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  • Zea mays  (193)
  • Springer  (193)
  • 2020-2024
  • 1990-1994  (193)
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  • 1
    Electronic Resource
    Electronic Resource
    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 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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  • 4
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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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  • 5
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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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  • 6
    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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  • 7
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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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  • 8
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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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  • 9
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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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  • 10
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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
    Source: Springer Online Journal Archives 1860-2000
    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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  • 11
    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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  • 12
    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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  • 13
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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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  • 14
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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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  • 15
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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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  • 16
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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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  • 17
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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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  • 18
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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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  • 19
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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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  • 20
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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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  • 21
    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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  • 22
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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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  • 23
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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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  • 24
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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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  • 25
    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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  • 26
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    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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  • 27
    ISSN: 1432-0789
    Keywords: Helianthus annuus ; Mycorrhiza ; Soil organic matter ; VAM response ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Vesicular-arbuscular mycorrhizal (VAM) fungi affect diverse aspects of plant form and function. Since mycorrhiza-mediated changes in host-plant responses to root colonization by different VAM fungi vary widely, it is important to assess each endophyte for each specific effect it can elicit from its host as part of the screening process for effectiveness. Three species of VAM fungi and a mixture of species were compared with non-VAM controls for their effects on soil organic matter contents and on nutrition and morphology in two varieties (native and hybrid) of corn (Zea mays L.) and one of sunflower (Helianthus annuus L.) in P-sufficient and N-deficient soil in pot cultures. Differences in soil organic matter due to the fungal applications were highly significant with all host plants. Native corn responded more to VAM colonization than the hybrid did; differences in treatments were significant in leaf area, plant biomass, and root: shoot ratio in the former, but not in the latter. Responses in the sunflower were similar to those in the native corn. Significant VAM treatment-related differences in shoot N and P contents were not reflected in shoot biomass, which was invariant. Correlations between plant or soil parameters and the intensity of VAM colonization were found only in soil organic matter with the native corn, in specific leaf area in the hybrid corn, and in plant biomass in the sunflower. The presence of the different endophytes and not the intensity of colonization apparently elicited different host responses.
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  • 28
    ISSN: 1432-0789
    Keywords: DRIS ; Helianthus annus ; Plant nutrition ; VAM ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Vesicular-arbuscular mycorrhizal (VAM) fungi improve plant growth in marginal soils. This study was conducted to determine the effects of three species of VAM fungi on plant nutrition in two cultivars of corn (Zea mays L.) and one of sunflower (Helianthus annus L.). Plants were grown in pot cultures under controlled (greenhouse) conditions in a soil high in K, Mg, and P, and low in Ca and N, and were supplied with amounts of VAM-fungal inocula in which equal numbers of infective propagules had previously been determined. Analysis of variance showed highly significant main effects and interactions due to both factors (plant and fungus) for N, P, Ca, and Mg. For K, only plant effects were significant (P〈0.043). The uptake of nutrients was selectively enhanced or inhibited by one or the other VAM fungus relative to non-VAM control plants. In sunflower, N concentration was markedly enhanced (73%) by the mixed inoculum of the three fungi, even though individual effects were not significant. Evaluation of leaf nutrient analyses by the Diagnosis and Recommendation Integrated System (DRIS) revealed the utility of this system to rank nutritional effects by VAM fungi in an order of relative nutrient deficiency. The DRIS therefore is seen as a useful tool in evaluating and selecting VAM fungi for the alleviation of specific nutrient disorders.
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  • 29
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    Biology and fertility of soils 15 (1993), S. 249-252 
    ISSN: 1432-0789
    Keywords: N fertilizer requirement ; Nitrification ; Zea mays ; N mineralization ; Lime ; Soil pH ; Nitrate-N
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The application of NH inf4 su+ -based fertilizers to soils slowly lowers soil pH, which in turn decreases nitrification rates. Under these conditions nitrification and N mineralization may be reduced. We therefore investigated the impact of liming fertilizer-acidified soils on nitrification and N mineralization. Soil samples were collected in the spring of 1987 from a field experiment, initiated in 1980, investigating N, tillage, and residue management under continuous corn (Zea mays L.). The pH values (CaCl2) in the surface soil originally ranged from 6.0 to 6.5. After 6 years the N fertilizer and tillage treatments had reduced the soil pH to values that ranged between 3.7 and 6.2. Incubation treatments included two liming rates (unlimed or SMP-determined lime requirement), two 15N-labeled fertilizer rates (0 or 20 g N m-2), and three replicates. Field-moist soil was mixed with lime and packed by original depth into columns. Labeled-15N ammonium sulfate in solution was surface-applied and columns were leached with 1.5 pore volumes of deionized water every 7 days over a 70-day period. Nitrification occurred in all pH treatments, suggesting that a ferilizer-acidified soil must contain a low-pH tolerant nitrifier population. Liming increased soil pH values (CaCl2) from 3.7 to 6.2, and increased by 10% (1.5 g N m-2) the amount of soil-derived NO3 --N that moved through the columns. This increase was the result of enhanced movement of soil-derived NO3 --N through the columns during the first 14 days of incubation. After the initial 14-day period, the limed and unlimed treatments had similar amounts of soil N leaching through the soil columns. Lime increased the nitrification rates and stimulated the early movement of fertilizer-derived NO3 --N through the soil.
