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  • gene expression  (307)
  • Triticum aestivum  (229)
  • Inversion
  • Rat
  • Springer  (593)
  • Conseil de l'Europe  (1)
  • 3
  • Wiley
  • 1995-1999  (594)
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Animal cognition 2 (1999), S. 123-129 
    ISSN: 1435-9456
    Keywords: Key words Face recognition ; Inversion ; Expertise ¶effect ; Matching-to-sample ;  Macaca mulatta
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract This study investigated the face inversion effect in rhesus monkeys (Macaca mulatta). Face stimuli consisted of ten black-and-white examples of unfamiliar rhesus monkey faces, brown capuchin faces, and human faces. Two non-face categories included ten examples of automobiles and abstract shapes. All stimuli were presented in a sequential matching-to-sample format using an automated joystick-testing paradigm. Subjects performed significantly better on upright than on inverted presentations of automobiles, rhesus monkey and capuchin faces, but not human faces or abstract shapes. These results are inconsistent with data from humans and chimpanzees that show the inversion effect only for categories of stimuli for which subjects have developed expertise. The inversion effect in rhesus monkeys does not appear to be face-specific, and should therefore not be used as a marker of specialized face processing in this species.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Entomologia experimentalis et applicata 91 (1999), S. 403-412 
    ISSN: 1570-7458
    Keywords: aphid ; Diuraphis noxia ; Triticum aestivum ; endosymbiont ; insect-plant interactions ; phloem ; amino acids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The feeding behavior of Diuraphis noxia Mordvilko (Homoptera: Aphididae) on susceptible hosts causes both ultrastructural and tissue level damage which may affect phloem composition. Genetic evidence suggests that endosymbiotic bacteria in most aphids overproduce limiting amino acids to benefit hosts but that D. noxia depends less on endosymbionts for these nutrients, possibly due to an enriched diet. To determine whether D. noxia feeding damage results in higher concentrations of essential amino acids, stylet exudates were analyzed from wheat (Triticum aestivum) damaged to different degrees. Comparison of samples from undamaged and damaged susceptible wheat revealed changes in amino acid composition and an increase in levels of essential amino acids, indicating a nutritionally enhanced ingesta. The changes in stylet exudates paralleled changes in leaf exudates, indicating that the effects are systemic. Feeding damage is not observed on a resistant wheat host, var. Halt, and leaf exudates from infested Halt did not show changes in amino acid composition. Mean relative growth of nymphs was significantly lower on Halt than on susceptible Arapahoe, indicating that Halt is a less suitable host. Both varieties show similar amino acid levels in non-infested samples, suggesting that D. noxia infestation does not enhance the phloem environment in Halt. This study provides evidence that aphid feeding can generate a nutritionally enhanced phloem diet.
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  • 3
    ISSN: 1570-7458
    Keywords: sieve element ; salivation ; aphid ; plant resistance ; wheat ; Sitobion fragariae ; Triticum aestivum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Extended sieve element salivation (E1 waveform in the electrical penetration graph) is a characteristic activity during early sieve element punctures, particularly in resistant plants. In order to explore a chemically-mediated mechanism of resistance associated with sieve element salivation, we compared the pattern of feeding behaviour of the aphid, Sitobion fragariae (Walker), on two cultivars of the wheat Triticum aestivum L., with different concentrations of hydroxamic acids (Hx). During 24 h of electronic monitoring, aphids dedicated over 50% of the total time to phloem ingestion from the sieve elements. Total time allocated to E1 in the experiment, time to first E1 within the experiment, time allocated to E1 before a sustained phloem ingestion (E2) and the contribution of sieve element salivation to the phloem phase (E1/[E1+E2]) were significantly higher in the high-Hx cultivar. The increased salivation in plants with higher contents of Hx suggests the existence, at least in this system, of a chemically-mediated sieve element constraint.
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  • 4
    ISSN: 1432-1890
    Keywords: Key words Glomus mosseae ; Hydroponics ; Nitrate uptake ; Root respiration ; Triticum aestivum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Oxygen and CO2 fluxes were measured in hydroponically grown mycorrhizal and non-mycorrhizal Triticum aestivum L. cv. Hano roots. The NO3 – uptake of the plants was used to estimate the amount of root respiration attributable to ion uptake. Plants were grown at 4 mM N and 10 μM P, where a total and viable mycorrhizal root colonisation of 48% and 18%, respectively, by Glomus mosseae (Nicol. and Gerd.) Gerd. and Trappe (BEG 107) was observed. The O2 consumption and NO3 – uptake rates were similar and the CO2 release was higher in mycorrhizal than in non-mycorrhizal wheat. This resulted in a significantly higher respiratory quotient (RQ, mol CO2 mol–1 O2) in mycorrhizal (1.27±0.13) than in non-mycorrhizal (0.79±0.05) wheat. As the biomass and N and P concentrations in mycorrhizal and non-mycorrhizal wheat were the same, the higher RQ resulted from the mycorrhizal colonisation and not differences in nutrition per se.
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  • 5
    ISSN: 1432-0789
    Keywords: Key words Phosphate-solubilizing microorganisms ; Mussoorie rock phosphate ; Vesicular-arbuscular mycorrhizae ; Triticum aestivum ; Nutrient-deficient soils
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract  The effect of inoculating wheat (Triticum aestivum L.) with the PO4 3–-solubilizing microorganisms (PSM) Bacillus circulans and Cladosporium herbarum and the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus sp. 88 with or without Mussoorie rock phosphate (MRP) amendment in a nutrient-deficient natural sandy soil was studied. In the sandy soil of low fertility root colonization by VAM fungi was low. Inoculation with Glomus sp. 88 improved root colonization. At maturity, grain and straw yields as well as N and P uptake improved significantly following inoculation with PSM or the VAM fungus. These increases were higher on combined inoculation of PSM and the VAM fungus with MRP amendment. In general, a larger population of PSM was maintained in the rhizosphere of wheat in treatments with VAM fungal inoculation and MRP amendment. The results suggest that combined inoculation with PSM and a VAM fungus along with MRP amendment can improve crop yields in nutrient-deficient soils.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology reporter 17 (1999), S. 371-383 
    ISSN: 1572-9818
    Keywords: epidermal peel ; extraction ; gene expression ; stomata ; tree tobacco
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Stomatal guard cells are critical for maintenance of plant homeostasis and represent an interesting cell type for studies of leaf cell differentiation and patterning. Here we describe techniques for the isolation of guard cell RNA and protein from blended epidermal peels of Nicotiana glauca. The RNA isolation procedure is a modification of the hot borate method, which is particularly well-suited for recalcitrant tissues. Protein was extracted by disrupting guard cell-enriched epidermis with a French® press. This system offers the following advantages: relatively high yield, low or no contamination by other cell types, fresh tissue as a source of RNA and protein rather than protoplasts, and a plant species that is readily transformable. These techniques will allow for cloning and analysis of genes expressed in guard cells, application of traditional biochemical techniques to guard cell proteins, as well as characterization of genetic manipulation of guard cell function in transgenic plants.
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  • 7
    Electronic Resource
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    Springer
    Nutrient cycling in agroecosystems 53 (1999), S. 157-175 
    ISSN: 1573-0867
    Keywords: Brassica napus ; Cicer arietinum ; current P ; Lens culinaris ; Lupinus albus ; Lupinus angustifolius ; P concentration response ; P content response ; Pisum sativum ; previous P ; sigmoid response ; single superphosphate ; Triticum aestivum ; Vicia faba ; yield response
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Phosphorus (P) is a major deficiency of soils of south-western Australia (WA). The fertilizer P requirements are not known for grain legumes being evaluated for neutral to alkaline, fine textured soils in WA. To rectify this, glasshouse and field experiments were undertaken to compare the responses of several grain legume species, wheat and canola to applications of single superphosphate and the results are reported in this paper. The glasshouse experiments measured responses of dried tops, harvested at 26 to 42 days after sowing, to P that was freshly-applied (current P) and previously-applied (previous P). Responses in the glasshouse were measured using yield, P concentration and P content (P concentration multiplied by yield) of oven dried tops of the following: wheat (Triticum aestivum), canola (Brassica napus), faba bean (Vicia faba), chickpea (Cicer arietinum), lentil (Lens culinaris), field pea (Pisum sativum), albus lupin (Lupinus albus) and narrow leaf lupin (Lupinus angustifolius). Field experiments in 1994 and 1995 compared seed (grain) yield responses of faba bean, chickpea, lentil, albus lupin and wheat to applications of current P. The P was banded (drilled) with the seed while sowing at 5 cm depth. Canola and wheat produced very large yield responses to increasing applications of current P. Responses were much smaller for albus lupin, faba bean and chickpea. Responses for lentil, narrow leaf lupin and field pea, fell in between responses of the small and large seeded species. Similar trends for responses were obtained as measured using yield, P concentration, or P content. For soils treated with previous P, similar trends were observed as for current P, but differences in yield responses between species were much less marked and the response curves tended to become more sigmoid. In the field experiments, grain yield responses to current P of albus lupin and chickpea were less than that for wheat. Relative to wheat, faba bean was the most responsive grain legume to applications of current P, with lentil producing similar responses to wheat in one experiment at a newly cleared, P deficient site.
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  • 8
    ISSN: 1432-1211
    Keywords: Key words Class II MHC sequence ; Rat ; Cloning ; RT-PCR ; Polymorphism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
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  • 9
    ISSN: 1573-4919
    Keywords: microbodies ; diabetes mellitus ; steroid hormone receptor ; β-oxidation ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract To determine whether the increased fatty acid β-oxidation in the peroxisomes of diabetic rat liver is mediated by a common peroxisome proliferation mechanism, we measured the activation of long-chain (LC) and very long chain (VLC) fatty acids catalyzed by palmitoyl CoA ligase (PAL) and lignoceryl CoA ligase and oxidation of LC (palmitic acid) and VLC (lignoceric acid) fatty acids by isotopic methods. Immunoblot analysis of acyl-CoA oxidase (ACO), and Northern blot analysis of peroxisome proliferator-activated receptor (PPAR-α), ACO, and PAL were also performed. The PAL activity increased in peroxisomes and mitochondria from the liver of diabetic rats by 2.6-fold and 2.1-fold, respectively. The lignoceroyl-CoA ligase activity increased by 2.6-fold in diabetic peroxisomes. Palmitic acid oxidation increased in the diabetic peroxisomes and mitochondria by 2.5-fold and 2.7-fold, respectively, while lignoceric acid oxidation increased by 2.0-fold in the peroxisomes. Immunoreactive ACO protein increased by 2-fold in the diabetic group. The mRNA levels for PPAR-α, ACO and PAL increased 2.9-, 2.8- and 1.6-fold, respectively, in the diabetic group. These results suggest that the increased supply of fatty acids to liver in diabetic state stimulates the expression of PPAR-α and its target genes responsible for the metabolism of fatty acids.
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  • 10
    ISSN: 1573-4919
    Keywords: regucalcin ; Ca2+-binding protein ; protein kinase C ; Ca2+signaling ; gene expression ; H4-II-E hepatoma cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The expression of hepatic Ca2+-binding protein regucalcin in the cloned rat hepatoma cells (H4-II-E) was investigated. The change in regucalcin mRNA levels was analyzed by Northern blotting using rat liver regucalcin complementary DNA (0.9 kb of open reading frame). Regucalcin mRNA was expressed in H4-II-E hepatoma cells. This expression was clearly stimulated in the presence of serum (10% fetal bovine serum). Bay K 8644 (2. 5 × 10-6 M), a Ca2+ channel agonist, significantly stimulated regucalcin mRNA expression in the absence or presence of 10% serum. Dibutyryl cyclic AMP (10-3 M) did not have a stimulatory effect on the regucalcin mRNA expression. The presence of phorbol 12-myristate 13-acetate (PMA; 10-6 M) or estrogen (10-8 M) caused a significant increase in regucalcin mRNA levels in the hepatoma cells cultured in serum-free medium, while insulin (5 × 10-9 M) or dexamethasone (10-6 M) had no effect. Bay K 8644-stimulated regucalcin mRNA expression in the hepatoma cells was completely blocked in the presence of trifluoperazine (10-5 M), an antagonist of calmodulin, or staurosporine (10-7 M), an inhibitor of protein kinase C. The stimulatory effect of PMA was clearly inhibited in the presence of stauroporine. The present study demonstrates that regucalcin mRNA is expressed in the transformed H4-II-E hepatoma cells, and that the expression is stimulated through Ca2+-dependent signaling factors.
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  • 11
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    Molecular and cellular biochemistry 199 (1999), S. 189-200 
    ISSN: 1573-4919
    Keywords: lung ; cancer ; urokinase ; receptor ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The urokinase-type plasminogen activator (uPA) interacts with its receptor (uPAR) to promote proteolysis as well as cell proliferation and migration. These functions contribute to the pathogenesis of neoplastic growth and invasiveness. Expression of uPAR in tumor extracts also inversely correlates with prognosis in many forms of cancer. In this study, we sought to determine if differences in uPAR expression were distinguishable between cultured human lung carcinoma and malignant mesothelioma subtypes. We also sought to determine if, as in malignant mesothelioma cells, uPAR expression is regulated at the posttranscriptional level in cultured malignant lung carcinoma cells. Using 125I-uPA binding and ligand blotting techniques, uPAR was expressed by phenotypically diverse lung carcinoma cell lines, including the H460, H157 and H1395 non-small cell lines and the H146 small cell lung carcinoma line. Increased uPAR expression was also detected in spindle-shaped (M33K) and epithelioid (M9K and MS-1) malignant mesothelioma cells. Selected mediators, including TGF-β, TNF-α, LPS and PMA, uniformly enhanced uPAR expression in each of the tumor cell lines. Steady state uPAR mRNA expression was determined by RNase protection assay and correlated directly with the changes in cell surface uPAR expression. By gel mobility shift and UV-cross linking assays, a uPAR mRNA binding protein (uPAR mRNABp) implicated in the posttranscriptional control of message stability, was identified in each of the cell lines. Expression of uPAR and its message in cultured lung carcinoma and malignant mesothelioma cells is similarly influenced by effectors present in the tumor microenvironment. Regulation of the uPAR message occurs at the posttranscriptional level in cultured small and non-small cell lung carcinoma cells as well as spindle-shaped and fibrous malignant mesothelioma cell lines. Posttranscriptional regulation of uPAR in all these cells involves the interaction of the uPAR mRNABp with uPAR mRNA, which promotes uPAR mRNA destabilization.
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  • 12
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    Molecular and cellular biochemistry 201 (1999), S. 111-123 
    ISSN: 1573-4919
    Keywords: complement factor I ; TPA ; protein kinase C ; gene expression ; Hep G2 cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract This study examined the role of the protein kinase C (PKC) signalling pathway in the regulation of expression of human complement factor I (CFI) gene. The production of CFI by Hep G2 cells was enhanced in a dose- and time-dependent fashion by 12-O-tetradecanoyl-1,2-phorbol 13-acetate (TPA), a potent PKC activator. 4α-phorbol didecanoate, an inactive phorbol ester, had no effect on CFI synthesis. The TPA-dependent increase in CFI secretion was correlated with an increase in CFI mRNA levels. Forskolin, a cAMP-inducing agent, augmented the TPA response. W7, an inhibitor of protein kinase A and genistein, an inhibitor of protein tyrosine kinase(s) both did not prevent the increase in CFI expression mediated by TPA. However, calphostin C, a specific inhibitor of PKC, abolished the TPA-induced increase in CFI mRNA levels. Down regulation of intracellular PKC levels by prior exposure of Hep G2 cells to a high concentration of TPA also blocked the increase in CFI mRNA levels induced by TPA suggesting that the TPA effects were mediated via activation of PKC. mRNA decay studies indicated that the half-life of CFI mRNA in TPA-induced cells was not significantly different from control. Nuclear run-on transcriptional assays on the other hand demonstrated that whereas the CFI gene is transcribed under basal conditions in Hep G2 cells, TPA induced a 3-4 fold increase in the transcription rate of CFI gene in 24 h. The transcription rate of GAPDH gene did not change, indicating that the effects were not general on gene transcription. Transient transfections of Hep G2 cells with chloramphenicol acetyltransferase reporter gene (CAT) constructs containing a series of sequential 5′ deletions of the CFI promoter and CAT assays showed that the sequence between -136 and -130, containing an AP-1 consensus sequence (TGAGTCA) was required for the TPA response. This observation was substantiated by the finding that mutation of this AP-1 site to TttaTCA or TtAtcCA abolished the TPA responsiveness. The enhancement of the activity of transfected chimeric CAT constructs by TPA was abrogated by calphostin C and by pyrrolidine dithiocarbamate (an inhibitor of NF-κB and AP-1 transactivation). These results indicate that TPA regulation of CFI gene requires PKC signalling and is mediated by via a TPA response element (TRE) in the CFI promoter region located at -136/-130 and involves the transactivation of AP-1 and NF-κB transcription factors. We suggest that PKC may be one of the intracellular pathways that control CFI gene expression and that cellular processes (involving growth factors, hormones, cytokines etc.) that activate PKC may upregulate the expression of the CFI gene.
