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  • Articles  (727)
  • wheat  (294)
  • Saccharomyces cerevisiae  (243)
  • Zea mays  (193)
  • Springer  (727)
  • 1990-1994  (727)
  • 1
    Electronic Resource
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    Springer
    Entomologia experimentalis et applicata 71 (1994), S. 177-180 
    ISSN: 1570-7458
    Keywords: aflatoxin ; Carophilus ; Zea mays ; corn ; plant resistance ; Coleoptera ; Nitidulidae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
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  • 2
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    Entomologia experimentalis et applicata 72 (1994), S. 17-23 
    ISSN: 1570-7458
    Keywords: plant varietal resistance ; armyworm ; Spodoptera exempta ; leaf extracts ; Zea mays ; feeding deterrent ; toxicity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Maize (Zea mays L.) leaf tissue of cv Bastille and cv Michoacan 12 was extracted with n-hexane. The extracts were bioassayed against 5th instar African armyworm,Spodoptera exempta (Walker)(Lepidoptera: Noctuidae), by feeding the larvae on agar based media or sucrose impregnated glass fibre discs. The hexane extract of the ‘resistant’ cv Bastille exhibited feeding deterrency and toxicity which were not shown by the ‘susceptible’ cv Michoacan 12. The hexane extract of cv Bastille was adsorbed onto silica gel, the solution filtered off and the adsorbed component taken up into ethyl acetate. Bioassay of these fractions indicated that the toxic and deterrent action was retained in the ethyl acetate fraction. Preparative thin layer chromatography of the ethyl acetate fraction isolated two biologically active constituents. These were both growth inhibitors and lethal by ingestion to the 5th instar African armyworm. Implications for resistance in maize varieties to insect pests are discussed.
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  • 3
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    Entomologia experimentalis et applicata 68 (1993), S. 219-229 
    ISSN: 1570-7458
    Keywords: Hymenoptera ; Aphidiidae ; Homoptera ; Aphididae ; Schizaphis graminum ; wheat ; tritrophic interactions ; learning ; host-habitat location
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of experience on the responsiveness of the aphidiid parasitoidLysiphlebus testaceipes (Cresson) (Hymenoptera: Aphidiidae) to host-associated cues was investigated using a wind-tunnel bioassay. Naive females were able to discriminate between uninfested wheat (Triticum aestivum L.) and wheat infested withSchizaphis gramimum (Rondani) (Homoptera: Aphididae), but oviposition experience significantly increased the parasitoid's propensity to respond to aphid-infested plants with upwind, targeted flight. The behavioural change associated with such experience was acquired rapidly (within five minutes) and persisted for at least 24 h. The parasitoid could be successfully conditioned to associate a novel odour with the presence of hosts, suggesting that the increase in response to aphid-infested plants which occurs as a result of experience is probably due to associative learning of olfactory cues from the plant-aphid complex.
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  • 4
    ISSN: 1572-8773
    Keywords: catalase ; copper resistance ; pH-dependent growth ; Saccharomyces cerevisiae ; superoxide dismutase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A strain of Saccharomyces cerevisiae has been adapted to increasing concentrations of copper at two different pH values. The growth curve at pH 5.5 is characterized by a time generation increasing with the amount of added copper. A significant decrease of cell volume as compared with the control is also observed. At pH 3 the cells grow faster than at pH 5.5 and resist higher copper concentrations (3.8 against 1.2 mm). Experimental evidence indicates that, after copper treatment, the metal is not bound to the cell wall, but is localized intracellularly. A significant precipitation of copper salts in the medium was observed only at pH 5.5. Increased levels of superoxide dismutase (SOD) activity were observed in copper-treated cells and which persisted after 20 subsequent inocula in a medium without added metal. On the contrary, catalase activity was not stimulated by copper treatment and, hence, not correlated with SOD levels. The mechanism of copper resistance, therefore, probably involves a persistent induction of SOD, but not of catalase, and it is strongly pH-dependent.
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  • 5
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    Cellular and molecular life sciences 50 (1994), S. 571-575 
    ISSN: 1420-9071
    Keywords: Ancient DNA ; archaeobotany ; carbonized grain ; DNA sequences ; glutenin alleles ; seed proteins ; Triticum ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have used hybridization analysis to detect ancient DNA in wheat seeds collected from three archaeological sites in Europe and the Middle East. One of these samples, carbonizedT. spelta dated to the first millennium BC, has yielded PCR products after amplification with primers directed at the leader regions of the HMW (high molecular weight) glutenin alleles. Sequences obtained from these products suggest that the DNA present in the Danebury seeds is chemically damaged, as expected for ancient DNA, and also indicate that it should be possible to study the genetic variability of archaeological wheat by ancient DNA analysis. Finally, we describe a PCR-based system that enables tetraploid and hexaploid wheats to be distinguished.
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  • 6
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    Cellular and molecular life sciences 48 (1992), S. 1162-1164 
    ISSN: 1420-9071
    Keywords: Polygodial ; warburganal ; antifungal activity ; Candida albicans ; Saccharomyces cerevisiae ; Pityrosporum ovale ; enhancing effect ; antioxidants ; vitamin C ; BHA ; anethole
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The antifungal activity of two drimane sesquiterpene dialdehydes, polygodial (1) and warburganal (2), alone and in combination with several other substances, was examined against three fungi,Candida albicans, Saccharomyces cerevisiae andPityrosporum ovale employing a broth dilution method. Anethole significantly synergized the activity of the two sesquiterpenoids againstC. albicans andS. cerevisiae however, it had only an, additive effect againstP. ovale. By contrast, two antioxidants, ascorbic acid (vitamin C) and BHA (butylated hydroxyanisole), noticeably enhanced the activity of the sesquiterpenoids againstP. ovale, but had no, effect againstC. albicans andS. cerevisiae.
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  • 7
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    Entomologia experimentalis et applicata 54 (1990), S. 29-36 
    ISSN: 1570-7458
    Keywords: Zea mays ; corn ; host plant resistance ; phenolics ; flavonoids ; hydroxamic acids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Description / Table of Contents: Résumé Les réactions de larves et adultes du nitidulidé C. hemipterus (L.), vecteur de champignons produisant la mycotoxine, aux composés phénoliques caractéristiques, aux flavonoïdes et aux acides hydroxamiques, métabolites secondaires qui provoquent la résistance du maïs (Zea mays L.) ont été examinées au cours d'expériences avec et sans choix. L'alimentation des adultes et des larves est généralement réduite par les acides coumarique et férulique et par la 6-méthoxy-2-benzoxazolinone dans des expériences sans choix; les insectes évitent généralement les aliments qui contiennent ces produits. Quoi qu'il en soit, les larves préfèrent consommer d'autres aliments contenant les autres phénoliques ou flavonoïdes examinés. Les adultes sont plus inconstants dans leur choix alimentaires, mais préfèrent souvent des aliments contenant de la quercetine. Ainsi, des programmes de sélection orientés contre les principaux ravageurs comme Heliothis zea (Boddie) ou Ostrinia nubilalis (Hübner), impliquant la sélection de plantes à teneur élevée en acides phénolique ou hydroxamique, augmentant probablement aussi la résistance aux nitidulidés.
    Notes: Abstract Selected secondary metabolites produced by maize (Zea mays L.) were tested for effects on larvae and adults of the dried-fruit beetle [Carpophilus hemipterus (L.)] in no-choice and choice assays. Feeding by adults and larvae was significantly reduced by ferulic acid and 6-methoxy-2-benzoxazolinone (MBOA) in no-choice assays. In choice assays, larvae and adults generally preferred diets with trans-cinnamic acid, quercetin, rutin, and thymol, but were repelled by diets with either ferulic acid or MBOA.
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  • 8
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    Journal of industrial microbiology and biotechnology 7 (1991), S. 131-135 
    ISSN: 1476-5535
    Keywords: Saccharomyces cerevisiae ; Jerusalem artichoke ; High-fructose syrup ; Ethanol ; Immobilized yeast cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The results from this study showed that Jerusalem artichoke juice can be used for the production of very enriched fructose syrup by selective conversion of glucose to ethanol in a continuous process using immobilized cells ofSaccharomyces cerevisiae ATCC 36859. The product contained up to 99% of the total carbohydrates as fructose compared to 76% in the feed. Using Jerusalem artichoke juice supplemented with some glucose a product was obtained with 7.5% w/v ethanol which made ethanol recovery economically favourable. It was found that some fructose was consumed in these continuous processes; the glucose/fructose conversion rate ratio was regulated by the glucose concentration in the product stream.
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  • 9
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    Journal of industrial microbiology and biotechnology 7 (1991), S. 181-189 
    ISSN: 1476-5535
    Keywords: Saccharomyces cerevisiae ; Torulaspora delbrueckii ; Aroma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Thirty-three fermentations of Pedro Ximénez grapes, collected in three degrees of ripeness, were carried out by inoculation with three types of inoculum: pure cultures ofSaccharomyces cerevisiae races and ofTorulaspora delbrueckii, indigenous yeasts, and mixed cultures of indigenous yeasts enriched with the pure cultures. By means of variance analysis 21 compounds were determined whose final concentrations in the wines significantly depended on the musts, the inocula or both. Eleven products that depended significantly on the inocula were subjected to a discriminant analysis in which most of the pure cultures gathered in a discriminant space area different from that occupied by the indigenous yeasts. The centroids corresponding to most of the mixed cultures were shifted to the central area of the discriminant space, moved away from their corresponding pure cultures and approached the indigenous yeasts. The results show a high similarity between the fermentations carried out with mixed cultures with the addedS. cerevisiae races and those fermentations carried out with the indigenous yeasts, with regard to those compounds which were significantly dependent on the inocula.
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  • 10
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    Mycorrhiza 5 (1994), S. 119-124 
    ISSN: 1432-1890
    Keywords: Key words Glomus mosseae ; Manganese uptake ; Root exudation ; Manganese reduction ; Mycorrhizal effect ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The influence of rhizosphere microorganisms and vesicular-arbuscular (VA) mycorrhiza on manganese (Mn) uptake in maize (Zea mays L. cv. Tau) plants was studied in pot experiments under controlled environmental conditions. The plants were grown for 7 weeks in sterilized calcareous soil in pots having separate compartments for growth of roots and of VA mycorrhizal fungal hyphae. The soil was left either uninoculated (control) or prior to planting was inoculated with rhizosphere microorganisms only (MO-VA) or with rhizosphere microorganisms together with a VA mycorrhizal fungus [Glomus mosseae (Nicol and Gerd.) Gerdemann and Trappe] (MO+VA). Mycorrhiza treatment did not affect shoot dry weight, but root dry weight was slightly inhibited in the MO+VA and MO-VA treatments compared with the uninoculated control. Concentrations of Mn in shoots decreased in the order MO-VA〉MO+VA〉control. In the rhizosphere soil, the total microbial population was higher in mycorrhizal (MO+VA) than nonmycorrhizal (MO-VA) treatments, but the proportion of Mn-reducing microbial populations was fivefold higher in the nonmycorrhizal treatment, suggesting substantial qualitative changes in rhizosphere microbial populations upon root infection with the mycorrhizal fungi. The most important microbial group taking part in the reduction of Mn was fluorescent Pseudomonas. Mycorrhizal treatment decreased not only the number of Mn reducers but also the release of Mn-solubilizing root exudates, which were collected by percolation from maize plants cultivated in plastic tubes filled with gravel quartz sand. Compared with mycorrhizal plants, the root exudates of nonmycorrhizal plants had two fold higher capacity for reduction of Mn. Therefore, changes in both rhizosphere microbial population and root exudation are probably responsible for the lower acquisition of Mn in mycorrhizal plants.
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  • 11
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    Biology and fertility of soils 18 (1994), S. 228-230 
    ISSN: 1432-0789
    Keywords: Fine root ; Root litter ; Biofertiliser ; Leucaena leucocephala ; Trigonella foenum-graecum ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The efficacy ofLeucaena leucocephala root litter as a natural biological fertiliser was assessed usingZea mays as a test plant. Up to 8% of the fine roots of the plants constituted root litter. This fine root litter was better than that ofTrigonella foenum-graecum at increasing the growth and productivity ofZea mays. The root litter increased the growth of maize shoots more than the growth of roots. This appears to be a general phenomenon when plant nutrients are insufficient, as in the present study.
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  • 12
    ISSN: 1432-0983
    Keywords: 2-oxoglutarate dehydrogenase ; Saccharomyces cerevisiae ; rad52-mediated chromosome loss
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Ogd1 mutants of Saccharomyces cerevisiae are deficient in mitochondrial 2-oxoglutarate dehydrogenase activity; they cannot grow on glycerol and produce an increased amount of organic acids during growth on glucose as substrate. Using gamma ray-induced rad52-mediated chromosome loss the ogd1 mutation can be assigned to chromosome IX. Tetrad analysis of crosses between ogd1 and other markers on chromosome IX revealed that the OGD1 gene maps on the left arm of this chromosome 1.9 cM from his5.