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  • 30
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    Biology and fertility of soils 17 (1994), S. 1-8 
    ISSN: 1432-0789
    Keywords: Ammonium excretion ; Azospirillum brasilense ; Auxine ; 2,4-Dichlor-phenoxy-acetic acid ; Nitrogen fixation ; Paranodulation ; Maize ; Zea mays ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Maize seedlings develop nodule-like tumour knots (para-nodules) along primary roots when treated with the auxin 2,4-dichlor-phenoxy-acetic acid (2,4-D). Inoculated NH 4 + -excreting Azospirillum brasilense cells were shown to colonize these tumours, mostly intracellularly, promoting a high level of N2 fixation when microaerophilic conditions were imposed. The nitrogenase activity inside the para-nodules was less sensitive to free O2 than in non-para-nodulating roots. Both light and electron microscopy showed a dense bacterial population inside intact tumour cells, with the major part of the cell infection along a central tumour tissue. The bacteria colonized the cytoplasm with a close attachment to inner cell membranes. In an auxin-free growth medium, young 2,4-D-induced para-nodules grew further to become mature differentiated root organs in which introduced bacteria survived with a stable population. These results provide evidence that gramineous plants are potentially able to create a symbiosis with diazotrophic bacteria in which the NH 4 + -excreting symbiont will colonize para-nodule tissue intracellularly, thus becoming well protected.
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  • 31
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    Biochemical genetics 28 (1990), S. 319-336 
    ISSN: 1573-4927
    Keywords: Zea mays ; Gramineae ; β-glucosidase ; null allelism ; immunoblotting ; electrophoresis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Maize β-glucosidase (β-d-glucoside glucohydrolase; EC 3.2.1.21) was extracted from coleoptiles of 15 maize genotypes (3 normals, 10 nulls, and 2 hybrids) in two fractions, the soluble and the insoluble. The enzyme activity was measured spectrophotometrically in the soluble fraction and also studied on zymograms after native gel electrophoresis and isoelectric focusing. The enzyme was purified from a normal genotype by anion-exchange chromatography and preparative electrophoresis. Antisera were raised in four rabbits, and the soluble and the insoluble extracts of each genotype were analyzed for a cross-reacting material by ELISA and immunoblotting. The results showed that extracts from both the normal and the null genotypes had β-glucosidase activity, and the activity measured spectrophotometrically was 2- to 10-fold higher in normals than in nulls. Zymograms of the null genotypes were devoid of distinct bands that were present in those of normals and hybrids from crosses between normals and nulls. Zymograms of both the normal and the null genotypes had a diffuse, smeared zone of activity at the cathodic end of native gels. A cross-reacting antigen was present in extracts of both genotypes when assayed by ELISA and a 60-kD polypeptide (β-glucosidase monomer) was detected by four different monospecific β-glucosidase antisera on Western blots by immunostaining. Moreover, six of seven null genotypes had a larger amount of their 60-kD polypeptide in the insoluble fraction than in the soluble fraction. These data show that both the null and the normal genotypes have similar amounts of the enzyme protein, but the enzyme occurs mostly as insoluble or poorly soluble polymers in nulls, and the monogenic inheritance reported for the null alleles of theglu locus is likely to be for a factor encoded by another locus which affects directly or indirectly the solubility of the enzyme by increasing its polymerization into large quaternary structures.
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  • 32
    ISSN: 1573-1561
    Keywords: Fall armyworm ; Spodoptera frugiperda ; Lepidoptera ; Noctuidae ; corn ; Zea mays ; plant-insect interaction ; amino acids ; herbivory ; feeding resistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The free amino acids have been shown by isolational work and choice bioassays to be more important than all other factors evaluated in defining leaf-feeding resistance of corn (Zea mays L.) to fall armyworm (FAW) [(Spodoptera frugiperda J.E. Smith)] larvae. 6-MBOA (6-methoxybenzoxazolinone) and maysin, toxins present in corn, were shown not to be significant factors for leaf-feeding resistance to first-instar FAW larvae because of their low concentrations in the whorl. Amino acid analysis showed that while the ratios of the essential amino acids in susceptible (S) and resistant (R) lines were similar, there were differences in the nonessential amino acids, particularly aspartic acid, which was higher in R lines. Also, the ratio of essential amino acids to nonessential amino acids was important, being too low in expressed whorl leaf juice (obtained from V8–V10 growth stage plants) to support larval growth, although juice was stimulatory in choice tests. The total protein content of whorls in S lines was about 15% higher than in R lines, but the significance of this difference is uncertain, because nutritional tests showed that larval growth increased with total protein only up to 12% protein. Sugars were only slightly stimulatory. Thus, the amino acids along with higher hemicellulose content of R lines, established by us earlier, appear to explain much of the basis of resistance in corn to larval leaf-feeding of the FAW.