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  • 13
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; gene expression ; Ca2+-ATPase ; brain microsomes ; aging
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The expression of calcium-binding protein regucalcin and its effect on the microsomal Ca2+-ATPase activity in rat brain tissues was investigated. The expression of regucalcin mRNA was demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) analysis in brain tissues using rat regucalcin-specific primers. Regucalcin concentration in the brain tissues was about 5 × 10-9 M as measured using enzyme-linked immunoadsorbent assay (ELISA), and this level was lowered with increasing age (50 weeks old). The presence of regucalcin (10-9 to 10-7 M) in the enzyme reaction mixture caused a significant decrease in Ca2+-ATPase activity in the brain microsomes of young rats (5 weeks old). Meanwhile, the enzyme activity was not significantly altered by the addition of calmodulin (1 or 50 μg/ml), calbindin (1 or 10 μg/ml), and S-100 A protein (5 or 25 μg/ml), which are other Ca2+-binding proteins in rat brain. The effect of regucalcin to inhibit microsomal Ca2+-ATPase activity was weakened in the brain of rats with increasing age (50 weeks old). The present study demonstrates that regucalcin is expressed in the brain, and that it can uniquely inhibit Ca2+-ATPase activity in the brain microsomes of rats. The findings suggest that regucalcin plays a role in the regulation of microsomal Ca2+-ATPase activity in rat brain tissues.
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  • 14
    ISSN: 1573-4919
    Keywords: mechanical stretch ; smooth muscle cells ; differential display ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Physical forces induce profound changes in cell phenotype, shape and behavior. These changes can occur in vascular structures as a result of pressure overload and their effects can be seen in atherosclerotic vessels in which smooth muscle cells have undergone hyperplastic and hypertrophic changes. At the molecular level, mechanical stimuli are converted into chemical ones and lead to modulation of gene expression and/or the activation of a new repertoire of genes whose encoded proteins help the cells to adapt to their microenvironment. In this study, we have used a two primer-based mRNA differential display technique to identify candidate mechano-responsive genes in pulmonary artery smooth muscle cells. As compared to the original method described by Liang and Pardee, this technique uses two arbitrary primers instead of an anchored oligo(dt) plus an arbitrary primer in the polymerase chain reaction. The chief advantages of these modifications are an increase in the efficiency of the amplification and in the identification of differentially expressed clones. Using this approach, we compared the pattern of expressed genes in cells cultured under static conditions with those in cells that were mechanically stretched (1 Hz) for 24 h in a well-defined in vitro mechanical system. Three candidate genes that showed reproducible differences were chosen for further characterization and cloning. One clone was under expressed in stretched cells and had a DNA sequence with 90% homology to the human fibronectin gene. Two other clones were highly expressed in stretched cells and had a 92% and a 83% sequence homology with human platelet-activating factor (PAF) receptor and rat insulin-like growth factor-I (IGF-I) genes respectively. Northern blot analysis confirmed low levels of fibronectin mRNA transcripts in stretched cells. In contrast, accumulation of PAF receptor mRNA occurred 30 min after mechanical stretch was initiated whereas IGF-I mRNA levels peaked at 8 h. Both mRNA levels were sustained for up to 24 h of mechanical stretching. These results demonstrate the usefulness of the two primer-based mRNA differential display that enabled us to identify and characterize alterations at the level of gene expression among matrix proteins, G-protein coupled receptors and growth factors, each of whose response to mechanical strain is different. A more complete understanding of these responses will provide further insight into the pathologic processes associated with hypertension and atherosclerosis.
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  • 15
    ISSN: 1432-1890
    Keywords: Key words Arbuscular mycorrhiza ; Hordeum vulgare ; Triticum aestivum ; Glomus intraradices ; Mycorrhiza-helper bacteria ; Secondary compounds
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Colonization of Hordeum vulgare L. cv. Salome (barley)and Triticum aestivum L. cv. Caprimus (wheat) roots by the arbuscular mycorrhizal fungus Glomus intraradices Schenck & Smith leads to de novo synthesis of isoprenoid cyclohexenone derivatives with blumenin [9-O-(2′-O-β-glucuronosyl)-β-glucopyranoside of 6-(3-hydroxybutyl)-1,1,5-trimethyl-4-cyclohexen-3-one] as the major constituent and to transient accumulation of hydroxycinnamate amides (4-coumaroylagmatine and -putrescine). Accumulation of these compounds in mycorrhizal wheat roots started 2 weeks after sowing together with the onset of arbuscule formation and proceeded with mycorrhizal progression. Highest levels were reached in 3- to 4-week-old secondary roots (root branches of first and higher order) characterized by the formation of vesicles. In the final developmental stages, the fungus produced massive amounts of spores, enclosing the stele of older root parts (older than 5 weeks) characterized by cortical death. In these root parts, the secondary compounds were detected in trace amounts only, indicating that they were located in the cortical tissues. Some rhizosphere bacteria tested, i.e. Agrobacterium rhizogenes, Pseudomonas fluorescens, and Rhizobium leguminosarum, markedly stimulated both fungal root colonization and blumenin accumulation, thus, acting as mycorrhiza-helper bacteria (MHB). Application of blumenin itself strongly inhibited fungal colonization and arbuscule formation at early stages of mycorrhiza development. This was associated with a markedly reduced accumulation of the hydroxycinnamate amides 4-coumaroylputrescine and -agmatine. The results suggest that both the isoprenoid and the phenylpropanoid metabolism are closely linked to the developmental stage and the extent of fungal colonization. Their possible involvement in the regulation of mycorrhiza development is discussed.
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  • 16
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    Theoretical and applied genetics 98 (1999), S. 472-477 
    ISSN: 1432-2242
    Keywords: Key words QTL ; Ear emergence time ; Plant height ; Recombinant substitution line ; Triticum aestivum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Chromosome 5A of wheat carries major gene loci for agronomic traits including the vernalization requirement (Vrn-A1) and ear morphology (Q). To determine whether the genetic variation for ear emergence time and plant height is attributable to either of these major genes as pleiotropic effects or independent QTL, we combined a RFLP map constructed from 120 recombinant substitution lines derived from a cross between ‘Chinese Spring’ (Cappelle-Desprez 5A) and CS(Triticum spelta 5A) with data collected from field trials over 3 years. For ear emergence time the main effects on flowering time were by Vrn-A1 and QEet.ocs-5A.1, the latter a QTL in the 28.6-cM Xcdo584/Q interval linked to Q by less than 10 cM. The CS(T. spelta 5A) allele at QEet.ocs-5A.1 contributed to an earlier ear emergence time by 2.7–6.0 days, which was approximately equal to the effects of Vrn-A1. For plant height, three QTLs were identified on the long arm and linked in repulsion. The CS(T. spelta 5A) allele at Vrn-A1 or closely linked to Xfba068 contributed to a height reduction of 3.5–6.1 cm, whereas both the Q allele and Qt.ocs-5A.1 allele within the Xcdo1088/Xbcd9 interval from CS(Cappelle-Desprez 5A) produced a shorter plant. When plant height was partitioned into culm length and ear length, the Vrn-A1 allele and CS(Cappelle-Desprez 5A) allele at QCl.ocs-5A.1 within the Xcd1088/Xbcd9 interval were found to contribute to a shorter culm. CS(T. spelta 5A) allele at q was a major determinant of a long ear, together with minor effects at QEl.ocs-5A.1 within the Xcdo1088/Xbcd9 interval.
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  • 17
    ISSN: 1432-2242
    Keywords: Key words Erysiphe graminis ; Powdery mildew resistance ; QTL ; Triticum aestivum ; Triticum spelta
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Powdery mildew is one of the major diseases of wheat in regions with a maritime or semi-continental climate and can strongly affect grain yield. The attempt to control powdery mildew with major resistance genes (Pm genes) has not provided a durable resistance. Breeding for quantitative resistance to powdery mildew is more promising, but is difficult to select on a phenotypic basis. In this study, we mapped and characterised quantitative trait loci (QTLs) for adult-plant powdery mildew resistance in a segregating population of 226 recombinant inbred lines derived from the cross of the Swiss wheat variety Forno with the Swiss spelt variety Oberkulmer. Forno possibly contains the Pm5 gene and showed good adult-plant resistance in the field. Oberkulmer does not have any known Pm gene and showed a moderate susceptible reaction. Powdery mildew resistance was assessed in field trials at two locations in 1995 and at three locations in 1996. The high heritability (h2=0.97) for powdery mildew resistance suggests that the environmental influence did not affect the resistance phenotype to a great extent. QTL analysis was based on a genetic map containing 182 loci with 23 linkage groups (2469 cM). With the method of composite interval mapping 18 QTLs for powdery mildew resistance were detected, explaining 77% of the phenotypic variance in a simultaneous fit. Two QTLs with major effects were consistent over all five environments. One of them corresponds to the Pm5 locus derived from Forno on chromosome 7B. The other QTL on 5A, was derived from the spelt variety Oberkulmer and did not correspond to any known Pm gene. In addition, five QTLs were consistent over three environments, and six QTLs over two environments. The QTL at the Pm5 locus showed a large effect, although virulent races for Pm5 were present in the mixture of isolates. Molecular markers linked with QTLs for adult-plant resistance offer the possibility of simultaneous marker-assisted selection for major and minor genes.
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  • 18
    ISSN: 1432-2242
    Keywords: Key words Aegilops tauschii ; Triticum aestivum ; Genetic mapping ; Molecular markers ; Agronomically important genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Aegilops tauschii is the diploid D-genome progenitor of bread wheat (Triticum aestivum L. em Thell, 2n=6x=42, AABBDD). A genetic linkage map of the Ae. tauschii genome was constructed, composed of 546 loci. One hundred and thirty two loci (24%) gave distorted segregation ratios. Sixty nine probes (13%) detected multiple copies in the genome. One hundred and twenty three of the 157 markers shared between the Ae. tauschii genetic and T. aestivum physical maps were colinear. The discrepancy in the order of five markers on the Ae. tauschii 3DS genetic map versus the T. aestivum 3D physical map indicated a possible inversion. Further work is needed to verify the discrepancies in the order of markers on the 4D, 5D and 7D Ae. tauschii genetic maps versus the physical and genetic maps of T. aestivum. Using common markers, 164 agronomically important genes were assigned to specific regions on Ae. tauschii linkage, and T. aestivum physical, maps. This information may be useful for map-based cloning and marker-assisted plant breeding.
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  • 19
    ISSN: 1432-2242
    Keywords: Key words Lodging ; Morphological traits ; QTL ; Triticum aestivum ; Triticum spelta
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Lodging can strongly affect both the grain yield and the quality of wheat. Lodging represents a quantitative trait and is difficult to assess on a phenotypic basis. Marker-assisted selection (MAS) could therefore become an important tool in breeding for lodging resistance. In this study, we mapped and characterised quantitative trait loci (QTLs) for lodging resistance, as well as morphological traits correlated with lodging, in a segregating population of 226 recombinant inbred lines derived from the cross of the lodging-resistant wheat variety Forno with the susceptible spelt variety Oberkulmer. Lodging, plant height, leaf width, leaf-growth habit, culm stiffness, culm swinging, culm thickness, days to ear emergence and days to flowering were assessed in field trials at two locations in 1996 and at one location in 1997. Additionally, at one location weight and length parameters were also assessed. Plant height and culm stiffness explained 77% of the phenotypic variance of lodging in a multiple regression model over all three environments. QTL analysis of lodging and morphological parameters was based on a genetic map containing 230 loci with 23 linkage groups (2469 cM). With the method of composite interval mapping nine QTLs for lodging resistance were detected, explaining 63% of the phenotypic variance in a simultaneous fit. Seven of these QTLs coincided with QTLs for morphological traits, reflecting the correlations between these traits and lodging. In our population the most efficient way to improve lodging resistance would be by a combination of indirect selection on plant height and culm stiffness together with MAS on the two QTLs for lodging resistance which did not coincide with QTLs for morphological traits.
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  • 20
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    Theoretical and applied genetics 98 (1999), S. 1248-1252 
    ISSN: 1432-2242
    Keywords: Key words Wheat ; Chromosome sorting ; Chromosome isolation ; Aneuploids ; Triticum aestivum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  We are reporting the successful isolation of wheat chromosome arm 1DS by flow cytometry. A chromosome suspension was prepared for the 1DS ditelosomic line and the normal ‘Chinese Spring’ (CS) by chopping 2-day-old root tip meristems, synchronized by hydroxyurea, in HEPES-magnesium sulfate buffer containing propidium iodide. Chromosomes were analyzed and sorted with a FACS Vantage flow cytometer and cell sorter. An extra peak was observed in the flow karyotype of the ditelosomic line that was absent in ‘CS’. The estimated size of chromosomes from the extra peak matched with the expected size of chromosome 1DS. Chromosomes from the putative 1DS peak were analyzed by both fluorescent microscopy and N-banding analysis. A total of 571 chromosomes from two separate experiments were analyzed, and all were observed to be telosomics except for 2 which were broken. About 82% of these telosomics showed the diagnostic N-band of 1DS, the remaining were unbanded and are probably also 1DS. This strategy can also be used to sort other wheat arms.
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  • 21
    ISSN: 1432-2242
    Keywords: Key words Transgenic wheat ; Triticum aestivum ; Tritordeum ; Transgene inheritance ; Marker gene stability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The stability and heritability of three marker genes was investigated in a population of twelve independent transgenic cereal lines (six wheat and six tritordeum). Integration patterns, inheritance of structural transgenes and inheritance of expression were analysed in the T0 and T1 generations for all 12 lines. Transmission and expression were analysed in the T2 generation for 9 lines and in the T3 generation for the six wheat lines. Inheritance of integration patterns was highly stable, and transmission of the transgenes and inheritance of their expression followed Mendelian ratios in the majority of lines. A gradual reduction in uidA expression was observed over three generations, which was not accompanied by a similar reduction in bar expression. Some unexpected phenomena associated with transgene inheritance were also observed and are discussed.
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  • 22
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    Cell & tissue research 296 (1999), S. 235-246 
    ISSN: 1432-0878
    Keywords: Key words Transplantation ; Parkinson’s disease ; CNS fetal development ; CNS differentiation ; Neurotrophic factors ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have previously shown that a combination of the cytokines interleukin (IL)-1, IL-11, leukemia inhibitory factor (LIF), and glial cell line-derived neurotrophic factor (GDNF) can convert rat fetal (E14.5) mesencephalic progenitor cells into tyrosine hydroxylase (TH)-immunoreactive (ir) neurons in vitro. The experiments described here characterize the mesencephalic progenitor cells and their cytokine-induced conversion into dopamine (DA) neurons. For all experiments, we used bromodeoxyuridine (BrdU)-ir cultures of (E14.5) mesencephalic progenitor cells that had been expanded at least 21 days. We first demonstrated that IL-1 induced DA neuron conversion in mesencephalic progenitors, but not in striatal progenitors (P〈0.001). Thus, these cells should be classified as lineage-restricted progenitors, and not omnipotent stem cells. To further characterize cell populations in these cultures, we used monoclonal antibodies against Hu (an early marker for neurons), growth-associated protein (GAP)-43 (a marker for neuronal process extension), TH (a marker for DA neurons), and glial fibrillary acidic protein (GFAP, a marker for astrocytes). We assessed (E14.5) mesencephalic progenitor cell cultures (plated at 125,000 cells/cm2) incubated in the cytokine mixture (described above) or in complete media (CM, negative control). Following 7 days incubation, GFAP-positive cells formed a nearly confluent carpet in both types of cultures. However, numbers of Hu-ir and GAP-43-ir cells in the cytokine-incubated cultures far exceeded those in CM-incubated controls (P=0.0003, P=0.0001, respectively), while numbers of TH-ir cells were 58-fold greater in the cytokine-incubated cultures versus CM-incubated controls. The TH phenotype persisted for 7 days following withdrawal of the differentiation media. Numerous double-labeled cells that were BrdU-ir and also TH-ir, or Hu-ir and also TH-ir, were observed in the cytokine-incubated cultures. These data suggest that cytokines ”drive” the conversion of progenitor cells into DA neurons.
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  • 23
    ISSN: 1432-0878
    Keywords: Key words Subcommissural organ ; Isograft ; Xenograft ; Reissner’s fiber ; Cerebrospinal fluid ; Rat ; Bovine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The subcommissural organ (SCO) secretes glycoproteins into the cerebrospinal fluid (CSF) that aggregate and form Reissner’s fiber (RF). The factors involved in this aggregation are not known. One factor may be the hydrodynamics of the CSF when flowing through the aqueduct. This hypothesis was tested by isografting rat SCO and xenografting bovine SCO into the lateral ventricle of rats. Xenografts were either fresh bovine SCO or explants cultured for 30 days before transplantation. The grafts were investigated by electron microscopy and immunocytochemistry using antibodies against RF glycoproteins, serotonin and the glucose transporter I. Maximal time of transplantation was 43 days for isografts and 14 days for xenografts. The isografts were not reinnervated but were revascularized; they secreted into the ventricle RF glycoproteins that became progressively packed into pre-RF and RF structures identical to those formed by the SCO in situ. RF was confined to the host ventricle and at its distal end the constituent proteins disassembled. Xenografts were neither reinnervated nor revascularized and secreted into the host ventricle a material that never formed an RF. These findings indicate that the CSF factor responsible for the formation of RF is species specific, and that this process does not depend on the hydrodynamics of the CSF. The blood vessels revascularizing the isografted SCO acquired the characteristics of the vessels irrigating the SCO in situ, namely, a tight endothelium displaying glucose transporter I, and a perivascular space containing long-spacing collagen, thus indicating that basal release of glycoproteins may also occur in the grafted SCO.