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  • 13
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Orotate phosphoribosyl transferase ; Nucleotide sequence-5-phosphoribosyl 1-pyrophosphate (5PRPP)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Orotate phosphoribosyl transferase (OPRTase) catalyses the transformation of orotate to OMP in the pyrimidine pathway. In the yeast Saccharomyces cerevisiae, the URA5 gene is known to encode this enzyme activity. In this paper we present the cloning and sequencing of a yeast gene, named URA10, encoding a second OPRTase enzyme. Comparison of the predicted amino acid sequences between URA5 and URA10 genes shows more than 75% similarity. These sequences have also been compared to those of Escherichia coli, Podospora anserina, Sordaria macrospora and Dictyostelium discoideum. Remarkable similarities in the primary structure of these proteins have been found. Gene disruption experiments revealed that URA10 gene expression is responsible for the leaky phenotype of a ura5 mutant. Assays of OPRTase activity in extracts from ura5 and ura10 mutants indicate that the URA10 product contributes only 20% of the total activity found in wild type cells.
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  • 14
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Mutants ; Farnesyl diphosphate synthetase ; Ergosterol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two yeast mutant strains auxotrophic for ergosterol and blocked in farnesyl diphosphate synthetase (EC 2.5.1.1) were isolated. Genetic analysis has shown that these mutant strains carry additional mutations in the ergosterol pathway besides erg20-1 and erg20-2 which affect FPP synthetase. The novel feature of these mutants is their ability to excrete prenyl alcohols (farnesol and geraniol). As geraniol is toxic for yeast cells, the above leaky mutations in FPP synthetase have to be associated with others in the sterol pathway, in order to slow down geraniol synthesis.
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  • 15
    ISSN: 1432-0983
    Keywords: Glucose oxidase ; Aspergillus ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We report the cloning of the Aspergillus niger glucose oxidase gene and its use to elevate glucose oxidase productivity in A. niger by increasing the gene dosage. In addition, the gene has been introduced into A. nidulans where it provides the novel capacity to produce glucose oxidase. A plasmid, in which DNA encoding the mature form of glucose oxidase was preceded by a Saccharomyces cerevisiae secretion signal, effected high-level production of extracellular glucose oxidase in this yeast.
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  • 16
    ISSN: 1432-0983
    Keywords: Schizosaccharomyces pombe ; Saccharomyces cerevisiae ; Argininosuccinate lyase ; Sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The complete nucleotide sequence of the ARG7 gene, coding for argininosuccinate lyase (EC 4.3.2.1), in the fission yeast (Schizosaccharomyces pombe) has been determined. It consists of an open reading frame of 461 codons. The deduced protein has a molecular weight of 51 200 Da. The gene is devoid of introns which is confirmed by the fact that it is expressed in Escherichia coli after spontaneous insertion of a bacterial sequence probably bearing a prokaryotic promoter. A perfect “TATA” box is found at-72 and the major transcription initiation site in Saccharomyces cerevisiae is located at-11 as shown by primer extension experiments. Comparison of the S. pombe lyase with related proteins from other organisms reveals an important degree of conservation except in the carboxyterminal part of the polypeptide. Additionally, a deletion removing 66 amino acids of the carboxy terminus yields an enzyme exhibiting some biological activity. A unique 1500 b transcript was found in S. cerevisiae when the intact gene was present, but the deleted version of the gene gave rise to at least three transcripts of 1800, 2800 and 3900 b.
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  • 17
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Pyrimidine salvage pathway ; Semi-dominant mutants ; FUR1 ; Uracil phosphoribosyl transferase ; Regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In Saccharomyces cerevisiae, the protein encoded by the FUR1 gene is absolutely required for the expression of uracil phosphoribosyl transferase activity. The occurrence of semi-dominant mutations for 5-fluorouracil-(5FU)-resistance at this locus led us to clone and sequence the semi-dominant fur 1–5 allele. A single point mutation, resulting in the substitution of arginine 134 for serine, is responsible for this mutant phenotype. The fur 1–5 allele is transcribed and expressed at the same level as the wild-type allele. But, in contrast with the wild-type, the UPR Tase activity of the fur 1–5 mutant strain is stimulated in vitro by UTP and does not, therefore, correspond to a loss of feedback of UPR Tase activity. We found that uracil, as a free base, induces a significative increase in transcription and UPR Tase activity in a wild-type strain as well as in uracil-overproducing mutants which principally explains the high efficiency of the pyrimidine salvage pathway in S. cerevisiae.
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  • 18
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    Current genetics 18 (1990), S. 401-403 
    ISSN: 1432-0983
    Keywords: Baking yeast ; Saccharomyces cerevisiae ; Dough leavening ; Benomyl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary To investigate the leavening ability of yeast in dough, chromosome loss was induced by benomyl treatment in YOY1037, a diploid between a baking strain and a laboratory strain, and its effect on the leavening ability was studied. When benomyl-treated cells were spread on plates with a dye indicator for ploidy, about 20% of the visible colonies were stained dark blue or dark purple; the rest stained pale blue, similar to the diploid YOY1037. Strains showing the MATα phenotype, and non-galactose fermenting strains, apparently having lost particular chromosomes, were observed only in those with darkcoloured colonies. Strains with dark-coloured colonies showed a wider range of leavening ability than did those with pale-coloured colonies.
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  • 19
    ISSN: 1432-0983
    Keywords: Xylitol dehydrogenase gene ; Pichia stipitis ; Saccharomyces cerevisiae ; Xylose utilization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A P. stipitis cDNA library in λgt11 was screened using antisera against P. stipitis xylose reductase and xylitol dehydrogenase, respectively. The resulting cDNA clones served as probes for screening a P. stipitis genomic library. The genomic XYL2 gene was isolated and the nucleotide sequence of the 1089 bp structural gene, and of adjacent non-coding regions, was determined. The XYL2 open-reading frame codes for a protein of 363 amino acids with a predicted molecular mass of 38.5 kDa. The XYL2 gene is actively expressed in S. cerevisiae transformants. S. cerevisiae cells transformed with a plasmid, pRD1, containing both the xylose reductase gene (XYL1) and the xylitol dehydrogenase gene (XYL2), were able to grow on xylose as a sole carbon source. In contrast to aerobic glucose metabolism, S. cerevisiae XYL1-XYL2 transformants utilize xylose almost entirely oxidatively.
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  • 20
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Centromere flanking sequences ; tRNA modification enzymes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Transcriptional analysis of the region flanking the left boundary of the centromere of chromosome VI revealed the presence of a gene immediately adjacent to CEN6. The transcription of the gene is directed toward the centromere, and nucleotide sequence analysis showed that the coding region terminates only 50 bp away from CEN6. Our results extend to chromosome VI the observation that centromere-flanking regions of S. cerevisiae are transcriptionally active. Disruption of the coding region of the gene showed that its product, whilst not essential for cell viability, is important for normal cell growth. The gene has been termed DEG1 (DEpressed Growth rate). Comparison of the deduced amino acid sequence of DEG1 with a protein sequence databank revealed homology with the enzyme tRNA pseudouridine synthase I of E. coli.
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  • 21
    ISSN: 1432-0983
    Keywords: Mutagen hyper-resistance ; Nitrogen mustard ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A screening of haploid yeast strains for enhanced resistance to nitrogen mustard (HN2) yielded a recessive mutant allele, hnm1, that conferred hyper-resistance (HYR) to HN2. Diploids, homo- or heterozygous for the HNM1 locus, exhibit normal wild-type like resistance while homozygosity for hnm1 leads to the phenotype HYR to HN2. The hnm1 mutation could be found in yeast strains proficient or deficient in different DNA repair systems. In these mostly HN2-sensitive haploid repair-deficient mutants, hnm1 acted as a partial suppressor of HN2 sensitivity. All isolated recessive mutations conferring hyper-resistance belonged to a single complementations group. The HYR to HN2 phenotype was maximally expressed in growing cells and was associated with reduced mutability by HN2. HNM1 most probably controls uptake of HN2 which would be impaired in the hnm1 mutants.
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  • 22
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; G418 resistance ; Gene cartridges ; Heterologous Gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Coding sequence cartridges for aminoglycoside phosphotransferase (APT) were isolated from bacterial transposon Tn903. When incorporated into a heterologous gene construction utilising the PGK1 promoter and terminator, the heterologous APT gene provided a G418-resistance determinant that functioned efficiently as a dominant marker for yeast in both multiple- and single-copy. Transformant colonies on selective medium appeared rapidly, within 36–48 h, and growth rate of the transformed cells was normal. A simple and highly sensitive radiolabelling assay for APT enzyme activity was developed for use with crude cell protein extracts. Enzyme activity units were equated to the amount of APT protein present in the cells, and the APT protein was shown to be stable in yeast. Heterologous APT expression was 130-fold reduced compared with homologous PGK1. This resulted from an estimated two-fold decrease in mRNA level and a 65-fold decrease in translation efficiency. The latter was unaffected by AUG sequence context change, but corresponded with a high frequency of minor codons in the APT-coding sequence. APT can be used as a semi-quantitative reporter of gene expression, whose useful features are in vivo detection via the G418-resistance phenotype and powerful cell-free assay.
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  • 23
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    Current genetics 19 (1991), S. 9-14 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Mevalonate kinase ; Ergosterol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The nucleotide sequence of the ERG12 gene, encoding mevalonate kinase, from Saccharomyces cerevisiae is presented. The longest open reading frame may code for a protein containing 443 amino acids with a deduced relative molecular mass of 48 500. The analysis of the nucleotide sequence reveals a complete identity with the yeast gene RAR1, isolated elsewhere by complementation of a rar1 mutation involved in the stability of plasmids with weak ARS. In addition, we show that mevalonate kinase is not a rate-limiting enzyme; however its sensitivity to FFP could be a key regulatory mechanism in the sterol pathway of yeast.
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  • 24
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Episomal plasmid ; Copy number control ; Plasmid maintenance ; Glycolytic enzyme levels
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary This study demonstrates how varying the promoter strength of an essential gene on a yeast 2μORI-STB YEp multicopy vector can influence vector copy levels. A phosphoglycerate kinase gene (PGK) on this plasmid was made essential for fermentative growth by transformation into a pgk - yeast strain. When in these PGK- transformants the requirement for PGK expression was the sole selective criterion for plasmid maintenance, PGK promoter activity was inversely related to vector copy levels. Plasmids with an efficiently-transcribed PGK gene were maintained at approximately one copy per cell, whereas those lacking the UAS that normally directs high basal PGK transcription levels were present at up to 10–15 copies. All cultures of these PGK+ transformants contained only a low proportion of pgk - cells. Since mitotic loss of the plasmid arrests growth through loss of a functional PGK allele, PGK confers high stability to the YEp vector in such a pgk - genetic background. In this system YEp vector levels are probably influenced by PGK transcription because high expression of PGK is needed in rapid fermentative growth. Remarkably, low plasmid PGK promoter activity caused PGK mRNA levels slightly higher than those found in yeast with normal PGK regulation. A higher plasmid copy number is therefore not the only factor counteracting the effects of low PGK transcription, and it is possible that PGK mRNA becomes more stable in response to inefficient PGK transcription.
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  • 25
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Sporulation ; Inessential genes ; Genome organization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The SPR6 gene of Saccharomyces cerevisiae encodes a moderately abundant RNA that is present at high levels only during sporulation. The gene contains a long open reading frame that could encode a hydrophilic protein approximately 21 kDa in size. This protein is probably produced by the yeast, because the lacZ gene of Escherichia coli is expressed during sporulation when fused to SPR6 in the expected reading frame. SPR6 is inessential for sporulation; mutants that lack SPR6 activity sporulate normally and produce viable ascospores. Nonetheless, the SPR6 gene encodes a function that is relevant to sporulating cells; the wild-type allele can enhance sporulation in strains that are defective for several SPR functions. SPR6 is located on chromosome V, 14.4 centimorgans centromere-distal to MET6.
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  • 26
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Nucleo-mitochondrial interactions ; Mitochondrial status ; Lycorine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In a previous paper we have shown that the alkaloid lycorine inhibits growth of rho +, mit - and rho -, strains of Saccharomyces cerevisiae, whereas strains devoid of mitochondrial DNA (rho o) are resistant to more than 200 μg/ml of the alkaloid. In this report we show that hypersuppressive petites are almost as resistant as rho o mutants, whereas isogenic rho - petites, which have retained tained longer segments of the genome, are sensitive to the drug.
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  • 27
    ISSN: 1432-0983
    Keywords: Schizosaccharomyces pombe ; Saccharomyces cerevisiae ; CaMV 35S promoter ; CaMV 35S terminator ; Heterologous expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Complementation of fission yeast mutants by plant genomic libraries could be a promising method for the isolation of novel plant genes. One important prerequisite is the functioning of plant promoters and terminators in Schizosaccharomyces pombe and Saccharomyces cerevisiae. Therefore, we studied the expression of the bacterial β-glucuronidase (GUS) reporter gene under the control of the Cauliflower Mosaic Virus (CaMV) 35S promoter and 35S terminator. We show here that S. pombe initiates transcription at exactly the same start site as was reported for tobacco. The 35S CaMV terminator is appropriately recognized leading to a polyadenylated mRNA of the same size as obtained in plant cells transformed with the same construct. Furthermore, the GUS-mRNA is translated into fully functional GUS protein, as determined by an enzymatic assay. Interestingly, expression of the 35S promoter in the budding yeast S. cerevisiae was found to be only moderate and about hundredfold lower than in S. pombe. To investigate whether different transcript stabilities are responsible for this enormous expression difference in the two yeasts, the 35S promoter was substituted by the ADH (alcohol dehydrogenase) promoter from fission yeast. In contrast to the differential expression pattern of the 35S promoter, the ADH promoter resulted in equally high expression rates in both fission and budding yeast, comparable to the 35S promoter in S. pombe. Since the copy number of the 35S-GUS constructs differs only by a factor of two in the two yeasts, it appears that differential recognition of the 35S promoter is responsible for the different transcription rates.
