highly folded and contains considerable amounts of both -helical and -sheet structures. No significant differences were detected in the secondary structure of the toxin either in solution or as a hydrated pellet.     

Age and Growth of the Whiskery Shark, Furgaleus macki, from Southwestern Australia

Age and growth of the whiskery shark, Furgaleus macki, from southwestern Australia were examined using vertebral ageing and tag-recapture data. The readability of bands on the vertebral centra varied markedly between individuals. Four readers were used to make band counts, with the most experienced reader having the lowest index of average percent error and the highest level of agreement with final counts. Marginal increment analysis indicated that opaque bands form in January. With parturition occurring from August to October, size data suggests that the first band is probably formed 15–17 months after birth. The age at maturity was estimated to be 4.5 years for males, and 6.5 years for females. The oldest male was 10.5 years, and oldest female was 11.5 years. Von Bertalanffy growth parameters for males were L =121.5cm fork length, K=0.423 year^–1, t ₀=–0.472 years, were L =120.7cm fork length, K=0.369 year^–1, t ₀=–0.544 years for females, and were L =118.1cm fork length, K=0.420 year^–1, t ₀=–0.491 years for combined sexes. Data from a tag recapture study were analysed using a maximum likelihood method to verify the estimates of growth parameters from vertebral ageing. Von Bertalanffy growth parameters from the tag recapture study were L =128.2cm fork length, K=0.288 year^–1, t ₀=–0.654 years. The two methods of estimating growth parameters produced similar results, with rapid growth until approximately 5 years of age, after which there was little increase in length.     

Voltage dependence of the Ca2+-activated K+ conductance of human red cell membranes is strongly dependent on the extracellular K+ concentration

Summary The conductance of the Ca²⁺-activated K⁺ channel (g _K(Ca)) of the human red cell membrane was studied as a function of membrane potential (V _m ) and extracellular K⁺ concentration ([K⁺]_ex). ATP-depleted cells, with fixed values of cellular K⁺ (145mm) and pH (7.1), and preloaded with 27 m ionized Ca were transferred, with open K⁺ channels, to buffer-free salt solutions with given K⁺ concentrations. Outward-current conductances were calculated from initial net effluxes of K⁺, correspondingV _m , monitored by CCCP-mediated electrochemical equilibration of protons between a buffer-free extracellular and the heavily buffered cellular phases, and Nernst equilibrium potentials of K ions (E _K) determined at the peak of hyperpolarization. Zero-current conductances were calculated from unidirectional effluxes of⁴²K at (V _m –E _K)0, using a single-file flux ratio exponent of 2.7. Within a [K⁺]_ex range of 5.5 to 60mm and at (V _m –E _K) 20 mV a basic conductance, which was independent of [K⁺]_ex, was found. It had a small voltage dependence, varying linearly from 45 to 70 S/cm² between 0 and –100 mV. As (V _m –E _K) decreased from 20 towards zero mVg _K(Ca) increased hyperbolically from the basic value towards a zero-current value of 165 S/cm². The zero-current conductance was not significantly dependent on [K⁺]_ex (30 to 156mm) corresponding toV _m (–50 mV to 0). A further increase ing _K(Ca) symmetrically aroundE _K is suggested as (V _m –E _K) becomes positive. Increasing the extracellular K⁺ concentration from zero and up to 3mm resulted in an increase ing _K(Ca) from 50 to 70 S/cm². Since the driving force (V _m –E _K) was larger than 20 mV within this range of [K⁺]_ex this was probably a specific K⁺ activation ofg _K(Ca). In conclusion: The Ca²⁺-activated K⁺ channel of the human red cell membrane is an inward rectifier showing the characteristic voltage dependence of this type of channel.     

Sisal plant regeneration via organogenesis

Callus was initiated from in vitro grown immature leaf and ex vitro grown mature leaf and rhizome explants of Agave sisalana Perr. ex. Engelm, on MS medium containing 2,4-D (9.05 M) and kinetin (4.6 M) or 2,4-D (9.05 M), kinetin (4.6 M) and CH (1000 mg l^–1) or mod. MS (NH₄NO₃, 1500 mg l^–1) containing 2,4-D (9.05 M) and kinetin (4.6 M). Light was essential for callus formation which, however, was different in three types of explants on three different media compositions. Increasing NH₄ ⁺had a negative impact while addition of CH had a positive impact on callus formation. Shoot regeneration from callus from CH-supplemented medium only was achieved for rhizome and immature leaf tissues. The highest rate of regeneration was obtained with BA (26.6 M) as the sole hormone. Shoot buds g^–1 callus varied according to BA concentrations. Shoot proliferation rate increased on half-strength MS medium containing BA (8.9 M). Microshoots developed on MS medium containing BA (2.22 M) and GA₃ (1.44 M) and finally rooted on MS medium containing IAA (11.42 M). Acclimatized rooted plantlets are growing satisfactorily in ex vitro. This is the first report on plant regeneration via organogenesis of A. sisalana.     

