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  • 1
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Thirteen cultivars of rice (Oryza sativa) were tested for plant regeneration from calli initiated from the scutella of mature seeds by water stress treatment using a high concentration of agarose, and examined for their response to Agrobacterium tumefaciens LBA4404, carrying a plasmid pTOK233, harboring genes for kanamycin resistance (nptII), hygromycin resistance (hpt) and β-glucuronidase (gus). Plant regeneration frequency was considerably increased in most of the cultivars when the calli were treated with water stress, as compared with untreated controls. In particular, the cultivars Dongjinbyeo, IR43, Nagdongbyeo and Sinseonchalbyeo showed an increased frequency of shoot regeneration. Expression of GUS was detected in all of the co-cultivated cultivars. Based on GUS expression at 3 days after co-cultivation with A. tumefaciens, three rice cultivars (Dongjinbyeo, Hwayoungbyeo and Nagdongbyeo) were judged highly susceptible to A. tumefaciens, while Milyang 23, Nonganbyeo and Samgangbyeo cultivars were weakly susceptible. Plantlets were readily regenerated when the hygromycin-resistant calli were transferred to a regeneration medium containing hygromycin. Intense blue staining was observed in GUS assays of leaf segments, roots and flower organs from regenerated plants. Stable integration and expression of the introduced hpt and gus genes were confirmed by Southern blot analysis of the transformants. Therefore, Dongjinbyeo and Nagdongbyeo cultivars proved to be both highly susceptible to A. tumefaciens and highly responsive to plant regeneration.
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  • 2
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A cDNA encoding RARF1 (rice ADP-ribosylation factor 1) was isolated from fungal elicitor-treated rice suspension culture cells by mRNA differential display. RARF1 transcripts accumulated in response to hydrogen peroxide (H2O2) and salicylic acid (SA) and rapidly in cells inoculated with an avirulent pathogen. Constitutively over-expressed RARF1 under the control of the cauliflower mosaic virus 35S promoter (CaMV 35S) triggered spontaneous induction of lesion mimics, induced an array of pathogenosis-related (PR) genes, reduced susceptibility to a fungal pathogen, and caused accumulation of SA. From these data, we deduced that RARF1 might be a component of various plant defence signalling pathways involved in inducing the expression of a subset of PR genes.
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  • 3
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A DNA fragment (pCHI5422) containing two genes encoding a 54-kDa and a 22-kDa chitinase was isolated from a cosmid DNA library of Serratia marcescens KCTC2172. The complete nucleotide sequence of pCHI5422 consisting of 4581 bp was determined. The nucleotide sequence of the 22-kDa chitinase consists of 681 bp of open reading frame encoding 227 amino acids and is located 1422 bp downstream of the translation termination codon of the 54-kDa chitinase sequence. The 54-kDa chitinase gene consisted of 1497 bp in a single open reading frame encoding 499 amino acids. The genes encoding the 54-kDa and 22-kDa chitinase were separately subcloned in Escherichia coli and the individual chitinases were expressed and purified from the culture broth using chitin affinity chromatography. When chitohexaose was used as substrate, the major product of the enzymatic reaction of both the 54-kDa and 22-kDa chitinases was a (GlcNAc)2 dimer with a minor amount of monomer. The specific activity of the 54-kDa and 22-kDa chitinases were 300 μM (min)−1 mg−1 and 17 μM (min)−1 mg−1 on the natural swollen chitin, respectively.
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  • 4
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A chitinase gene (pCHI52) encoding the 52-kDa chitinase was isolated from a Serratia marcescens KCTC2172 cosmid library. This chitinase gene consists of 2526 bp with an open reading frame that encodes 485 amino acids. Escherichia coli harboring the pCHI52 gene secreted not only a 52-kDa but also a 35-kDa chitinase into the culture supernatant. We purified both 52-kDa and 35-kDa chitinases using a chitin affinity column and Sephacryl-S-300 gel filtration chromatography. We determined that the 17 N-terminal amino acid sequences of the 52-kDa and the 35-kDa chitinase are identical. Furthermore, a protease obtained from S. marcescens KCTC2172 cleaved the 52-kDa chitinase into the 35-kDa protein with chitinase activity. These results suggest that the 35-kDa chitinase derives from the 52-kDa chitinase by post-translational proteolytic modification. The optimal reaction temperature of 45°C and the optimal pH of 5.5 were identical for both enzymes. The specific activities of the 52-kDa and 35-kDa chitinases on natural swollen chitin were 67 μmol min−1 mg−1 and 60 μmol min−1 mg−1, respectively.