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  • 28
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Mitochondria ; Intron-encoded proteins ; Recombination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The respiratory competency of a yeast strain devoid of mitchondrial introns is quite normal. However, it may be asked whether intron-encoded proteins participate in metabolisms other than those of mitochondrial introns. Using strains without mitochondrial introns we have answered two questions. The first was: does the absence of intron-encoded proteins abolsh mitochondrial recombination? The second was: do mitochondrial introns and intron-encoded proteins play a part in mitochondrial DNA rearrangements induced by ethidium bromide (rho- production)? We have shown that the introns and intron-encoded proteins are not essential essential components of either phenomenon.
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  • 29
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    Current genetics 18 (1990), S. 23-27 
    ISSN: 1432-0983
    Keywords: Protein translocation ; Saccharomyces cerevisiae ; Peroxisomes ; Overexpression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Import of proteins into organelles usually requires a cis-acting targeting signal. Analysis of various hybrid proteins, consisting of mouse DHFR and parts of catalase A from Saccharomyces cerevisiae, revealed that fusion proteins containing the N-terminal 126 amino acids, or less, of catalase A remain in the cytosol whereas fusion proteins containing 140, or more, N-terminal amino acids of catalase A form large aggregates inside the cell. These protein bodies, which lack a surrounding membrane, copurified with peroxisomes on cell fractionation. The peroxisomal targeting signal of catalase A does not reside at the C-terminus or at the N-terminus.
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  • 30
    ISSN: 1432-0983
    Keywords: Chloroplast DNA ; Junction between small single copy and inverted repeat regions ; Zea mays ; Grammineae ; ndhH gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The junctions JSA and JSB between the two inverted repeat regions IRA and IRB and the small single copy region of the maize chloroplast DNA have been identified by DNA sequencing. The JSA junction coincides with the initiation codon of the ndhH gene which is encoded by the adjacent region of the small single copy region. A comparison with the plastomes of rice, rye, tobacco and liverwort shows that linkage of this junction with the ndhH gene is specific for gramminean species. The amino acid sequences deduced from the ndhH genes show conserved histidine and cysteine residues which are likely to form a metal-binding domain.
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  • 31
    ISSN: 1432-1432
    Keywords: Thiolase ; Peroxisome evolution ; Bootstrap analysis ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The thiolase family is a widespread group of proteins present in prokaryotes and three cellular compartments of eukaryotes. This fact makes this family interesting in order to study the evolutionary process of eukaryotes. Using the sequence of peroxisomal thiolase from Saccharomyces cerevisiae recently obtained by us and the other known thiolase sequences, a phylogenetic analysis has been carried out. It shows that all these proteins derived from a primitive enzyme, present in the common ancestor of eubacteria and eukaryotes, which evolved into different specialized thiolases confined to various cell compartments. The evolutionary tree obtained is compatible with the endosymbiotic theory for the origin of peroxisomes.
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  • 32
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    Journal of molecular evolution 38 (1994), S. 363-368 
    ISSN: 1432-1432
    Keywords: Saccharomyces cerevisiae ; 2-μm circle ; DNA sequencing ; Horizontal transmission ; Site-specific recombination ; Selfish DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We compared the nucleotide substitution pattern over the entire genome of two unique variants of the 6,300-bp selfish DNA (2 μm) plasmid in Saccharomyces cerevisiae. The DNA sequence of the left-unique region is identical among 2-μm variants, while the right-unique region shows substantial divergence. This chimeric pattern cannot be explained by neutral or Darwinian selection models. We propose that horizontal transmission of the 2-μm plasmid coupled with a directed, polarized gene conversion maintains the DNA sequence of the left-unique region, whereas the right-unique region is subject to random drift and Darwinian selection.
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  • 33
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    Mycopathologia 109 (1990), S. 177-182 
    ISSN: 1573-0832
    Keywords: Helminthosporium maydis ; Zea mays ; Green islands/infection sites ; cytokinin activity ; pathogenicity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Green islands/infection sites recorded higher cytokinin activity than surrounding tissue as well as non-inoculated tissue. This activity in infected areas increased with time of incubation while in tissue surrounding the green islands and non-inoculated tissue, cytokinin activity decreased with time of incubation. The culture filtrate extracts of H. maydis had cytokinin activity which increased with growth of the fungus. Cytokinin activity of thin-layer Chromatographic fractions from tissue and culture filtrate extracts revealed that a major portion of the activity was confined to Rf zone 0.6 to 0.8 which co-chromatographed with zeatin and zeatin riboside. Presence of zeatin and zeatin riboside in tissue and culture filtrates was confirmed by high performance liquid chromatography. Cytokinin substances, such as zeatin and zeatin riboside, increase at infection sites with growth of the pathogen suggesting they may be involved in the pathogenicity of H. maydis on maize.
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  • 34
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    Mycopathologia 117 (1992), S. 157-161 
    ISSN: 1573-0832
    Keywords: Phyllachora maydis ; Monographella maydis ; Coniothyrium phyllachorae ; Zea mays ; tarspot complex
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The tarspot complex is caused by the interaction of Phyllachora maydis and Monographella maydis. Coniothyrium phyllachorae, possibly a mycoparasite, is found in older ascostromata of P. maydis, which always appears first causing tarspot. M. maydis follows and is responsible for the damaging “fisheye” symptom. The fisheye symptom is always associated with a tarspot in the center of the lesion, whereas 12 to 20% of the Phyllachora ascostromata remained free of M. maydis. Inoculations of maize leaves with the Microdochium anamorph of the Monographella (usually produced in lesions) failed to produce infections. Some infections with M. maydis were, however, obtained under unusual conditions in the field. Inoculations onto tarspots in the laboratory were unsuccessful, but in field experiments inoculations with conidia of M. maydis enhanced severity of the tarspot complex. Fisheye symptoms of the complex naturally appear 2 to 7 days after the manifestation of P. maydis. This is followed a week later by the appearance of M. maydis which became predominant in the lesions and is associated with empty perithecia of P. maydis. In the early stages of the tarspots pycnidia of the anamorph of P. maydis, Linochora sp., could occasionally be observed. Ascomata of M. maydis were rare in the field. Of the 36 genetic materials of CIMMYT tested, 30 developed the fisheye symptom, 4 tarspots only and 2 remained free of symptoms
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  • 35
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    Mycopathologia 119 (1992), S. 181-190 
    ISSN: 1573-0832
    Keywords: aflatoxin B1 ; electron microscopy ; in vitro ; immature maize embryo ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immature maize (Zea mays L.) embryos were treated with aflatoxin B1 concentrations, ranging from 0.1 μg ml−1 to 25 μg ml−1. Below 5 μg ml−1 aflatoxin B1, root and shoot elongation was not significantly inhibited. Ultrastructurally, root tip cells showed little deterioration, except a possible diffused clearing in mitochondria and plastids. As the toxin concentration was increased above 5 μgml−1, shoot, and particularly root elongation, was progressively inhibited. Associated with this, there was an apparent decrease in the ribosome population. Furthermore, membranes, particularly the vacuolar membrane, became abnormal and vacuolar distension occurred. At 20 and 25 μg ml−1, these effects were exacerbated, and mitochondria and plastid structure was disrupted. At these concentrations, there was evidence of a disruption in lipid metabolism. The results are discussed in the context of known aflatoxin effects on cellular control mechanisms and ultrastructure in animal systems.
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  • 36
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    Mycopathologia 111 (1990), S. 181-189 
    ISSN: 1573-0832
    Keywords: mycotoxin ; ochratoxin ; Penicillium ; storage ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Eleven-kilogram parcels of HY-320 wheat, a cultivar of the new Canada Prairie Spring class, were kept at 15 and 19% initial moisture contents (IMC) in simulated storage in a Manitoba farm granary for 60 weeks to determine biotic and abiotic changes and mycotoxin production. Ochratoxin A reached a maximum of 0.24 ppm by week 20 in the 19% IMC wheat, but was absent in the 15% IMC wheat; no other mycotoxins were detected. Temperature, moisture content, O2 and CO2 levels, fat acidity values, seed germination, microfloral incidence and abundance, and the presence of other mycotoxins were monitored. Principal component analysis of all variables showed that the first principal components accounted for 32–41% of the system variability, and contained the ochratoxin A variable. Ochratoxin A was produced in moist grain that had decreased seed germination andAltermaria activity, and high fungal activity byPenicillium andAspergillus versicolor. Compared to other stored cereals previously studied, HY-320 wheat would be ranked in a low-risk category for mycotoxin formation, based on the ochratoxin A levels observed.
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  • 37
    ISSN: 1573-0832
    Keywords: Nystatin ; amphotericin B ; amphotericin B methyl ester ; polyene antibiotics ; yeast ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Saccharomyces cerevisiae was cultured under anaerobiosis in semi-complete medium to which either palmitoleic or oleic acid was added. Cells were grown at 20 °C or 30 °C. The levels of total lipids, total sterols, and phospholipids were higher in cells grown at 20 °C than at 30 °C. The effects of nystatin (NYS), amphotericin B (AMB), and amphotericin B methyl ester (AME) were evaluated by determining cell viability and liberation of intracellular compounds. The loss of cell viability is higher in the first 30 minutes of incubation with the drugs and is the same regardless of the type of cells obtained. Low molecular weight compounds and ions such as K+ are liberated a few minutes after incubation with the drugs whereas proteins and substances absorbing at 260 nm are liberated later. Phosphate liberation comes after K+ and before compounds of higher molecular weights.
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  • 38
    ISSN: 1573-0832
    Keywords: Penicillium griseofulvum ; patulin ; wheat
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    Topics: Biology , Medicine
    Notes: Abstract Sixty-four wheat samples from Spanish flour factories were screened for patulin and patulin-producing moulds. None of them was found to contain any patulin, whereas samples experimentally contaminated with this toxin proved it to be highly unstable. On the other hand, Penicillium griseofulvum was the only in vitro patulin-producing species found (19 samples). Mould growth in the samples was investigated by using yeast-sucrose medium (YES) and high-performance liquid chromatography (HPLC) to measure the amounts of toxin produced during 40 day's incubation at 20 and 28°C. The highest yield rate of patulin was obtained between the 20th and 30th day of incubation; such a rate, however, was very low throughout the vigorous growth phase, during the first 20 days of incubation. The more appropriate temperature for incubation and patulin production was 28 °C. We also investigated the influence of other incubation conditions in the yield and found stationary dark cultures to be more efficient that shaken or fermentation cultures in YES medium. The best patulin yield achieved was 11.9 mg in the culture broth and 6.3 mg in the mycelium from 100 ml of medium.
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  • 39
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    Nutrient cycling in agroecosystems 40 (1994), S. 175-183 
    ISSN: 1573-0867
    Keywords: Nitrogen response ; on-farm research ; risk ; probability ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In many developing countries, fertilizer recommendations must be made in the absence of plant and soil analyses. When the region is variable in terms of soils, weather, and magnitude of response to fertilizer, a recommendation is likely to involve a high degree of risk for the farmer. Quantification of such risk is key to developing appropriate recommendations for the farmer. However, most methodologies generally used in analyzing fertilizer trials do not allow adequate quantification, especially as a continuous function, of the risk associated with a given recommendation. Three years of on-farm trials conducted in the High Valley of Mexico were used to evaluate different methodologies for generating N fertilization recommendations and their associated risk for wheat (Triticum aestivum) production in the absence of soil tests. When the traditional approach, using average yield responses or separate trial results, was used, an economic optimum was identified, but it was not possible to quantify the associated risk. In contrast, however, by using a combination of response surface methodology and simple probability analysis, the risk associated with any given recommendation was developed, even under the highly variable conditions of the study zone. The approach uses a treatment difference matrix (developed using average yield differences between a treatment and the zero N (0N) check) and its associated standard deviation over locations. From the matrix, an equation (being a function of N rate and relative grain:N price ratios) was developed that shows the probability of outperforming the 0N check for the economic optimum rate.
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  • 40
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    Nutrient cycling in agroecosystems 29 (1991), S. 163-172 
    ISSN: 1573-0867
    Keywords: Superphosphate ; residual value ; sandy soil ; leaching of phosphorus ; lupins ; barley ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In a field experiment on deep, yellow, sandy soil near Badgingarra, Western Australia, the residual value of superphosphate applied one and two years previously was measured relative to freshly-applied superphosphate using yields of narrow-leafed lupin (Lupinus angustifolius), barley and wheat. In addition, soil samples were collected for measurement of bicarbonate-extractable soil P. This was also used to estimate the residual value of the superphosphate. For lupins and wheat, and for bicarbonate-extractable soil P, the residual value decreased with increasing level of application. For barley grain, the residual value was not significantly affected by the level of application. The decrease in residual value of superphosphate with increasing level of application is attributed to increased leaching of applied phosphorus (P) down the profile of the sandy soils as the level of application increases. This may reduce subsequent plant yields due to the delay in seedling roots reaching the P in the soil during the crucial early stages of plant growth. For lupins, the relationship between yield and the level of superphosphate applied was markedly sigmoidal. The relationship for wheat and barley was exponential. Consequently, at suboptimal levels of P application, lupins required about two to three times more P than wheat or barley to produce the same yield. However, lupins required less P to achieve near-maximum yield.