Respiratory stimulus in Rhizobium sp. by legume lectins

High molecular weight lectins (> 100 kDa) from seeds of the legumes Canavalia brasiliensis (CnBr), Cratylia floribunda (CFL), Phaseolus vulgaris (PHA) and Vatairea macrocarpa (VML), temporarily stimulate the respiration of Rhizobium tropici-CIAT899 and R. etli-CFN42. These stimulants were significant (P < 0.05) in bacterial suspensions (> 2.85 mg dry biomass ml^–1), having at least 6200 molecules of lectins per bacteria. The VML (20 g ml^–1), induced specific O₂ demand of 2.3–2.5 M O₂ min^–1 mg dry biomass^–1, in CFN42 and CIAT899, respectively. However, CnBr, CFL and PHA induced smaller demands of O₂ (5×), in both strains. The order of affinities of the lectins was approximately VML > PHA > CFL > CnBr, with regard to respiratory stimuli in CIAT899 strain. The co-administration of 10 g VML ml^–1 and 9.8 M galactose, in CIAT899 suspensions, reduced the respiratory stimuli significantly in relation to the treatment with VML alone. These respiratory stimuli, induced by the lectins, increase the significance of the interaction lectin × Rhizobium in terms of bacterial physiology. Its understanding could be important in relation to bacterial symbiotic behaviour.     

Enzymic synthesis of the trisaccharide core region of the carbohydrate chain ofN-glycoprotein

Transmannosylation from mannotriose (Man1-4Man1-4Man) to the 4-position at the nonreducing end N-acetylglucosaminyl residue ofN,N-diacetylchitobiose was regioselectively induced through the use of -d-mannanase fromAspergillus niger. The enzyme formed the trisaccharide Man1-4GlcNAc1-4GlcNAc (3.7% of the enzyme-catalysed net decrease ofN,N-diacetylchitobiose) from mannotriose as a donor andN,N-diacetylchitobiose as an acceptor. Mannobiose (Man1-4Man) was also shown to be useful as a donor substrate for the desired trisaccharide synthesis.Abbreviations Man d-mannose - (M _n) (n=1–5) -linkedn-mer of mannose - GlcNAc₂ 2-acetamido-2-deoxy--d-glucopyranosyl-(1–4)-2-acetamido-2-deoxy-d-glucose     

Organ-specific modulation of gene expression in transgenic plants using antisene RNA

We have shown leaf-specific inhibition GUS gene expression in transgenic Nicotiana plants using an antisense RNA with a 41-base homology spanning the translation start codon of the gene. GUS was expressed from the nominally constitutive 35S promoter and the antisense RNA was expressed from the light-regulated ca/b promoter of Arabidopsis thaliana. A range of GUS inhibition from 0 to 100% was obtained by screening a small population of transgenic plants and the specific levels of inhibition observed were stably inherited in two generations. An antiGUS gene dosage effect was observed in plants which were homozygous for antiGUS. RNA detection results suggest that duplex formation with the 41 base pair antiGUS RNA destabilized the GUS mRNA and that an excess of antisense. RNA was not required. Our results demonstrate the potential of antisense RNA as a strategy for obtaining plant mutants, especially down mutations in essential genes where only a short 5 sequence of the mRNA is required. They also suggest that the position effect on gene expression could be used in conjunction with an antisense RNA strategy to provide a versatile approach for crop improvement.     

Studies on adenosine triphosphate transphosphorylases. XVIII. Synthesis and preparation of peptides and peptide fragments of rabbit muscle ATP-AMP transphosphorylase (adenylate kinase) and their nucleotide-binding properties

