Meristem transformation of sunflower via Agrobacterium

Summary For transformation of sunflower (Helianthus annuus L. cv. Zebulon), shoot apical meristems were dissected from seeds and cocultivated with a disarmed Agrobacterium tumefaciens strain harboring a binary vector carrying genes encoding GUS- and NPTII-activity. The influence of the media conditions, the time of cocultivation and the stage of the developing seed on shoot development and meristem transformation was analysed. Transformants were selected by their ability to grow on kanamycin. Transformation was confirmed by assays for GUS and NPTII. GUS-positive shoots were rooted on rockwool and transferred to soil. Transformation of shoot meristem cells occurred at low frequencies. Chimaeric expression of the two genes was observed in transformed plants. Integration of the foreign DNA in the sunflower genome was confirmed with the polymerase chain reaction.Abbreviations GUS ß-Glucuronidase - NPTII Neomycin phosphotransferase II     

In vitro study of lipid metabolism in the mealworm larval fat body

Lipid metabolism in Tenebrio larval fat body has been studied in vitro. Lipid release required the presence of diluted hemolymph in the incubation medium. This time-dependent release of lipid was strongly stimulated in a dose-dependent manner by Tenebrio corpora cardiaca (CC) extracts or synthetic adipokinetic hormone (AKH I). Furthermore, some glycerol was released when larval fat body was incubated without hemolymph, and this phenomenon was also dose dependent for added CC extracts. Lipid synthesis was estimated in vitro by following the incorporation of radioactivity from [6-¹⁴C] glucose into fatty acids. Lipogenesis occurred in the absence of added carbohydrates in the medium, but it was stimulated by the addition of glucose, and especially trehalose (10 mg ml^?1). Intestinal insulin-like peptide (ILP) also stimulated in vitro lipogenesis in a dose-dependent fashion. We conclude that lipolytic and lipogenetic activities of larval mealworm fat body in vitro are effectively under hormonal control.     

Actin-binding proteins are conserved from slime molds to man

DNA clones encoding the actin-binding proteins alpha-actinin and severin from Dictyostelium discoideum were isolated and sequenced. Comparisons of the deduced amino acid sequences with proteins from other species showed striking similarities at distinct regions. The F-actin cross-linking molecule alpha-actinin carries two characteristic EF-hand structures highly homologous to the Ca2+-binding loops of proteins from the calmodulin superfamily. An N-terminal region that is conserved in alpha-actinin from D. discoideum and vertebrates is also related to parts of the dystrophin sequence and might represent the F-actin binding site. Severin, gelsolin, villin, and fragmin share homologous sequences that are believed to participate in the severing activity of these proteins.     

Effect of the Urtica dioica agglutinin on germination and cell wall formation of Phycomyces blakesleeanus Burgeff

The lectin from stinging nettle rhizomes, Urtica dioica agglutinin (UDA), did not affect the evolution of wet and dry weight, protein, nucleic acid, ATP, cAMP and glycerol content during early germination of Phycomyces blakesleeanus spores. However, earlier investigations established a strongly reduced mycelial growth of several phytopathogenic fungi by this small plant lectin. Total uptake and incorporation of radioactive precursors showed no differences between UDA or control hyphae, but UDA significantly altered the distribution patterns of [¹⁴C]-glucose incorporated into the walls of Phycomyces blakesleeanus (more label was recovered in the chitin fraction). Moreover, a small but significant stimulation of chitin synthase and a similar inhibition of chitin deacetylase was found in cell wall preparations. These observations could lead to a better understanding of plant-pathogen interrelationships and to a further elucidation of cell wall structure in fungi.Abbreviations GlcNAc N-Acetylglucosamine - PDB potato dextrose broth - PMM Phycomyces minimal medium - UDA Urtica dioica agglutinin - TEA tri-ethyl-amine - DAB 1,4-diaminobutanone     

Tetrastylis ovalis: A second case of bat-pollinated passionflower (Passifloraceae)

The floral biology ofTetrastylis ovalis and its pollination by bats was studied in southeastern Brazil. The flowers ofT. ovalis seem more fitted to bat-pollination than those ofPassiflora mucronata, the other known chiropterophilous passionflower. Two species of bats were observed visiting flowers ofT. ovalis: the phyllostomidsGlossophaga soricina andRhinophylla pumilio. Due to the attributes ofTetrastylis, its presumed primitiveness and a better defined bat-pollination syndrome we suggest that chiropterophily evolved independently in the former genus and inPassiflora.     

A 2.6 kb intron separates the signal peptide coding sequence of an anther-specific protein from the rest of the gene in sunflower

Summary Metabolism of sulfonylurea herbicides by Streptomyces griseolus ATCC 11796 is carried out via two cytochromes P-450, P-450_SU1 and P-450_SU2. Mutants of S. griseolus, selected by their reduced ability to metabolize a fluorescent sulfonylurea, do not synthesize cytochrome P-450_SU1 when grown in the presence of sulfonylureas. Genetic evidence indicated that this phenotype was the result of a deletion of > 15 kb of DNA, including the structural genes for cytochrome P-450_SU1 and an associated ferredoxin Fd-1 (suaC and suaB, respectively). In the absence of this monooxygenase system, the mutants described here respond to the presence of sulfonylureas or phenobarbital in the growth medium with the expression of only the suhC,B gene products (cytochrome P-450_SU2 and Fd-2), previously observed only as minor components in wild-type cells treated with sulfonylurea. These strains have enabled an analysis of sulfonylurea metabolism mediated by cytochrome P-450_SU2 in the absence of P-450_SU1, yielding an in vivo delineation of the roles of the two different cytochrome P-450 systems in herbicide metabolism by S. griseolus.     

