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91.
Christoph Theurer Hans-Joachim Treumann Thomas Faust Ursula May Wolfgang Kreis 《Plant Cell, Tissue and Organ Culture》1994,38(2-3):327-335
The glycosylation and deglycosylation of cardiac glycosides was investigated using cell suspension cultures and shoot cultures, both established from Digitalis lanata EHRH. plants, as well as isolated enzymes. Shoots were capable of glucosylating digitoxigenin, evatromonoside, digiproside, glucodigitoxigenin and digitoxin. Suspension cultured Digitalis cells glucosylated all the substrates mentioned but digiproside, whereas the UDP-glucosedependent cardinolide glucosyltransferase isolated from that source did not accept digitoxigenin and digiproside as substrates. It is concluded that at least three different glucosyltransferases are involved in cardiac glycoside formation in Digitalis. Similar experiments carried out with glucosylated cardenolides which were administered to cultured cells, shoots and a cardenolide -glucosidase isolated from young leaves revealed that at least two different glucosidases occur in Digitalis lanata, albeit in different tissues or during different phases of development. The biotransformation of glucoevatromonoside was investigated using unlabelled compound and [14C-glucose]-glucoevatromonoside synthesized enzymatically. After 7 d of incubation almost no radioactivity could be recovered from the cardenolide fraction, indicating that the terminal glucose of glucoevatromonoside was now incorporated into volatile, hydrophilic and insoluble compounds. Since, on the other hand, large amounts of cardenolides were found in the experiments with unlabelled glucoevatromonoside it is assumed that steady state or pool size regulation is achieved by the coordinated action of a cardenolide glucosidase and a glucosyltransferase.Abbreviations Acdox
D-acetyldigitoxose
- dgen
digoxigenin
- dox
D-digitoxose
- dten
digitoxigenin
- dtl
D-digitalose
- fuc
D-fucose
- gten
gitoxigenin
- qun
D-quinovose
- CGH
cardenolide 16-O-glucohydrolase
- DFT
UDP-fucose:digitoxigenin 3-O-fucosyltransferase
- DGT
UDP-glucose:Digitoxin 16-O-glucosyltransferase
- DQT
UDP-quinovose:digitoxigenin 3-O-quinovosyltransferase 相似文献
92.
Indole-3-acetic acid (IAA), fusicoccin and weak acids all lower the cytoplasmic pH (pHi) and induce elongation growth of maize (Zea mays L.) coleoptiles. Gibberellic acid (GA3) also induces elongation growth and we have used confocal laser scanning microscopy to study the effects of GA3 on pHi employing the pH-indicator dyes, 2,7-bis(2-carboxyethyl)-5-(and-6) carboxyfluorescein and carboxy-semi-naphthorhodafluor-1. We confirm that GA3 induces growth significantly in light-grown but only slightly or not at all in dark-grown coleoptiles. The growth induced by IAA treatment was similar in light- and dark-grown coleoptiles. The pHi decreased by up to 0.6 units during the first 7 min of GA3 or IAA treatment of both light- and dark-grown coleoptiles. Gibberellic acid inhibited IAA-induced growth of dark-grown coleoptiles. Hence, in dark-grown coleoptiles GA3 may activate either directly or indirectly reactions that interfere with the signalling pathway leading to elongation growth. The possible role of pHi in growth is discussed.Abbreviations ABA
abscisic acid
- AM
acetoxymethyl ester
- BCECF
2,7-bis(2-carboxyethyl)-5-(and-6) carboxyfluorescein
- [Ca2+]i
cytoplasmic free calcium
- GA(n)
gibberellin A(n)
- GA3
gibberellic acid
- IAA
indole-3-acetic acid
- PGR
plant growth regulator
- pHi
cytoplasmic pH
- Pipes
piperazine-N,N-bis[2-ethanesulfonic acid]
- Snarf-1
carboxy-semi-naphthorhodafluor-1
We thank Dr R. King (CSIRO, Canberra) for providing the GA1 and T. Phillips for processing the photographic material. H.R. Irving was supported by an Australian Research Council Research Fellowship and the work was supported by an Australian Research Council grant. 相似文献
93.