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  • 5
    Electronic Resource
    Electronic Resource
    s.l. ; Stafa-Zurich, Switzerland
    Solid state phenomena Vol. 124-126 (June 2007), p. 671-674 
    ISSN: 1662-9779
    Source: Scientific.Net: Materials Science & Technology / Trans Tech Publications Archiv 1984-2008
    Topics: Physics
    Notes: Hydrophobic thin film silica aerogels were synthesized by ambient pressure drying methodfrom silicic acid which was prepared from sodium silicate (water glass) solution. The pH value of thesilica precursor sol was adjusted to make a spinable sol, and gel films were coated on a glass substrateby dip coating technique. Aerogel-like thin films with the thickness of about 1 μm could besuccessfully fabricated by repeating the dip coating process three times
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  • 6
    Electronic Resource
    Electronic Resource
    s.l. ; Stafa-Zurich, Switzerland
    Key engineering materials Vol. 277-279 (Jan. 2005), p. 481-486 
    ISSN: 1013-9826
    Source: Scientific.Net: Materials Science & Technology / Trans Tech Publications Archiv 1984-2008
    Topics: Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Notes: We have chosen the GPR method for investigating the inner structure of a stone pagoda in a non-destructive way. The selection of a suitable source frequency of the GPR antenna is the key because the main frequency of its pulse controls the resolution and the depth of the investigation. Through theoretical consideration and numerical simulation, we found that 500 to 800 MHz is suitable for a field model with a foundation part of 3 to 4 m. To image the inner structure we selected 3-D Kirchhoff prestack depth migration technique used in seismic processing. We have used themodified migration algorithm for the source and receiver configurations of common offset GPR data. To verify the scheme, we calculated the synthetic data using the 3-D FDTD algorithm and applied the migration technique to it. Through these experiments, we confirmed that the 3-D Kirchhoff prestack depth migration technique is a very powerful tool to image the inside of a stone pagoda with high resolution. We have also applied the technique to the field data of the foundation of a five-story stone pagoda at Jeongnim temple site in Buyeo City, Korea. Based on the 3-D migrated images, we inferred that the structure of the foundation of this pagoda seemed to be preserved quite well and the thickness of the outer wall was about 0.5 m
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  • 7
    Electronic Resource
    Electronic Resource
    s.l. ; Stafa-Zurich, Switzerland
    Materials science forum Vol. 544-545 (May 2007), p. 1053-1056 
    ISSN: 1662-9752
    Source: Scientific.Net: Materials Science & Technology / Trans Tech Publications Archiv 1984-2008
    Topics: Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Notes: In order to prevent the irreversible collapse of the silica aerogel backbone upon the dryingof the liquid phase of silica wet gel derived from water glass based silicic acid, we tried to strengthenthe aerogel back bone by aging silica wet gels in water and TEOS/ethanol solutions. Although agingof silica wet gels in water has been shown to grow the neck between silica particles of the aerogelbackbone, it is hard to obtain a crack-free aerogel monolith. On the other hand, the mechanicalstability of silica aerogels was improved significantly by aging the wet gel in TEOS/ethanol solutionswith different TEOS content
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  • 8
    ISSN: 1432-2048
    Keywords: Key words: Differential display ; Functional analysis ; Fungal elicitor treatment ; Oryza (rab-GDIs) ; Pathogen signaling ; Rab-specific GDP dissociation inhibitor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. By using the mRNA differential display approach to isolate defense signaling genes active at the early stage of fungal infection two cDNA fragments with high sequence homology to rab-specific GDP-dissociation inhibitors (GDIs) were identified in rice (Oryza sativa L.) suspension cells. Using polymerase-chain-reaction products as probes, two full-length cDNA clones were isolated from a cDNA library of fungal-elicitor-treated