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  • 33
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    Journal of chemical ecology 18 (1992), S. 1227-1237 
    ISSN: 1573-1561
    Keywords: Western corn rootworm ; Diabrotica virgifera virgifera ; Coleoptera ; Chrysomelidae ; bacteria ; carbon dioxide ; pheromone ; semiochemicals ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract FemaleDiabrotica virgifera virgifera LeConte were allowed to choose between oviposition substrates that were and those that were not associated with potential sources of semiochemicals. Females deposited over five times more eggs on moist towelettes that were treated with homogenates of female abdomens than on towelettes treated with distilled water. Similar results were obtained when screening separated the homogenates from the towelettes, indicating that odors alone could elicit the response. In contrast, females did not choose towelettes that had previously been used for oviposition or towelettes containing eggs over unused towelettes. Further tests with homogenates of abdomens and a bacteriostatic agent (sorbate) indicated that the females were probably responding to bacterial odors rather than an oviposition-enhancing pheromone. Four strains of bacteria were isolated from a homogenate of female abdomens; females deposited 4 to 16 times more eggs on substrates with odors of the bacteria than on substrates with odors of uninoculated nutrient agar. In no-choice tests, bacterial odors did not increase the number of eggs deposited per female beetle; however, in choice tests with dishes that tended to retain any beetles that entered, there were more eggs per female (but not more beetles) after 24 hr in dishes with bacterial odors than in those without the odors. Females also chose dishes with odors of excised maize (Zea mays L.) roots or elevated levels of carbon dioxide over “control” dishes.
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  • 34
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    Biochemical genetics 28 (1990), S. 319-336 
    ISSN: 1573-4927
    Keywords: Zea mays ; Gramineae ; β-glucosidase ; null allelism ; immunoblotting ; electrophoresis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Maize β-glucosidase (β-d-glucoside glucohydrolase; EC 3.2.1.21) was extracted from coleoptiles of 15 maize genotypes (3 normals, 10 nulls, and 2 hybrids) in two fractions, the soluble and the insoluble. The enzyme activity was measured spectrophotometrically in the soluble fraction and also studied on zymograms after native gel electrophoresis and isoelectric focusing. The enzyme was purified from a normal genotype by anion-exchange chromatography and preparative electrophoresis. Antisera were raised in four rabbits, and the soluble and the insoluble extracts of each genotype were analyzed for a cross-reacting material by ELISA and immunoblotting. The results showed that extracts from both the normal and the null genotypes had β-glucosidase activity, and the activity measured spectrophotometrically was 2- to 10-fold higher in normals than in nulls. Zymograms of the null genotypes were devoid of distinct bands that were present in those of normals and hybrids from crosses between normals and nulls. Zymograms of both the normal and the null genotypes had a diffuse, smeared zone of activity at the cathodic end of native gels. A cross-reacting antigen was present in extracts of both genotypes when assayed by ELISA and a 60-kD polypeptide (β-glucosidase monomer) was detected by four different monospecific β-glucosidase antisera on Western blots by immunostaining. Moreover, six of seven null genotypes had a larger amount of their 60-kD polypeptide in the insoluble fraction than in the soluble fraction. These data show that both the null and the normal genotypes have similar amounts of the enzyme protein, but the enzyme occurs mostly as insoluble or poorly soluble polymers in nulls, and the monogenic inheritance reported for the null alleles of theglu locus is likely to be for a factor encoded by another locus which affects directly or indirectly the solubility of the enzyme by increasing its polymerization into large quaternary structures.
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  • 35
    ISSN: 1573-5028
    Keywords: chloroplast genome ; operon organization ; ribosomal protein ; rpl32 ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract By sequencing the rpl32 gene, we have characterized the apparent complete set of the RP genes in Zea mays plastid genome. Key data for these 21 genes (total of 26 gene copies) and the proteins encoded by them are presented, and the operon organization is discussed on the basis of available transcription data. A nomenclature for the inferred 13 operons is suggested.