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  • 41
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    Nutrient cycling in agroecosystems 31 (1992), S. 319-329 
    ISSN: 1573-0867
    Keywords: Superphosphate ; fertilizer effectiveness ; residual value ; lupins ; oats ; wheat ; grain production ; bicarbonate-extractable soil phosphorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In a field experiment on a deep pale-yellow sand in a 600 mm per annum rainfall Mediterranean environment of south-western Australia, six levels of phosphorus (P) as superphosphate (O up to 546 kg P ha−1) were applied once only, to the soil surface, before sowing lupins (Lupinus angustifolius). The lupins were grown in a continuous arable cropping rotation with, in successive years, oats (Avena sativa), wheat (Triticum aestivum), lupins. Five such rotations were started in the experiment from 1985 to 1989. The experiment continued until the end of 1990. The relationship between lupin seed (grain) yields and the level of P applied was measured in the year of P application for five successive years (1985 to 1989). The relationship had the same general form but it varied between years, largely due to different maximum yields (yield plateaux) in each year. The residual value of superphosphate applied three years previously was measured for lupins on two occasions (1988 and 1989) relative to superphosphate applied in the current year. The residual values was different in the two years. The superphosphate applied three years previously was about 30% as effective as freshly applied superphosphate in 1988, and 12% as effective in 1989. At each harvest, the relationship between grain yield and the P concentration in the grain differed for different species. However, for each species at each harvest, the relationship was similar regardless of when the P was applied in the previous years. Thus each species had the same internal efficiency of P use curve, and yields varied only with P concentration in tissue. Bicarbonate-extractable soil P was determined on soil samples taken in mid-July of 1989 and 1990. These soil test values were related to grain yields at harvest. The relationship between yield and soil test values had the same general form but varied for different species within years and for each species between years. It also varied for each species within years depending on the year the P was applied.
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  • 42
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    Nutrient cycling in agroecosystems 31 (1992), S. 331-340 
    ISSN: 1573-0867
    Keywords: Superphosphate ; residual value ; wheat ; lupins ; soil test for phosphorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In a field experiment on a sandplain soil in a low rainfall (326 mm per annum) Mediterranean environment of south-western Australia, the effectiveness of superphosphate applied in 1986 was measured in three subsequent years relative to freshly-applied superphosphate each year, using grain (seed) yields of wheat (Triticum aestivum) and lupins (Lupinus angustifolius). The wheat and lupins were grown in rotation and both crops were grown each year starting in 1986. Bicarbonate-soluble phosphorus was determined on soil samples taken in mid June from where the P treatment was applied in 1986 only. These soil test values were related to the grain yields produced that year. For each level of superphosphate applied in 1986, soil test values decreased with increasing time from application. The relationship between grain yield and soil test values had the same general form within each year for both plant species, but varied between years. For both species, the effectiveness of superphosphate decreased by about 70–80% between the year of application and the first and second years after application, and by a further approximate 10% in the third year. The relationship between grain yield and the level of superphosphate applied became sigmoidal by 1989.
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  • 43
    ISSN: 1573-0867
    Keywords: nitrogen fertilizer rate ; electroultrafiltration ; wheat ; soil test
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The objective of the investigation was to examine whether there exist relationships between the optimum nitrogen fertilizer rate for winter wheat and soil nitrogen fractions extracted by electroultrafiltration (EUF) from autumn samples of the upper soil layer (0–30 cm). Optimum nitrogen fertilizer rates were derived from grain yield curves of field trials carried out with increasing nitrogen fertilizer rates on 19 different sites in 1985/86 and 1986/87. Most soils were luvisols derived from loess, two soils were brown earths and one a pararendzina. Total Nitrogen fertilizer rates were 0, 40, 80, and 120 kg N/ha applied twice before ear emergence. The final nitrogen rate at ear emergence was the same for all treatments, namely 60 kg N/ha. Optimum nitrogen fertilizer rates were derived from the grain yield curve fitted to a modified Mitscherlich equation. The optimum nitrogen fertilizer rates were correlated with the nitrogen fractions extracted by EUF. The regression equation thus obtained showed that NO 3 - , the organic N fraction (EUF Norg), and the EUF Norg-quotient each had a highly significant impact on the optimum nitrogen fertilizer rate. The higher the amounts of EUF-N extracted the lower the optimum nitrogen rate. Substituting the EUF Norg-fraction for total nitrogen concentration in the upper soil layer gave a poorer relationship between the optimum nitrogen fertilizer rate and the soil data. In absolute terms the EUF Norg-fraction had by far the greatest impact on calculating the optimum nitrogen fertilizer rate. The investigation shows that the EUF method is a suitable technique for the determination of available soil nitrogen from which optimum nitrogen fertilizer rates can be derived for winter wheat cultivated under soil and climatic conditions typical for cereal growing areas in central Europe.
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  • 44
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    Nutrient cycling in agroecosystems 40 (1994), S. 207-214 
    ISSN: 1573-0867
    Keywords: available P ; citrate insoluble P ; phosphorus sources ; triple superphosphate ; Triticum aestivum ; water soluble P ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A greenhouse study was conducted to determine if soil pH affects the requirement for water-soluble P and the tolerance of water-insoluble impurities in TSP fertilizers. Two commercial TSP fertilizers were selected to represent a range in phosphate rock sources and impurities. Phosphate fertilizer impurities were isolated as the water-washed fraction by washing whole fertilizers with deionized water. TSP fertilizers with various quantities of water-soluble P (1.2 to 99% water-soluble P) were simulated by mixing the water-washed fertilizer fractions or dicalcium phosphate (DCP) with reagent-grade monocalcium phosphate (MCP). The fertilizers were applied to supply 40 mg AOAC available P kg−1 to a Mountview silt loam (fine-silty, siliceous, thermic Typic Paleudults). Wheat (Triticum aestivum (L.)) was harvested at 49 and 84 days after planting. Soil pH values at the final forage harvest were 5.4±0.16 and 6.4±0.15. At a soil pH of 5.4, the TSP fertilizers required only 37% water-soluble P to reach maximum yields while at pH 6.4 the fertilizers required 63% water-soluble P. Results of this study show that higher levels of water -insoluble P can be tolerated in TSP fertilizers when applied to acid soils. Phosphorus uptake was not affected by soil pH, but for the mixtures containing the fertilizer residues the source having the lowest level of Fe and Al had a higher relative agronomic effectiveness.
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    Nutrient cycling in agroecosystems 21 (1990), S. 163-166 
    ISSN: 1573-0867
    Keywords: Critical level of Zn ; alkaline soils ; Zn-deficiency ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Field experiments were conducted at 32 locations, chosen for their wide range in DTPA extractable Zn, to determine the critical deficiency level of Zn for predicting response of wheat to Zn application. Soil application of 5.6 kg Zn ha−1 significantly increased the grain yield in deficient soils. Soil extractable Zn was significantly related with per cent grain response and absolute grain yield. Both the graphical and statistical methods of Cate and Nelson indicated the critical level to be 0.75 mg kg−1 soil of DTPA extractable Zn. This level gave a predictability value of 82 per cent.
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  • 46
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    Nutrient cycling in agroecosystems 22 (1990), S. 147-159 
    ISSN: 1573-0867
    Keywords: Sulfate ; elemental sulfur ; leaching ; ryegrass ; wheat ; greenhouse experiments ; split application
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Three factorial experiments with four replications were conducted in a greenhouse to examine the effectiveness of gypsum, elemental sulfur (ES powder) and three S containing N fertilizers, viz., ammonium sulfate (AS), urea + ES, and Ureas (20% AS and 80% urea). All experiments were conducted twice in different years. In the first experiment with uncropped soil, the effects of soil type, leaching rate (2.3 and 6.9 mm water per day) and urea addition on sulfate leaching losses were studied. Leaching losses decreased in the order Ureas 〉 ammonium sulfate (AS) 〉 gypsum ≫ urea + ES. Increasing the leaching rate greatly increased sulfate losses from both soils. Losses were greater in the sandy Typic Hapludoll than in the clayey Oxic Paleustalf. Sulfate adsorption was found to decrease strongly with rising the pH in both soils. Hydrolysis of urea temporarily raised the pH of the soil, thereby increasing the sulfate leaching losses. In the second experiment the effects of S rate (0–65 mg per kg soil), split application and leaching rate (0 and 2.3 mm per day) on sulfate leaching losses and ‘apparent S recovery’ (ASR) by three successive cuts of ryegrass (Lolium perenne L.) were studied. Herbage yield more than doubled when S was applied. The effectiveness of the sulfate fertilizers was greater when S was split-applied than given all at once. With split applications the ASR decreased in the order: Ureas 〉 AS 〉 gypsum 〉 urea + ES 〉 ES powder. ES fertilizers were least effective, because the oxidation rate of ES to sulfate was clearly too slow. In the third experiment the effects of S rate (0–40 mg per kg soil) and split application on sulfate leaching losses and ASR in the grain of wheat (Triticum aestivum L.) were studied under leaching conditions (2.3 mm per day). Grain yield increased strongly due to S application. Split application greatly increased the effectiveness of the sulfate fertilizers and appeared to be an effective tool in satisfying the S need of the crop under leaching conditions. Again, ES fertilizers were least effective, because the oxidation rate of ES was too slow to meet the S demand of the crop. In all experiments leaching losses of sulfate from the ES fertilizers were smaller than from the sulfate fertilizers.
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  • 47
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    Nutrient cycling in agroecosystems 22 (1990), S. 97-107 
    ISSN: 1573-0867
    Keywords: Superphosphate ; placement depth ; banding ; relative effectivenesss ; lupins ; wheat ; field experiment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In a field experiment on a sandplain soil in a low rainfall (326mm per annum) Mediterranean environment of south-western Australia, seven levels of single superphosphate, 0, 7.5, 10, 14, 19.5, 30 and 39 kg P ha−1, were placed at either 3, 5, 7, 9, 11 or 13 cm depth before sowing wheat (Triticum aestivum) at 3 cm. In a separate treatment, superphosphate was drilled with the seed (the normal practice). In the second year, the plots were sown with lupins (Lupinus angustifolius) at 3 cm depth with no additional superphosphate. In three separate treatments, superphosphate at 0, 14 and 39 kg P ha−1, was drilled with the lupin seed (the normal practice) on plots that had received no superphosphate in the first year. Yields of wheat and lupins were used as a measure of the effectiveness of the superphosphate placement treatments relative to the effectiveness of superphosphate drilled with seed of wheat (year 1) or lupins (year 2), to give relative effectiveness (RE) values in each of the two years. In the first year the RE of superphosphate was increased by about 20% when the fertilizer was placed 5 to 9 cm deep in the soil. In the second year, the RE of superphosphate for producing lupin grain was increased by about 30–60% where the fertilizer had been placed 5–13 cm deep in the previous year compared with freshly drilled 3 cm deep. The yield of wheat or lupins was closely related to the P content of plant tissue; each relationship was independent of the depth or year of superphosphate application.
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    Nutrient cycling in agroecosystems 26 (1990), S. 229-235 
    ISSN: 1573-0867
    Keywords: Grass ; wheat ; nitrogen nutrition ; dilution curve ; mineral content ; mineral removal ; phosphorus ; potassium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The important effect of nitrogen in changing the patterns of mineral content and mineral removal is analysed for grass swards and wheat. Different models are proposed; accumulated dry matter developed throughout a growing period is shown to be an excellent reference for assessing the evolution of the plant mineral content and the mineral removal the growing crop. Applications in diagnosing mineral nutrition status and optimising fertilizer use are proposed and discussed.
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    Nutrient cycling in agroecosystems 39 (1994), S. 77-82 
    ISSN: 1573-0867
    Keywords: Acidic subsurface layer ; application rate ; North Carolina phosphate rock ; placement method ; time of application ; triple superphosphate ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Using soils with an acidic subsurface layer, three glasshouse experiments were carried out to evaluate the effect of placement method and application rate of triple superphosphate (TSP) and North Carolina phosphate rock (NCPR) on dry matter (DM) yields. Time of application of NCPR on DM yield response of wheat was also studied. For Experiment 1, soil was collected in depth intervals of 0–2; 4–6; 6–8; and 8–10 cm from a red earth (chromic luvisol). The treatments included two P sources (TSP and NCPR), three placement methods (broadcast, banded or mixed into the subsurface layer, 6–8 cm), and six application rates. In this P deficient soil with an acidic subsurface layer, there was relatively little effect of application method of TSP on wheat yield responses. The maximum dry matter yield responses for broadcast, band and mix application methods was 30, 42 and 50 %, respectively. Responses to NCPR broadcast, band and mix methods were 20, 9 and 44 %, respectively. Mixing NCPR into to acidic subsurface layer produced yields similar to those from TSP although a higher application rate of P as NCPR was needed to achieve this outcome. Treatments for Experiments 2 and 3 were time of application of NCPR (0 and 30 days before sowing) and rate of application of NCPR (0 and 40 mg P/pot). In Experiment 2 (same soil as Experiment 1) application of NCPR prior to sowing, resulted in higher Colwell P concentration than when applied at sowing, but time of application had no effect on final DM yields. Experiment 3 used a red podzolic (chromic luvisol) soil which had a lower P-status, was more acid and had a lower exchangeable Ca2+ concentration than the red earth. Application of NCPR prior to sowing resulted in lower DM yield than when it was applied prior to sowing.