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  • 24
    ISSN: 1432-0878
    Keywords: Key words Axonal transport ; Purkinje cell ; Organotypic culture ; Microinjection ; Antimitotic drugs ; Cytoskeleton ; Dendritic transport ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Axonal and dendritic transport in single Purkinje neurons of cerebellar slice cultures was quantified as single transport distances. Examination of the cells within a vital tissue was regarded as being an approach to the in situ condition. The Purkinje cells were organotypically integrated in the in vitro tissues and extended long axonal projections connecting synapses to the target neurons. The tracer horseradish peroxidase (HRP) was applied via microinjection to the somata of the Purkinje cells and the injected neurons were incubated thereafter for defined time-intervals. The tracer was transported anterogradely into the neuron processes. The measurements on both the axonal and the dendritic transport of microinjected HRP revealed continuous transportation with increasing times of postincubation. This transport was reduced by the use of microtubule-depolymerizing drugs. The axonal transport of the tracer was either retarded in colchicine-treated cells or continuously reduced for up to 50% in vinblastine-treated neurons. Thus, a correlation of axonal transport to the microtubules was demonstrated. The dendrites were filled with the tracer after 60 min of postincubation. Dendritic transport was reduced by the use of vinblastine, and not significantly by colchicine. The results strongly support the dependence of neuronal transport on microtubules as a component of the cytoskeleton.
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  • 25
    ISSN: 1432-0878
    Keywords: Key words Neurofilament ; Basket cell ; Pinceau ; Golgi apparatus ; Calcium binding protein ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract NCS-1 (neuronal calcium sensor) is a recently characterized member of a highly conserved neuron-specific family of calcium-binding proteins, which also includes frequenin and recoverin. The cellular and subcellular distributions of NCS-1 in the rat nervous system were investigated using light- and electron-microscopic immunohistochemistry. NCS-1 immunoreactivity was localized to neuronal cell bodies and axons throughout the brain and spinal cord but not to glial cells. The most intense labeling was observed in myelinated axons, the axonal ramifications of the basket cell in the cerebellar cortex, and large neurons in the brainstem and pons. These same structures were also characterized by heavy labeling for neurofilament protein, as determined by double-labeling experiments. Most axon terminals were unlabeled or only lightly labeled. The most remarkable subcellular staining occurred in the perikarya where intense labeling was associated with the membranes of the trans saccules of the Golgi apparatus. The widespread distribution of NCS-1 indicates that it may be active in a variety of calcium-dependent neuronal functions, whereas the specific subcellular localization to the Golgi apparatus and neurofilament-rich structures suggests a specialized role in calcium regulated protein trafficking and cytoskeletal interactions.
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  • 26
    ISSN: 1432-0878
    Keywords: Key words Glial cell line-derived neurotrophic factor ; GDNF ; Ret ; GDNFR-α ; Brain-derived neurotrophic factor ; BDNF ; NT-3 ; NT-4 ; trk receptors ; Thyroid tissue ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Levels of mRNA for neurotrophins (brain-derived neurotrophic factor, BDNF; neurotrophin 3, NT-3; neurotrophin 4, NT-4) and their receptors (trkA, trkB, trkC) and for glial cell line-derived neurotrophic factor (GDNF) and its receptors (ret, GDNFR-α) were measured in rat thyroid tissue by ribonuclease protection assays. In thyroid tissue the NT-3 mRNA level was threefold lower and the NT-4 mRNA level sixfold higher than those detected in adult rat hippocampus, while BDNF mRNA was undetectable. Very low levels of mRNA for truncated trkB and trkC receptors and no catalytic trkA, trkB or trkC were found. In conclusion NT-3 and NT-4, but not the corresponding functional receptors, are expressed in the thyroid tissue. Therefore, it is unlikely that these factors serve a direct local autocrine or paracrine function in thyroid cell types, and a target-derived mode of action on neurons innervating the thyroid tissue is suggested. An opposite result has been found for the neurotrophic factor GDNF: thyroid tissue showed a high level of transcripts for the GDNF receptor subunits (GDNFR-α and Ret), while GDNF mRNA was undetectable. The in situ hybridization analysis of GDNFR-α and ret mRNA revealed an interesting difference in the cell distribution of these transcripts: ret mRNA is selectively expressed in a subpopulation of cells scattered in the follicular epithelium and in the interfollicular spaces, while GDNFR-α expression is more homogeneous and widespread, including the more abundant cell type of the thyroid gland: the follicular cell. Double-labeling in situ hybridization/immunocytochemistry experiments, with a specific marker (calcitonin), showed that parafollicular cells express ret but not GDNFR-α. This differential distribution of the GDNF receptor components (GDNFR-α and ret) may reflect a peculiar biological role in intercellular communication in the thyroid gland.
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  • 27
    ISSN: 1432-0878
    Keywords: Key words Water channel protein ; Aquaporin ; AQP5 ; Rat ; Salivary glands ; Immunolocalization ; Secretory stimulation ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Aquaporin-5 (AQP5) is a water channel protein and is considered to play an important role in water movement across the plasma membrane. We raised anti-AQP5 antibody and examined the localization of AQP5 protein in rat salivary and lacrimal glands by immunofluorescence microscopy. AQP5 was found in secretory acinar cells of submandibular, parotid, and sublingual glands, where it was restricted to apical membranes including intercellular secretory canaliculi. In the submandibular gland, abundant AQP5 was also found additionally at the apical membrane of intercalated duct cells. Upon stimulation by isoproterenol, apical staining for AQP5 in parotid acinar cells tended to appear as clusters of dots. These results suggest that AQP5 is one of the candidate molecules responsible for the water movement in the salivary glands.
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  • 28
    ISSN: 1871-4528
    Keywords: Solanum tuberosum L. ; tuberisation ; extensin ; acyl carrier protein thioesterase ; high mobility group protein ; gene expression ; plant development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary In screening to isolate a full-length copy of a previously isolated cDNA clone, a further three cDNAs were also isolated from a library prepared from sub-apical swelling-stolon tissue of potato (Solanum tuberosum L.). Sequence analysis showed these clones to be similar to extensin-like protein genes, acyl carrier protein thioesterase genes and high mobility group protein genes, respectively. A further cDNA, isolated by subtractive hybridisation, was similar to a tomato cDNA previously isolated on the basis of its down-regulation following nematode infection. While all the newly isolated genes were expressed in swelling stolons, for most, maximal expression was seen to be in stem tissue. Possible roles for these genes in the development of potato plants are discussed, as is the significance of gene expression in stems and stolons to the process of tuberisation.
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  • 29
    ISSN: 1573-9368
    Keywords: transgenic mice ; prolactin ; mammary gland ; gene expression ; Stat5 ; β-globin insulator
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In order to establish a possible correlation between in vitro prolactin induction and the transcriptional activity of mammary gene promoters in transgenic mice, a functional Stat5-binding site was created by means of site-directed mutagenesis at position −70 on a 560 bp murine α-lactalbumin promotor linked to a CAT reporter gene. Surprisingly, the wild-type promoter was constitutively active in vitro and could not be induced by prolactin. Introducing the proximal Stat5 site abolished this constitutive activity and resulted in prolactin dependence in both CHO-K1- and HC11-transfected cells. In transgenic mice, both the frequency of lines expressing the transgene and the prevalence of mid to late pregnancy expression were increased.
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  • 30
    ISSN: 1573-9368
    Keywords: low molecular weight glutenin promoter ; particle bombardment ; transgene expression ; Triticum aestivum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transgenic wheat plants from specific cultivars can now be routinely engineered in many laboratories. However, our understanding of the factors controlling transgene expression and stability in wheat compared to other cereals is rather limited. Only a few promoters have been tested in transgenic wheat, and relatively little is known of their relative activities and expression parameters. In the present study, the spatial and temporal properties of one heterologous constitutive promoter and one seed‐specific wheat promoter were investigated. We generated constructs with the reporter gene gusA (β‐glucuronidase) driven by: (a) the constitutive maize ubiquitin‐1 (ubi‐1) promoter, and (b) two different‐sized fragments of the seed‐specific low molecular weight glutenin (LMWG1D1) promoter from wheat. The activities of all three promoter constructs were comparable in endosperm tissue. A detailed analysis of spatial and temporal properties of the promoters is described. Heat shock treatment of transgenic plants carrying the ubi‐1: gusA construct resulted in a significant elevation in the levels of GUS activity. The inheritance of transgene expression levels and stability was evaluated over four generations, as a function of transgene integration patterns and copy number.
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  • 31
    ISSN: 1618-2545
    Keywords: BLB light ; conidiogenesis ; Fusarium globosum ; subtropical Japan ; Triticum aestivum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The second report ofFusarium globosum is based on strains isolated from wheat in Ishigaki Island, Okinawa, in subtropical Japan. Morphological features of the Japanese isolates are described. These strains show different reactions in conidiogenesis to light conditions. Under continuous BLB light, falcate sporodochial conidia are typically induced, but production of aerial globose conidia is suppressed. In two of the strains, clavate conidia became longer under BLB light. Application of both, complete darkness and continuous BLB light, is recommended as standard light conditions to cultureFusarium isolates.
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  • 32
    ISSN: 1573-0778
    Keywords: CHO cells ; gene expression ; kinetic model ; protein secretion ; transcription ; translation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The elevation of expression levels for secreted glycoproteins by gene amplification in mammalian cells shows a saturation behavior at high levels of gene amplification. At high expression levels a drop in the secretion efficiency for the recombinant protein occurs (Schröder and Friedl, 1997), coinciding with the appearance of misfolded protein in the cell. In this communication we investigated whether additional limitations exist at the levels of transcription and translation. Four Chinese hamster ovary (CHO) cell lines expressing different amounts of human antithrombin III (ATIII) were used as a model system. A tenfold increase in the ATIII cDNA copy number from the lowest to the highest producing cell line coincided with a 38-fold increase in ATIII mRNA levels, and an 80-fold increase in the amount of intracellular ATIII levels. The data was analyzed using a simple kinetic model. The following conclusions were derived: I. The transcriptional activity for the recombinant protein is not saturated. II. Translation itself is not saturated either, but may be downregulated as secretion efficiency drops. III. Two explanations for the previously reported drop in secretion efficiency for the recombinant protein with increasing expression level are possible: A. Protein degradation is an alternative fate for translated ATIII and the fraction of ATIII degraded after translation increases as expression level is increased. B. Translation is downregulated as the secretory apparatus becomes exhausted to maintain cell viability.
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  • 33
    ISSN: 1573-0778
    Keywords: cardiogenesis ; cell differentiation ; gene expression ; mouse embryonic stem cells ; myogenesis ; neurogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Embryonic stem cells, totipotent cells of the early mouse embryo, were established as permanent cell lines of undifferentiated cells. ES cells provide an important cellular system in developmental biology for the manipulation of preselected genes in mice by using the gene targeting technology. Embryonic stem cells, when cultivated as embryo-like aggregates, so-called ‘embryoid bodies’, are able to differentiate in vitro into derivatives of all three primary germ layers, the endoderm, ectoderm and mesoderm. We established differentiation protocols for the in vitro development of undifferentiated embryonic stem cells into differentiated cardiomyocytes, skeletal muscle, neuronal, epithelial and vascular smooth muscle cells. During differentiation, tissue-specific genes, proteins, ion channels, receptors and action potentials were expressed in a developmentally controlled pattern. This pattern closely recapitulates the developmental pattern during embryogenesis in the living organism. In vitro, the controlled developmental pattern was found to be influenced by differentiation and growth factor molecules or by xenobiotics. Furthermore, the differentiation system has been used for genetic analyses by ‘gain of function’ and ‘loss of function’ approaches in vitro.
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  • 34
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    Cytotechnology 30 (1999), S. 71-83 
    ISSN: 1573-0778
    Keywords: gene expression ; HEK293(EBNA) cells ; serum-free ; transient transfection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract In order to establish a simple and scaleable transfection system we have used the cationic polymer polyethylenimine (PEI) to study transient transfection in HEK293 and 293(EBNA) cells grown in serum-free suspension culture. The transfection complexes were made directly within the cell culture by consecutively adding plasmid and PEI (direct method). Alternatively, the DNA-PEI transfection complexes were prepared in fresh medium (1/10 culture volume) and then added to the cells (indirect method). The results of this study clearly show that the ratio of PEI nitrogen to DNA phosphate is very important for high expression levels. The precise ratio is dependent on the DNA concentration. For example, using 1 μg/ml DNA by the indirect method, the ratio of optimal PEI:DNA was about 10–13:1. However, the ratio increases to 33:1 for 0.1–0.2 μg/ml DNA. By testing several different molecular weights of the polycationic polymer we could show that the highest transfection efficiency was obtained with the PEI 25 kDa. Using PEI 25 kDa the indirect method is superior to the direct addition because significantly lower DNA concentrations are needed. The expression levels of the soluble human TNF receptor p55 are even higher at low DNA compared to 1 μg/ml plasmid. The EBV-based pREP vectors gave better transient gene expression when used in 293(EBNA) cells compared to HEK293 cells in suspension culture. No differences in expression levels in the two cell lines were observed when the pC1 (CMV)-TNFR was used. In conclusion, PEI is a low-toxic transfection agent which provides high levels of transient gene expression in 293(EBNA) cells grown in serum-free suspension culture. This system allows highly reproducible, cost-effective production of milligram amounts of recombinant proteins in 2–5 l spinner culture scale within 3–5 days. Fermentor scale experiments, however, are less efficient because the PEI-mediated transient tranfection is inhibited by conditioned medium.
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  • 35
    ISSN: 1573-0778
    Keywords: cell cycle ; CHO ; flow cytometry ; gene expression ; synchronisation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Understanding the relationships between cell cycle and protein expression is critical to the optimisation of media and environmental conditions for successful commercial operation of animal cell culture processes. Using flow cytometry for the analysis of the early phases of synchronised batch cultures, the dependency of product expression on cell cycle related events has been evaluated in a recombinant CHO cell line. Although the production of recombinant protein is initially found to be cell cycle related, the maximum specific protein productivity is only achieved at a later stage of the exponential phase which also sees a maximum in the intracellular protein concentration. Subsequent work suggests that it is the batch phase/medium composition of cultures which is the major determinant of maximum specific productivity in this cell line. Furthermore the effect of the positive association between S phase and specific productivity is subordinate to the effect of batch phase/medium composition on the specific productivity of batch cultures.
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  • 36
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    European journal of plant pathology 105 (1999), S. 519-533 
    ISSN: 1573-8469
    Keywords: genome ; gene expression ; mollicute ; recombination ; transposition ; virus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Spiroplasmas are members of the Class Mollicutes, wall-less prokaryotes having a high adenosine–thymidine content in their small genomes. Spiroplasma citri is a plant pathogen that inhabits phloem. Like other phytopathogenic spiroplasmas and the related phytoplasmas, it is transmitted from plant to plant by phloem-feeding leafhoppers that serve as alternate hosts for the spiroplasma as well as vectors. Genetic information in spiroplasmas is carried on a circular chromosome, on plasmids and/or in virus genomes. A picture emerging from recent research on the S. citri genome is one of frequent and often extensive variation, resulting from a number of different mechanisms. Expansion and contraction events must continually be occurring in about equal proportions so that the net genome size varies within defined boundaries. Particularly impressive are large changes in genome size that can occur in only a few generations. As with most organisms, genetic variation in S. citri results from variation in extrachromosomal DNA content, changes due to DNA replication and repair processes and changes due to recombination. The implied flux of genetic information into and out of the S. citri genome should be beneficial to the bacterium, allowing it, with its small genome size, to adapt to new environments.
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  • 37
    ISSN: 1573-4935
    Keywords: Mucin ; lung cancer ; gene expression ; secretion ; lung adenocarcinoma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Mucins comprise an important class of tumor-associated antigens. The objectives of the present study were (a) to establish an in vitro model system using human non-small cell lung adenocarcinoma cell lines NCIH650 and NCIH2077 (b) provide evidence that these cell lines secrete mucin in culture conditions and (c) investigate the effects of select secretagogues on mucin secretion. The cell lines were established in ACL-4 medium containing several growth factors and retinoic acid and 5% fetal calf serum. The high molecular weight glycoconjugates secreted in the culture medium were purified by ammonium sulfate precipitation and Superose 6 and Superose 12 FPLC chromatography. The purified high molecular weight glycoconjugate fraction and the carcinoma cells were shown to have mucin by dot blot, Western blot and immunohistochemical analysis, respectively, using specific antibodies to purified major mucin, HTM-1. Also, incorporation experiments with mucin precursor 3H-glucosamine demonstrated that the cells indeed synthesize high molecular weight mucins. The effects of secretagogues such as, 8-bromocyclic AMP, ionomycin, phorbol-12-myristate-13-acetate and neutrophil elastase on mucin secretion were also investigated. Only 8-bromocyclic AMP and neutrophil elastase influenced mucin secretion. These studies provided strong evidence that the lung adenocarcinoma cell lines secrete high molecular weight mucins in culture conditions and only two of the four tested secretagogues significantly increased mucin secretion. Thus, this in vitro model system may be useful in determining alterations in mucin structure, if any, in lung adenocarcinomas as well as in studying the regulation of mucin gene expression.