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  • 36
    ISSN: 1573-5028
    Keywords: geminivirus ; agroinfection ; Zea mays ; large intergenic region (LIR)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Maize streak virus as well as other geminiviruses contain a potential hairpin structure with the conserved sequence TAATATTAC in the loop. We assessed the possible involvement of this structure in replication and symptom induction of the virus. A series of insertion and deletion mutants were analyzed by agroinfection. Deletion of the hairpin or insertions in the conserved sequence abolished symptom development. Viral DNA could not be detected in the infected tissue. However, a mutant with a point mutation in the ‘conserved’ sequence, isolated after inoculation of maize plants with an insertion mutant, was able to replicate and to induce symptoms.
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  • 37
    ISSN: 1573-5028
    Keywords: gene-specific probes ; glutamine synthetase ; transcript accumulation ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The maize genome has been shown to contain six glutamine synthetase (GS) genes with at least four different expression patterns. Noncoding 3′ gene-specific probes were constructed from all six GS cDNA clones and used to examine transcript levels in selected organs by RNA gel blot hybridization experiments. The transcript of the single putative chloroplastic GS2 gene was found to accumulate primarily in green tissues, whereas the transcripts of the five putative GS1 genes were shown to accumulate preferentially in roots. The specific patterns of transcript accumulation were quite distinct for the five GS1 genes, with the exception of two closely related genes.
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  • 38
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    Plant molecular biology 24 (1994), S. 35-49 
    ISSN: 1573-5028
    Keywords: acetyl CoA carboxylase ; cDNA Cloning ; herbicide ; nucleotide sequence ; purification ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Acetyl CoA carboxylase (EC 6.4.1.2) in plants is a chloroplast-localized, biotin-containing enzyme that catalyses the carboxylation of acetyl CoA to malonyl CoA, the first committed step of the fatty acid biosynthesis pathway. Acetyl CoA carboxylase is the target site for the monocotyledon-specific aryloxy-phenoxypropionate and cyclohexanedione groups of herbicides. We have purifed a herbicide-sensitive acetyl CoA carboxylase from maize leaves to homogeneity (specific activity 7 μmol min-1 mg-1), separating it during the purification from a minor herbicide-resistant acetyl CoA carboxylase. The purified enzyme is a dimer of 230 kDa subunits. Antibodies raised to the purified acetyl CoA carboxylase detected three cross-reacting clones in a maize leaf cDNA expression library, each having an insert of 4–4.5 kb. Restriction analysis and sequencing showed that the cDNAs were derived from two different transcripts. Comparison of the deduced amino acid sequences with those of chicken and yeast acetyl CoA carboxylases confirmed that both types encoded acetyl CoA carboxylase, corresponding to the C-terminal half of the enzyme. The overall identity of the maize and chicken sequences was 37% (58% similarity) but for some shorter regions was much higher. Analysis of six other acetyl CoA carboxylase clones recovered from the maize cDNA library showed four belonged to one type and two to the other. The nucleotide sequence similarity between the two types of cDNA was approximately 95% in the coding region but considerably less in the 3′-untranslated region. Northern blot analysis of maize RNA showed a single band of 8.2–8.5 kb for acetyl CoA carboxylase mRNA. Southern blot hybridisations indicated that there are probably no more than two genes in maize for acetyl CoA carboxylase. The possible significance of two different cDNAs for acetyl CoA carboxylase is discussed.
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  • 39
    ISSN: 1573-5028
    Keywords: Opaque-2 and opaque-2 genes ; allelic diversity ; Opaque-2 proteins ; transcriptional activator ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The expression of the various members of the zein multigene family in maize endosperm is controlled by different regulatory loci. One of these loci, Opaque-2, coding for a bZIP transcriptional factor, controls the expression of a subset of zein genes. Analysis of genomic DNA from plants carrying wild-type (O2) or mutant o2 alleles shows specific DNA restriction patterns that correlate with transcript types and their various gene products. Northern and western analyses show the presence in different wild types of a 1.7 kb transcript coding for different sizes of normal O2 proteins that migrate as doublets in the 68–72 kDa range. Among the various o2 mutants analysed we showed the occurrence of various null-transcript alleles, the presence of alleles with a normal size transcript which, however, produce a different-sized o2 protein, and a mutant producing both a normal size transcript and a longer transcript, but generating only a single o2 product migrating around 40 kDa. Analysis of other mutations (o7, fl2) known to affect zien polypeptide synthesis shows no interference of these mutations in the expression of the O2 gene products. The overall results indicate the occurrence of micro heterogeneity in the O2 wild-type genes and a broad spectrum of o2 mutations, both producing different sizes of O2 or o2 proteins. A nomenclature of the O2 and o2 genes based on the RFLP, transcripts and products of the various alleles is presented.