Two peptide fragments, derived from the head and tail of rabbit muscle myokinase, were found to possess remarkable and specific ligand-binding properties (Hamadaet al., 1979).By initiating systematic syntheses and measurements of equilibrium substrate-binding properties of these two sets of peptides, or portions thereof, which encompass the binding sites for (a) the magnesium complexes of the nucleotide substrates (MgATP^2– and MgADP^–) and (b) the uncomplexed nucleotide substrates (ADP^3– and AMP^2–) of rabbit muscle myokinase, some of the requirements for binding of the substrates to ATP-AMP transphosphorylase are being deduced and chemically outlined. One requirement for tight nucleotide binding appears to be a minimum peptide length of 15–25 residues. In addition, Lys-172 and/or Lys-194 may be involved in the binding of AMP.The syntheses are described as a set of peptides corresponding to residues 31–45, 20–45, 5–45, and 1–45, and a set of peptides corresponding to residues 178–192, 178–194, and 172–194 of rabbit muscle adenylate kinase. The ligand-binding properties of the first set of synthetic peptides to the fluorescent ligands: MgATP/ATP and MgADP/ADP are quantitatively presented in terms of their intrinsic dissociation constants (K_d) and values ofN (maximal number of moles bound per mole of peptide); and compared with the peptide fragment MT-I (1–44) obtained from rabbit muscle myokinase (Kubyet al., 1984) and with the native enzyme (Hamadaet al., 1979). In addition, the values ofN andK_d are given for the second set of synthetic peptides to the fluorescent ligands AMP and ADP as well as for the peptide fragments MT-XII(172–194) and CB-VI(126–194) (Kuby et al., 1984) and, in turn, compared with the native enzyme.A few miscellaneous dissociation constants which had been derived kinetically are also given for comparison (e.g., theK _i for AMP and the value of obtained for the native enzyme) (Hamada and Kuby, 1978), and theK'_d measured for Cr³⁺ and the synthetic peptide I_1–45 (Fryet al., 1985b).Paper XVII of this series is Kubyet al. (1983).     

Isolation and characterization of α-amylase derived from starchgrownClostridium acetobutylicum ATCC 824

Summary An extracellular -amylase was purified to homogeneity from the culture supernatant ofClostridium acetobutylicum ATCC 824 grown in synthetic medium containing starch by using a combination of ammonium sulfate fractionation, anion exchange chromatography and HPLC-gel filtration. The molecular weight of the 160-fold purified -amylase was determined by SDS-PAGE to be 61 kDa. HPLC analysis of end-products of enzyme activity on various substrates indicated that the enzyme acted specifically in an endo-fashion on the -1,4-glucosidic linkages. Enzyme activity was optimal over a pH range of 4.5–5.0 and temperature of 55°C, but was rapidly inactivated at higher temperatures. Addition of calcium chloride (2–5 mM) increased -amylase activity by ca. 20%, while the addition of 19 g ml^–1 of acarbose (a differential inhibitor of amylases) resulted in 50% inhibition. TheV _max andK _m of -amylase were 2.17 mg min^–1 and 3.28 mg ml^–1 on amylose, and 1.67 mg min^–1 and 1.73 mg ml^–1 on soluble starch, respectively.     

The high maltose-producing α-amylase of Penicillium expansum

Summary An -amylase capable of producing exceptionally high levels of maltose (74%) from starch has been identified from a strain of Penicillium expansum. The enzyme is produced extracellularly and was purified to homogeneity by starch adsorption and Sephadex gel filtration chromatography. P. expansum -amylase has a pH optimum of 4.5 and is stable in the pH range of 3.6–6.0. Other properties include a temperature optimum of 60° C, a molecular weight of 69 000 and an isoelectric point of 3.9. The most outstanding feature of the P. expansum enzyme is its ability to yield 14% more maltose and 17.1% less maltotriose than a currently used commercial enzyme. This may be partly explained by the greater affinity of this new enzyme for maltotriose (K _m=0.76 mM) relative to the commerical enzyme, Fungamyl (K _m=2.9 mM). The enzyme reported here is unique among fungal -amylases in being able to produce such high levels of maltose and its physicochemical properties suggest that it has potential for commercial development.     

Fractionation of the enzymes of the barley aleurone layer: Evidence for a soluble mode of enzyme release

Summary Aleurone cells of barley (Hordeum vulgare L.) contain microbodies as determined by histochemical localization with diaminobenzidine. These microbodies can be isolated from both water and gibberellic acid (GA₃) treated cells and identified on sucrose density gradients as glyoxysomes on the basis of their buoyant densities (1.25 g cm^-3) and their enzyme complement. Fractionation of aleurone layer homogenates by differential centrifugation after varying times of exposure to GA₃, however, does not indicate the presence of a discrete secretory vesicle containing either -amylase or -1,3-glucanase. Cytological evidence also suggests that at least -1,3-glucanase is not released from these cells by means of a discrete secretory vesicle.Supported by National Science Foundation Grant GB-27468. The assistance of Dr. William Breidenbach in the assay of glyoxysomal enzymes is gratefully acknowledged.     