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  • 50
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    Nutrient cycling in agroecosystems 31 (1992), S. 43-49 
    ISSN: 1573-0867
    Keywords: Superphosphate ; nitrogen fertilizer ; take-all ; (Gaeumannomyces graminis) ; wheat ; Triticum aestivum L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Wheat was grown continuously in soil amended with 5 levels of superphosphate and with 4 levels of urea at 3 sites. The incidence and severity of take-all, caused byGaeumannomyces graminis var.tritici, declined with increasing rates of application of both superphosphate and urea. In both years, the severity of take-all on plants receiving neither superphosphate nor urea was about 40% while at the highest level of superphosphate and urea supply the take-all severity was approximately halved at 22%. There was an increase in grain yield in response to applied superphosphate and urea to the highest level of each nutrient. There was also an increase in the 1,000-kernal weights with superphosphate and urea fertilizer application.
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  • 51
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    Nutrient cycling in agroecosystems 32 (1992), S. 27-36 
    ISSN: 1573-0867
    Keywords: Phosphorus requirement ; wheat ; oats ; barley ; lupins ; triticale ; superphosphate ; grain yield ; phosphorus in grain
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The phosphorus (P) requirement for grain production of different crop species (oats (Avena sativa), barley (Hordeum vulgare), triticale (xTriticosecale), narrow-leafed lupins (Lupinus angustifolius), and sandplain lupins (L. cosentinii) was compared with wheat (Triticum aestivum) in five field experiments on different lateritic soils in south-western Australia. Seven or eight levels of superphosphate were applied at the start of each experiment. The amount of P required to produce 70% (four experiments) or 90% (one experiment) of the maximum yield was used to compare P requirements. Large differences in the P requirements of the species were obtained. On P deficient soil in 3 experiments, oats required from 50 to 70% less P than wheat, but required 40% more P on a soil with a long history of superphosphate applications. Compared with wheat, in the year of P application, barley required 50% less P in one experiment, had similar P requirements in two experiments, and required 80% more P in another experiment. In the years after P application, barley required 20% less P in two experiments. On an acidic soil triticale required from 50% to 70% less P than wheat, but on less acidic soil it required 100% more P. In the year of P application, narrow-leafed lupins required 800% more P than wheat in one experiment, and 30% more P in the other experiment. In the year of P application, sandplain lupins required 70% less P than wheat in one experiment.
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  • 52
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    Nutrient cycling in agroecosystems 32 (1992), S. 185-194 
    ISSN: 1573-0867
    Keywords: Residual P ; P fixation ; Bray P ; Olsen P ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Nineteen soils from the south east of the Province of Buenos Aires (Argentina) that had been fertilized with moderate amounts of P (10–40 kgP/ha) during the last 10 years were used to investigate the effect of time on the decline of P availability as measured by three soil tests (Bray 1, Bray 2, Olsen) and the null-point method. Differences in rates of P decline among soils and chemical methods were characterized by an exponential coefficient for time (b 2) in equations which describe the changes of the added P retained by the soil (Pr =ac b1 t b2). The rate of decline of P for the nineteen soils calculated for the soil test methods was ordered decreasingly as: null-point 〉 Olsen 〉 Bray 1 〉 Bray 2. The ability of the chemical methods for assessing the residual value of P for wheat growth (RV) was tested in a pot experiment on seven of the soils that differed in their individual rates of reaction with P. Differences between soils in the rate of reaction with P as measured in the laboratory by the null-point method and by the Olsen test were reflected in different residual values for P fertilizer for wheat plants. Thus the value ofb 2 for these methods was well correlated with the observed residual values. The soil properties commonly associated with the retention of P were not related to the value ofb 2 suggesting that more than one soil property may be involved in the measure ofb 2. The exponent for timeb 2 may be used as an index of the ability of the soil test to reflect the decline of P availability with time.
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  • 53
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    Nutrient cycling in agroecosystems 39 (1994), S. 11-18 
    ISSN: 1573-0867
    Keywords: Copper fertiliser ; nitrogen fertiliser ; residual effectiveness ; Triticum aestivum L. ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The residual effectiveness of copper (Cu) applied 18 to 21 years previously was estimated for grain yield of wheat. In one field experiment, current levels of Cu fertiliser were applied and its effectiveness was compared to that of the same level of Cu applied previously. The effects of nitrogen (N) fertiliser on the Cu concentration in the youngest emerged blade and in the grain, as well as the effects of N levels on the grain yield of wheat, were also studied. Where the recommended level of Cu fertiliser had been applied previously, its residual effectiveness depended on the soil type. On the grey sands over clay and gravelly sands over clay, the residual Cu would last approximately 20 years where wheat is grown in rotation with a legume crop (Lupinus augustifolius L.) and where N fertiliser is applied at high levels (92 kg N ha−1). On the yellow brown sandy earths of the Newdegate district, the residual value was in excess of 30 years. When Cu levels in the soil are marginal, high levels of N applied to wheat crops grown on stubbles of legume crops (high soil N) could suffer from induce Cu deficiency which could reduce grain production. Critical concentrations of Cu in the youngest emerged blade of less than 1.2 mg Cu kg−1 at Gs50–59 would indicate Cu deficiency. Cu concentrations of less than 1.1–1.2 mg Cu kg−1 in the grain suggest that the wheat crop is marginally supplied with Cu. In both situations, Cu fertiliser needs to be applied before the next crop.
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  • 54
    ISSN: 1573-0867
    Keywords: Cochliobolus sativus ; common root rot ; Penicillium bilaji ; phosphorus fertilization ; tillering ; wheat
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    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A strain ofPenicillium bilaji Chalabuda (PB) has recently been commercially released as a seed inoculant to increase phosphorus (P) uptake by wheat (Triticum aestivum L.). The purpose of this study was to compare the effects of drill applied P (15 kg P ha−1) with PB seed inoculation on early growth, development, P uptake, and grain yield of ‘Stoa’ spring wheat at four sites in North Dakota. Fertilization with P consistently enhanced early season growth, main stem development, tillering and P uptake. Seed inoculation with PB had little or no effect on these traits. Phosphorus fertilization tended to increase common root rot severity (CRR, incited byCochliobolus sativus (Ito & Kurib) Drechs.), while PB inoculation had no effect. Grain yields were significantly increased by P fertilization at one location. Inoculation with PB also increased grain yield at this location. The reason why PB inoculation increased yield at this location is not evident, as plant growth and P uptake were not enhanced earlier in the season. Averaged across all four sites, PB inoculation increased wheat yields 66 kg ha−1, which is similar to averaged yield responses reported from the Prairie Provinces of Canada.
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  • 55
    ISSN: 1572-9699
    Keywords: 2-Deoxy-D-glucose transport ; polyphosphate ; Saccharomyces cerevisiae ; sugar phosphorylation
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    Topics: Biology
    Notes: Abstract The role of polyphosphate in 2-deoxy-D-glucose transport was studied in yeast cells, pulse-labeled with [32P]orthophosphate, by comparing the concentrations and specific activities of polyphosphate, orthophosphate and 2-dGlc-phosphate. When 2-dGlc transport was measured under aerobic conditions, it appeared that polyphosphate replenished the orthophosphate pool, indicating that polyphosphate has, at least mainly, an indirect role in sugar phosphorylation. Also in cells with a reduced respiratory capacity, due to a treatment with antimycin A, no direct role for polyphosphate in 2-dGlc transport could be detected. Under these conditions, only a very limited breakdown of polyphosphate occurred, probably because of the small decrease in the orthophosphate concentration.
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    Antonie van Leeuwenhoek 62 (1992), S. 35-46 
    ISSN: 1572-9699
    Keywords: introns ; pre-mRNA splicing ; RNA processing ; Saccharomyces cerevisiae ; yeast genetics
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    Topics: Biology
    Notes: Abstract The occurrence of introns in nuclear precursor RNAs (pre-mRNAs) is widespread in eukaryotes, and the splicing process that removes them is basically the same in yeasts as it is in higher eukaryotes. Splicing takes place in a very large, multi-component complex, the spliceosome, and biochemical studies have been complicated by the large number of splicing factors involved. This review describes how genetic approaches used to study RNA splicing inSaccharomyces cerevisiae have complemented the biochemical studies and led to rapid advances in the field.
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    Cellular and molecular life sciences 46 (1990), S. 193-200 
    ISSN: 1420-9071
    Keywords: Saccharomyces cerevisiae ; protein toxin ; yeast toxin precursor ; protease processing ; lectin ; (1→6)-β-D-glucan ; receptor ; resistant mutants ; spheroplasts ; ion-permeable channels ; site-directed mutagenesis ; toxin functional domains
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    Topics: Biology , Medicine
    Notes: Summary The K1 killer toxin ofSaccharomyces cerevisiae is a secreted, virally-coded protein lethal to sensitive yeasts. Killer yeasts are immune to the toxin they produce. This killer system has been extensively examined from genetic and molecular perspectives. Here we review the biology of killer yeasts, and examine the synthesis and action of the protein toxin and the immunity component. We summarise the structure of the toxin precursor gene and its protein products, outline the proteolytic processing of the toxin subunits from the precursor, and their passage through the yeast secretory pathway. We then discuss the mode of action of the toxin, its lectin-like interaction with a cell wall glucan, and its probable role in forming channels in the yeast plasma membrane. In addition we describe models of how a toxin precursor species functions as the immunity component, probably by interfering with channel formation. We conclude with a review of the functional domains of the toxin structural gene as determined by site-directed mutagenesis. This work has identified regions associated with glucan binding, toxin activity, and immunity.
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    Cellular and molecular life sciences 46 (1990), S. 1016-1017 
    ISSN: 1420-9071
    Keywords: In vitro absorption ; calcium ; wheat ; Bengal gram
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The in vitro absorption of calcium from the duodenum was significantly less in a group of rats fed on a wheat diet than in a group fed a wheat and Bengal gram (70∶30) diet.
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    Cellular and molecular life sciences 46 (1990), S. 971-972 
    ISSN: 1420-9071
    Keywords: Zea mays ; inhibitor of the oxidation of catechin ; root release
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    Topics: Biology , Medicine
    Notes: Summary When the specific activities of the catechin oxidases (catechin as the substrate) which were released from the roots of the seedlings of alfalfa, tomato, wheat, lettuce and corn were compared, it was found that the oxidizing activity was absent from the root exudate of corn seedlings. A 6.3 kDa protein was purified from the root exudate of corn seedlings and in the presence of this protein, the oxidation of catechin was inhibited. This inhibitor is responsible for the inability of the root exudate of corn seedlings to oxidize catechin.
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  • 60
    ISSN: 1432-2048
    Keywords: Ammonium compartmentation ; Cytoplasm ; Vacuole ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We show that the pH dependence of the base-catalysed exchange rate of the ammonium ion provides a basis for discriminating between the cytoplasmic and vacuolar pools of ammonium in plant tissues. In vivo, 14N-nuclear magnetic resonance spectra were recorded with and without 1H-decoupling and information on the subcellular distribution of NH 4 + was obtained from a lineshape analysis of the 1H-coupled spectrum. We applied this method to maize (Zea mays L.) root tissues and found that: (i), the cytoplasmic ammonium concentration was low, which was in accord with the large activity of glutamine synthetase present in the roots; and (ii), inhibition of glutamine synthetase with methionine sulphoximine increased the cytoplasmic ammonium concentration, and led to the appearance of ammonium in the xylem sap.
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  • 61
    ISSN: 1432-203X
    Keywords: wheat ; rye ; embryogenesis ; growth ; tissue culture
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    Topics: Biology
    Notes: Summary The influence of the short arm of rye chromosome 1 (1RS) from Secale cereale var. Imperial on the growth and differentiation of callus cultures from wheat Triticum aestivum var. Chinese Spring immature embryos was analysed. This chromosome arm was found to stimulate both embryogenesis and the rate of growth of calli. Recombinant lines carrying segments of 1RS were used to delineate the regions of 1RS responsible for the tissue culture effects. The enhancement of embryogenesis and the stimulation of growth were shown to be associated with two distinct genetic regions of the chromosome arm; the former is located between the centromere and the Sec 1 locus, while the latter is situated in the immediate vicinity of the Sec 1 locus.
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  • 62
    ISSN: 1432-2048
    Keywords: Anthocyanin ; Cold stress ; mRNA ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Changes in anthocyanin content and transcript abundance for genes whose products function in general phenylpropanoid metabolism and the anthocyanin pathway were monitored in maize (Zea mays L.) seedlings during short-term, low-temperature treatment. Anthocyanin and mRNA abundance in sheaths of maize seedlings increased with the severity and duration of cold. Anthocyanin accumulation was found in all tested lines that were genotypically capable of any anthocyanin production. Within 24 h of transferring 7-d maize (B37N) seedlings to 10° C, phenylalanine ammonia-lyase (Pal) (EC 4.3.1.5)-homologous and chalcone synthase (C2) (EC 2.3.1.74) transcript levels increased at least 8- and 50-fold, respectively, and 4-coumarate:CoA ligase (4Cl) (EC 6.2.1.12)-homologous and chalcone isomerase (Chi) (EC 5.5.1.6)-homologous transcripts increased at least 3-fold over levels in unstressed plants. Time-course studies showed thatPal (EC 4.3.1.5) andC2-transcript levels remained relatively constant for the first 12 h of cold stress, dramatically increased over the next 12 h, and declined to pretreatment levels within 2 d of returning coldstressed seedlings to ambient (25° C) temperature. Transcripts4Cl (EC 6.2.1.12) andChi (EC 5.5.1.6) increased in abundance within 6 h of cold stress, exhibited no further increase over the next 36 h, and declined to pretreatment levels upon returning seedlings to 25° C. Transcripts homologous to two regulatory (R, C1) and three structural (A1,A2, andBz2) anthocyanin genes increased at least 7- to 10-fold during cold treatment, exhibiting similar kinetics of accumulation as forPal (EC 4.3.1.5) andC2 transcripts. Transcripts encoded byBz1, the anthocyanin structural gene for UDP:glucose-flavonol glucosyltransferase (EC 2.4.1.91), were relatively abundant in control tissues and exhibited only a transient increase during the cold period. Our studies suggest that the genes of the anthocyanin biosynthetic pathway can be consideredcor (Cold-Regulation) genes, and because this pathway is well defined, it is an excellent subject for characterizing plant molecular responses to low temperatures.