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  • 38
    ISSN: 1573-904X
    Keywords: adriamycin ; doxorubicin ; HPMA copolymer ; apoptosis, multidrug resistance ; gene expression ; signal transduction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Purpose. To study peculiarities and the mechanism of the anticancer effect of free and HPMA copolymer-bound ADR in sensitive and resistant human ovarian carcinoma cells. Methods. Sensitive A2780 and ADR resistant A2780/AD cells were exposed to different doses of drugs during 12, 24, 36, 48, 60, and 72 hours. Cell viability, drug accumulation, apoptosis, cellular metabolism, lipid peroxidation, DNA content and gene expression were studied. Results. HPMA copolymer-bound ADR (P(GFLG)-ADR) possessed a comparable cytotoxicity to free ADR when comparison was based on intracellular concentrations. While free ADR up-regulated genes encoding ATP driven efflux pumps (MDR1, MRP), P(GFLG)-ADR overcame existing pumps and down regulated the MRP gene. Free ADR also activated cell metabolism and expression of genes responsible for detoxification and DNA repair. P(GFLG)-ADR down-regulated HSP-70, GSr-π, BUDP, Topo-IIα, β, and TK-1 genes. Apoptosis, lipid peroxidation and DNA damage were significantly higher after exposure to P(GFLG)-ADR, as reflected by simultaneous activation of p53, c-fos in A2780 cells) or c-jun (A2780/AD) signaling pathways and inhibition of the bcl-2 gene. Differences between free ADR and P(GFLG)-ADR increased with the time of incubation and drug concentration. Conclusions. P(GFLG)-ADR overcame drug efflux pumps, more significantly induced apoptosis and lipid peroxidation, inhibited DNA repair, replication, and biosynthesis when compared to free ADR.
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  • 39
    ISSN: 1573-9104
    Keywords: Bioavailability ; Magnesium ; Hijiki ; Sodium alginate ; Rat ; Wakame
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The bioavailability of magnesium from Wakame and Hijiki, and the effects of alginic acid on absorption of dietary magnesium were examined in five groups of rats fed either control, Wakame, Hijiki, AW (containing the same amount of alginate as in the Wakame) and AH (containing the same amount of alginate as in the Hijiki) diets, and animals fed a low magnesium diet (LMg) (twentieth amount of magnesium in the original mineral mixtures as the control). Food intake and body weight gain were decreased by adding sodium alginate to the diets. A large amount of calcium accumulated only in the kidneys of the rats fed the LMg diet. Serum magnesium concentration decreased only in the LMg group. The magnesium content in the defatted left femurs did not differ between the control and Wakame fed animals and also among the animals eating Wakame, Hijiki and AW diets. The breaking force of the right femurs did not differ among all the groups except the LMg group. The ratio of apparent magnesium absorption (%) of the control, LMg, Wakame, Hijiki, AW and AH groups was 82.2, 72.7, 66.9, 50.8, 69.3 and 54.2 in the first experimental period, and was 75.3, 52.1, 57.7, 46.9, 62.6 and 60.5 in the second experimental period, respectively. It was clear that the bioavailability of magnesium in the Wakame fed rats was higher than in those eating the Hijiki. Large amounts of sodium alginate lowered magnesium absorption from the diet.
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  • 40
    ISSN: 1573-5028
    Keywords: UV-B ; soybean ; chalcone synthase ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract By developing gene-specific RT-PCR and using filters to allow transmission down to 290 nm (UV-B+) or blocking all radiation below 320 nm (UV-B−), the effect of UV-B+ and UV-B− light on expression of each of the presently known seven members of soybean chalcone synthase (CHS) gene family in dark-grown seedlings was analyzed. Dark expression was detectable already in 18 h dark-germinating embryos, with progressive increases on successive days, suggesting that chs belongs to a class of genes expressed very early during germination, and that the expression at this stage is either constitutive or induced by non-light-dependent factors present in the seed or made available following imbibition. Exposure of 18 h dark-germinating embryos to UV-B− or to UV-B+ light did not lead to an increase in chs signal. However, the 24 h dark-germinating embryos showed a distinct effect of UV-B+, interestingly coinciding with the stage when the head of seedlings was in the process of being pushed up above ground by stem elongation, suggesting the possibility of a developmental switch modulating the appearance of UV-B response. The response to UV-B− was most prominent in chs1 and almost silent in chs2, while the up-regulation by UV-B+ was most prominent in chs5 and chs6 and much less so in chs2. Interestingly, chs2 was noted to be the only member of the Gmchs gene family devoid of H-box, raising the possibility that the H-box may be a good indicator of the photo-inducibility of a chs gene.
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  • 41
    ISSN: 1573-5028
    Keywords: amphidiploid genome structure ; gene expression ; glutamine synthetase ; multigene family ; nitrogen assimilation ; oilseed rape
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In the amphidiploid genome of oilseed rape (Brassica napus) the diploid ancestral genomes of B. campestris and B. oleracea have been merged. As a result of this crossing event, all gene loci, gene families, or multigene families of the A and C genome types encoding a certain protein are now combined in one plant genome. In the case of the multigene family for glutamine synthetase, the key enzyme of nitrogen assimilation, six different cDNA sequences were isolated from leaf and root specific libraries. One sequence pair (BnGSL1/BnGSL2) was characterized by the presence of amino- terminal transit peptides, a typical feature of all nuclear encoded chloroplast proteins. Two other cDNA pairs (BnGSR1-1/BnGSR1-2 and BnGSR2-1/BnGSR2-2) with very high homology between each other were found in a root specific cDNA library and represent protein subunits for cytosolic glutamine synthetase isoforms. Comparative PCR amplifications of genomic DNA isolated from B. napus, B. campestris and B. oleracea followed by sequence–specific restriction analyses of the PCR products permitted the assignment of the cDNA sequences to either the A genome type (BnGSL1/BnGSR1- 1/BnGSR2-1) or the C genome type (BnGSL2/BnGSR1-2/BnGSR2-2). Consequently, the ancestral GS genes of B. campestris and B. oleracea are expressed simultaneously in oilseed rape. This result was also confirmed by RFLP (restriction fragment length polymorphism) analysis of RT-PCR products. In addition, the different GS genes showed tissue specific expression patterns which are correlated with the state of development of the plant material. Especially for the GS genes encoding the cytosolic GS isoform BnGSR2, a marked increase of expression could be observed after the onset of leaf senescence.
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  • 42
    ISSN: 1573-5028
    Keywords: defense ; gene expression ; leaf senescence ; nitrilase ; pathogen-free ; salicylic acid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To determine the range of gene activities associated with leaf senescence, we have identified genes that show preferential transcript accumulation during this developmental stage. The mRNA levels of a diverse array of gene products increases during leaf senescence, including a protease, a ribosomal protein, two cinnamyl alcohol dehydrogenases, a nitrilase and glyoxalase II. Two of the genes identified are known to be pathogen-induced. The senescence specificity of each gene was determined by characterization of transcript accumulation during leaf development and in different tissues. The increased expression of nitrilase in senescent leaves is paralleled by an increase in free indole-3-acetic acid (IAA) levels. Additionally, we have demonstrated that the induction of defense-related genes during leaf senescence is pathogen-independent and that salicylic acid accumulation is not essential for this induction. Our data indicate that the induction of certain genes involved in plant defense responses is a component of the leaf senescence program.
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  • 43
    ISSN: 1573-5028
    Keywords: gene expression ; gibberellin ; H1 histone ; H2B histones ; leaf ; Lycopersicon esculentum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract After differential screening we isolated cDNA clones encoding a histone H1 (leH1) and three variants of histone H2B (leH2B-1, -2 and -3) from the gibberellin (GA)-deficient mutant of tomato (gib-1). The deduced polypeptide of leH1 is 271 amino acids long and exhibits the typical tripartite structure of histones H1. The full-length cDNA clone leH2B-1 encodes for a protein of 142 amino residues and shows the tripartite organization of histones H2B. The histones leH1 and leH2B, which show no tissue specificity, are developmentally expressed in the leaf. The mRNA accumulation was higher in organs which contain meristematic tissue and/or which have a high proportion of actively cycling cells. In the leaf of the gib-1 mutant we demonstrated GA-enhanced histone leH1 and leH2B expression which was not observed in the wild type. GAs of the early-13-hydroxylated pathway (GA1 and GA3) caused most enhanced transcription compared to GAs of the early-non-hydroxylation pathway (GA4 and GA9). Application of GA to the mutant increased histone expression that could correlate with enhanced DNA replication in leaf tissue. Increased chromosome replication may indicate that there is a higher rate of cell division and/or increase of endopolyploidy which both may be dependent on cell elongation induced by GAs.
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  • 44
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    Plant molecular biology 39 (1999), S. 933-943 
    ISSN: 1573-5028
    Keywords: cloning ; fruit development ; gene expression ; pea ; polyamine ; spermidine synthase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two cDNAs from young pea fruits coding for functional spermidine synthases (EC 2.5.1.16) were isolated. The corresponding genes were named psSPDSYN1 and psSPDSYN2. Both cDNAs complemented spe3Δ gene when introduced into the Y480 strain of Saccharomyces cerevisiae, which is a null mutant for the spermidine synthase gene. psSPDSYN1 and psSPDSYN2 are regulated differentially. psSPDSYN1 is up-regulated early after fruit set whereas psSPDSYN2 is expressed later. Spermidine synthase activity was detected in pea ovaries, and correlates with the pattern of expression of psSPDSYN1. In the pea plant, psSPDSYN1 is highly expressed in actively growing tissues, whereas the highest level of psSPDSYN2 mRNA was detected in fully elongated stem.
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  • 45
    ISSN: 1573-5028
    Keywords: chitin oligomer ; chitinase ; elicitor ; gene expression ; rice ; signal transduction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Expression patterns of chitinase transcripts induced by N-acetylchitooligosaccharide elicitor were analyzed by northern blot hybridization in order to reveal a signal transduction pathway leading to the activation of class I chitinase genes (Cht-1 and Cht-3), which may play an important role in producing N-acetylchitooligosaccharide elicitor. The transcription level of both genes was enhanced in response to N-acetylchitooligosaccharides larger than pentaose at subnanomolar concentrations. These structure and dose dependencies were consistent not only with those for a 75 kDa high-affinity binding protein for N-acetylchitooligosaccharide elicitor in the plasma membrane, but also with other series of cellular responses including phytoalexin production and the expression of elicitor-responsive genes (EL2, EL3). Therefore, the elicitor signal to evoke these cellular responses including the activation of the chitinase genes could be common and transmitted into cells through the 75 kDa protein. However, the signal transduction pathway for the activation of the chitinase gene appeared to diverge from those for the other elicitor-responsive genes shortly after the signal perception. It was shown that the induction of chitinase expression by N-acetylchitooligosaccharide would require protein phosphorylation, but not de novo protein synthesis. The oxidative burst was demonstrated not to be necessary for transcriptional induction of the all four elicitor-responsive genes (Cht, PAL, EL2, EL3) by N-acetylchitooligosaccharide.
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  • 46
    ISSN: 1573-5028
    Keywords: cytochrome b5 ; fruit and flower development ; gene expression ; in situ hybridisation ; olive
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We report the characterisation of two cytochrome b5 genes and their spatial and temporal patterns of expression during development in olive, Olea europaea. A PCR-generated probe, based on a tobacco cytochrome b5 sequence, was used to isolate two full-length cDNA clones (cytochrome b5-15 and cytochrome b5-38) from a library derived from 13 WAF olive fruits. The cDNAs encoded proteins of 17.0 and 17.7 kDa, which contained all the characteristic motifs of cytochromes b5 from other organisms and exhibited 63% identity and 85% similarity with each other. The olive cytochrome b5-15 cDNA was then used as a probe for more detailed analysis. Southern blotting revealed a gene family of at least 4–6 members while northern blotting and in situ hybridisation showed a highly specific pattern of gene expression. Very low levels of cytochrome b5 mRNA were detected in tissues characterised by high rates of lipid accumulation, such as young expanding leaves, maturing seeds and ripening mesocarp. The cytochrome b5 genes were not induced at 6 °C and their response to ABA was relatively slow compared with fatty acid desaturase genes. In contrast, high levels of cytochrome b5 gene expression were found in young fruits at the pattern formation (globular/heart) stage of embryogenesis and in vascular and transmitting tissues of male and female reproductive organs. The data are consistent with a major role for cytochrome b5 in developmental processes related to plant reproduction in addition to being an electron donor to microsomal desaturases.
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  • 47
    ISSN: 1573-5028
    Keywords: Amaranthus hypochondriacus) ; gene expression ; trypsin inhibitor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We previously isolated and sequenced the major trypsin inhibitor from Amaranthus hypochondriacus seeds. This amaranth trypsin inhibitor (AmTI) is a 69 amino acid protein with high homology to members of the potato-1 inhibitor family. This paper describes the cloning and expression of a cDNA encoding this trypsin inhibitor in various vegetative tissues of the amaranth plant during seed development and imbibition, and investigates the possible induction of AmTI expression by wounding. We obtained a 393 bp cDNA sequence with an open reading frame corresponding to a polypeptide with 76 amino acid residues. With the exception of one residue (Ser-41), the polypeptide agrees with the amino acid sequence previously reported, plus 7 more residues at the N-terminus. These N-terminal residues are thought to be part of the signal used for intracellular sorting. The organ specificity of AmTI gene expression was investigated by northern analysis, showing that mRNA corresponding to AmTI genes was present in stems of plants growing under normal conditions. The kinetics of accumulation of the AmTI-mRNA, protein, and inhibitory activity during seed development and imbibition was determined. AmTI-mRNA accumulation reached a maximum at 14 days after anthesis (daa) and then gradually decreased, being barely detectable 36 daa. The AmTI protein accumulation followed the same profile as the inhibitory activity, both were delayed with respect to the mRNA. The maximum level was observed 22 daa, and then gradually decreased until a steady state was reached as seed maturation proceeded. Upon imbibition, a gradual decrease in AmTI protein and inhibitory activity was shown; however, an AmTI transcript was detected 24 h after imbibition. In contrast to representative members of the potato I family, this inhibitor was not inducible by wounding of leaves.
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  • 48
    ISSN: 1573-5028
    Keywords: embryo ; gene expression ; Glycine max ; oxidoreductase ; seed coat ; testa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The soybean Ep gene encodes an anionic peroxidase enzyme that accumulates in large amounts in seed coat tissues. We have isolated a second peroxidase gene, Prx2, that is also highly expressed in developing seed coat tissues. Sequence analysis of Prx2 cDNA indicates that this transcript encodes a cationic peroxidase isozyme that is far removed from Ep in peroxidase phylogeny. To determine the expression patterns for these two peroxidases in developing seeds, the abundance and localization of the Ep and Prx2 transcripts were compared by in situ hybridization. Results show the expression of Ep begins in a small number of cells flanking the vascular bundle in the seed coat, spreads to encircle the seed, and then migrates to the hourglass cells as they develop. Expression of Prx2 occurs throughout development in all cell layers of the seed coat, and is also evident in the pericarp and embryo. Nonetheless, the Ep-encoded enzyme accounts for virtually all of the peroxidase activity detected in mature seed coats. The Prx2 enzyme is either insoluble in a catalytically inactive form, or is subject to degradation during seed maturation.
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  • 49
    ISSN: 1573-5028
    Keywords: ABRE ; embryogenesis ; G-box ; gene expression ; maize ; protein-DNA interaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transcription of the rab28 gene from maize is induced in late embryo development and in response to abscisic acid. We have studied the regulation of the activity of the rab28 promoter in embryos. Two abscisic acid-responsive elements (ABREs) were necessary for expression in embryos of transgenic Arabidopsis and in transient transformation in maize embryos. In vivo footprinting showed that there was protein binding to the ABREs and to other cis elements in the promoter in young embryos before expression of rab28. This shows that the rab28 promoter is in an open chromatin structure before developmental activation. The ABREs are important for the induction and have protein binding in young embryos. Nuclear proteins extracted from embryos before activation of rab28 bound to the ABREs in band shift assays. A complex with different mobility was formed between nuclear proteins and the ABREs after induction of rab28 suggesting a modification of the ABRE-binding factor or an exchange of proteins. The footprints on the ABREs were unaltered by induction with abscisic acid or during developmental activation of rab28. These results indicate that constitutive binding of transcription factor(s) on the ABRE is central in embryonic regulation of the rab28 gene.
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  • 50
    ISSN: 1573-5036
    Keywords: genotypic variation ; Secale cereale ; Triticum aestivum ; Triticum durum ; zinc efficiency ; zinc deficiency
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Effect of zinc (Zn) nutritional status on uptake of inorganic 65Zn was studied in rye (Secale cereale, cv. Aslim), three bread wheat (Triticum aestivum, cvs. Dagdas, Bezostaja, BDME-10) and durum wheat (Triticum durum, cv. Kunduru-1149) cultivars grown for 13 days in nutrient solution under controlled environmental conditions. The cultivars were selected based on their response to Zn deficiency and to Zn fertilization in calcareous soils under field conditions. When grown in Zn-deficient calcareous soil in the field, the rye cultivar had the highest, and the durum wheat the lowest Zn efficiency. Among the bread wheats, BDME-10 showed higher susceptibility to Zn deficiency and Bezostaja and Dagdas were less affected by Zn deficiency. Similarly to field conditions, in nutrient solution visual Zn deficiency symptoms (i.e. necrotic lesions on leaf blade) appeared to be more severe in Kunduru-1149 and BDME-10 and less severe in rye cultivar Aslim. Under Zn deficiency, shoot concentrations of Zn were similar between all cultivars. Cultivars with adequate Zn supply did not differ in uptake and root-to-shoot translocation rate of 65Zn, but under Zn deficiency there were distinct differences; rye showed the highest rate of Zn uptake and the durum wheat the lowest. In the case of bread wheat cultivars, 65Zn uptake rate was about the same and not related to their differential Zn efficiency. Under Zn deficiency, rye had the highest rate of root-to-shoot translocation of 65Zn, while all bread and durum wheat cultivars were similar in their capacity to translocate 65Zn from roots to shoots. When Zn2+ activity in uptake solution ranged between 117 p M and 34550 pM, Zn-efficient and Zn-inefficient bread wheat genotypes were again similar in uptake and root-to-shoot translocation rate of 65Zn. The results indicate that high Zn efficiency of rye can be attributed to its greater Zn uptake capacity from soils. The inability of the durum wheat cultivar Kunduru-1149 to have a high Zn uptake capacity seems to be an important reason for its Zn inefficiency. Differential Zn efficiency between the bread wheat cultivars used in this study is not related to their capacity to take up inorganic Zn.