rice, and designated as OsGDI1 and OsGDI2. The deduced amino acid sequences of the isolated cDNAs exhibited substantial homology to Arabidopsis rab-GDIs. Northern analysis revealed that transcripts detected with the 3′-gene-specific DNA probes accumulated to high levels within 30 min after treatment with a fungal elicitor derived from Magnaporthe grisea. The functionality of the OsGDIs was demonstrated by their ability to rescue the Sec19 mutant of Saccharomyces cerevisiae which is defective in vesicle transport. The proteins, expressed in Escherchia coli, cross-reacted with a polyclonal antibody prepared against bovine rab-GDI. Like bovine rab-GDI, the OsGDI proteins efficiently dissociated rab3A from bovine synaptic membranes. Using the two-hybrid system, it was shown that the OsGDIs specifically interact with the small GTP-binding proteins belonging to the rab subfamily. The specific interaction was also demonstrated in vitro by glutathione S-transferase resin pull-down assay.
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  • 9
    ISSN: 1573-5044
    Keywords: morphology ; Oryza sativa cv. Taipei 309 ; plant regeneration ; rice protoplast culture ; somatic variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Rice protoplasts were cultured using 4 different culture procedures such as agarose embedding (AE) without feeder cells and the use of filter membranes (MEM), one layer of nylon mesh (MS1), or a double layer of nylon mesh (MS2) with the inclusion of Lolium multiflorum as feeder cells. The protoplast plating efficiency was highest on the MEM, followed by MS2, MS1 and AE. However, plant regeneration frequencies were highest for MS1, followed by MS2, MEM and AE. The protoclonal plants differed in the morphology of leaves, flowers, spikelets, and panicles in comparison to seed-derived plants. They varied in almost every phenotypic characters evaluated. In many cases, the variation was significantly different in characteristics such as plant height, flag leaf length and width and ratio, and in panicle characteristics such as panicle length, number of primary branches, and number of spikelets per panicle. The number of seeds per panicle was greatly reduced in protoclonal plants when compared with seed-derived control plants. The seeds showed also significant differences in grain length and width in comparison to the control plants. Among the 4 groups of protoclonal plants derived from the 4 different culturing procedures themselves, there were also variations in almost all the phenotypic characteristics assessed.
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  • 10
    ISSN: 1573-5028
    Keywords: class II chitinase ; dephosphorylation ; defense response ; elicitors ; Oryza sativa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Among the four classes of chitinase, a class II chitinase had not yet been reported for rice. We have isolated and characterized a class II acidic chitinase, Rcht2, from rice (Oryza sativa L. cv. Cheongcheongbyeo). The protein consists of a single polypeptide chain of 261 amino acid residues and includes a putative signal sequence of 29 amino acids at its N-terminus. It has a calculated molecular mass of 27 642 Da and an isoelectric point of 5.56. The Rcht2 chitinase lacks the cysteine-rich and hinge domains in the N-terminal region of the protein, which is the criterion for its classification as a class II chitinase. Comparison of the genomic and the cDNA sequence revealed that the coding region of Rcht2 consist of three exons of 301, 112, and 370 bp separated by two introns of 89 and 984 bp. In suspension-cultured rice cells, the transcript level of Rcht2 was dramatically increased by treatment with both glycol chitin and fungal elicitor. The application of protein phosphatase 1 and 2A inhibitors, calyculin A and okadaic acid, effectively abolished the induction of Rcht2 in response to fungal elicitor. In contrast, the activation of Rcht2 transcript was not inhibited by both cycloheximide and protein kinase inhibitors. These results demonstrate that protein dephosphorylation events play a crucial role in the elicitor-mediated induction of Rcht2 in rice cells, while de novo protein synthesis is not required for induction.
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