Regulatory elements mediating transcription from the Drosophila melanogaster actin 5C proximal promoter.

The major cytoskeletal actin gene of Drosophila melanogaster, the actin 5C gene, has two promoters, the proximal one of which controls constitutive synthesis of actin in all growing tissues. To locate regulatory elements required for constitutive activity of the proximal promoter, mutants of this promoter were fused to the bacterial chloramphenicol acetyltransferase gene and assayed for transient expression activity in cultured Drosophila embryonic Schneider line 2 cells. An essential regulatory element has been located 313 base pairs upstream from the cap site. Deletion of this element lowered expression to one-third of the wild-type level. The element has the sequence AAGTTGTAGTTG, as shown by protein-binding footprinting with the reagent methidiumpropyl-EDTA-Fe(II). This element is probably not a general one, since it was not detected in a search of the published 5'-flanking sequences of 27 Drosophila genes. In addition to this regulatory element, there are five GAGA elements in the actin 5C proximal promoter, some or all of which are essential for the promoter activity as shown by an in vivo competition assay. Although this promoter has no classical TATA element, there is an essential promoter region about 35 base pairs upstream from the cap site that could be a TATA surrogate. The promoter also shows sequences homologous to the alcohol dehydrogenase factor 1-binding site and to the core of the vertebrate serum response element, but mutations of these sites did not affect promoter activity in transient expression assays.     

Induction of freezing tolerance in microspore-derived embryos of winter Brassica napus

Freezing tolerance was induced in microspore derived embryos of winter Brassica napus cv. Jet neuf by the addition of ABA or mefluidide to the culture media during embryogenesis. Survival after freezing was estimated by culture of frozen-thawed embryos to plantlets. A higher freezing tolerance (50% survival at –15°C) was induced when 50 M ABA or 3.2 M mefluidide was incorporated initially into the medium during embryogenesis at 25°C followed by culture at 2°C for 3 weeks. When embryos were induced in the absence of ABA or mefluidide and maintained at 2°C for even as long as 12 weeks a lower degree of freezing tolerance (10% survival at –15°C) was obtained. Plants regenerated from embryos hardened maximally by a combination of either ABA or MFD with low temperature did not require further vernalization for flowering.Abbreviations ABA abscisic acid - MFD mefluidide - 2,4-D 2,4-dichlorophenoxyacetic acid - LT₅₀ killing temperature for 50% of the embryos     

Seasonal variation of calcium,magnesium, potassium,and manganese contents in xylem sap of beech(Fagus sylvatica L.) in a 35-year-old limestone beech forest stand

Summary In a 35-year-old calcareous beech forest stand five beech trees (Fagus sylvatica L.) were felled every 2 weeks, and xylem sap was obtained by means of water displacement from the lowest trunk sections, each 100 cm in length. From mid-October 1988 to mid-October 1989 a total of 130 trees were investigated. The seasonal variations of the Ca, Mg, K and Mn contents, as well as those of pH, show four characteristic phases. Additionally, distribution of the mineral contents along the trunk was studied in four trees. The seasonal increase and decrease of xylem sap mineral contents along the trunk is shown for the characteristic phenophases. The Ca, Mg, K, and Mn contents of xylem saps were determined by means of atomicabsorptionspectrophotometry.     

A heparin-binding protein involved in inhibition of smooth-muscle cell proliferation.

A heparin-binding protein was isolated from bovine uteri and purified to homogeneity. This protein appears as a double band of approx. 78 kDa in SDS/polyacrylamide-gel electrophoresis and has an isoelectric point of 5.2. The binding of heparin to this protein is saturable. No other glycosaminoglycan from mammalian tissue, such as hyaluronic acid, chondroitin sulphate, dermatan sulphate or keratan sulphate, binds to the 78 kDa protein. Dextran sulphate binds in a non-saturable fashion. Certain heparan sulphate polysaccharide structures are required for binding to the 78 kDa protein. Some proteoheparan sulphates, such as endothelial cell-surface proteoheparan sulphate, show only weak interaction with the 78 kDa protein in contrast with a basement-membrane proteoheparan sulphate from HR-9 cells. Antibodies against the 78 kDa protein inhibit binding of proteoheparan [35S]sulphate from basement membranes to smooth-muscle cells. Conventional antibodies, Fab fragments and some monoclonal antibodies, inhibit smooth-muscle cell proliferation in a similar range as that observed for heparin. The protein was detected in a variety of tissues and cells but not in blood cells. A possible role of this protein as a receptor for heparin or heparan sulphate and its function in the control of the arterial wall structure are discussed.     

A new recombinant procoagulant protein derived from the cDNA encoding human factor VIII

We have constructed new B domain deletion derivatives of human factor VIII (FVIII) by manipulating the cDNA using recombinant DNA techniques. One of these new derivatives, FVIII delta II, in which amino acids 771(pro)-1666(asp) have been deleted, no longer contains the protease cleavage site at amino acid position 1648(arg)-1649(glu) known to be involved in the initial step of FVIII processing. We have expressed this molecule in both baby hamster kidney (BHK) 21 cells using the vaccinia virus (VV) expression system and have established Chinese hamster ovary (CHO) derived permanent cell lines expressing either recombinant (r)FVIII or FVIII delta II. The characteristics of FVIII delta II have been compared to those of rFVIII and/or plasma derived (pd) FVIII. FVIII delta II has the following properties: (i) it exhibits FVIII procoagulant activity; (ii) it is expressed at 5-fold higher levels than is the complete molecule in comparable systems; (iii) it migrates for the most part as a single major band on SDS-PAGE, in contrast to the complete molecule; (iv) it is activated to a greater extent by thrombin than is either rFVIII or pdFVIII; and (v) it retains the ability to bind von Willebrand factor (vWf).     