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  • 63
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    Plant cell reports 11 (1992), S. 567-570 
    ISSN: 1432-203X
    Keywords: Microspore ; Electroporation ; Transformation ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The ability to deliver and detect reporter gene activity in maize microspores was tested. Tested expression vectors contained the chloramphenicol acetyl transferase (CAT) gene and one of the following promoter-intron combinations: 1) cauliflower mosaic virus (CaMV 35S), 2) CaMV 35S + maize alcohol dehydrogenase 1 intron 6 (Adh1-I6), 3) maize alcohol dehydrogenase 1 + intron 1 (Adh1-I1), or 4) maize ubiquitin 1 + intron 1 (Ubiq 1-I1) promoter + intron. The expression vectors were delivered into maize microspores using electroporation or polyethylene glycol (PEG). Both methods were effective for delivering free DNA into microspores. Although all four promoters were active in maize protoplasts, only two promoters were active in maize microspores. The CaMV 35S and the Adh1 promoters did not promote gene expression in maize microspore. The CaMV 35S + Adh1-I6 and Ubiq1-I1 promoters produced high levels of CAT activity in maize microspores.
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  • 64
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    Sexual plant reproduction 3 (1990), S. 109-115 
    ISSN: 1432-2145
    Keywords: Zea mays ; Ear initials ; Kinetin ; Gibberellic acid ; Male and female flowers ; Development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Young ears of maize were cultured in two different liquid media containing either kinetin (KN) or kinetin + gibberellic acid (KN + GA3) in order to manipulate stamen and gynoecium development. In KN medium, stamens developed and gynoecia aborted in the flowers of the cultured immature ears. In the KN + GA3 medium, however, ovaries with silks developed and stamens aborted. These differential morphological events were recorded with SEM photomicrographs at regular intervals after excision of ear inflorescences. In addition, the mitotic activity in the developing or aborting organs was determined over a 75-h period. It increased from 6% to 14% in developing organs (i.e. stamens in KN medium, and gynoecia in KN + GA3 medium) and gradually decreased to 1% in the degenerating organs (i.e. gynoecia in KN medium, and stamens in KN + GA3 medium) by 45 h of culture. The mitotic activity reached zero in degenerating flower organs by 75 h of culture. Whether these differential sensitivities to the exogenously applied members of these two plant growth regulator classes are unique to our in vitro system or reflect a more general control feature of in vivo inflorescences must await further clarification.
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    Sexual plant reproduction 3 (1990), S. 160-169 
    ISSN: 1432-2145
    Keywords: Pollen ; In vitro germination ; Microinjection ; In vitro fertilization ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The manipulation of single pollen grains of maize was studied. The effects of delivering substances both locally to the grain wall, tube or tip by a microcapillary and directly into the pollen grain by microinjection, and single grain pollination were investigated. Germination was induced by adding small amounts of water locally to the grains with either a microcapillary or with a waterdelivering emulsion without any other ingredients in the medium. The grains were overlayered by mineral or silicone oil so that tube growth proceeded without the grains bursting. There was no apparent penetration of high-molecular-weight substances (FITC-dextran, ethidium bromide labelled DNA) into the living grain either before or after pollination. Neither could the penetration of these substances be detected in both dry, viable and hydrated grains, tubes and tube tips, with or without treatment with Triton X-100 and dimethyl sulfoxide. By microinjection, however, the delivery of high-molecular-weight substances into grains was possible. Such injected grains successfully pollinated stigmas of cultured ear segments. Pollination with pore-injected grains was most efficient (mean 26%). No difference in fertilization rates between mass pollination (mean 41%) and single grain pollination (mean 39%) could be found. A mean fertilization rate of 29% could be obtained after microinjection. Seedlings developed 3 weeks after being pollinated by means of the in vitro pollination and fertilization method.
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    Sexual plant reproduction 5 (1992), S. 224-226 
    ISSN: 1432-2145
    Keywords: Zea mays ; Maize ; Polyembryony
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two types of cleavage polyembryony are described in the inbred line VIR 17 of maize. Suspensorial embryony was observed to occur spontaneously. Typical cleavage of the zygotic proembryo occurred spontaneously, but could also be induced by treating the developing caryopses with 2,4-Dichlorophenoxyacetic acid (2,4-D) on the second day after pollination. 2,4-D was active as a decorelative factor also evoking the expression of totipotency in individual proembryonal cells.
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    Sexual plant reproduction 4 (1991), S. 12-16 
    ISSN: 1432-2145
    Keywords: In vitro fertilization ; Egg cell ; Sperm cell ; Electrofusion ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Electrofusion-mediated in vitro fertilization of maize using single sperm and egg cells was performed. Sperm cells were released from pollen grains after rupture of the latter by osmotic shock in the fusion medium (0.55 M mannitol). Egg cells were isolated by enzyme treatment (pectinase, pectolyase, hemicellulase, and cellulase) followed by mechanical isolation. The conditions generally used for the electrical fusion of protoplasts of somatic cells were also applied to the protoplasts of gametic cells of maize. Electrofusion was performed with single pairs of gametes under microscopic observation. The mean fusion frequency was 79%. Isolated egg cells of maize showed protoplasmic streaming during 22 days of culture, but they did not divide. However, after fusion of the sperm with the egg cells, these fused cells did develop, with a mean division frequency of 83%, and grew to multicellular structures. Egg cells and fusion products were cultivated with a maize feeder-cell system.
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    Sexual plant reproduction 5 (1992), S. 227-231 
    ISSN: 1432-2145
    Keywords: Zea mays ; Maize ; Pollen-tube growth regulation ; In vitro pollination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In vitro pollen-tube growth in maize was studied using an in vitro pollination system. In the ‘cut-silk’ method, ovaries with silks were placed on medium in vitro, whereafter the silk was cut and the upper part of the silk was pollinated. Pollen tubes were not able to bridge the space between the two silk parts. Even when silk parts were tightly connected, pollen tubes still were not able to pass the cut ends and reach the lower silk part. Pollen-tube growth rates and the direction of tube growth were not influenced by the presence or absence of an ovary. Prepollination did not have any influence on pollen-tube growth rate. Measurements of pollen-tube growth rate also showed that there was no ‘population effect’, i.e. growth rate was not stimulated by pollination with an excess of pollen grains. We found that the direction in which maize pollen grew was determined only by the positioning of the silk hairs.
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    Sexual plant reproduction 6 (1993), S. 239-243 
    ISSN: 1432-2145
    Keywords: Zea mays ; Sperm cell ; Protein synthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Sperm cells are thought to be quiescent in pollen and activated upon pollen germination. To test this hypothesis, protein, RNA and DNA synthesis were assessed in Zea mays sperm cells at different times after isolation from pollen. Protein synthesis changed with time; while some proteins were found to be constitutive in both 0 and 24 h cells, others were synthesized and some disappeared. Overall, the number of proteins detected at 24 h doubled compared with freshly isolated cells. Incorporation of [3H]leucine in 24 h cells was about 50 times that in freshly isolated cells, and that of [5, 6-3H]uridine, about 7 times. Very low incorporation of [6-3H]thymidine into the cells was detected; there was no difference between freshly isolated and 24 h cells. It is possible that the differences in synthetic activity between freshly isolated and 24-h-old cells might correspond to sperm cell activation during pollen tube growth. If so, these metabolic changes may play an important role in fertilization.
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    Plant cell reports 11 (1992), S. 535-539 
    ISSN: 1432-203X
    Keywords: Zea mays ; in vitro culture ; in vitro pollen ; pollen germination ; fertilization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Maturation of maize pollen was obtained in male reproductive structures cultured in vitro. Immature tassels containing microspores at the mid-uninucleate to late-binucleate stage of development were excised and spikelets, anthers, and/or isolated microspores were cultured on a medium capable of supporting pollen maturation. Microspore mitosis, culminating in the production of starch-filled, trinucleate pollen capable of germination, was observed after 7–15 days, depending on the genotype and stage at which the cultures were initiated. Up to 100%, 70%, and 20% of the cultured spikelets, anthers, and isolated microspores, respectively, produced mature pollen, which germinated, however, at different frequencies (i.e., spikelets, 50–70%; anthers, 5–10%; microspores, 〈1%). Mature kernels were produced following fertilization with pollen from cultured spikelets and anthers. These procedures provide methods for the in vitro manipulation of a significant phase of the maize life cycle.
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  • 71
    ISSN: 1432-203X
    Keywords: Zea mays ; Matrix-associated ; DNA ; repetitive sequences ; DNA loops
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In order to elucidate some features of the topological organization of DNA within the plant nucleus, DNA fragments involved in the attachment of the DNA loops to the nuclear matrix in maize were studied. The matrix-associated DNA from dry embryo and meristematic cells after extensive digestion with DNase I and high salt treatment was about 2% of the total DNA, sized within the range of 50 and 250 bp. This DNA was found to be enriched in repetitive DNA sequences, both for nuclei from dry embryo and meristematic cells. The loop size of the DNA in cells of Zea mays appeared to be between 5 and 25 kbp.
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  • 72
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    Plant cell reports 12 (1993), S. 564-568 
    ISSN: 1432-203X
    Keywords: Zea mays ; In vitro culture ; Isolated microspores ; Pollen development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary An in vitro method to simulate pollen development was developed in maize (Zea mays L.). Microspores at the late uninucleate to early binucleate stage were isolated and cultured under various conditions. Cell viability, starch content and the formation of the three nuclei as found in normal mature pollen were monitored during the course of the culture. Media composition was modified in order to promote starch accumulation and frequency of mitosis, while maintaining the viability of the microspores. Under the best conditions, up to 12% of the microspores matured in vitro into trinucleate, starch-filled viable pollen grains which were unable to germinate or produce seeds. At different stages during in vitro maturation, proteins patterns were analyzed and compared with their in vivo equivalent and the patterns were only partially similar.
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  • 73
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    Plant cell reports 12 (1993), S. 607-611 
    ISSN: 1432-203X
    Keywords: Transient expression ; Particle bombardment ; Tassel primordia ; In vitro culture ; Anthers ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Maize (Zea mays L.) tassel primordia were used as a target for particle bombardment, to assess the possibility of introducing foreign DNA into male reproductive structures. Transient expression of the β-glucuronidase gene (GUS) or anthocyanin marker genes (C1 and B-Peru) driven by the CaMV 35S promoter was obtained in tassel primordia 24h after bombardment. Gold particles coated with DNA reached stamen primordia tissues, which eventually form the anthers and pollen. Bombarded tassels were also cultured in vitro and GUS activity was detected in the vascular tissue of mature anthers that developed within 4 weeks. This new approach represents a preliminary step toward pollen mediated transformation.
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  • 74
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    Plant cell reports 12 (1993), S. 648-651 
    ISSN: 1432-203X
    Keywords: chromosome doubling ; Zea mays ; tissue culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Anther-derived calli of corn were treated with 10 μM pronamide for 2, 3 and 4 days. The ploidy level of the calli was then evaluated using flow cytometry, at different times after the treatment. Untreated haploid calli did not change in ploidy level for 97 days but by 466 days, there were up to 50% diploid or higher ploidy cells thus showing that spontaneous doubling may occur during corn calli subculture with this genotype. Pronamide treatment did increase the percentage of diploid and tetraploid cells and by 466 days, all of the lines showed an additional change toward higher ploidy levels. This change may be due to spontaneous chromosome doubling or to differential cell cycle times of cells with different ploidy levels. The ploidy level of plants regenerated from the cultures was determined by counting the guard cell chloroplast numbers and the correlation with the ploidy level of the cultures was r2=0.84. These studies show that pronamide treatments can increase haploid maize callus chromosome numbers and that spontaneous chromosome doubling can occur with time in maize callus.
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  • 75
    ISSN: 1432-0983
    Keywords: Trans-kingdom conjugation ; DNA integration ; Saccharomyces cerevisiae ; Escherichia coli
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary IncQ-derived conjugative shuttle vectors, which carried the yeast gene URA3 and/or the yeast autonomously replicating sequence (ARS1), were constructed. Both the ars-plus plasmid pAY205 and the ars-less plasmid pAY201 were successfully transmitted from E. coli to S. cerevisiae by the action of mob and tra. In this trans-kingdom conjugation, plasmid pAY205 could replicate and be retained in transconjugants. Plasmid pAY201 caused the formation of “micro-colonies” of abortive transconjugants due to its transient expression and rapid disappearance. Nevertheless, one per about 103 colonies caused by transmitted pAY201 plasmids were uncurable by integration into the homologous region of a yeast chromosome. Analyses by restriction enzyme mapping and Southern hybridization indicate that this integration is primarily caused by a double crossover during conjugation and not by a single reciprocal recombination.