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  • 40
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    Plant molecular biology 25 (1994), S. 909-916 
    ISSN: 1573-5028
    Keywords: heat shock 70 kDa protein ; multigene family ; polymerase chain reaction ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In this study we have analysed the multigene family coding for the cytoplasmic heat shock 70 kDa proteins (hsp70) inZea mays. Fully degenerate primers were used in a polymerase chain reaction (PCR) to amplify selected regions of the hsp70 genes. Sequence and Southern blot analysis reveals that at least three highly conserved genes exist in maize. In addition, amplification reveals the presence of a conserved intron in all genes examined. Expression analysis shows that the hsp70 genes studied represent members of the inducible and constitutive families. The results obtained may indicate that there are subfamilies of cytoplasmic hsp70 genes expressed in higher plants.
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  • 41
    ISSN: 1573-5028
    Keywords: cortex ; developmental regulation ; in situ hybridization ; organ-specific gene expression ; roots ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A near full-length cDNA clone (pZRP3) corresponding to an mRNA that accumulates specifically in roots of maize was isolated. The ZRP3 mRNA is ca. 600 nucleotides in length. The amino acid sequence of the predicted polypeptide is rich in leucine (16%), proline (11%), and cysteine (8.5%). The zrp3 gene appears to be expressed exclusively in roots, whereas other ZRP3-related genes are expressed in additional organs of the maize plant. In situ hybridization shows that ZRP3 mRNA accumulation is largely confined to the cells of the cortical ground meristem. Furthermore, accumulation of this mRNA occurs within a distinct subset of cortical cells, the inner three to four cell layers.
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  • 42
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    Plant molecular biology 20 (1992), S. 1159-1168 
    ISSN: 1573-5028
    Keywords: DNA polymerase ; germination ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three different DNA polymerase activities can be resolved by passing a protein extract from 24 h imbibed maize axes through DEAE-cellulose. These activities have been numbered 1, 2 and 3, according to their elution order. One of them, DNA polymerase 2, elutes at 100–120 mM phosphates. This enzyme was further purified by passing it through Heparin-Sepharose, Sephacryl S-300 and DNA cellulose. Purification was nearly 5000-fold. The enzyme needs Mg2+, is stimulated by K+, has an optimum pH of 7.0 and its optimum temperature is 30–37 °C. Specific inhibitors for different types of polymerases, such as aphidicolin, dideoxythymidine triphosphate and N-ethyl maleimide, gave intermediate values of inhibition, making impossible the definition of the type of enzyme purified by its inhibitory pattern. SDS-PAGE indicated the presence of several bands of molecular masses of 28–40, 56 and 15 kDa. Most of these bands could be visualized when proteins from crude extracts were analyzed by western blot, using an antibody against calf thymus DNA polymerase α. A high molecular mass (around 500 kDa) was calculated by western blot of native gels using the same antibody. Finally, specific activity of this enzyme increased 100-fold during maize germination whereas polymerase 3 virtually did not increase. Furthermore, immunoprecipitation experiments with the antipolymerase α-antibody showed a decrease in DNA polymerase activity by 70%. The possibility that polymerase 2 is a replicative enzyme is discussed.
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  • 43
    ISSN: 1573-5028
    Keywords: allergens ; gene expression ; microsporogenesis ; pectate lyase ; pollen ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cDNA clone (Zm58.1) was isolated by differential screening from a cDNA library made to mature Zea mays pollen, and shown to be pollen-specific by RNA blot analysis. When this partial-length clone was used to probe a genomic library, a similar but distinct pollen-specific genomic clone (68% sequence identity) was isolated (Zm58.2). The putative proteins coded for by these two clones show sequence homology to several flower-expressed gene products from various plant species, including known pollen allergens from short ragweed (Ambrosia artemisiifolia), and to pectate lyases from the plant pathogenic bacteria Erwinia spp. The two genes map to different chromosomes.
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  • 44
    ISSN: 1573-4927
    Keywords: alcohol dehydrogenase ; maize ; Zea mays ; mutation ; cDNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The homodimeric alcohol dehydrogenase gene product of maize (Zea mays L.)Adh1-1S1108 mutation was purified and compared with the parentalAdh1-1S enzyme. The mutant alcohol dehydrogenase activity had pH optima and substrate specificity similar to those of the parental enzyme, but exhibited somewhat increased and decreasedK mvalues for acetaldehyde and NADH, respectively. The mutant enzyme was also markedly less stable than the enzyme from parental tissues to temperatures as low as 50°C. Sequence analysis of a polymerase chain reaction (PCR)-generated cDNA clone revealed a G-to-C mutation at position 406 and a C-to-T mutation at position 974. These would result in residue 103 of each protein subunit being changed from an alanine to a proline and residue 292 being changed from an alanine to a valine. Whether one or both of these changes in primary sequence is responsible for the altered substrate affinities and stability is not yet understood.