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  • 51
    ISSN: 1573-5028
    Keywords: alanine aminotransferase ; gene expression ; GUS expression ; promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cDNA clone encoding alanine aminotransferase (AlaAT) has isolated from randomly sequenced clones derived from a cDNA library of maturing rice seeds by comparison to previously identified genes. The deduced amino acid sequence was 88% and 91% homologous to those of the enzymes from barley and broomcorn millet (Panicum miliaceum), respectively. Using this cDNA as a probe, we isolated and sequenced the corresponding genomic clone. Comparison of the sequences of the cDNA and the genomic gene revealed that the coding region of the gene was interrupted by 14 introns 66 to 1547 bp long. Northern and western blotting analyses showed that the gene was expressed at high levels in developing seeds. When the 5′-flanking region between −930 and +85 from the site of initiation of transcription was fused to a reporter gene for β-glucuronidase (GUS) and then introduced into the rice genome, histochemical staining revealed strong GUS activity in the inner endosperm tissue of developing seeds and weak activity in root tips. Similar tissue-specific expression was also detected by in situ hybridization. These results suggest that AlaAT is involved in nitrogen metabolism during the maturation of rice seed.
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  • 52
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 39 (1999), S. 161-169 
    ISSN: 1573-5028
    Keywords: expansin ; fruit growth ; fruit softening ; gene expression ; Lycopersicon esculentum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract cDNA clones encoding homologues of expansins, a class of cell wall proteins involved in cell wall modification, were isolated from various stages of growing and ripening fruit of tomato (Lycopersicon esculentum). cDNAs derived from five unique expansin genes were obtained, termed tomato Exp3 to Exp7, in addition to the previously described ripening-specific tomato Exp1 (Rose et al. (1997) Proc Natl Acad Sci USA 94: 5955–5960). Deduced amino acid sequences of tomato Exp1, Exp4 and Exp6 were highly related, whereas Exp3, Exp5 and Exp7 were more divergent. Each of the five expansin genes showed a different and characteristic pattern of mRNA expression. mRNA of Exp3 was present throughout fruit growth and ripening, with highest accumulation in green expanding and maturing fruit, and lower, declining levels during ripening. Exp4 mRNA was present only in green expanding fruit, whereas Exp5 mRNA was present in expanding fruit but had highest levels in full-size maturing green fruit and declined during the early stages of ripening. mRNAs from each of these genes were also detected in leaves, stems and flowers but not in roots. Exp6 and Exp7 mRNAs were present at much lower levels than mRNAs of the other expansin genes, and were detected only in expanding or mature green fruit. The results indicate the presence of a large and complex expansin gene family in tomato, and suggest that while the expression of several expansin genes may contribute to green fruit development, only Exp1 mRNA is present at high levels during fruit ripening.
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  • 53
    ISSN: 1573-5028
    Keywords: cDNA cloning ; fruit ripening ; gene expression ; non-climacteric fruit ; wild strawberry (Fragaria vesca L.)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Wild strawberry (Fragaria vesca L.) is an attactive model system for studying ripening in non-climacteric fruit, because of its small diploid genome, its short reproductive cycle, and its capacity for transformation. We have isolated eight ripening-induced cDNAs from this species after differential screening of a cDNA library. The predicted polypeptides of seven of the clones exhibit similarity to database protein sequences, including acyl carrier protein, caffeoyl- CoA 3-O-methyltransferase, sesquiterpene cyclase, major latex protein, cystathionine γ-synthase, dehydrin and an auxin- induced gene. A ninth cDNA clone that was constitutively expressed is predicted to encode a metallothionein-like protein. None of these proteins appear to be directly related to events generally associated with ripening such as cell wall metabolism or the accumulation of sugars and pigments, rather, their putative functions are indicative of the wide range of processes upregulated during fruit ripening.
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  • 54
    ISSN: 1573-5028
    Keywords: cowpea (Vigna unguiculata L.) ; drought ; gene expression ; lipid degradation ; phospholipase D
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Phospholipase D, a major lipid-degrading enzyme in plants, was studied in two cultivars of Vigna unguiculata L.Walp, differing in their tolerance to drought (cv. EPACE-1, drought-tolerant, and cv. 1183, drought-susceptible). Enzymatic activities, measured with 14C-PC as substrate, increased when plants were submitted to water stress, the increase being much higher in the drought-sensitive cultivar. A 2911 bp cDNA encoding a putative phospholipase D (VuPLD1) was isolated from a cDNA library prepared from V. unguiculata leaves. The deduced amino acid sequence (809 residues) shows 85.5% identity and 91.3% similarity to that of PLD from Ricinus communis. The expression of the VuPLD1 gene in the leaves is differently modulated by water deficit, depending on the intensity of stress and the tolerance or sensitivity of the plants. In the drought-susceptible V. unguiculata cv. 1183, it readily increased under water stress, reaching maximum values at mild water deficit (−1.5 MPa). In the drought-tolerant cv. EPACE-1, VuPLD1 mRNA remained low throughout the whole drought treatment. Dehydration of leaves led to a dramatic increase in transcript level in both cultivars. Changes in protein amounts semi-quantified by immunoblotting correlated well with variations in transcript steady-state level. Taken together, these results showed that phospholipase D in cowpea plants is essentially regulated at the transcriptional level, and that gene expression is strongly stimulated even by moderate water deficit in the drought-sensitive plant. On the contrary, the drought-tolerant plant presents a remarkable stability of PLD gene expression in conditions of water stress.
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  • 55
    ISSN: 1573-5028
    Keywords: reproductive development ; gene expression ; subtractive hybridization ; cauliflower
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Using the meristems of the cauliflower curd as a source of tissue and a series of subtractive hybridizations and amplification reactions, we have constructed a cDNA library highly enriched in cDNAs expressed in reproductive meristems. The analysis of a sample of 250 clones from this library identified 22 cDNA clones corresponding to genes specifically expressed in these cauliflower meristems. Apart from two clones that corresponded to APETALA1, and two other ones showing similarity to different aminoacyl-tRNA synthetases, the remaining clones showed no similarity to any sequence in the databases and may correspond to novel genes. One of these clones, BoREM1, was further characterized and found to correspond to a gene encoding a protein with features of regulatory proteins that follows a expression pattern very similar to the LEAFY transcripts.
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  • 56
    ISSN: 1573-5028
    Keywords: ammonium ; gene expression ; glutamine synthetase ; nodules ; positive element ; promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In order to identify important promoter elements controlling the ammonium-regulated expression of the soybean gene GS15 encoding cytosolic glutamine synthetase, a series of 5′ promoter deletions were fused to the GUS reporter gene. To allow the detection of positive and negative regulatory elements, a series of 3′ deletions were fused to a −90 CaMV 35S promoter fragment placed upstream of the GUS gene. Both types of construct were introduced into Lotus corniculatus plants and soybean roots via Agrobacterium rhizogenes-mediated transformation. Both spectrophotometric enzymatic analysis and histochemical localization of GUS activity in roots, root nodules and shoots of transgenic plants revealed that a strong constitutive positive element (SCPE) of 400 bp, located in the promoter distal region is indispensable for the ammonium- regulated expression of GS15. Interestingly, this SCPE was able to direct constitutive expression in both a legume and non- legume background to a level similar to that driven by the CaMV 35S full-length promoter. In addition, results showed that separate proximal elements, located in the first 727 bp relative to the transcription start site, are essential for root- and root nodule-specific expression. This proximal region contains an AAAGAT and two TATTTAT consensus sequences characteristic of nodulin or nodule-enhanced gene promoters. A putative silencer region containing the same TATTTAT consensus sequence was identified between the SCPE and the organ-specific elements. The presence of positive, negative and organ-specific elements together with the three TATTTAT consensus sequences within the promoter strongly suggest that these multiple promoter fragments act in a cooperative manner, depending on the spatial conformation of the DNA for trans-acting factor accessibility.
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  • 57
    ISSN: 1573-5028
    Keywords: embryo-abundant cDNAs ; gene expression ; gymnosperm ; Picea glauca ; somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Six somatic embryogenesis-associated cDNAs (PgEMB2, 6, 7, 8, 24 and 34) from white spruce (Picea glauca (Moench) Voss) somatic embryos have been characterized. Transcript accumulation during somatic embryo development and subsequent germination related to these genes, indicated that they were developmentally regulated. The transcripts related to clones PgEMB2, 6, 24 and 34 were also detected during zygotic embryo development, but transcripts of clones PgEMB7 and 8 were not. PgEMB24 had a similar gene expression pattern to spruce Em-like late embryo abundant (lea) gene, but other clones had no similarities in gene expression to either spruce lea-like or storage protein genes. Abscisic acid, a stimulator for spruce somatic embryo maturation, did not obviously affect gene expression corresponding to these cDNAs. The predicted proteins are distinguishable from known LEA proteins based on analyses of hydropathy plots, amino acid compositions and deduced protein structures. The similarities of the spruce cDNAs, and protein sequences predicted from these cDNAs, to other sequence data are described.
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  • 58
    ISSN: 1573-5028
    Keywords: gene expression ; GT-1 ; PR-1a ; PR proteins ; salicylic acid-induced ; transcription factors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Infection of Nicotiana tabacum Samsun NN with tobacco mosaic virus (TMV) results in a hypersensitive plant response and leads to systemic acquired resistance (SAR). The induction of SAR is mediated by the plant hormone salicylic acid (SA) and is accompanied by the induced expression of a number of genes including the pathogenesis-related (PR) gene 1a. Previously, it has been found that TMV infection and SA treatment resulted in a reduction of binding of nuclear protein GT-1 to far-upstream regions (−902 to −656) of the PR-1a gene. To test if GT-1 is a negative regulator of PR-1a gene expression, the effects of mutations in the seven putative GT-1 binding sites in this region were studied in vitro using dimethyl sulfate interference footprinting and band shift assays. This showed that at least one of the seven sites is indeed a GT-1 binding site. However, when tested in transgenic plants, the mutations did not result in constitutive expression of the chimeric PR-1a/GUS transgene, while inducible expression after SA treatment was decreased. The results suggest that binding of GT-1-like proteins to far-upstream PR-1a promoter regions indeed influences gene expression. A possible model for GT-1's mode of action in PR-1a gene expression is discussed.
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  • 59
    ISSN: 1573-5028
    Keywords: aquaporin ; gene expression ; growth ; Oryza sativa ; plasma membrane intrinsic protein (PIP) ; rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Membrane intrinsic proteins facilitate movement of small molecules often times functioning as water channels. We have identified two genes from rice which encode proteins with characteristic features of plasma membrane intrinsic proteins (PIP). They possess six membrane-spanning domains, an NPA repeat, overall high sequence homologies and characteristic C- and N-terminal hallmark motifs which allowed assignment of OsPIP1a to the PIP1 subfamily and of OsPIP2a to the PIP2 subfamily. OsPIP1a and OsPIP2a showed similar but not identical expression patterns. The two genes were expressed at higher levels in seedlings than in adult plants and expression in the primary root was regulated by light. In internodes of deepwater rice plants which were induced to grow rapidly by submergence, transcript levels were slightly induced in the intercalary meristem (IM) and slightly reduced in the elongation zone (EZ) after 18 h. In internodes of GA-induced excised stem sections transcript levels transiently declined in the IM and EZ after 1 h and subsequently recovered to elevated levels after 18 h. GA also induced OsPIP expression in non-growing tissue after 18 h. In the IM of submergence-induced stem sections transcript levels remained constitutive. The different growth-promoting treatments showed no direct correlation between growth rate and OsPIP gene expression in dividing or expanding cells. In fact, treatment of excised stem sections with ABA or drought stress induced similar changes in OsPIP expression in the growing zone during the first 6 h as GA did. We conclude that regulation of OsPIP1a and OsPIP2a expression is not primarily controlled by growth. GA-induced growth may however change the water status of cells which in turn results in altered PIP abundance.
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  • 60
    ISSN: 1573-5028
    Keywords: ACC synthase ; chilling ; Citrus sinensis ; ethylene ; gene expression ; peel
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Diurnal change in the temperature below or above 12.5 °C hastens the degreening of citrus peel and elicits the phytohormone ethylene production in citrus fruit. Ethylene triggers the degradation of chlorophyll and synthesis of carotenoids in citrus peel. To investigate if ethylene is required for the degreening of citrus peel elicited by low temperatures, we studied the chilling-regulated gene expression of ACC synthase, one of the key enzymes catalyzing ethylene biosynthesis. We isolated and characterized a chilling-inducible 1-aminocyclopropane-1-carboxylate synthase (ACC synthase) gene, CS-ACS1, and a chilling-repressible gene, CS-ACS2, from citrus peel. The CS-ACS1 transcript 1.7 kb in length encodes a polypeptide of 483 amino acids (M r 54 115, pI 6.63), whereas the CS-ACS2 transcript of 1.8 kb encodes a polypeptide of 477 amino acids (M r 53 291, pI 6.72). Both genes showed a rapid but transient induction (within 2.4 h) of transcripts upon rewarming after the chilling (4 °C) treatment. After 24 h of incubation at room temperature, CS-ACS1 mRNA diminished to an undetectable level, whereas the CS-ACS2 mRNA regained its basal level of expression attained prior to the chilling treatment. Chilling-induced ethylene production and ACC accumulation were also observed upon rewarming. Both genes were also induced by the wound stress (excision). The protein synthesis inhibitor cycloheximide super-enhances the accumulation of both ACS transcripts at room temperature. Molecular analysis of the 3.3 kb genomic DNA of CS-ACS1 revealed that this gene consists of three introns and four exons. The intron 3 is exceptionally large (1.2 kb) and shares significant homology with mitochondrial DNA, supporting the intron-late theory.
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  • 61
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    Plant molecular biology 41 (1999), S. 351-361 
    ISSN: 1573-5028
    Keywords: chromatin ; gene expression ; high-mobility-group protein HMG1 ; HMGe ; protein stability ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The nuclear HMG1 proteins of higher plants are small non-histone proteins that have DNA-bending activity and are considered architectural factors in chromatin. The occurrence of the chromosomal HMG1 proteins, HMGa, HMGc1/2 and HMGd, in various maize tissues was analyzed, and in the course of these studies a novel HMG1 protein, now termed HMGe, was identified. Purification and characterization of HMGe (Mr 13 655) and cloning of the corresponding cDNA revealed that it displays only moderate similarity to other members of the plant HMG1 protein family. The five maize HMG1 proteins could be detected in kernels, leaves, roots and suspension culture cells, indicating that these proteins can be expressed simultaneously and occur relatively ubiquitously. However, the various HMG1 proteins are present in significantly different quantities with HMGa and HMGc1/2 being the most abundant HMG1 proteins in all tissues tested. Furthermore, the relative amounts of the various HMG1 proteins differ among the tissues examined. The HMG1 proteins were found to be relatively stable proteins in vivo, with HMGc1/2, HMGd and HMGe having a half-life of ca. 50 h in cultured cells, while the half-life of the HMGa protein is ca. 65 h. Collectively, these findings are compatible with the concept that the different plant HMG1 proteins might act as general architectural proteins in concert with site-specific factors in the assembly of certain nucleoprotein structures involved in various biological processes.
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  • 62
    ISSN: 1573-5028
    Keywords: alkaloids ; gene expression ; Nicotiana tabacum ; nicotine ; putrescine N-methyltransferase ; tobacco gene evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The structure and nuclear genomic organization of the gene family encoding putrescine N-methyltransferase (PMT), the key enzyme in diverting polyamine metabolism towards the biosynthesis of nicotine and related alkaloids, was examined in Nicotiana tabacum. Five genes encoding PMT are present in the N. tabacum genome and all are expressed. The complete coding region and immediate 5′- and 3′- flanking regions were characterized for four members of the gene family and the Exon 1 region of the fifth member of the family was determined. Comparison of the nucleotide and deduced amino acid sequences of the N. tabacum PMT genes with those of presumed progenitor species, N. sylvestris, N. tomentosiformis and N. otophora, revealed that three members of the N. tabacum PMT gene family were most similar to the three genes present in N. sylvestris, whereas the two remaining PMT genes were similar to PMT genes present in N. tomentosiformis and N. otophora genomes, respectively. These data are consistent with an evolutionary origin of N. tabacum resulting from a cross involving N. sylvestris and an introgressed hybrid between N. tomentosiformis and N. otophora. The five PMT genes present in N. tabacum are expressed in the roots of wild-type plants, but not in other organs. The steady-state level of all five PMT transcripts is transiently increased in roots following topping (removal of the floral meristem), although the maximum level of induction for the individual transcripts varies considerably. In contrast to wild-type plants, no increase in PMT transcript levels was observed in a low-alkaloid (nic1nic2) mutant of Burley 21. These data support a role for nic1 and nic2 in the global regulation of alkaloid formation in tobacco and provide for the first time molecular confirmation of the presumed origin of cultivated tobacco.
