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301.
Jost R Altschmied L Bloem E Bogs J Gershenzon J Hähnel U Hänsch R Hartmann T Kopriva S Kruse C Mendel RR Papenbrock J Reichelt M Rennenberg H Schnug E Schmidt A Textor S Tokuhisa J Wachter A Wirtz M Rausch T Hell R 《Photosynthesis research》2005,86(3):491-508
The treatment of Arabidopsis thaliana with methyl jasmonate was used to investigate the reaction of 2467 selected genes of primary and secondary metabolism by
macroarray hybridization. Hierarchical cluster analysis allowed distinctions to be made between diurnally and methyl jasmonate
regulated genes in a time course from 30 min to 24 h. 97 and 64 genes were identified that were up- or down-regulated more
than 2–fold by methyl jasmonate, respectively. These genes belong to 18 functional categories of which sulfur-related genes
were by far strongest affected. Gene expression and metabolite patterns of sulfur metabolism were analysed in detail, since
numerous defense compounds contain oxidized or reduced sulfur. Genes encoding key reactions of sulfate reduction as well as
of cysteine, methionine and glutathione synthesis were rapidly up-regulated, but none of the known sulfur-deficiency induced
sulfate transporter genes. In addition, increased expression of genes of sulfur-rich defense proteins and of enzymes involved
in glucosinolate metabolism was observed. In contrast, profiling of primary and secondary sulfur metabolites revealed only
an increase in the indole glucosinolate glucobrassicin upon methyl jasmonate treatment. The observed rapid mRNA changes were
thus regulated by a signal independent of the known sulfur deficiency response. These results document for the first time
how comprehensively the regulation of sulfur-related genes and plant defense are connected. This interaction is discussed
as a new approach to differentiate between supply- and demand-driven regulation of the sulfate assimilation pathway. 相似文献
302.
The maize (Zea mays L.) glyceraldehyde-3-phosphate dehydrogenase gene 4 (GapC4) promoter confers anaerobic gene expression in tobacco (Nicotiana tabacum L.), potato (Solanum tuberosum L.) and Arabidopsis thaliana (L.) Heynh. Here we have investigated its expression in hybrid poplar (Populus tremula × P. alba). Our results show that the promoter is not expressed in leaves and stems under normoxic conditions while anaerobiosis induces reporter gene expression in leaves up to a level observed for the STLS-1 promoter from potato that is shown to confer leaf-specific gene expression in transgenic poplar. Anaerobic induction is cell autonomous and requires a CO2 atmosphere and light. As in tobacco, the GapC4 promoter in poplar is wound inducible. The induction by CO2
and light may reflect a natural situation because flooding, a natural cause of anaerobiosis, is often accompanied by high CO2 concentrations in the floodwater. Our results show that the GapC4
promoter is suitable as an anaerobic reporter and as an inducible gene expression system in poplar.Abbreviations CaMV
cauliflower mosaic virus
- GapC4
glyceraldehyde-3-phosphate dehydrogenase gene 4
- GUS
-glucuronidase
- 4-MU
methylumbelliferone
- STLS-1
stem- and leaf-specific promoter 1 相似文献
303.
The tetrahydropyranopterin structure of the sulfur-free and metal-free molybdenum cofactor precursor
Santamaria-Araujo JA Fischer B Otte T Nimtz M Mendel RR Wray V Schwarz G 《The Journal of biological chemistry》2004,279(16):15994-15999
The molybdenum cofactor (Moco), a highly conserved pterin compound coordinating molybdenum (Mo), is required for the activity of all Mo-dependent enzymes with the exception of nitrogenase. Moco is synthesized by a unique and evolutionary old multi-step pathway with two intermediates identified so far, the sulfur-free and metal-free pterin derivative precursor Z and molybdopterin, a pterin with an enedithiolate function essential for Mo ligation. The latter pterin component is believed to form a tetrahydropyranopterin similar to the one found for Moco in the crystal structure of Mo as well as tungsten (W) enzymes. Here we report the spectroscopic characterization and structure elucidation of precursor Z purified from Escherichia coli overproducing MoaA and MoaC, two proteins essential for bacterial precursor Z synthesis. We have shown that purified precursor Z is as active as precursor Z present in E. coli cell extracts, demonstrating that no modifications during the purification procedure have occurred. High resolution electrospray ionization mass spectrometry afforded a [M + H]+ ion compatible with a molecular formula of C10H15N5O8P. Consequently 1H NMR spectroscopy not allowed structural characterization of the molecule but confirmed that this intermediate undergoes direct oxidation to the previously well characterized non-productive follow-up product compound Z. The 1H chemical shift and coupling constant data are incompatible with previous structural proposals and indicate that precursor Z already is a tetrahydropyranopterin system and carries a geminal diol function in the C1' position. 相似文献
304.