Immunocytochemical demonstration of the neurosecretory systems containing putative moult-inhibiting hormone and hyperglycemic hormone in the eyestalk of brachyuran crustaceans

Summary By use of antisera raised against purified moultinhibiting (MIH) and crustacean hyperglycemic hormone (CHH) from Carcinus maenas, complete and distinct neurosecretory pathways for both hormones were demonstrated with the PAP and immunofluorescence technique. By double staining, employing a combination of silver-enhanced immunogold labelling and PAP, both antigens could be visualized in the same section. Immunoreactive structures were studied in Carcinus maenas, Liocarcinus puber, Cancer pagurus, Uca pugilator and Maja squinado. They were only observed in the X-organ sinus gland (SG) system of the eyestalks and consisted of MIH-positive perikarya, which were dispersed among the more numerous CHH-positive perikarya of the medulla terminalis X-organ (XO). The MIH-positive neurons form branching collateral plexuses adjacent to the XO and axons that are arranged around the CHH-positive central axon bundle of the principal XO-SG tract. In the SG, MIH-positive axon profiles and terminals, clustered around hemolymph lacunae, are distributed between the more abundant CHH-positive axon profiles and terminals. Colocalisation of MIH and CHH was never observed. The gross morphology of both neurosecretory systems was similar in all species examined, however, in U. pugilator and M. squinado immunostaining for MIH was relatively faint unless higher concentrations of antiserum were used. Possible reasons for this phenomenon as well as observed moult cycle-related differences in immunostaining are discussed.     

An empirical model of primary productivity (14C) using mesocosm data along a nutrient gradient

The two parameters of the hyperbolic tangent equation, P_m and, were estimated from in situ vertical profiles of primary productionusing mesocosm data along a nutrient gradient. The parameters,derived from 4-h (around noon) ¹⁴C incubations, were used togetherwith the photosynthesis-light curve and hourly solar radiationdata to calculate daily primary production rates (P_d). Approximately40% of the daily production occurred in the 4 h around noon.Considering parameter uncertainty, there was no indication ofan increase in variation in production with increased nutrientloading, nor did biomass-specific P-I parameters increase. Annualproduction ranged from 82 to 901 g C m^–2 year^–1and was highest in the highest nutrient treatment tank. Dailyproductivity ranged from 0.02 to 9.1 g C m^–2 day^–1and was significantly correlated, in all treatments, with acomposite parameter BI₀/k (where B is phytoplankton biomass;I₀ is daily radiation and k is the extinction coefficient).Linear regressions of P_d against BI₀/k indicated that much ofthe variability (86%) in productivity was explained by lightavailability and phytoplankton biomass. Two approaches for predictingproductivity were compared: (i) predicting production directlyfrom environmental variables (i.e. BI₀/k) and (ii) predictingthe parameters of the P-I curve from environmental variablesand using these to calculate daily production.     

Auxins,ABA and gibberellin-like activity in abscising and non-abscising flowers and pods of Vicia faba L.

Abscission probability varies among floral positions within inflorescences of Vicia faba L. Flowers from proximal positions have a greater chance to develop into mature pods than flowers from more distal positions which normally abscise either as older flowers or as young pods. In three field experiments with the indeterminate single stem variety Herz-Freya, changes in the contents of extractable auxins, abscisic acid (ABA) and gibberellins in flowers and pods during their development, and their possible influence on abscission were investigated.Inflorescences at different positions along the stem were divided into the two proximal and the remaining fruits. The content of all three hormones was at a low level during flower development, increased greatly in parallel with dry matter accumulation in the young pods, and then decreased to maturity. The first hormone to increase in the fruits was auxin and this took place when abscission from the distal positions began. ABA and gibberellins at this time were still at a low level. This ontogenic course of hormone production was very similar in fruits of both positions within an inflorescence, but in flowers and young pods from proximal positions, auxin content in most inflorescences was greater than in those from the abscising distal positions. No such positional differences were observed with ABA and gibberellins. Decapitation of the plants reduced flower and pod drop from the remaining reproductive nodes. Although decapitation resulted in less abscission among distal flowers and young pods from these nodes, it did not affect the ontogenic course of auxin and ABA production in these fruits.     

The stimulating effect of a cold, dark pretreatment on the etioplast/chloroplast transformation of angiosperms. I. The stimulating effect of cold predarkening on different stages of greening under white light

Schönbohm, E., Stute, U., Thienhaus, P. and Werner, U. 1988. The stimulating effect of a cold, dark pretreatment on the etioplast/chloroplast transformation of angiosperms I. The stimulating effect of cold predarkening on different stages of greening under white light. - Physiol. Plant. 72: 541–546.
The etioplast/chloroplast transformation in angiosperms is controlled by light; most of the processes are mediated by phytochrome. We have shown that in the primary leaves of etiolated seedlings of wheat ( Triticum aestivum L. cv. Kolibri), fire-bean ( Phaseolus multiflorus L. cv. Preisgewinner) and in the cotyledons of etiolated sun flower seedlings ( Helianthus annuus L. cv. macrocarpa) the chlorophyll accumulation in the phase after the end of the lag phase can be greatly stimulated by a cold predarkening period. This effect is not necessarily coupled with a red preirradiation. Furthermore the lag phase can be dramatically shortened by the cold, dark pretreatment, whereas the amount of photoconvertible protochlorophyll(ide) in the darkness remains unaffected by the cold, dark pretreatment. The stimulating effect of a cold, predarkening period on greening is fully reversible by a warm, dark phase that is placed between the cold period and the onset of the continuous white light phase. These findings cannot be generalized: We could demonstrate that in the tropical plant Momordica charantia greening under white light was not affected by different temperature pretreatments during predarkening. The stimulating effect of a cold, predarkening period on greening is assumed to have ecological relevance.