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  • 63
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    Plant molecular biology 41 (1999), S. 425-433 
    ISSN: 1573-5028
    Keywords: D1 protein ; gene expression ; psbA genes ; redox regulation ; Synechococcus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three psbA genes encode two different forms of the photosystem II reaction centre protein D1 in Synechococcus sp. PCC 7942. The psbAI gene encoding D1 protein form I (D1:1) is mainly expressed under low growth light conditions while the psbAII and psbAIII genes, encoding D1 protein form II (D1:2), are induced under stress conditions (e.g. high light or low temperature). In this paper we show that psbAII/III genes can be rapidly induced even under low growth light conditions by adding the thiol reductant (DTTred) to Synechococcus cell culture, at a concentration that does not affect cell growth or photosynthetic activity. Similar induction of psbAII/III genes was obtained by illuminating the cells with photosystem I light. In both instances psbAI gene down-regulation coincided with the up-regulation of psbAII/III genes. DTTred-induced exchange in transcript pools was subsequently followed by an exchange of D1:1 for D1:2 at the protein level. Thiol oxidants, iodosobenzoic acid or diamide, reverted the effects of DTTred on psbA gene expression. Thiol oxidants and the thiol-modifying agent N-ethylmaleimide also totally prevented high-light induction of psbAII/III genes. These data strongly suggest that the up-regulation of psbAII/III genes that occurs under stress conditions is mediated by production of thiol reductants, whereas the expression of the psbAI gene is sustained by the more oxidizing conditions that prevail during the steady-state growth of cells.
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  • 64
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    Plant and soil 215 (1999), S. 65-72 
    ISSN: 1573-5036
    Keywords: barley ; boron deficiency ; Hordeum vulgare ; Triticum aestivum ; variation ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Responses of a range of barley (Hordeum vulgare L.) genotypes to boron (B) deficiency were studied in two experiments carried out in sand culture and in the field at Chiang Mai, Thailand. In experiment 1, two barley genotypes, Stirling (two-row) and BRB 2 (six-row) and one wheat (Triticum aestivum L.) genotype, SW 41, were evaluated in sand culture with three levels of applied B (0, 0.1 and 1.0 μM B) to the nutrient solution. It was found that B deficiency depressed flag leaf B concentration at booting, grain number and grain yield of all genotypes. In barley Stirling, B deficiency also depressed number of spikes plant-1, spikelets spike-1 and straw yield. However, no significant difference between genotypes in flag leaf B concentration was found under low B treatments. Flag leaf B concentration below 4 mg kg-1 was associated with grain set reduction and could, therefore, be used as a general indicator for B status in barley. In experiment 2, nine barley and two wheat genotypes were evaluated in the field on a low B soil with three levels of B. Boron levels were varied by applying either 2 t of lime ha-1 (BL), no B (B0) or 10 kg Borax ha-1 (B+) to the soil prior to sowing. Genotypes differed in their B response for grain spike-1, grain spikelet-1 and grain set index (GSI). The GSI of the B efficient wheat, Fang 60, exceeded 90% in all B treatments. The B inefficient wheat SW 41 and most of the barley genotypes set grain normally (GSI 〉80%) only at the B+. In B0 GSI of the barley genotypes ranged from 23% to 84%, and in BL from 19% to 65%. Three of the barley with severely depressed GSI in B0 and BL also had a decreased number of spikelets spike-1. In experiment 3, 21 advanced barley lines from the Barley Thailand Yield Nursery 1997/98 (BTYN 1997/98) were screened for B response in sand culture with no added B. Grain Set Index of the Fang 60 and SW 41 checks were 98 and 65%, respectively, and GSI of barley lines ranged between 5 and 90%. One advanced line was identified as B efficient and two as moderately B efficient. The remaining lines ranked between moderately inefficient to inefficient. These experiments have established that there is a range of responses to B in barley genotypes. This variation in the B response was observed in vegetative as well as reproductive growth. Boron efficiency should be considered in breeding and selection of barley in low B soils.
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  • 65
    ISSN: 1573-5036
    Keywords: ammonium assimilation ; Amanita muscaria ; carbon allocation ; ectomycorrhiza ; gene expression ; Picea abies ; sugar transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The symbiosis (ectomycorrhiza, ECM) between roots of trees and shrubs of boreal and temperate forest ecosystems and soil fungi is essential for water and nutrient acquisition of the plants. The functionality of ECM is largely dependent on the ability of the host plant to supply photoassimilates to the fungus via the symbiotic interface. Based on sterile in vitro and non-sterile pot experiments, we review data which gives evidence that hexoses are supplied to the fungus by the host plant (mainly glucose and fructose), and that these sugars, at least in part, control development and function of ECM by interfering with fungal gene expression. We further show that any factor which reduces hexose allocation to the host–fungus interface will adversely affect ECM development. As an example, we address the impact of increased supply of nitrogen on the biochemistry of plant–fungus interaction and discuss potential consequences on host performance.
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  • 66
    ISSN: 1573-5028
    Keywords: bundle sheath cell ; C4 photosynthesis ; gene expression ; PEP carboxykinase ; prokaryotic expression ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We isolated a full-length cDNA that encodes ATP-dependent phosphoenolpyruvate carboxykinase (EC 4.1.1.49, PCK) from leaves of maize, an NADP-malic enzyme type C4 plant. The mRNA was specifically and rather abundantly expressed in bundle sheath cells in accordance with the recent finding of cell-type-specific localization of PCK protein in maize, which has been detected with antibodies against cucumber PCK protein. The predicted protein had an N-terminal extension, which is characteristic of plant PCKs. The transcript level was much higher in the daytime than at night in 14-day old seedlings. However, in 42-day old plants the extent of diurnal change decreased. The maize PCK was expressed in Escherichia coli with the pET32 plasmid and purified to homogeneity. Through digestion with enterokinase, two types of enzyme were prepared; one with an intact N-terminus and the other lacking its N-terminal 77 amino acid residues due to over-digestion. The truncated protein had about 2-fold higher specific activity than the intact one, and was inhibited by 3-phosphoglycerate (3-PGA) with an I0.5 of 17.5 mM. In contrast, the intact protein was almost insensitive to 3-PGA. These results strongly suggest that the intact N-terminal extension may be involved in the regulation of PCK activity in vivo through some modification such as reversible phosphorylation.
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  • 67
    ISSN: 1573-5028
    Keywords: calcium-dependent protein kinase ; gene expression ; immunoassays ; light regulation ; rice ; seed development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We investigated the spatial and temporal expression patterns of two rice calcium-dependent protein kinases (CDPKs), OsCDPK2 and OSCDPK11, using isoform-specific antisera. Bands of the expected molecular sizes for OsCDPK2 (59 kDa) and OsCDPK11 (61 kDa) were detected on western blots. OsCDPK2 and OsCDPK11 mRNA and protein levels increased in unison during flower development. However, at the onset of seed development, the protein expression profiles diverged significantly. OsCDPK2 protein was expressed at low levels during early seed development, but increased to high levels that were maintained in later stages (20 days after fertilisation, DAF). Conversely, OsCDPK11 protein levels were high at the beginning of seed development, but fell rapidly from 10 DAF onwards. This decrease in the level of OsCDPK11 protein was associated with the abundant synthesis of a truncated mRNA species. OsCDPK2 expression was also closely associated with light perception. OsCDPK2 protein was barely detectable in green leaves exposed to light, but levels increased sharply when plants were shifted to darkness. Initially, this increase reflected a rapid elevation in the levels of OsCDPK2 mRNA, which was normally located in the mesophyll. Conversely, OsCDPK11 mRNA and protein levels were unaffected by light. These data strongly indicate that two rice CDPK isoforms have different functions in seed development and in response to light in leaves.
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  • 68
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    Euphytica 108 (1999), S. 91-95 
    ISSN: 1573-5060
    Keywords: amylose content ; granule-bound starch synthase ; null alleles ; Triticum aestivum ; Wx loci
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The Wx locus controls amylose synthesis in the cereal endosperm. Hexaploid wheat (Triticum aestivum L.) has the three Wx loci on chromosomes 7A ( Wx-A1), 4A (Wx-B1) and 7D (Wx-D1). To verify the effects of null alleles on reducing amylose content and determine the amylose synthesis capacity of each Wx gene independently and accurately, we produced eight possible types of recombinant lines carrying different null alleles at the Wx loci under the ‘Chinese Spring’ genetic background. Amylose content varied from 0% of the waxy ‘Chinese Spring’ to 25% of the ‘Chinese Spring’ normal type. The reducing effect of the single null alleles was the largest in Wx-B1b, and there was no significant difference between Wx-A1b and Wx-D1b. More than 3% reductions in amylose content were detected in the double null types. The results of the double null lines further demonstrated that for the capacity of amylose synthesis, Wx-B1a predominates and produces 21–22% amylose, followed by Wx-D1a (20–21%) and Wx-A1a (15–18%). These significant differences were partly correlated with variation in the amounts of the Wx proteins produced by different Wx genes. However, comparisons of the double null lines with the single null or normal lines indicated that amylose content was not linearly proportional to the number of the Wx genes, suggesting that the Wx genes act in an epistatic manner.
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    Euphytica 106 (1999), S. 117-123 
    ISSN: 1573-5060
    Keywords: differential display ; gene expression ; heterosis ; hybrid wheat ; seedling leaf
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Differential display of mRNA was used to analyze the differences of gene expression in seedling leaves between heterotic hybrid/nonheterotic hybrid and their parental inbreds in order to study the molecular basis of heterosis in wheat. The results indicated that patterns of gene expression in hybrids differ significantly from their parents. Both quantitative and qualitative differences were observed. The quantitative differences include gene over-expression, gene under-expression in hybrid and dominant expression of highly-expressed parental genes in hybrids. The qualitative differences include silencing in hybrids of genes expressed either in male or female parent, and silencing in hybrids of genes expressed in both parents. Expression in hybrid of genes only expressed either in male or female parent was also observed. It was also found that some genes expressed at high level in heterotic hybrid were underexpressed or expressed at low level in nonheterotic hybrid. One differentially expressed cDNA fragment 4B was cloned and sequenced after being confirmed through Northern blot analysis. Homology search in GenBank proved that the cDNA fragment is a new sequence. The selection of primers for differential RNA display in wheat and the relationship between wheat heterosis and alteration of gene expression in hybrids as compared to their parental inbreds were also discussed.
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    Plant growth regulation 28 (1999), S. 129-132 
    ISSN: 1573-5087
    Keywords: dormancy ; Lactuca sativa ; lettuce seeds ; Brassica napus ; rapeseed ; combustion products ; Salix viminalis ; Themeda triandra ; Triticum aestivum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In darkness, dormancy was imposed on seeds of lettuce (Lactuca sativa L. cv. Grand rapids) by high temperature and on seeds of oilseed rape (Brassica napus L. cv. Apex) by osmotic stress using polyethylene glycol (PEG 8000). In both cases, dormancy was broken by incubating the seeds in aqueous extracts of combustion products from Salix viminalis wood chips or Themeda triandra leaves. Dormancy of rapeseed, but not lettuce, was also broken by a solution of smoke from burnt straw of Triticum aestivum. The greatest stimulation from burnt vegetation was achieved with an aqueous extract of pyrolysed willow wood chips, which had been subjected to temperatures of up to 800 °C during combustion in a down-draught gasifier. This suggests that some biologically active substances obtained from combustion of plant tissues are highly heat-stable.
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    Plant growth regulation 29 (1999), S. 47-76 
    ISSN: 1573-5087
    Keywords: cold ; chill ; freezing ; gene expression ; signal transduction pathways
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Temperature is expected to affect all plant processes. Consistent with this, expression of a large number of specific mRNAs and proteins is up-regulated during cold-acclimation. Their possible functions are outlined, encompassing a wide range of processes, some possibly related to other winter stresses besides cold. Some of the cold-responsive sequences are associated with constitutive or stress-related metabolism, others are probably protective, some may influence freezing, and many are of as yet unknown function. While many of the sequences code for intracellular proteins, a significant number code for apoplastic proteins with a variety of possible functions. Transient influx of calcium into the cytosol appears to be a key step in the response to cold, and also to many other stresses and signals. Similarly, the cold-responsive promoter element identified so far is also responsive to drought and salt. However, how cold is sensed, and any cold-specific aspects of the cold signal transduction pathway, are, so far, unknown. What is clear is that, at least in the model plant Arabidopsis thaliana, cold-, desiccation-, salt- and ABA-triggered signal transduction pathways, and possibly others, run partly in parallel and partly intersect. This may be partly explained by a need for an integrated winter-response. The total number of genes which are cold-responsive and the quantity of resources which this implies are used in this way, indicate that many must have a positive role in acclimation. Experiments which modify membrane lipid unsaturation or solute accumulation, achieved by transformation of plants to express exotic or heterologous genes or by other means, confirm that these factors affect chill- or freezing-tolerance. A transgenic test has shown that one cold-up-regulated gene of previously unknown function contributes to freezing-tolerance, but the small effect re-emphasises the probably cumulative nature of the contributions of many cold-up-regulated sequences to acclimation. On the other hand, mutant analysis indicates some genes may make a comparatively larger contribution. Transformation of alfalfa to overexpress a superoxide dismutase gene increased cold-tolerance and drought-resistance and demonstrated that improvements in field-survival of stresses is possible by transgenic means. Over-expression of a transcription factor, CBF1, conferred freezing-tolerance on Arabidopsis, showing that manipulation of the signal transduction pathway could be an important method for modifying cold-tolerance.
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  • 72
    ISSN: 1573-5087
    Keywords: cortical fibre cell ; gibberellic acid ; spring wheat ; Triticum aestivum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The role of gibberellic acid (GA) in differentiation and secondary cell-wall deposition of fibre cells of spring wheat (Triticum aestivum) culms was studied using applications of GA and chlormequat (a GA biosynthesis inhibitor). In certain genotypes, higher GA levels may increase the number of cortical fibre cell files by changing cell fate from parenchyma to fibre, and induce thicker secondary cell-walls.
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    Genetic resources and crop evolution 46 (1999), S. 469-475 
    ISSN: 1573-5109
    Keywords: foliar blight ; germplasm ; resistance ; Triticum aestivum ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract One thousand three hundred and eighty-seven spring wheat germplasm (Triticum aestivum L.) lines belonging to the Indian and CIMMYT wheat programmes were evaluated for their tolerance to foliar blight disease for three consecutive years i.e., from 1994 to 1997. Disease severity at six different growth stages, beginning from tillering to late milk stage, was recorded. None of the genotypes showed immunity to the disease. Of 43 lines showing resistant reaction, a major proportion (25) was represented by CIMMYT material. Comparatively, Indian germplasm lines tended to be more susceptible at more advanced growth stages. Area Under Disease Progress Curve (AUDPC) and Apparent Infection Rate (r) values of resistant lines were much lower than those of susceptible ones, but lower AUDPC in some of the resistant lines did not correspond to a lower 'r' value. Most of the resistant lines were derived from Seri, Myna, Bau, kauz, Hork 's' and Aegilops tauschii Coss.
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  • 74
    ISSN: 1573-5168
    Keywords: fish ; gene expression ; GH ; GtH ; gonad ; growth factors ; RT-PCR ; salmonid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In this study, Northern blot analysis of RNA from trout testis revealed a single transcript of insulin-like growth factor II (IGF-II) around 4.7 kb. The cellular distribution of IGF-II mRNA was studied and quantified in different testicular cells enriched populations by RT-PCR. IGF-II mRNA appears to be expressed in all cellular types tested: spermatogonia A and B, primary spermatocytes, spermatids and secondary spermatocytes and Sertoli cells. A significantly higher expression of IGF-II was found in premeiotic germ cells. The levels of IGF-II mRNA appear to be higher than those of IGF-I in immature trout testis, as judged from the semi-quantitative RT-PCR results. These data suggest that in addition to IGF-I, IGF-II may play a role in testicular physiology in fish. The hormonal regulation of IGF-I and IGF-II gene expression was investigated both in vitro and in vivo using RT-PCR approach. Gonadotropin (GtH) added to testicular explants increased IGF-II mRNA levels but had no effect on IGF-I. No statistically significant effect was observed with androgens. In vivo, GH and pituitary extracts resulted in an 8 fold and 2-3 fold increase in both IGF-I and IGF-II mRNA levels, respectively. Taken together, our study suggests that IGF-I and IGF-II may act as local mediators of GH and GtHs in fish testis. Moreover, our results imply that in fish testicular cells, IGFs are potential paracrine/autocrine regulators inside the spermatogenic compartment and can act directly on germ cells to stimulate their proliferation.
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    Journal of bioenergetics and biomembranes 31 (1999), S. 467-473 
    ISSN: 1573-6881
    Keywords: Uncoupling proteins ; fatty acids ; skeletal muscle ; brown adipose tissue ; obesity ; thermogenesis ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract The recently discovered uncoupling protein 3 (UCP3) is highly homologous to the mitochondrialinner membrane protein UCP1, which generates heat by uncoupling the respiratory chainfrom oxidative phosphorylation. The thermogenic function of UCP1 protects against cold andregulates the energy balance in rodents. We review in vitro studies investigating the uncouplingactivity of UCP3 and in vivo studies, which address UCP3 gene expression in brown adiposetissue and skeletal muscle under various metabolic conditions. The data presented are, for themost, consistent with an uncoupling role for UCP3 in regulatory thermogenesis. We alsodiscuss mediators of UCP3 regulation and propose a potential role for intracellular fatty acidsin the mechanism of UCP3 modulation. Finally, we hypothesize a role for UCP3 in themetabolic adaptation of the mitochondria to the degradation of fatty acids.