Sex pheromone of the citrus mealybug Planococcus citri: synthesis and optimization of trap parameters 总被引:1,自引:0,他引:1
Zada A Dunkelblum E Harel M Assael F Gross S Mendel Z 《Journal of economic entomology》2004,97(2):361-368
A simple synthesis of the pheromone of the citrus mealybug, Planococcus citri (Risso) (Hemiptera: Pseudococcidae), has been developed. Various factors affecting capture of males have been assessed to optimize the trap design and to develop a lure with high efficacy and longevity. Male capture was the same with the racemic and chiral pheromone; technical pheromone (85% purity) was statistically as attractive as pure pheromone (97%). A special formulation was used to determine the actual release rate of the pheromone under field conditions as related to male capture. Generally, plate traps caught more males than delta traps, and large traps caught more than small ones. The effects of aging on the performance of three types of rubber dispensers were evaluated. It was found that the American dispenser displayed the most consistent trapping performance and could be used for monitoring for at least 16 wk with a load of 200 microg of pheromone. The dose-response of the males to sex pheromone was tested within the range of 25-1,600 microg. 相似文献
305.
A fast and highly efficient short-term in vitro regeneration system was developed for barley (Hordeum vulgare L.) based on readily available explants. Clumps of multiple shoots and buds suitable for transformation were obtained 9–10 weeks after culture initiation from model and current commercial cultivars. Meristematic shoot segments (MSSs) excised from mature embryo-derived seedlings and subsequently cultured on MS-based medium containing 2 mg/l Picloram and 3 mg/l thidiazuron (TDZ) differentiated up to ten multiple shoots after 3–4 weeks with no or very little callus formation. Sectors of the already multiplied shoot clumps were further multiplied on proliferation-maintenance medium containing 2 mg/l Picloram and 2.5 mg/l TDZ. Biweekly subcultures resulted in a continuous process of multiplication of these highly differentiating green sectors without any loss of morphogenic potential. The differentiated small shoots and shoot buds gave rise to normal shoots on medium with 0.1 mg/l Picloram and 1 mg/l TDZ. After rooting on basal medium with 0.5 mg/l or 1 mg/l IBA the plants were transferred to soil and showed normal growth and fertility compared to the seed-grown plants. All of the genotypes tested formed multiple shoots. The percentage of relative MSS multiplication was 63–83%, and the average number of multiplied shoots per MSS ranged from 16 to 34 among the genotypes after 9–11 weeks.Abbreviations BAP
6-Benzylaminopurine
- 2,4-D
2,4-Dichlorophenoxyacetic acid
- Dicamba
3,6-Dichloro-2-methoxybenzoic acid
- IBA
Indole-3-butyric acid
- MSS
Meristematic shoot segment
- NAA
-Naphthaleneacetic acid
- Picloram
4-Amino-3,5,6-trichloropicolinic acid
- TDZ
Thidiazuron 相似文献
306.
Nowak K Luniak N Meyer S Schulze J Mendel RR Hänsch R 《Plant biology (Stuttgart, Germany)》2004,6(1):65-73
The jellyfish (Aequorea victoria) green fluorescent protein (GFP) and its variants (CFP [cyan] and YFP [yellow]) were successfully used as a vital marker system for the transformation of hybrid poplar (Populus tremula x P. alba). Our results show that, in this woody plant, fluorescent proteins can be expressed: (i) transiently in protoplasts after PEG-mediated transformation, as well as in leaf cells after particle bombardment, and (ii) stably in callus cells and plants after Agrobacterium-mediated transformation. For these studies, we constructed vectors permitting easy recloning of any promoter fragments of interest. Confocal laser scanning microscopy was used both for visualization and differentiation between the different colours of the GFP variants and autofluorescence of chlorophyll and lignified xylem vessels. Peroxisomes were chosen as target organelles for GFP translocation via the peroxisomal targeting sequence PTS1 because this allowed us to concentrate the fluorochrome in the small volume of a few peroxisomes, giving a bright fluorescence over background noise. 相似文献
307.