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  • 45
    ISSN: 1573-4927
    Keywords: alcohol dehydrogenase ; maize ; Zea mays ; mutation ; cDNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The homodimeric alcohol dehydrogenase gene product of maize (Zea mays L.)Adh1-1S1108 mutation was purified and compared with the parentalAdh1-1S enzyme. The mutant alcohol dehydrogenase activity had pH optima and substrate specificity similar to those of the parental enzyme, but exhibited somewhat increased and decreasedK mvalues for acetaldehyde and NADH, respectively. The mutant enzyme was also markedly less stable than the enzyme from parental tissues to temperatures as low as 50°C. Sequence analysis of a polymerase chain reaction (PCR)-generated cDNA clone revealed a G-to-C mutation at position 406 and a C-to-T mutation at position 974. These would result in residue 103 of each protein subunit being changed from an alanine to a proline and residue 292 being changed from an alanine to a valine. Whether one or both of these changes in primary sequence is responsible for the altered substrate affinities and stability is not yet understood.
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  • 46
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    Plant molecular biology 16 (1991), S. 919-923 
    ISSN: 1573-5028
    Keywords: Zea mays ; maize ; embryogenesis ; abscisic acid ; seed maturation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have isolated by differential hybridization a cDNA, termed Emb564, which is complementary to an 800 nt embryo-specific transcript in Zea mays. The expression of Emb564 can be enhanced by exogenous abscisic acid (ABA) and the effect of ABA on the accumulation of Emb564 transcript appears to be confined to embryos at the early and mid embryonic stages. In addition, Emb564 is expressed at low levels in ABA-deficient but not in ABA non-responsive embryos. Genomic analysis suggested that the Emb564 mRNA is encoded by a single gene. Sequence analysis showed that Emb564 exhibits extensive similarities with several known ABA-inducible genes.
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  • 47
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    Plant molecular biology 18 (1992), S. 847-849 
    ISSN: 1573-5028
    Keywords: cDNA ; nucleotide sequence ; glycine-rich proteins ; chemical stress ; Zea mays
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  • 48
    ISSN: 1573-5028
    Keywords: Zea mays ; Mutator ; controlling element ; genomic DNA sequence
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    Topics: Biology
    Notes: Abstract The 4942 bp DNA sequence of Zea mays transposon MuA2 was determined. Previous evidence indicated MuA2 controls activity of the Mu1 transposon located in the mutable allele a1-mum2. MuA2 contains two large, ATG-initiated open reading frames (ORFs) of 612 and 232 codons, respectively, located on opposite strands. MuA2 produces two transcripts, each containing one of these ORFs. Four different tandem direct repeat sequences are located downstream of the 612 codon ORF. The restriction map of MuA2 is identical to that of transposon MuR1, which also is known to regulate mutability of a1-mum2. Furthermore, except for a single nucleotide, MuA2 is identical to the Mutator element Mu9.
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  • 49
    ISSN: 1573-5028
    Keywords: promoter ; electroporation ; protoplasts ; transient assay ; Agrobacterium ; Ti plasmid ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Regulatory elements controlling transcriptional activity of the mannopine synthase 2′ promoter (mas 2′) were defined by analysis of deletion mutants in transient expression assays in maize protoplasts. Deletion of the region between −305 and −290 containing sequence similarity to the octopine synthase (ocs) promoter element reduced activity by 67% compared to wild type activity. Less than 1% of the activity remained in 5′ deletions downstream of −153. Inclusion of various heterologous enhancer-like sequences immediately upstream of position −325 increased activity by up to 7.5-fold. Insertion of the −325 to −275 sequence alone, or in combination with heterologous enhancer-like elements, restored activity of some of the 5′-deletion mutants. Restoration of activity was not obtained with mutants deleted past position −127. Our results suggest that a single class of nuclear proteins from maize interact with high affinity at elements designated mas b (−306 to −275; mas 1′ element), d (−127 to −108), and e (−82 to −39; mas 2′ element) as well as the 20 bp element from the ocs promoter. Although the binding site at mas d only appears to accommodate a single protein, this element has the potential to make a weak, but positive, contribution to the activity of the mas 2′ promoter. The binding of nuclear proteins could not be demonstrated at mas a and c, both of which showed limited homology to the ocs element. Mutational evidence suggested that mas a and c may also contribute to mas 2′ transcription.
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  • 50
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    Plant molecular biology 25 (1994), S. 343-353 
    ISSN: 1573-5028
    Keywords: cytochrome P450 ; flower development ; meristem-specific gene ; Zea mays
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    Topics: Biology
    Notes: Abstract Four cDNA clones exhibiting preferential hybridization to transcripts present in developing maize tassels were isolated by differential screening. One of these cDNA clones hybridizes to transcripts detectable only in the shoot apex. The abundance of this transcript is significantly higher in developing inflorescence apices than in vegetative apices. DNA sequence analysis of a 2107 nucleotide cDNA clone corresponding to this transcript revealed that the transcript encodes a polypeptide of 547 amino acids, with a molecular mass of 58.4 kDa. This polypeptide shares significant sequence similarity with members of the cytochrome P450 monooxygenase gene superfamily, including the conserved C-terminal domains typical of the cytochrome P450 monooxygenases.