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  • 76
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Transcriptional activator ; Oxidative stress ; Glutathione
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The PAR1/SNQ3 gene of S. cerevisiae, which increases resistance to iron chelators in multi-copy transformants, is identical to the YAP1 gene, a yeast activator protein isolated as a functional homologue of the human c-jun oncogene by binding specifically to the AP-1 consensus box. The observed H2O2-sensitivity of par1 mutants has been attributed to an increased sensitivity to reduced oxygen intermediates. Accordingly, par1 mutants did not survive an elevated oxygen pressure and were very sensitive to menadione and methylviologene, two chemicals enhancing the deleterious effects of oxygen. The specific activities of enzymes involved in oxygen detoxification, such as superoxide dismutase, glucose 6-phosphate dehydrogenase and glutathione reductase, were decreased in par1 mutants and increased after PAR1 over-expression. As in the case of oxygen detoxification enzymes, the cellular levels of glutathione were similarly affected. These observations indicate that PAR1/YAP1/SNQ3 is involved in the gene regulation of certain oxygen detoxification enzymes. The finding that H2O2 promotes DNA-binding of human c-jun is consistent with a similar function for PAR1/YAP1/SNQ3 and c-jun in cellular metabolism.
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  • 77
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Mitochondrial trp-tRNA synthetase ; Nuclear mutation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The conditional respiratory-deficient Saccharomyces cerevisiae mutant pet-ts2281 was complemented by an yeast genomic DNA library. The gene thus isolated was sequenced and proved to be identical to the known MSW1 sequence encoding mitochondrial tryptophanyl-tRNA synthetase (Myers and Tzagoloff 1985). Compared to the wild-type, the ts2281 mutant allele of MSW1 contained a single T→C transition leading to a Leu→Ser replacement at position 294 of the protein sequence. In addition to this mutational alteration, our sequence data for the wild-type gene differ from the originally published MSW1 sequence at five other DNA positions which affect two locally restricted regions of the polypeptide chain. As expected, at the non-permissive temperature ts2281 cells are specifically defective in mitochondrial trp-tRNA formation and, thus, in overall mitochondrial protein synthesis. In addition, the patterns of cytochrome b mRNA maturation intermediates were distinctly different in ts2281 and wild-type yeast cells. The mutational effect of the observed amino-acid substitution in ts2281 is discussed in terms of weakened hydrogen bonding in the C-terminal half of the MSW1-encoded protein.
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  • 78
    ISSN: 1432-0983
    Keywords: Glucoamylase ; Gene cloning ; Hormoconis resinae ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cDNA coding for glucoamylase P of Hormoconis resinae was cloned using a synthetic oligonucleotide probe coding for a peptide fragment of the purified enzyme and polyclonal anti-glucoamylase antibodies. Nucleotide-sequence analysis revealed an open reading frame of 1848 base pairs coding for a protein of 616 amino-acid residues. Comparison with other fungal glucoamylase amino-acid sequences showed homologies of 37–48%. The glucoamylase cDNA, when introduced into Saccharomyces cerevisiae under the control of the yeast ADC1 promoter, directed the secretion of active glucoamylase P into the growth medium.
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  • 79
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    Current genetics 26 (1994), S. 95-99 
    ISSN: 1432-0983
    Keywords: Translational fidelity ; Paromomycin ; Stuttering ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Missense errors in the translation of mRNAs in Saccharomyces cerevisiae were screened by looking for charge heterogeneity of proteins on two-dimensional gels resulting from the substitution of charged and neutral amino acids. No such mistranslation was detected in wild-type yeast strains grown in the presence of the translational error-inducing antibiotic paromomycin. However, paromomycin-induced mistranslation of a heterologous mRNA, encoding human phosphoglycerate kinase expressed in yeast, was seen. We suggest that the combination of error-prone translation of a heterologous mRNA, and growth in the presence of paromomycin, leads to an accumulation of mistranslated proteins that can be detected by two-dimensional gel electrophoresis.
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  • 80
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Dynamin ; Mitochondria ; GTP binding protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The isolation and characterization of MGM1, and yeast gene with homology to members of the dynamin gene family, is described. The MGM1 gene is located on the right arm of chromosome XV between STE4 and PTP2. Sequence analysis revealed a single open reading frame of 902 residues capable of encoding a protein with an approximate molecular mass of 101 kDa. Loss of MGM1 resulted in slow growth on rich medium, failure to grow on non-fermentable carbon sources, and loss of mitochondrial DNA. The mitochondria also appeared abnormal when visualized with an antibody to a mitochondrial-matrix marker. MGM1 encodes a dynamin-like protein involved in the propagation of functional mitochondria in yeast.
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  • 81
    ISSN: 1432-0983
    Keywords: ABC superfamily ; Multidrug resistance ; Saccharomyces cerevisiae ; YDR1 gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A multidrug resistance gene, YDR1, of Saccharomyces cerevisiae, which encodes a 170-kDa protein of a member of the ABC superfamily, was identified. Disruption of YDR1 resulted in hypersensitivity to cycloheximide, cerulenin, compactin, staurosporine and fluphenazine, indicating that YDR1 is an important determinant of cross resistance to apparently-unrelated drugs. The Ydr1 protein bears the highest similarity to the S. cerevisiae Snq2 protein required for resistance to the mutagen 4-NQO. The drug-specificity analysis of YDR1 and SNQ2 by gene disruption, and its phenotypic suppression by the overexpressed genes, revealed overlapping, yet distinct, specificities. YDR1 was responsible for cycloheximide, cerulenin and compactin resistance, whereas, SNQ2 was responsible for 4-NQO resistance. The two genes had overlapping specificities toward staurosporine and fluphenazine. The transcription of YDR1 and SNQ2 was induced by various drugs, both relevant and irrelevant to the resistance caused by the gene, suggesting that drug specificity can be mainly attributed to the functional difference of the putative transporters. The transcription of these genes was also increased by heat shock. The yeast drug-resistance system provides a novel model for mammalian multidrug resistance.
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  • 82
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    Current genetics 20 (1991), S. 167-171 
    ISSN: 1432-0983
    Keywords: Glycolysis ; Repetitive elements τ/δ ; Promoter ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In this study we report on the complete nucleotide sequence of the yeast phosphoglycerate mutase gene (GPM1) and its essential 5′ and 3′ non-coding regions. The transcriptional start points were determined by S1-mapping and sequencing of a cDNA clone. Several sequences identified as important for transcriptional regulation in yeast promoters are present upstream of the transcription start point. 3′ to the coding region we sequenced a composite repetitive element which, apparently, originated from a recombination between a delta-and a tau-element. Finally, we mapped the GPM1 gene 13 cM distal to fas1 on chomosome XI.
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  • 83
    ISSN: 1432-0983
    Keywords: Growth control ; Genetic mapping ; Molecular cloning ; Nucleo-mitochondrial interaction ; Saccharomyces cerevisiae ; Viability of petites
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The PEL1 gene of Saccharomyces cerevisiae is essential for the cell viability of mitochondrial petite mutants, for the ability to utilize glycerol and ethanol on synthetic medium, and for cell growth at higher temperatures. By tetrad analysis the gene was assigned to chromosome III, centromere proximal of LEU2. The PEL1 gene has been isolated and cloned by the complementation of a pel1 mutation. The molecular analysis of the chromosomal insert carrying PEL1 revealed that this gene corresponds to the YCL4W open reading frame on the complete DNA sequence of chromosome III. The putative Pel1 protein is characterized by a low molecular weight of approximately 17 kDa, a low codon adaptation index, and a high leucine content.
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  • 84
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Papaver somniferum L. ; ARS ; Mitochondrial DNA
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract The minimal fragment of mitochondrial DNA from Papaver somniferum L. (poppy) able to promote autonomous plasmid replication in the yeast Saccharomyces cerevisiae was sequenced. Sequence analysis of the 917-bp MK4/8 DNA fragment revealed a high AT content, and the presence of two 12-bp sequences differing from the ARS core consensus of S. cerevisiae only by a T and C insertion, respectively. The mitochondrial insert contains a further six 11-bp sequences with one mismatch to the S. cerevisiae core consensus, more then 20 related sequences with two base pair exchanges, numerous direct and inverted repeats, and many copies of a sequence motif called the ARS box. The original 4.2-kb mitochondrial DNA fragment, as well as the minimal 917-bp subfragment in vector pFL1-E (a variant of YIP5, lacking an origin of replication in yeast), were then tested for their ability to replicate autonomously in another fungus, Kluyveromyces lactis.
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  • 85
    ISSN: 1432-0983
    Keywords: 2-Oxoglutarate dehydrogenase ; Molecular cloning ; Saccharomyces cerevisiae ; Sequencing ; Suppressor ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The activity of mitochondrial 2-oxoglutarate dehydrogenase in S. cerevisiae can be impaired either by the ogd1 or the kgd1 mutation. The OGD1 gene and two suppressor genes were isolated by complementation of the ogd1 mutant. The complementation of the kdg1 mutant by the OGD1 gene, an allelism test, and meiotic mapping, revealed that the ogd1 and kgd1 mutations are allelic. The two mutations were differentiated by the cloned suppressor gene which was able to partially complement ogd1, but not kgd1. The molecular analysis of the suppressor gene revealed its identity with the natural tRNA CAG Gln gene found in the upstream region of URA10.
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  • 86
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Aminoacyl-tRNA synthetase mutant ; PGK overexpression ; In vivo misreading
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The hts1.1 temperature-sensitive histidinyl-tRNA synthetase mutation enables Saccharomyces cerevisiae to be starved for His-tRNAHis by upshift to the non-permissive temperature of 38°C. If yeast behaves similarly to bacterial and mammalian cells, this lack of His-tRNAHis should greatly enhance misreading at histidine codons (CAU/CAC) by Gln-tRNAGln, resulting in substitution of the neutral amino acid glutamine in place of histidine, a basic amino acid. Such misreading causes the isoelectric point (pI) of proteins to shift to lower values, and is readily detectable as “stuttering” on two-dimensional (2D) protein gels. By gel analysis of pulse-labelled proteins of hts1.1 yeast cells that were overexpressing phosphoglycerate kinase (PGK), our study sought to detect this specific translational error in PGK protein. It was not detected by this relatively sensitive technique, indicating that missense errors due to glutamine insertion at histidine codons do not occur in yeast at the readily-detectable level found in bacterial and mammalian cells.
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  • 87
    ISSN: 1432-0983
    Keywords: Yeast ; Saccharomyces cerevisiae ; Sporulation mutants ; Reporter genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Reporter genes consisting of sporulation-specific promoters fused to lacZ were used as markers to monitor the sporulation pathway of the yeast Saccharomyces cerevisiae. Strains transformed with these lacZ gene fusions expressed β-galactosidase (assayable on plates using the substrate 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside, X-gal) in a sporulation-dependent manner. Mutagenesis experiments performed on transformed strains resulted in the recovery of a number of novel sporulation mutants. Three classes of mutants were obtained: those which overexpressed the reporter gene under sporulation conditions, those which did not express the gene under any conditions, and those which expressed the gene in vegetative cells not undergoing sporulation. On the basis of the blue colony-colour produced in the presence of X-gal these have been described as superblue, white, and blue vegetative mutants, respectively. These were further characterised using earlier reporter genes and other marker systems. This study established that the multicopy reporter plasmids chosen do not interfere with sporulation; they are valid tools for monitoring the pathway and they provide a way to isolate mutations not readily selected by other markers.
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  • 88
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    Current genetics 24 (1993), S. 461-464 
    ISSN: 1432-0983
    Keywords: Chromosome fragmentation ; MEL gene family ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nine members, MEL2–MEL10, of the MEL gene family coding for α-galactosidase were physically mapped to the ends of the chromosomes by chromosome fragmentation. Genetic mapping of the genes supported the location of all the MEL genes in the left arm of their resident chromosomes.
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  • 89
    ISSN: 1432-0983
    Keywords: Yeast ; Saccharomyces cerevisiae ; Transformation ; Plasmid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have compared a number of procedures for the transformation of whole cells of the yeast Saccharomyces cerevisiae and assessed the effects of dimethylsulphoxide (DMSO) or ethanol, both of which have been reported to enhance transformation efficiency. We find that simplified methods benefit from the addition of one of these compounds, and although differences are observed between strains as to the more beneficial reagent, peak transformation efficiency is, in general obtained with 10% DMSO or 10% EtOH. Increases of between six- and 50-fold are observed, despite a reduction in cell viability, and at this concentration the two compounds are not additive in their effects. The optimum level appears to depend on a balance between improved DNA uptake and reduced cell viability. As a result of this work we present a straightforward and rapid transformation procedure.