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    Photosynthesis research 59 (1999), S. 1-7 
    ISSN: 1573-5079
    Keywords: leaf gas exchange ; ozone ; Pisum sativum ; Rubisco ; Triticum aestivum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Previous studies have shown that short exposure of plants to high doses of ozone decreases subsequent photosynthesis; initially by reducing carboxylation capacity. This study tests the hypothesis that this is also the primary cause of loss of photosynthetic capacity in leaves affected by development under a low level of ozone. Triticum aestivum and Pisum sativum plants were exposed from germination to ozone in air (80 nmol mol-1 for 7 hours per day, for 18 days. Leaves that had completed lamina expansion at this time were free of visible injury and light absorptance was unaffected. However, some significant changes in photosynthetic gas exchange were evident. Photosynthetic CO2 uptake at light saturation was decreased significantly by 35% in T. aestivum but was unchanged in P. sativum. The reduction in photosynthesis of T. aestivum was accompanied by a 31% decline in the maximum velocity of carboxylation measured in vivo. Decreased stomatal conductance did not contribute to this reduction of photosynthesis because there was no significant change in the stomatal limitation to CO2. Processes directly dependent upon photochemical reactions; that is, the quantum yield of CO2 uptake and capacity for regeneration of ribulose 1,5-bisphosphate were not affected by O3 fumigation in either species. This suggests that for wheat, the quantitative cause of decreased photosynthetic rate in vivo is a decrease in the quantity of active ribulose-1,5- bisphosphate carboxylase-oxygenase.
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    Boundary layer meteorology 91 (1999), S. 495-504 
    ISSN: 1573-1472
    Keywords: Evaporation ; Heat storage ; Marsh ; Inversion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Geosciences , Physics
    Notes: Abstract Concurrent measurements of the surface energy balance components (net radiation, heat storage, and sensible and latent heat fluxes) were made in three communities (open water, Phragmites australis, Scirpus acutus) in a wetland in north-central Nebraska, U.S.A., during May-October, 1994. The Bowen ratio – energy balance method was used to calculate latent and sensible heat fluxes. This paper presents results from the open water area. The heat stored in water (G) was found to play a major role in the energy exchange over the water surface. During daytime, G consumed 45–60% of R n , the net radiation (seasonally averaged daytime G was about 127 W m−2). At night, G was a significant source of energy (seasonally averaged nighttime G was about -135 Wm−). The diurnal pattern of latent heat flux (λ E) did not follow that of R n . On some days, λ E was near zero during midday periods with large R n . The diurnal variability in λ E seemed to be significantly affected by temperature inversions formed over the cool water surface. The daily evaporation rate (E) ranged from 2 to 8 mm during the measurement period, and was generally between 70 and 135% of the equilibrium rate.
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  • 78
    ISSN: 1573-1561
    Keywords: Aphid ; experience ; probing behavior ; sieve elements ; salivation ; cell punctures ; hydroxamic acids ; EPG ; Sitobion fragariae ; Triticum aestivum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract We hypothesized that aphids after previous exposure to hydroxamic acids (Hx), a family of secondary plant compounds deleterious to aphids, are able to reduce their subsequent exposure to them. This hypothesis was tested by evaluating the time to produce salivation into a sieve element (SSE) by the aphid Sitobion fragariae on seedlings of two wheat cultivars of Triticum aestivum differing in their concentration of Hx. The total time to produce a first SSE was significantly longer in the high-Hx cultivar; however, the subsequent, second SSE (first SSE after interruption of probing) in this cultivar was significantly reduced, reaching the level observed in the low-Hx plants. Therefore, a strategy to reduce the exposure to secondary compounds was observed only in the second SSE in high-Hx plants. When the experimental plant was replaced by a new unattacked plant after the first SSE, aphids did not change the behavior described, thus excluding an aphid-induced plant susceptibility. The number of cell punctures and accumulated duration was not affected by previous exposure to Hx, either in low or high Hx cultivars. Total time and pathway time but not cell punctures, seem to be the variables affected by previous exposure to Hx.
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  • 79
    ISSN: 1573-0867
    Keywords: bicarbonate-extractable soil phosphorus ; Lupinus angustifolius ; residual value ; Triticum aestivum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Decreases in Colwell bicarbonate soil test P in the years after applying single (ordinary) superphosphate, and the residual value of superphosphate, was measured in a long-term field experiment on a duplex (texture contrast) soil (sand over lateritic ironstone gravel clay sand at 10–15 cm), at Wongan Hills, Western Australia, typical of many soils used to grow crops in Western Australia. Ten levels of P (0–91 kg P ha-1) were applied once only in late May to different plots in different years from 1988 to 1993. Wheat (Triticum aestivum), or lupin ( Lupinus angustifolius)) were sown in late May of each year, when the P treatments applied that year were banded (drilled) with the seed. Soil samples were collected each June to measure soil test P. Seed (grain) yields of the crops were measured each December. The residual value (RV) of P applied in previous years was calculated relative to P applied in the current year, using grain yields (RVyield) and soil test P (RVsoil). Soil test P measured on soil samples collected in June was related to yields measured in December that year to provide soil P test calibrations. Relative to P applied in the current year, soil test P decreased by between 15 to 30% for P applied one year previously, by 25 to 30% for P applied three years previously, and by 60 to 70% for P applied six years previously. Soil test P was affected by spatial variation, and it also varied in the different years, for P applied in the current year, one year previously, two years previously, etc. Compared with P applied in the current year, mean RVyield determined in the different years decreased by about 40% one year after P application, followed by a further 20% decrease for P applied two years previously, followed by a further 20% decrease for P applied three to five years previously. Relative to current P, RVsoil decreased by about 25% one year after P application, followed by a further 20% for P applied two years previously, followed by a further 10% for P applied three years ago, and followed by a further 6% for P applied four and five years ago. As measured in the different years, the soil P test calibration varied between years for P applied one, two etc. years previously. This was so even when the same cultivar of wheat was grown at the same site in different years.
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  • 80
    ISSN: 1573-0603
    Keywords: Dichloromethylene diphosphate ; Hepatic stellate cell isolation ; Liposome ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Hepatic perisinusoidal cell population consists of hepatic stellate cells, Kupffer cells, endothelial cells, and Pit cells. These cells are isolated by enzymic digestion and purified by density gradient centrifugation. With isolation of stellate cells, conventional method is unable to eliminate the contamination of Kupffer cells because the densities of these two cells are similar. We report here an improved method for isolation of highly purified hepatic stellate cells, using dichloromethylene diphosphate (CL2MDP), which has selective cytotoxicity of Kupffer cells. Three days after the single intravenous administration of liposome-encapsulated CL2MDP, the Kupffer cells disappeared almost completely from the liver. Following Percoll density gradient centrifugation, the purity of the hepatic stellate cells exceeded 98% without any contamination of the Kupffer cells. Kupffer cells are reported to affect the physiological functions of stellate cells. The availability of highly purified stellate cells will facilitate the investigation of their functions in primary culture.
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  • 81
    ISSN: 1573-5087
    Keywords: flooding ; kinetin ; leaf relative water content ; membrane stability ; Triticum aestivum ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Wheat plants, 22d. old, were exposed to wide range of soil water osmotic potential (Ψs = 0 to −1.2 MPa) induced by NaCl and CaCl2 treatments in combination with roots maintained under aerobic (drained at field capacity) or nonaerobic (flooded) conditions in the soil, and sprayed with 10 mg L−1 kinetin solution. In drained plants, not receiving kinetin, increased soil salinity resulted in appreciable inhibition of shoot growth and reduction in chlorophyll (Ch1.), soluble sugars (SS) contents and grain yield. Shoot growth, Ch1. content, soluble sugars and grain yield were significantly lower for flooded plants than unflooded analogues over the entire Ψs range. Both salinity and waterlogging synergize to increase Na+, Ca+ and Cl− accumulation in shoot tissues and to decrease the stability of leaf membranes to either dehydration (40% polyethylene glycol 6000) or heat (51 °C) stress. The ratio of K+/Na+ transported to shoots under aerobic and anaerobic conditions decreased progressively on salinization. The association between the internal mineral element concentrations was largely affected by kinetin treatment. Kinetin application ameliorated the deleterious effects of salinity and oxygen deficiency. It reduced Na+, Ca2+ and Cl− accumulation and improved K+ uptake under salinity and waterlogging stresses. Increased K+/Na+ ratio helped the plants to avoid Na+ toxicity and enhanced shoot growth and grain yield. Kinetin also reduced membrane injury by dehydration and heat stresses and improved the water status of plants under both aerobic and anaerobic conditions. The effects of single factors (Soil salinity ‘Ψs’, soil waterlogging ‘WL’ and Kinetin ‘Kin’) and their interactions (Ψs × WL, Ψs × Kin, WL × Kin and Ψs × WL × Kin) were shown by analysis of variance to be statistically significant for most parameters tested. Calculation of the coefficient of determination (η+) led to three important findings. (1) Salinity (Ψs) was dominant in affecting leaf relative water content (RWC), shoot dry mass, grain yield, stability of leaf membranes to dehydration stress and the contents of Na+, Ca2+, Mg2+ and Cl−. (2) Kinetin (Kin) had a dominant effect on the stability of leaf membranes to heat stress as well as on chlorophyll and soluble sugars contents. (3) The share of waterlogging (WL) was dominant for K+ content. It can be concluded that kinetin application helped wheat plants to grow successfully in the areas subjected to combined effects of salinity and oxygen deficiency, such as in salt marshes.
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  • 82
    ISSN: 1573-5109
    Keywords: Aegilops tauschii ; hairy auricles ; hairy leaf sheath ; inheritance ; Triticum aestivum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Inheritance of genes for hairy auricles and hairy leaf sheath of Ae. tauschii in hexaploid wheat backgrounds (synthetic hexaploid wheat and common wheat varieties) was analyzed. The results indicated that hairy auricles and hairy leaf sheath of Ae. tauschii can be transferred and are expressed in hexaploid wheat. In a synthetic hexaploid wheat ('Ae. tauschii' 188) hairy auricles was proved to be controlled by a single dominant gene derived from Ae. tauschii, which was different from the Pa gene located on chromosome 4BS of common wheat. The hairy leaf sheath phenotype of 'Altar 84/Ae. tauschii 188' was also controlled by a single dominant gene derived from Ae. tauschii, which is obviously different from the Hls gene in T. dicoccoides. We suggest to designate the Ae. tauschii genes for hairy auricles and hairy leaf sheath as Pa2 and Hls2, respectively; such genes could be used as useful genetic markers in common wheat.
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  • 83
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    Plant cell, tissue and organ culture 59 (1999), S. 151-154 
    ISSN: 1573-5044
    Keywords: Ne1 ; Ne2 ; necrotic hybrid ; Triticum aestivum ; viable seeds
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Hybrid necrosis in wheat is a problem for gene transfer in wheat breeding. Hybrid necrosis occurs due to dominant complementary interaction of two genes, Ne1 and Ne2. A cross between wheat (Triticum aestivum L.) varieties C306 (drought tolerant, Ne1 carrier) and WL711(high yielding, Ne2 carrier) produced necrotic F1 hybrids, which died before ear emergence and produced no seeds. To overcome the problem of hybrid necrosis, ears enclosed in the leaf sheath were taken and cultured to maturity in liquid medium containing 5% sucrose and 0.04% glutamine. The necrotic hybrids produced only a few seeds per ear compared to parents, but individual grain weight was similar in the hybrid and the parents. The F1 ear culture study has been repeated for three years and F2 seeds obtained. In 1996–97, the cultured ears of F1 hybrids produced 62 seeds, of which only 52 showed germination and were grown under normal field conditions. Out of the 52 seeds, 50% were non-necrotic and showed segregation for various physiological traits. The results reveal that hybrids ears had the potential to form viable seeds. Culturing of wheat ears before ear emergence and production of viable F2 seeds from necrotic hybrids is a simple and efficient method for overcoming the problem of hybrid necrosis.
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  • 84
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    Euphytica 105 (1999), S. 125-131 
    ISSN: 1573-5060
    Keywords: disease resistance ; inheritance ; Karnal bunt ; Neovossia ; Tilletia indica ; Triticum aestivum ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Inheritance of resistance to Neovossia indica was studied in a Triticum aestivum line HD 29. To overcome the influence of environment on disease expression, the study was conducted by extensive evaluation of advanced generation (F8) recombinant inbred lines (RILs) developed by single seed descent from the cross WL 711 (susceptible) × HD 29 (resistant. The results suggested that HD 29 possesses three major genes for resistance to isolated Ni7 and two genes for resistance to isolate Ni8. One of the two genes controlling resistance to Ni8 is common with one of the genes conferring resistance to Ni7. These observations have important implications in breeding for Karnal bunt resistance.
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  • 85
    ISSN: 1573-5060
    Keywords: C-banding ; chromosome additions ; chromosome substitution ; Elymus tsukushiense ; homoeology ; in situ hybridization ; RFLP ; Roegneria kamoji ; Triticum aestivum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Elymus tsukushiense Honda (syn. Roegneria kamoji C. Koch) (2n = 6x = 42, StsStsHtsHtsYtsYts) is a hexaploid species, distantly related to bread wheat Triticum aestivum L. em Thell (2n = 6x = 42, AABBDD). Apart from the delineation of evolutionary relationships, this species is a potential source of resistance to scab, a devastating disease of wheat caused by Fusarium graminearum Schw. A standard C-banded karyotype was established identifying all 21 chromosome pairs of E. tsukushiense. By using C-banding and genomic in situ hybridization analyses, three wheat-E. tsukushiense chromosome addition lines, one ditelosomic addition line, and one disomic substitution line were identified in BC2 progenies from wheat × E. tsukushiense hybrids. Twenty DNA markers specific for the seven homoeologous groups of the Triticeae were used to determine the homoeology of the added E. tsukushiense chromosomes. The E. tsukushiense chromosomes in the addition lines NAU702, NAU703, and NAU701 were identified as belonging to homoeologous groups 1, 3, and 5, and thus, were designated as 1Ets#1, 3Ets#1, and 5Ets#1, respectively. NAU751 was identified as a disomic substitution line with chromosome 3A of wheat replaced by chromosome 3Ets#1. Line NAU702 has a high level of resistance to scab and will be used in chromosomal engineering and development of improved wheat germplasm for scab resistance breeding.
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  • 86
    ISSN: 1573-5060
    Keywords: 1BL/1RS ; bread-making quality ; prolamins ; wheat-rye translocations ; Triticum aestivum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The allelic variation of prolamin loci was studied in three F2 progenies from three crosses between the 1BL/1RS cultivar Triana and Yécora Rojo, Pavón and Florence Aurora, cultivars without the translocation. According to the 1:2:1 theoretical proportions observed in the allelic variants of the Glu-B3/Gli-B1 loci of the parent without the translocation, the inheritance as a block of the rye chromosome arm was confirmed. A group of F3-F4 recombinant lines, developed from these crosses was evaluated using the SDS-sedimentation test and the mixograph and alveograph tests. The presence of the 1BL/1RS translocation was not associated with significantly lower grain protein content values or with the optimum mixing time in the mixograph of the genotypes. The effect of the 1BL/1RS translocation on most of the quality parameters was highly dependent on the genetic pool. Significant increases in gluten strength and better mixing properties associated with the presence of some alleles of the Glu-A1, Glu-A3/ Gli-A1 and Gli-D2 loci were detected. The additivity and the interaction of prolamin gene effects with the rye translocation in the 1BL/1RS lines and its possible use in plant breeding are discussed.
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  • 87
    ISSN: 1573-5060
    Keywords: Blumeria graminis ; powdery mildew ; QTL ; RFLPs ; Triticum aestivum ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A segregating population of doubled-haploid lines issued from the cross between the wheat (Triticum aestivum L. em. Thell) cultivars Courtot, resistant to several isolates of powdery mildew (Blumeria graminis DC. f. sp. tritici Em. Marchal), and Chinese Spring (susceptible) was used to map Mlar, a gene carried by Courtot and conferring resistance to this pathogen. The assignation of Mlar using monosomic lines of Courtot was confirmed by the mapping analysis. Mlar was located on the short arm of the chromosome 1A, in the vicinity of the locus XGli-A5 coding for storage proteins. This result was in accordance with those demonstrating that Mlar was an allele of the Pm3 locus (Pm3g), a gene also involved in the resistance to powdery mildew.
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  • 88
    ISSN: 1573-5060
    Keywords: disease assessment ; maturity ; resistance ; Septoria tritici ; Triticum aestivum ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Nineteen cultivars, with large differences in heading date, were evaluated for their response to septoria tritici blotch in two experimental setups in Njoro, Kenya. Due to the more or less constant temperatures during the growing season and the overhead irrigation applied the epidemic conditions were similar over the whole observation period for the early and late cultivars. In experiment 1 the cultivars were assessed for disease severity at the same moment irrespective of the developmental stage, while in experiment 2 the cultivars were assessed at the same developmental stage. Measured at the same time, the disease severity was highest in the early maturing cultivars and lowest in the late maturing cultivars (r = –0.78). When assessed at the same development stage the disease build up was independent of heading date (r = –0.10) but strongly dependent on resistance level. There were no indications that early heading cultivars were more susceptible than late heading cultivars.