Werner Landolt Madeleine Günthardt-Goerg Ilse Pfenninger Christoph Scheidegger 《Trees - Structure and Function》1994,8(4):183-190
Summary Cuttings of hybrid poplar (Populus × euramericana var. Dorskamp) were exposed to ozone (80 g/m3 from 2100 hours to 0700 hours, 180 g/m3 from 0700 hours to 2100 hours) for 3 months. Ozone reduced the starch content in leaves and stem bark, whereas starch granules accumulated in bundle sheath cells along small leaf veins. At the same time, sucrose and inositol content increased in the leaves. Mesophyll cells in the vicinity of the stomata were injured first, and droplet-like material appeared on their walls. In the sieve plates of fumigated trees, the pores showed a higher degree of narrowing than those of the control treatment. Cell collapse in the leaves was accompanied by water loss and an increase in air space. In the stems, the ozone treatment led to a reduced radial width, particularly in the xylem tissue. These results are discussed in relation to reduced or inhibited phloem loading and ozone-induced drought stress. The plants injured by ozone showed quite distinct patterns of metabolite responses as well as enzyme activities (PEP- and RubP-carboxylase) in the leaves from the top to the bottom. There were also remarkable differences in the reaction of sucrose and inositol between leaves and stem bark. Future research should therefore increasingly follow a whole-plant approach for a better understanding of complex plant reactions. 相似文献
94.
95.
Liang Ru Shi Dirk Eichelbauer Franz Borchard Herbert Jürgens Ulrich Göbel E. Marion Schneider 《Cancer immunology, immunotherapy : CII》1994,38(3):208-213
A selection of 16 monoclonal antibodies has been produced against a fresh Ewing's sarcoma (ES) tumor mixed with a permanent ES cell line. The majority of antibodies identify an 80-kDa molecule, which is not detected on healthy tissues except on certain cultured monocytes. One antibody recognize the CD2 ligand MIC2 and 2 antibodies (numbers 13 and 16) define a higher-molecular-mass antigen. Antibody 16 is also expressed on mesenchymal fibroblasts of bone marrow or fetal origin. Tumorspecific antigen expression is potentially linked to the chromosome 22 abnormality decribed in Ewing's sarcoma. 相似文献
96.
In the existing genetic sexing strains for the medfly, Ceratitis capitata, male recombination leads to breakdown of the sexing mechanism under mass rearing conditions. The rate of breakdown depends on the recombination frequency and on the fitness of the recombinants. We have tested two different sexing genes, white pupa and a temperature sensitive lethal, in combination with the translocation T(Y;5)30C. Both sexing strains broke down, although at very different rates. In the case of the white pupa strain, 3.5% recombinants were observed after rearing the strain for 15 generations. The second strain, utilizing white pupa and the temperature sensitive lethal as selectable markers, already reached a comparable level after six generations and was broken down completely in the ninth generation. In these strains the frequency of recombination is high because the breakpoint of T(Y;5)30C and the sexing gene(s) are far apart. To remedy the situation, we have isolated four new translocations with breakpoints located closer to the sexing genes. Mass rearing was simulated for several generations with strains based on these translocations and no breakdown was observed under the conditions used. 相似文献
97.
Summary The chloroplast ribosomal intron of Chlamydomonas reinhardtii encodes a sequence-specific DNA endonuclease (I-CreI), which is most probably involved in the mobility of this intron. Here we show that I-CreI generates a 4 by staggered cleavage just downstream of the intron insertion site. The I-CreI recognition sequence is 19–24 by in size and is located asymmetrically around the intron insertion site. Screening of natural variants of the I-CreI recognition sequence indicates that the I-CreI endonuclease tolerates single and even multiple base changes within its recognition sequence. 相似文献
98.