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  • 51
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    Plant molecular biology 25 (1994), S. 565-568 
    ISSN: 1573-5028
    Keywords: chromatin ; high-mobility-group (HMG) proteins ; protein stability ; Zea mays
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    Topics: Biology
    Notes: Abstract Chromosomal non-histone high-mobility-group (HMG) proteins represent essential components of eukaryotic chromatin and have also been isolated from a variety of plants. In maize, studies on structure and function of the two larger of the four major HMG proteins have recently been performed and are now extended by analysis of theirin vivo stability using pulse-chase experiments in a cell suspension culture. The half-life of the analyzed HMGa and HMGb proteins was found to be 65 h or more than 78 h, respectively.
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  • 52
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    Plant molecular biology 25 (1994), S. 817-828 
    ISSN: 1573-5028
    Keywords: B-Peru ; germinal revertants ; Mutator ; tandem duplication ; unequal recombination ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Theb locus ofZea mays encodes a transcriptional activator of the anthocyanin biosynthetic pathway. TheB-Peru allele is expressed in the aleurone layer of the seed, which results in dark purple pigmentation of this tissue. An unstableMutator-inducedB-Peru mutant allele,b-Perum220, displays weak, variable pigment and a high germinal reversion rate not characteristic of otherMutator insertions. Characterization of relevant regions ofb-Perum220 revealed aMu2 element insertion in one copy of a 534 bp sequence. This 534 bp sequence is tandemly triplicated in the progenitorB-Peru allele, upstream of theB-Peru transcription start site. In addition to theMu2 insertion, theb-Perum220 allele contains a newly formed large tandem duplication of 4.0 kb, which includes the promoter region and the first three exons of theB-Peru gene. TheMu2 element does not reside at any of the duplication breakpoints. The molecular study of eleven independent germinal revertants revealed five structural classes including structures in which the 4.0 kb tandem duplication is partially or completely deleted, theMu2 element is partially or completely deleted, or a combination of these events has occurred. We hypothesize that most of the revertants arose by unequal recombination between the duplicated regions. Based on these structural analyses, models are discussed to explain the reducedb gene expression inb-Perum220.
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  • 53
    ISSN: 1573-5028
    Keywords: primary response ; ferredoxin NADP+ oxidoreductase ; nitrate ; cycloheximide ; Zea mays ; roots
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    Topics: Biology
    Notes: Abstract To more fully understand the biochemical and molecular events which occur in plants exposed to nitrate, cDNAs whose accumulation was enhanced in nitrate- and cycloheximide-treated maize (Zea mays L. W64A × W182E) roots were isolated. The 340 bp Zmrprn 1 (for Zea mays root primary response to nitrate) cDNA also hybridized with a probe enriched for nitrate-induced sequences, and was characterized further. Sequence analysis of a near full-length cDNA (Zmrprn 1A) showed strong homology (〉90% amino acid identity) with a root ferredoxin-NADP+ oxidoreductase (FNR) of rice, and 45–50% amino acid identity with leaf FNR genes. When expressed in Escherichia coli, the Zmrprn 1A cDNA produced a protein with NADPH: ferricyanide reductase activity, consistent with the enzymatic properties of an FNR. The Zmrprn 1 cDNA hybridized with a 1.4 kb transcript which was expressed in the maize root primary response to nitrate. That is, mRNA levels in roots increased rapidly and transiently in response to external nitrate, and low levels of nitrate (10 μM) induced transcript accumulation. The accumulation of the Zmrprn 1 transcript was not prevented by cycloheximide, indicating that the cellular factor(s) required for expression were constitutively present in maize roots. The Zmrprn 1 mRNA accumulated specifically in response to nitrate, since neither K+ nor NH4 + treatment of roots caused transcript accumulation. Maize leaves had about 5% of the transcript level found in roots, indicating a strong preference for expression of Zmrprn 1 in roots. Analysis of maize genomic DNA indicated the presence of only a single gene or very small gene family for the Zmrprn 1. Together, the data indicate that Zmrprn 1A encodes a nitrate regulated maize root FNR.