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  • 90
    ISSN: 1432-0983
    Keywords: Psoralen ; DNA repair mutants ; Gene conversion ; Recombination ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The influence of the DNA repair genePSO3 on photoactivated psoralen-induced meiotic recombination, gene conversion, reverse mutation, and on survival, was assayed in diploid strains ofSaccharomyces cerevisiae homozygous for the wild-type or thepso3-1 mutant allele. Sporulation was normal in thepso3-1 diploid. Wild-type and mutant strains had the same sensitivity to photoactivated monofunctional psoralen (3-CPs+UVA) in meiosis-uncommitted and meiosis-committed stages. The mutant showed higher sensitivity to photoactivated bifunctional psoralen (8-MOP+UVA) during all stages of the meiotic cycle. Mutation induction by 3-CPs+UVA or 8-MOP+UVA in meiosis-committed cells revealed no significant differences between wild-type and thepso3-1 mutant. The status of thePSO3 gene has no influence on the kinetics of induction of gene conversion and crossing-over after 3-CPs+UVA treatment in meiosis-committed cells: gene conversion was blocked while recombination was induced. After treatment with 8-MOP+UVA gene conversion was also blocked in both strains while crossing-over could only be observed in meiosis-committed wild-type cells.
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  • 91
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    Current genetics 25 (1994), S. 180-183 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; In-vivo cloning ; Non-replicative vectors ; Homologous recombination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have devised a new strategy to clone DNA sequences from an yeast autonomously-propagating plasmid into a non-autonomous integrative vector by in-vivo recombination. The method consists of a first step in which the replicative plasmid carrying the DNA fragment of interest forms a co-integrate with the non-replicative plasmid by an induced in-vivo reciprocal exchange accompanied by gene conversion. The dimeric plasmid obtained is then purified and cut with an appropriate restriction enzyme and ligated independently to obtain the two intact monomeric plasmids, the original autonomous plasmid plus the new non-autonomous plasmid carrying the subcloned DNA fragment. The dimeric co-integrate can also serve as substrate for a second in-vivo reciprocal exchange that produces new autonomous plasmids carrying the desired DNA fragment. The technique considerably expands the applications of in-vivo cloning in yeast by complementing three important characteristics of previously published methods: (1) it can be used to clone into non-propagating vectors; (2) co-transformation experiments are not required; and (3) the intermediate co-integrate can be used to generate new types of autonomously-propagating plasmids directly. These characteristics are independent of whether the DNA insert is flanked by appropriate restriction sites or whether it does, or does not, express a detectable phenotype in yeast. The method is particularly useful for the cloning of large DNA fragments and can be used for plasmids from organisms other than yeasts.
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  • 92
    ISSN: 1432-0983
    Keywords: 1,3-β-glucanase genes ; Saccharomyces cerevisiae ; Chromosomal mapping ; Genetic mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The map position of three 1,3-β-glucanase-encoding genes in S. cerevisiae has been determined following conventional meiotic and mitotic mapping combined with recombinant DNA techniques. EXG1, EXG2 and SSG1 were localized to chromosomes XII, IV and XV, respectively, by hybridizing the cloned genes to Southern blots of chromosomes sepaated by pulsed-field gel electrophoresis, in conjunction with the rad52-1-dependent chromosome-loss mapping technique. Meiotic tetrad analyses further localized the EXG1 gene 6.1 centimorgans centromere-proximal to CDC25 on the right arm of chromosome XII. EXG2 was positioned between LYS4 and GCN2 on the right arm of chromosome IV, at distances of 6.2 centimorgans from LYS4 and 4.9 centimorgans from GCN2. Finally, the SSG1 locus mapped on the right arm of chromosome XV, about 8.2 centimorgans to the centromere-proximal side of HIS3.
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  • 93
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Hydrostatic pressure ; Tetraploidy ; Homozygous diploid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Hydrostatic pressure and a dye plate method were used to investigate the direct induction of tetraploids or homozygous diploids from the industrial diploid or haploid yeast Saccharomyces cerevisiae. Above 200 MPa, hydrostatic pressure greatly inactivated the strains HF399s1 (α haploid), P-540 (a/α diploid), and P-544 (a/α diploid). At the same time, when pressure-treated cells of these strains were spread on a dye plate, some of the visible colonies were stained red/blue or dark blue (variant colonies); the rest stained violet, similar to colonies originating from diploid cells or haploid cells that were not pressure-treated. In addition, above 100 MPa, the formation of variant colonies increased with increasing pressure, and maximized (1x10-1) at 200 and 250 MPa, respectively. The size of almost all variant cells from P-544, P-540, and HF399s1 was visibly increased compared with that of untreated cells and the measured cellular DNA content of P-540 and HF399s1 was double that of untreated cells. Furthermore, based on random spore analysis and mass-matings, induced variants in the diploid strains were found to be tetraploid with an a/a/α/α genotype at the mating-type locus or, in the haploid strains, homozygous diploid with an α/α genotype. From these results we conclude that pressure treatment in combination with a dye plate is a useful method for strain improvement by direct induction of tetraploids or homozygous diploids from industrial strains whether diploid or haploids.
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  • 94
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    Current genetics 25 (1994), S. 291-298 
    ISSN: 1432-0983
    Keywords: Cytochrome c 1 ; Cytochrome c 1 heme lyase ; GRF2p ; Glucose repression ; HAPp ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In this paper we examine the expression of the Saccharomyces cerevisiae CYT2 gene, which encodes cytochrome c 1 heme lyase. This enzyme is required for covalent attachment of heme to apocytochrome c 1, a subunit of the mitochondrial respiratory chain. Transcription of the 1-kb CYT2 mRNA initiates at four prominent sites at a distance of 52–225 bp in front of the AUG start codon. The level of CYT2 mRNA is not influenced by the presence or absence of oxygen or of heme, but it is subject to carbonsource control. The concentration of the CYT2 mRNA is significantly reduced in glucose-grown cells as compared to cells grown under non-repressing conditions. Neither the HAPp activator proteins nor MIG1p, a repressor protein involved in glucose repression, seem to mediate this effect.
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  • 95
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; recA gene expression ; UV radiation ; Mitotic gene conversion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of the Escherichia coli RecA protein on mitotic recombination in the diploid D7 strain of Saccharomyces cerevisiae damaged by UV radiation was investigated. The D7 strain was transformed by two modified versions of the pNF2 plasmid: one, containing the ADH-1 promoter, and the other containing the recA gene tandemly arranged behind the ADH-1 promoter region. Immunological analysis proved the presence of the 38-kDa RecA protein in D7/pNF2ADHrecA transformants. We observed a positive effect of recA gene expression on mitotic gene conversion, mainly at higher doses of UV radiation. The results indicate that a RecA-like activity could participate in steps preceeding mitotic conversion events in yeast.
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  • 96
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    Current genetics 26 (1994), S. 15-20 
    ISSN: 1432-0983
    Keywords: Cell-division cycle ; Mitochondrial genome ; Nuclear mutation ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In former studies it was found that the ERV1 gene is essential for cell viability and for the biogenesis of functional mitochondria. A temperature-sensitive nuclear mutant exhibits a severe reduction in all the mitochondrial transcripts. Elimination of the gene leads to growth arrest after a few cell divisions. The putative gene product bears the characteristics of a regulatory factor since it has low expression rate and a high content of charged amino acids. In this study it is further verified that the ERV1 gene alone is responsible for the observed cellular and mitochondrial defects. The 5′ region of the gene is analysed by DNA deletions and complementation studies. Expression of the gene under the control of the GAL1-10 promoter in a disruption strain of ERV1 allows a more detailed specification of its influence on mitochondrial and cellular functions. Immediate and complete loss of mitochondrial genomes is observed after the promoter has been shut off, whereas the yeast cells are still able to grow for a limited time under these conditions. Analysis of the cells by in-vivo DNA flurorescence demonstrates a specific arrest in the cell-division cycle as the terminal phenotype. To further characterize the temperature-sensitive allele of ERV1 the mutated gene has been isolated and sequenced. A single point mutation which leads to the exchange of a single amino acid is found in the reading frame.
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  • 97
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Nuclear gene ; Mitochondria ; Mitochondrial ribosomal protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The nuclear gene MRP-L13 of Saccharomyces cerevisiae, which codes for the mitochondrial ribosomal protein YmL13, has been cloned and characterized. It is a single-copy gene residing on chromosome XI. Its nucleotide sequence was found to be identical to that of the previously reported ORF YK105. A comparison of the predicted protein sequence of the MRP-L13 gene product and the actual N-terminal amino-acid sequence of the isolated YmL13 protein indicated that the mature protein is preceded by a mitochondrial signal peptide of 86 amino-acid residues, which is the longest among all known mitochondrial ribosomal proteins of S. cerevisiae. No sequence similarity was found to any other ribosomal protein in the current databases. The transcription of MRP-L13 was found to be repressed in the presence of glucose. Its protein product is not strictly essential for mitochondrial functions, but disruption of the gene by insertion of LEU2 noticeably affected cellular growth on non-fermentable carbon sources.
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  • 98
    ISSN: 1432-0789
    Keywords: Vigna unguiculata ; Zea mays ; Nutrient competition ; Intercropping ; Nitrate depletion ; N2 fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The use of N and P by mixed and by sole cropping (crop rotation) of maize and cowpeas were compared in a field experiment on an Alfisol at the Nyankpala Agricultural Experiment Station in the northern Guinea Savanna of Ghana, using two levels of N (0 and 80 kg N ha-1 year-1 as urea) and P application (0 and 60 kg P ha-1 year-1 as Volta phosphate rock). Maize grain yields were significantly reduced in the mixed cropping system. This yield difference became smaller with the application of N and P fertilizer. The N and P concentrations in maize ear leaves at silking indicated that a deficiency in N and P contributed to the maize yield depression in mixed cropping. Competition for soil and fertilizer N between maize and cowpeas was suggested by: (1) A similarity in total N uptake between the two cropping systems; (2) efficient use of soil nitrate by the cowpeas; and (3) low N2 fixation by the cowpeas, calculated with the aid of an extended-difference method. In general, N2 fixation was low, with the highest values in the sole cropping (53 kg ha-1) and a substantial reduction in the mixed cropping system. The application of N fertilizer further reduced N2 fixation. This was substantiated by nodule counts. The lower N2 fixation in the mixed cropping system was only partly explained by the lower density of cowpeas in this system. In addition, dry spells during the cropping season and shading by the maize component could have reduced the nodulation efficiency. No N transfer from the legume/rhizobium to the non-legume crop was observed. Impaired P nutrition in the mixed compared with the sole-cropped maize might have been due to less P mobility in the soil. This was indicated by lower soil moisture contents in the topsoil under mixed cropping, especially during the dry year of 1986. The results show that mixed cropping of maize and cowpeas did not lead to improved use of soil and fertilizer N and P or to an enhanced N2 fixation. On the contrary, an annual rotation of maize and cowpeas was clearly superior.
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  • 99
    ISSN: 1432-0789
    Keywords: Antifungal activity ; Saccharomyces cerevisiae ; Phytopathogenic fungi ; Heterocyclic non-protein amino acid ; Pisum sativum ; Constitutive plant defence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary β-(Isoxazolin-5-on-2-yl)-alanine (βIA), a heterocyclic non-protein amino acid from root extracts and root exudates of pea seedlings, acts as a potent growth inhibitor of several eukaryotic organisms, including yeasts, phytopathogenic fungi, unicellular green algae, and higher plants. The antibiotic effect on baker's yeast was reversed by l-methionine, l-cysteine, and l-homocysteine. Phytopathogenic fungi such as Botrytis cinerea, Pythium ultimum, and Rhizoctonia solani grown on agar containing βIA were inhibited in the growth of mycelia or in the production of sclerotia. In contrast, no significant inhibition of either Gram-positive or Gram-negative bacteria was observed. Rhizobium leguminosarum, the compatible microsymbiont of Pisum spp., and Rhizobium meliloti were able to tolerate up to 2.9 mM βIA (500 ppm) without any effect on the growth rate. Bradyrhizobium japonicum even gave a positive chemotactic response to βIA. The ecological significance of βIA as a preformed plant protectant during the seedling stage of Pisum spp. and other βIA-containing legumes is discussed.
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  • 100
    Electronic Resource
    Electronic Resource
    Springer
    Biology and fertility of soils 13 (1992), S. 142-146 
    ISSN: 1432-0789
    Keywords: A value ; Common bean ; N remobilization ; Soil N balance ; Atom% 15N excess ; Phaseolus vulgaris ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The common bean (Phaseolus vulgaris L.) is generally regarded as a poor N2 fixer. This study assessed the sources of N (fertilizer, soil, and fixed N), N partitioning and mobilization, and soil N balance under field conditions in an indeterminate-type climbing bean (P. vulgaris L. cv. Cipro) at the vegetative, early pod-filling, and physiological maturity stages, using the A-value approach. This involved the application of 10 and 100 kg N ha-1 of 15N-labelled ammonium sulphate to the climbing bean and a reference crop, maize (Zea mays L.). At the late pod-filling stage (75 days after planting) the climbing bean had accumulated 119 kg N ha-1, 84% being derived from fixation, 16% from soil, and only 0.2% from the 15N fertilizer. N2 fixation was generally high at all stages of plant growth, but the maximum fixation (74% of the total N2 fixed) occurred during the interval between early (55 days after planting) and late podfilling. The N2 fixed between 55 and 75 days after planting bas a major source (88%) of the N demand of the developing pod, and only about 11% was contributed from the soil. There was essentially no mobilization of N from the shoots or roots for pod development. The cultivation of common bean cultivars that maintain a high N2-fixing capacity especially during pod filling, satisfying almost all the N needs of the developing pod and thus requiring little or no mobilization of N from the shoots for pod development, may lead to a net positive soil N balance.
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