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  • 89
    ISSN: 1573-5060
    Keywords: allelic variation ; glutenin subunit ; Japanese Norin variety ; seed storage protein ; Triticum aestivum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Seed storage proteins of 131 Japanese Norin wheat (Triticum aestivum) varieties were fractionated by sodium dodecyl sulfate polyacrylamide gel electrophoresis to determine allelic make-up in varieties at each of three loci that control high-molecular-weight (HMW) glutenin subunits. Three alleles were identified at the Glu-A1 locus, six at the Glu-B1 locus and five at the Glu-D1 locus. Twenty-four different, major glutenin HMW subunits were identified and each contained three to five subunits and seventeen different glutenin subunit patterns were observed for 19 subunits in the 131 Japanese Norin varieties. Fourteen alleles were identified by comparison of subunit mobility with that previously found in hexaploid wheat. Japanese Norin varieties showed a specific pattern of allelic variation in glutenin HMW subunits, different from that of Chinese and other country common wheats in allelic frequency at Glu-1 loci.
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  • 90
    ISSN: 1573-5028
    Keywords: gene expression ; Gossypium hirsutum ; plant defence response ; Verticillium dahliae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Verticillium wilt is a vascular disease of cotton (Gossypium spp.) caused by the fungal pathogen Verticillium dahliae. To begin to understand the molecular mechanisms of the disease response in cotton cultivars that display superior wilt tolerance, such as Gossypium hirsutum cv. Sicala V-1, a cDNA library was constructed with mRNA isolated from root tissue of Sicala V-1, 24 h after inoculation with V. dahliae. The library was screened by a differential screening technique which was successful in identifying differences in gene expression between uninfected and V. dahliae-infected G. hirsutum root tissue. Among the differentially expressed clones, 51% represented up-regulated genes which had the potential to be involved in the defence response of G. hirsutum. The temporal expression patterns of nine suspected defence response genes were examined by northern blot analysis at several time intervals after inoculation with V. dahliae. The rapid increase in mRNA transcripts corresponding to each of these clones upon infection suggests a role for these genes in the defence response of G. hirsutum. Genes not previously associated with the defence response of the cotton plant, such as those for a 14-3-3-like protein and pathogenesis-related (PR) proteins, have been identified together with presumably novel genes, for which a definite function could not be ascribed.
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  • 91
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    Plant molecular biology 39 (1999), S. 1063-1071 
    ISSN: 1573-5028
    Keywords: cell cycle ; gene expression ; in vitro fertilization ; maize ; zygote ; embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Differential screening of cDNA libraries of unfertilized egg cells and in vitro zygotes of maize resulted in the isolation of more than 50 different genes whose expression is up- or down-regulated after in vitro fertilization (IVF). Amoung these genes, we identified a cDNA encoding the eukaryotic translation initiation factor eIF-5A. This highly conserved factor is thought to be necessary for selective mRNA stabilization and translation. It is also the only known protein that contains the unusual amino acid hypusine which is required for biological activity. High transcript amounts are stored in the egg cell, which is, in terms of metabolism, relatively inactive. Upon fertilization transcript amounts decrease, in contrast to metabolically inactive embryos in which the transcript cannot be detected and transcript levels increase upon germination. The expression pattern during the first embryonic cell cycle is also different from that observed during the somatic cell cycle: egg cells in the G0 phase contain high transcript levels, while arrested suspension cells contain few transcripts. In the somatic cell cycle, eif-5A is strongly induced during the G1 phase and transcripts are continuously degraded during the S, G2 and M phases until new induction during the G1 phase of the next cycle. eif-5A, a member of a small gene family in maize, is expressed in most maize tissues investigated. Based on our results, we suggest that the unfertilized egg cell of maize, although relatively inactive regarding its metabolism, is prepared for selective mRNA translation that is quickly triggered after fertilization. We also suggest that the regulation of eif-5A in the first embryonic cell cycle is different from the somatic cell cycle.
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  • 92
    ISSN: 1573-5028
    Keywords: aquaporin ; canola ; gene expression ; germination ; priming ; salt and osmotic stresses
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two aquaporin genes were isolated from a cDNA library of canola (Brassica napus L.). The first aquaporin, BnPIP1 of 1094 bp, encoding a putative polypeptide of 287 amino acids with a predicted molecular mass of 30.4 kDa and a pI of 7.8, belongs to the family of plasma membrane intrinsic protein (PIPs) aquaporins. The B. napus aquaporin showed 85–94% identity to the Arabidopsis thaliana PIPs. ABA priming of seed induced high levels of BnPIP1 transcript which remained after subsequent re-drying of the seed. The second aquaporin, Bnγ-TIP2 of 1020 bp, encoded a putative polypeptide of 253 amino acids with a predicted molecular mass of 25.8 kDa and a pI of 5.8. Bnγ-TIP2 showed 83–90% identity to γ-TIP genes from a variety of plant species. Bnγ-TIP2 was expressed only when radicle protrusion occurred in either untreated or primed seeds. Seeds primed with PEG or ABA germinated earlier and showed a higher final percentage of germination than unprimed seed, particularly under salt and osmotic stresses at low temperature. Transcripts of both BnPIP1 and Bnγ-TIP2 genes were present earlier during germination of primed seeds than non-primed seed. From these results, we conclude that BnPIP1 is related to the water transportation required for enzymatic metabolism of storage nutrients at the early stages of canola seed germination whereas Bnγ-TIP2 expression is related to cell growth associated with radicle protrusion. Priming induced the expression of BnPIP1 but had no effect on Bnγ-TIP2.
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  • 93
    ISSN: 1573-5028
    Keywords: Brassica napus ; cauliflower mosaic virus ; epidermis ; gene expression ; light induction ; lipid transfer protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract cDNA and genomic clones encoding Brassica napus non-specific lipid transfer proteins (LTP) were isolated and sequenced. The encoded amino acid sequences were very similar to those reported previously for LTPs from B. napus and other species. Sequence information indicates that B. napus contains an LTP gene family. The 5′-flanking region of one gene, designated BnLTP, was fused to GUS and the fusion introduced into Arabidopsis. LTP transcripts and BnLTP-Gus expression were present predominantly in the epidermis of leaf and stem, consistent with the hypothesised function of LTPs in the deposition of cuticular or epicuticular waxes. However, GUS activity was detected in other tissues, including lateral root initials, anthers, stigmas and vascular tissues, which may suggest additional functions. LTP transcript levels in B. napus and Arabidopsis and BnLTP-GUS expression in transgenic Arabidopsis were stimulated by blue and red light but not UV-B. BnLTP promoter activity was also stimulated upon viral infection, at a time when the virus had spread systemically. No increase in expression was observed in response to cold or wounding.
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  • 94
    ISSN: 1573-5028
    Keywords: 1-aminocyclopropane-1-carboxylate synthase ; auxin ; brassinosteroid ; ethylene ; gene expression ; hypocotyl ; promoter ; transgenic tobacco ; Vigna radiata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Indole-3-acetic acid (IAA) markedly increased ethylene production by inducing the expression of three 1-aminocyclopropane-1-carboxylate (ACC) synthase cDNAs (pVR-ACS1, pVR-ACS6 and pVR-ACS7) in mung bean hypocotyls. Results from nuclear run-on transcription assay and RNA gel blot studies revealed that all three genes were transcriptionally active displaying unique patterns of induction by IAA and various hormones in etiolated hypocotyls. Particularly, 24-epibrassinolide (BR), an active brassinosteroid, specifically enhanced the expression of VR-ACS7 by a distinct temporal induction mechanism compared to that of IAA. In addition, BR synergistically increased the IAA-induced VR-ACS6 and VR-ACS7 transcript levels, while it effectively abolished both the IAA- and kinetin-induced accumulation of VR-ACS1 mRNA. In light-grown plants, VR-ACS1 was induced by IAA in roots, and VR-ACS6 in epicotyls. IAA- and BR-treatments were not able to increase the VR-ACS7 transcript in the light-grown tissues. These results indicate that the expression of ACC synthase multigene family is regulated by complex hormonal and developmental networks in a gene- and tissue-specific manner in mung bean plants. The VR-ACS7 gene was isolated, and chimeric fusion between the 2.4 kb 5′-upstream region and the β-glucuronidase (GUS) reporter gene was constructed and introduced into Nicotiana tabacum. Analysis of transgenic tobacco plants revealed the VR-ACS7 promoter-driven GUS activity at a highly localized region of the hypocotyl-root junction of control seedlings, while a marked induction of GUS activity was detected only in the hypocotyl region of the IAA-treated transgenic seedlings where rapid cell elongation occurs. Although there was a modest synergistic effect of BR on the IAA-induced GUS activity, BR alone failed to increase the GUS activity, suggesting that induction of VR-ACS7 occurs via separate signaling pathways in response to IAA and BR. A scheme of the multiple regulatory pathways for the expression of ACC synthase multigene family by auxin and BR is presented.
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  • 95
    ISSN: 1573-5036
    Keywords: boron ; genotypic difference ; sterility ; Triticum aestivum ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Two pot experiments at the Plant Environment Laboratory (PEL), Reading, UK investigated sterility, boron (B) accumulation and B partitioning of wheat cultivars grown with limited B in the growing medium. The first experiment evaluated nine cultivars of spring wheat with diverse field responses to low available soil B, supplied with or without 20 μM B. A second experiment examined the response of a susceptible (SW-41) and a tolerant (Fang-60) cultivar to B-deficiency. These cultivars were supplied with either 20 μM B from sowing to flag leaf emergence and no added B thereafter, or 20 μM B from sowing to maturity. When B was not supplied in the nutrient solution, the number of grains ranged from 4 per ear (cv. BL-1135) to 32 per ear (cv. BL-1249) and sterility of competent florets ranged from 39% to 93%. Boron concentration in the flag leaf at anthesis did not differ greatly when the growing medium contained limited B, but differences between cultivars were evident when B was unlimited. Tolerance of B-deficiency was not related to the B concentration in the flag leaf. Some cultivars produced viable pollen and set grains while others failed to do so at similar B concentrations in the flag leaf. The two contrasting cultivars did not differ much in their pattern of B partitioning when B supply was restricted from flag leaf emergence onwards. Similarly, little evidence was found that the tolerant cultivars translocated B from their leaves, roots or stems when the supply in the growing medium was restricted. The proportion of total B partitioned in different organs was the same irrespective of B supply and cultivar. On average, leaves contained 68% of the total B content in the whole plant compared to 16% in the roots, 10% in the ears and only 6% in the stems. Tolerant or susceptible cultivars of wheat could not be distinguished based on the B concentration and B content of the flag leaf.
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  • 96
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    Plant cell, tissue and organ culture 57 (1999), S. 207-210 
    ISSN: 1573-5044
    Keywords: biolistics ; gene expression ; haploid ; transformation ; Triticum aestivum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Using the PDS-1000/He Biolistic® Particle Delivery System, the microprojectile travel distance, rupture disk pressure and DNA/gold particle concentrations were assessed in order to optimise short and longer-term β-glucuronidase reporter gene expression in microspore-derived embryos of wheat. The effects were also evaluated of using sterile filter paper to support explants and treatment with a high osmoticum medium (0.2 M mannitol/0.2 M sorbitol or 0.4 M maltose). In the optimised procedure, wheat microspore-derived embryos (MDEs), were placed on filter paper and incubated on medium containing 0.4 M maltose, for 4 h pre- and 45 h post-bombardment. Five μl pAHC25 (0.75 mg ml-1 in TE buffer) was precipitated onto 25 μl gold particles (60 mg ml-1 in sterile water), using 20 μl spermidine (0.1 M) and 50 μl CaCl2 (2.5 M). The particles were centrifuged and resuspended in 75 μl absolute ethanol prior to the preparation of 6 macrocarriers. A microprojectile travel distance of 70 mm, a rupture pressure of 1300 p.s.i., and a vacuum of 29′′ Hg were employed. Maltose at 0.4 M in the support medium was the most important factor influencing GUS activity in bombarded tissues. GUS activity, 1 day post-bombardment, reached 52 ± 17 GUS-positive foci/MDE (mean ± s.e.m, n=3), with 17 ± 4 foci/MDE at 15 days, giving a 3.0-fold increase (p〈0.05) compared to expression in MDEs bombarded on medium without a high osmoticum treatment.
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  • 97
    ISSN: 1573-6822
    Keywords: cytokines ; gene expression ; in vitro human epidermis ; acute and chronic skin irritation ; predictivity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract An in vitro human reconstructed epidermis model (SkinEthic) used for screening acute and chronic skin irritation potential was validated against in vivo data from skin tolerability studies. The irritation potential of sodium lauryl sulfate (SLS), calcipotriol and trans-retinoic acid was investigated. The in vitro epidermis-like model consists of cultures of keratinocytes from human foreskin on a polycarbonate filter. The modulation of cell viability, the release and gene expression of proinflammatory cytokines, interleukins 1α and 8, and morphological changes were evaluated during 3 days as endpoints representative for an inflammatory reaction. The cumulative irritation potential of the topical products was evaluated in a human clinical study by visual scoring and biophysical measurement of inflammatory skin reaction after repeated 24 h applications over 3 weeks under Finn chamber patches. All topical products that were nonirritating in the human study were noncytotoxic and did not induce cytokine expression in the in vitro acute model (day 1 exposure). All irritating controls exhibited specific cell viability and cytokine patterns, which were predictive of the in vivo human data. The ranking of mild to moderate skin irritation potential was based on the lack of cytotoxicity and the presence of cytokine patterns including gene expression specific for each irritant, using the chronic in vitro model (up to 3 days exposure). The human reconstructed epidermis model SkinEthic was shown to be a reliable preclinical tool predicting the irritation potential of topical products. Moreover, it is a useful model in a two-step tiered strategy for screening acute and chronic irritation potential for the selection of vehicles for new topical drugs.
    Type of Medium: Electronic Resource
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  • 98
    Electronic Resource
    Electronic Resource
    Springer
    Cellular and molecular neurobiology 19 (1999), S. 93-108 
    ISSN: 1573-6830
    Keywords: focal cerebral ischemia ; cerebral infarction ; penumbra ; gene expression ; stress response ; inflammatory reaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract 1. We review the biochemical and molecular changes in brain with developing cerebral infarction, based on recent findings in experimental focal cerebral ischemia. 2. Occlusion of a cerebral artery produces focal ischemia with a gradual decline of blood flow, differentiating a severely ischemic core where infarct develops rapidly and an area peripheral to the core where the blood flow reduction is moderate (called penumbra). Neuronal injury in the penumbra is essentially reversible but only for several hours. The penumbra area tolerates a longer duration of ischemia than the core and may be salvageable by pharmacological agents such as glutamate antagonists or prompt reperfusion. 3. Upon reperfusion, brain cells alter their genomic properties so that protein synthesis becomes restricted to a small number of proteins such as stress proteins. Induction of the stress response is considered to be a rescue program to help to mitigate neuronal injury and to endow the cells with resistance to subsequent ischemic stress. The challenge now is to determine how the neuroprotection conferred by prior sublethal ischemia is achieved so that rational strategies can be developed to detect and manipulate gene expression in brain cells vulnerable to ischemia. 4. Expansion of infarction may be caused by an apoptotic mechanism. Investigation of apoptosis may also help in designing novel molecular strategies to prevent ischemic cell death. 5. Ischemia/reperfusion injury is accompanied by inflammatory reactions induced by neutrophils and monocytes/macrophages infiltrated and accumulated in ischemic areas. When the role of the inflammatory/immune systems in ischemic brain injury is revealed, new therapeutic targets and agents will emerge to complement and synergize with pharmacological intervention directed against glutamate and Ca2+ neurotoxicity.
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  • 99
    ISSN: 1573-2932
    Keywords: Ponderosa pine ; elevated CO2 ; growth ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract Biochemical and gene expression changes in response to elevated atmospheric CO2 were investigated in five maternal half-sibling breeding families of Ponderosa pine. Seedlings were grown in a common garden located at Lawrence Livermore National Laboratory, in open-topped chambers (OTC) for two years. Chamber atmospheres were maintained at ambient, ambient + 175 μL L-1, CO2, or ambient + 350 μL L-1CO2. Growth measurements showed significant increases in stem volumes and volume enhancement ratios in three of the five families studied when grown under elevated CO2. Biochemical and gene expression studies were undertaken to gain a mechanistic understanding of these phenotypic responses. Biochemical studies focused on sucrose phosphate synthase (SPS) specific activities at increase CO2 levels. Kinetic evaluations of SPS showed an increase in VMax. Specific SPS probes revealed increases in the transcriptional levels of one SPS gene with exposure to increasing CO2. RT-PCR differential gene displays showed that overall only a small fraction of visualized gene transcripts responded to elevated CO2 (8-10%). There were also significant differences between the gene expression patterns of the different families, some of which correlated with alterations in growth at elevated CO2 levels.
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  • 100
    ISSN: 1573-9058
    Keywords: diurnal variation ; photoinhibition ; photosystem 2 reaction centres ; state transition ; trans-thylakoid membrane proton gradient ; Triticum aestivum ; xanthophyll cycle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Non-photochemical quenching of chlorophyll fluorescence (qN) and its three components (qNf, qNm, and qNs) in the flag leaves of wheat grown in the field were studied by a fluorometer PAM-2000 on clear days. The diurnal variation patterns of qN in just fully extended (JFEL) and aging leaves (AL) were similar, but qNm declined markedly in JFEL while it remained at a relatively high level in AL under strong sunlight at noon. Furthermore, at midday qNf was higher than qNs in JFEL, but much lower in AL. The results show the relative contributions of different mechanisms in preventing the photosynthetic apparatus from photodamage change during leaf development.
    Type of Medium: Electronic Resource
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