Identification and biochemical characterization of Arabidopsis thaliana sulfite oxidase. A new player in plant sulfur metabolism 总被引:9,自引:0,他引:9
Eilers T Schwarz G Brinkmann H Witt C Richter T Nieder J Koch B Hille R Hänsch R Mendel RR 《The Journal of biological chemistry》2001,276(50):46989-46994
In mammals and birds, sulfite oxidase (SO) is a homodimeric molybdenum enzyme consisting of an N-terminal heme domain and a C-terminal molybdenum domain (EC ). In plants, the existence of SO has not yet been demonstrated, while sulfite reductase as part of sulfur assimilation is well characterized. Here we report the cloning of a plant sulfite oxidase gene from Arabidopsis thaliana and the biochemical characterization of the encoded protein (At-SO). At-SO is a molybdenum enzyme with molybdopterin as an organic component of the molybdenum cofactor. In contrast to homologous animal enzymes, At-SO lacks the heme domain, which is evident both from the amino acid sequence and from its enzymological and spectral properties. Thus, among eukaryotes, At-SO is the only molybdenum enzyme yet described possessing no redox-active centers other than the molybdenum. UV-visible and EPR spectra as well as apparent K(m) values are presented and compared with the hepatic enzyme. Subcellular analysis of crude cell extracts showed that SO was mostly found in the peroxisomal fraction. In molybdenum cofactor mutants, the activity of SO was strongly reduced. Using antibodies directed against At-SO, we show that a cross-reacting protein of similar size occurs in a wide range of plant species, including both herbacious and woody plants. 相似文献
308.
The molybdenum cofactor biosynthetic protein Cnx1 complements molybdate-repairable mutants, transfers molybdenum to the metal binding pterin, and is associated with the cytoskeleton
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Schwarz G Schulze J Bittner F Eilers T Kuper J Bollmann G Nerlich A Brinkmann H Mendel RR 《The Plant cell》2000,12(12):2455-2471
Molybdenum (Mo) plays an essential role in the active site of all eukaryotic Mo-containing enzymes. In plants, Mo enzymes are important for nitrate assimilation, phytohormone synthesis, and purine catabolism. Mo is bound to a unique metal binding pterin (molybdopterin [MPT]), thereby forming the active Mo cofactor (Moco), which is highly conserved in eukaryotes, eubacteria, and archaebacteria. Here, we describe the function of the two-domain protein Cnx1 from Arabidopsis in the final step of Moco biosynthesis. Cnx1 is constitutively expressed in all organs and in plants grown on different nitrogen sources. Mo-repairable cnxA mutants from Nicotiana plumbaginifolia accumulate MPT and show altered Cnx1 expression. Transformation of cnxA mutants and the corresponding Arabidopsis chl-6 mutant with cnx1 cDNA resulted in functional reconstitution of their Moco deficiency. We also identified a point mutation in the Cnx1 E domain of Arabidopsis chl-6 that causes the molybdate-repairable phenotype. Recombinant Cnx1 protein is capable of synthesizing Moco. The G domain binds and activates MPT, whereas the E domain is essential for activating Mo. In addition, Cnx1 binds to the cytoskeleton in the same way that its mammalian homolog gephyrin does in neuronal cells, which suggests a hypothetical model for anchoring the Moco-synthetic machinery by Cnx1 in plant cells. 相似文献
309.
The crystal structure of human ornithine transcarbamylase (OTCase) complexed with carbamoyl phosphate (CP) and L-norvaline (NOR) has been determined to 1.9-A resolution. There are significant differences in the interactions of CP with the protein, compared with the interactions of the CP moiety of the bisubstrate analogue N-(phosphonoacetyl)-L-ornithine (PALO). The carbonyl plane of CP rotates about 60 degrees compared with the equivalent plane in PALO complexed with OTCase. This positions the side chain of NOR optimally to interact with the carbonyl carbon of CP. The mixed-anhydride oxygen of CP, which is analogous to the methylene group in PALO, interacts with the guanidinium group of Arg-92; the primary carbamoyl nitrogen interacts with the main-chain carbonyl oxygens of Cys-303 and Leu-304, the side chain carbonyl oxygen of Gln-171, and the side chain of Arg-330. The residues that interact with NOR are similar to the residues that interact with the ornithine (ORN) moiety of PALO. The side chain of NOR is well defined and close to the side chain of Cys-303 with the side chains of Leu-163, Leu-200, Met-268, and Pro-305 forming a hydrophobic wall. C-delta of NOR is close to the carbonyl oxygen of Leu-304 (3.56 A), S-gamma atom of Cys-303 (4.19 A), and carbonyl carbon of CP (3.28 A). Even though the N-epsilon atom of ornithine is absent in this structure, the side chain of NOR is positioned to enable the N-epsilon of ornithine to donate a hydrogen to the S-gamma atom of Cys-303 along the reaction pathway. Binding of CP and NOR promotes domain closure to the same degree as PALO, and the active site structure of CP-NOR-enzyme complex is similar to that of the PALO-enzyme complex. The structures of the active sites in the complexes of aspartate transcarbamylase (ATCase) with various substrates or inhibitors are similar to this OTCase structure, consistent with their common evolutionary origin. 相似文献
310.