Floral morphogenesis in Anagallis: Scanning-electron-micrograph sequences from individual growing meristems before,during, and after the transition to flowering

Non-destructive scanning electron microscopy allows one to visualize changing patterns of individual cells during epidermal development in single meristems. Cell growth and division can be followed in parallel with morphogenesis. The method is applied here to the shoot apex of Anagallis arvensis L. before, during, and after floral transition. Phyllotaxis is decussate; photoperiodic induction of the plant leads to the production of a flower in the axil of each leaf. As seen from above, the recently formed oval vegetative dome is bounded on its slightly longer sides by creases of adjacent leaf bases. The rounded ends of the dome are bounded by connecting tissue, horizontal bands of node cells between the opposed leaf bases. The major growth axis runs parallel to the leaf bases. While slow-growing at the dome center, this axis extends at its periphery to form a new leaf above each band of connecting tissue. Connecting tissue then forms between the new leaves and a new dome is defined at 90° to the former. The growth axis then changes by 90°. This is the vegetative cycle. The first observed departure from vegetative growth is that the connecting tissue becomes longer relative to the leaf creases. Presumably because of this, the major growth axis does not change in the usual way. Extension on the dome continues between the older leaves until the axis typically buckles a second time, on each side, to form a second crease parallel to the new leaf-base crease. The tissue between these two creases becomes the flower primordium. The second crease also delimits the side of a new apical dome with the major axis and growth direction altered by 90°. During this inflorescence cycle the connecting tissue is relatively longer than before. Much activity is common to both cycles. It is concluded that the complex geometrical features of the inflorescence cycle may result from a change in a biophysical boundary condition involving dome geometry, rather than a comprehensive revision of apical morphogenesis.Abbreviation SEM scanning electron microscopy, micrograph Use of the SEM facility of Professor G. Goffinet, Institute of Zoology, University of Liège, is greatly appreciated. We thank Dr. R. Jacques, C.N.R.S., Le Phytotron, Gif-sur-Yvette, France, for providing the experimental material, and Mr. Philippe Ongena for expert photography. Support was from grants from the U.S. Department of Agriculture and National Science Foundation as well as from the Fonds National de la Recherche Scientifique, Fonds de la Recherche Fondamentale et Collective, and the Action de Recherche Concertée of Belgium.     

Stimulation by auxin of phospholipase A in membrane vesicles from an auxin-sensitive tissue is mediated by an auxin receptor

Microsomal vesicles were prepared from zucchini (Cucurbita pepo L.) hypocotyls containing radioactive phosphatidylethanolamine or phosphatidylcholine, and these lipids were used as substrates by phospholipase A which is activated by auxins. Phospholipase D and phospholipase C hydrolysed the same substrates but were not influenced by auxin. Phospholipase A was activated by the auxins indolyl-3-acetic acid, 2,4-dichlorophenoxyacetic acid and, to a lesser extent, by -naphthaleneacetic acid whereas the weak auxins 2,3-dichlorophenoxyacetic acid and -naphthaleneacetic acid were almost inactive. This hormone specificity was also found in growth tests with etiolated zucchini hypocotyls. Phospholipase A activation by auxin was blocked by a polyclonal antibody against the maize auxin-binding protein. We propose that phospholipase A activation is a primary reaction in the signal transduction leading from hormone-binding to the growth response.Abbreviations IAA indolyl-3-acetic acid - 2,3-D, 2,4-D 2,3- and 2,4-dichlorophenoxyacetic acid - -NAA; -NAA - and -naphthaleneacetic acid This work was supported by the Deutsche Forchungsgemeinschaft. We thank D. Klämbt (Botanical Institute, University of Bonn, FRG) for a generous gift of polyclonal antibody (IgG fraction) against auxin-binding protein and U. Kutschera (Botanical Institute, University of Bonn, FRG) for advice with the growth tests.     

Calcium dependence of the contractile response of the aorta in the rat, rabbit and guinea pig and in the human uterine artery

The influence of extracellular Ca2+ on the contraction produced by noradrenaline (NA) (3 x 10(-6) M), KCl (60 mM) and BaCl2 (30 mM) on human uterine arteries (AUH) and aortic strips from rats, rabbits and guinea-pigs have been studied. The vessels were cut spirally and incubated in Krebs solution containing 2.5 mM Ca2+ (KN), 0 mM Ca2+ (K-0Ca) or 0 mM Ca2+ + 3 mM EDTA (K-EDTA). Both phases (fast and slow) of the response of aortic strips to NA and of the AUH to NA, KCl and BaCl2 were significantly smaller in solutions without Ca2+. Only in rabbit aortic strips the slow phase was significantly more reduced than the fast phase. Overall, the contractions of the rat aortic strips were most resistant to the absence of extracellular Ca2+. These results confirm the variability of the responses of blood vessels from different vascular beds and species to the removal of extracellular Ca2+.