Esterases in histochemistry and ultrahistochemistry

Synopsis The histochemical identification of individual esterases is a problem that has not yet been overcome. Inhibitors and different substrates reveal different patterns of distribution. 8-hydroxyquinoline acetate is a useful substrate in ultrahistochemistry. There is evidence of a relationship between esterase distribution and function.ACTH adrenocorticotropic hormone - 5Bri–O-2 5-bromoindoxyl acetate - 5Br–4ClI–O-2 5-bromo-4-chloro indoxyl acetate - cAMP cyclic adenosine monophosphate - DFP di-isopropyl-fluorophosphate - hCG human chorion gonadotropin - HS-2/4 thiol acetate/butyrate - I-O-2/4 indoxyl acetate/butyrate - N-O-2/3/4 -naphthyl acetate/propionate/butyrate - N-O-2 -naphthyl acetate - N-S-2/9 -naphthyl thiolacetate/nonanoate - NAS-O-2 naphthol AS acetate - NASD-O-2 naphthol AS-D acetate - 4NP-O-2/3 p-nitrophenyl acetate/propionate - 4NP-S-2 p-nitrophenyl thiol acetate - P-O-2 phenyl acetate - Q-O-2/4 8-hydroxyquinoline acetate/butyrate - Q-S-2/4 8-mercaptoquinoline acetate/butyrate - TBA-S-2/9 -thiolbenzanilide acetate/nonanoate - TSH thyroid-stimulating hormone     

Identification of a glutathione-S-transferase effector domain for inhibition of jun kinase, by molecular dynamics

We have recently found that the glutathione-S-transferase -isozyme (GST-), a cellular detoxification enzyme, potently and selectively inhibits activation of jun protein by its upstream kinase, jun kinase (JNK). This newly identified regulatory activity of GST- is strongly inhibited by a group of agents that inhibit its enzymatic activity. Since loss of enzymatic activity in general does not correlate with loss of regulatory activity, it is likely that inhibitor binding induces changes in the structure of one or more domains of GST that block its interaction with JNK. To identify regions of GST that change conformation on the binding of inhibitors, we have performed molecular dynamics calculations on GST- to compute its average structure in the presence and absence of the inhibitor, glutathione sulfonate. Superposition of the two average structures reveals that several regions change local structure depending upon whether the inhibitor is bound or not bound. Two of these regions, residues 36–50 and 194–201, are highly exposed. We have synthesized peptides corresponding to these two segments and find that the 194–201 sequence strongly inhibits the ability of GST- to block the in vitro phosphorylation of jun by JNK. These results suggest that this region of GST- is critical to its functioning as a newly discovered regulator of signal transduction.     

The responsiveness of Avena fatua aleurone protoplasts to gibberellic acid

The sensitivity to gibberellic acid (GA₃) of aleurone protoplasts isolated from a single harvest of an inbred line of Avena fatua seed that had been after-ripened over anhydrous CaCl₂ at 25±2°C and 4±2°C for three years was assessed. Protoplasts isolated from aleurones of seed stored at 25°C produced substantially more -amylase in response to 10^–7 M GA₃ than those isolated from aleurones of seed stored at 4°C. The apparent difference in responsiveness does not appear to be due to a change in the duration of the lag phase between addition of GA₃ and the production of -amylase. The dose response of aleurone protoplasts to GA₃, measured as -amylase production, is complex and appears to have three phases. Protoplasts from seed stored at both temperatures respond appreciably to 10^–14 M GA₃. With increasing concentrations of GA₃, up to 10^–9 M, -amylase production increases similarly in protoplasts from both lots of seed, reaching a level approximately 2.7–3.8 times greater than when no GA₃ is applied. GA₃-induced -amylase production increases markedly as the concentration is raised from 10^–9 M to 10^–6 M, and the response then appears to be saturated. Over this part of the response curve protoplasts from the two seed lots differ markedly in their responsiveness to GA₃. Those from seed stored at 25°C produce considerably more -amylase, >130-fold higher than the minus GA₃ control, than those from seed stored at 4°C, <35-fold higher than the minus GA₃ control. This apparent difference in the responsiveness of aleurone protoplasts to GA₃ could be correlated with the loss of embryo dormancy in seed stored at 25°C. Seed stored at 4°C retained the dormancy characteristics present immediately after harvesting.     

Relative positions of homologous chromosomes or groups in male and female metaphase figures

Summary This study investigates statistically, with computer assistance, the square distances between chromosome centromeres in homologous pairs or groups after circularizing transformation. By the method described here, it is possible to eleminate all subjective measurments and just use the coordinates x _i and y _i of the centromeres. The values obtained for homologous chromosomes are shown to be specific but not always small. Low square distance values occur in greater number for the chromosomes that are most frequently involved in aneuploïdies. This is true for acrocentric chromosomes which, moreover, tend to lie close together significantly more often in female than in male mitoses; it is also true for group 17–18 especially in males, and of XX in female mitoses. Furthermore, we find significantly low square distances in chromosome pair 1.