Alwin Krmer Jan Pudil Heinz Frank Franz Oesch Handruedi Glatt 《Mutation research》1993,290(2):165-174
Trans-stilbene oxide, trans-β-methylstyrene, 7,8-oxide, trans-β-ethylstyrene, 7,8-oxide, trans-β-propylstyrene 7,8-oxide and 4-fluorochalcone oxide were investigated for genotoxic activity in bacterial and mammalian cells, in the absence of external xenobiotic-metabolising systems. All compounds strongly enhanced the frequency of sister-chromatid exchanges (SCE) in cultured human lymphocytes. None of them was mutagenic in Salmonella typhimurium (reversion of the his− strains TA98, TA100 and TA104). The limit of detection was 1/20,000 to 1/106 of the activity of the positive control, benzo[a]pyrene 4,5-oxide, depending on the compound and the bacterial strain. Trans-β-methylstyrene 7,8-oxide and 4-fluorochalcone oxide were additionally tested for induction of SCE and gene mutations in the same target cells, namely Chinese hamster V79 cells. Their influence on the level of SCE was similar to that observed in human lymphocytes, whilst gene mutations (at the hprt locus) were not induced. The four investigated styrene oxide derivatives are known to be excellent substrates for a mammalian enzyme, cytosolic epoxide hydrolase (cEH). 4-Fluorochalcone oxide is a potent selective inhibitor of this enzyme and is structurally similar to the investigated styrene oxide derivatives. These properties of the test compounds however cannot explain the observed discrepancies in the results, since the genetic end point (SCE versus gene mutations) was decisive, and SCE were induced in cEH-proficient human lymphocytes as well as in cEH-deficient V79 cells. 相似文献
99.
Uwe Ludewig Christoph Lorra Olaf Pongs Stefan H. Heinemann 《European biophysics journal : EBJ》1993,22(4):237-247
The members of the RCK family of cloned voltage-dependent K+ channels are quite homologous in primary structure, but they are highly diverse in functional properties. RCK4 channels differ from RCK1 and RCK2 channels in inactivation and permeation properties, the sensitivity to external TEA, and to current modulation by external K+ ions. Here we show several other interesting differences: While RCK1 and RCK2 are blocked in a voltage and concentration dependent manner by internal Mg2+ ions, RCK4 is only weakly blocked at very high potentials. The single-channel current-voltage relations of RCK4 are rather linear while RCK2 exhibits an inwardly rectifying single-channel current in symmetrical K+ solutions. The deactivation of the channels, measured by tail current protocols, is faster in RCK4 by a factor of two compared with RCK2. In a search for the structural motif responsible for these differences, point mutants creating homology between RCK2 and RCK4 in the pore region were tested. The single-point mutant K533Y in the background of RCK4 conferred the properties of Mg2+ block, tail current kinetics, and inward ion permeation of RCK2 to RCK4. This mutant was previously shown to be responsible for the alterations in external TEA sensitivity and channel regulation by external K+ ions. Thus, this residue is expected to be located at the external side of the pore entrance. The data are consistent with the idea that the mutation alters the channel occupancy by K+ and thereby indirectly affects internal Mg2+ block and channel closing.Abbreviations TEA
tetraethylammonium
- EGTA
Ethylene glycol-bis (-aminoethyl ether) N,N,N,N-tetraacetic acid
- 2S3B model
2-site 3-barrier model
Correspondence to: S. H. Heinemann 相似文献
100.
Summary Polyclonal antibodies were produced against the highly purified enzymes L-hydantoinase, hydantoin-racemase and L-N-carbamoylamino acid amidohydrolase of Arthrobacter aurescens DSM 3747. In order to exploit these antibodies for basic research (molecular biology) or bioengineering (process development), the serological properties had to be characterized. Both, the hydantoinase- and carbamoylase-antibodies were observed to be monofunctional, whereas the hydantoin-racemase-antibody was found to be additionally specific against the L-hydantoinase. Monospecificity was realized after affinity chromatography. Investigations on serological crossreactions with several linear- and cyclic amidases (e.g. hydantoinases) as well as hydantoin-racemases are demonstrated in this paper.Deticated to Prof. Dr. Klaus Mosbach on the occation of his 60th birthday. 相似文献