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  • 54
    ISSN: 1573-5028
    Keywords: glutamine synthetase genes ; regulation ; nitrate ; ammonium ; Zea mays
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    Notes: Abstract The responses of the five cytosolic-type glutamine synthetase (GS1) genes of maize to treatment of hydroponically grown seedlings with 10 mM KNO3 or 10 mM NH4Cl were analyzed. Non-coding 3′ gene-specific hybridization probes and radioanalytic imaging were used to quantitate individual gene transcript levels in excised roots and shoots before treatment and at selected times after treatment. Genes GS1−1 and GS1−2 exhibited distinct organ-specific responses to treatment with either nitrogen source. The GS1−1 transcript level increased over three-fold in roots, but changed little if any in shoots. In contrast, the GS1−2 transcript level increased over two-fold in shoots, but decreased in roots after treatment. Increased transcript levels were evident at 4 h after treatment with either nitrogen source, with maximum accumulations present at 8 h after treatment with ammonium and at 10–12 h after treatment with nitrate. The GS1−3 gene transcript level showed little or no change after treatment with either nitrogen source. The GS1−4 gene transcript level remained constant in shoots of treated seedlings, whereas in roots, it exhibited relatively minor, but complex responses to these two nitrogen sources. The GS1−5 gene transcript is present in very small amounts in seedlings, making it difficult to analyze its response to metabolites in young plants. These results provide support for the possibility that different cytosolic GS genes of maize play distinct roles in nitrogen metabolism during plant growth and differentiation.
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  • 55
    ISSN: 1573-5028
    Keywords: ribosomal protein ; rps12 ; rpl20 ; clpP ; chloroplast genome ; Zea mays
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  • 56
    ISSN: 1573-5028
    Keywords: cycloheximide ; leaf ; nitrate induction ; nitrate reductase transcript ; root ; scutellum ; Zea mays
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    Notes: Abstract The nitrate induction of NADH:nitrate reductase mRNA in maize roots, scutella and leaves was investigated in the presence and absence of inhibitors of protein synthesis. In the absence of inhibitors, nitrate treatment caused a fairly rapid (2 to 3 h) increase in the level of the nitrate reductase transcript in all tissues. When cytoplasmic protein synthesis was inhibited by cycloheximide, nitrate reductase mRNA was induced by nitrate in all tissues to levels equal to or greater than those found with nitrate treatment alone. Treatment of maize tissues with cycloheximide in the absence of nitrate had only a small effect on the accumulation of the nitrate reductase mRNA. Inhibition of organellar protein synthesis with chloramphenicol also had little or no effect on nitrate-induced nitrate reductase mRNA accumuiation in roots and scutella, but did appear to partially inhibit appearance of transcript in leaves. Excision of scutella in the absence of nitrate was sufficient to cause some accumulation of the nitrate reductase transcript. Since cytoplasmic protein synthesis was not required for expression of nitrate reductase transcripts, induction of these transcripts by nitrate is a primary response of maize to this environmental signal. Thus, it appears that the signal transduction system mediating this response is constitutively expressed in roots, scutella and leaves of maize.
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  • 57
    ISSN: 1573-5028
    Keywords: Arabidopsis thaliana ; ATPase ; cpn60 ; developmental regulation ; molecular chaperones ; Zea mays
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    Notes: Abstract Mitochondria contain a nuclear-encoded heat shock protein, HSP60, which functions as a chaperonin in the post-translational assembly of multimeric proteins encoded by both nuclear and mitochondrial genes. We have isolated and sequenced full-length complementary DNAs coding for this mitochondrial chaperonin in Arabidopsis thaliana and Zea mays. Southern-blot analysis indicates the presence of a single hsp60 gene in the genome of A. thaliana. There is a high degree of homology at the predicted amino acid levels (43 to 60%) between plant HSP60s and their homologues in prokaryotes and other eukaryotes which indicates that these proteins must have similar evolutionarily conserved functions in all organisms. Northern- and western-blot analyses indicate that the expression of the hsp60 gene is developmentally regulated during seed germination. It is also heat-inducible. Developmental regulation of the (β-subunit) of F1-ATPase, an enzyme complex that is involved in the cyanide-sensitive mitochondrial electron transport system, indicates that imbibed embryos undergo rapid mitochondrial biogenesis through the early stages of germination. Based on the functional role of HSP60 in macromolecular assembly, these data collectively suggest that the presence of higher levels of HSP60 is necessary during active mitochondrial biogenesis, when the need for this protein is greatest in assisting the rapid assembly of the oligomeric protein structures.
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  • 58
    ISSN: 1573-5028
    Keywords: pollen-specific gene expression ; promoter analysis ; transient assays ; Tradescantia ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have previously reported the isolation and characterization of a gene (Zm 13) from Zea mays which shows a pollen-specific pattern of expression. Stably transformed tobacco plants containing a reporter gene linked to portions of the Zm 13 5′ flanking region show correct temporal and spatial expression of the gene. Here we present a more detailed analysis of the 5′ regions responsible for expression in pollen by utilizing a transient expression system. Constructs containing the β-glucuronidase (GUS) gene under the control of various sized fragments of the Zm13 5′ flanking region were introduced into Tradescantia and Zea mays pollen via high-velocity microprojectile bombardment, and monitored both visually and with a fluorescence assay. The results suggest that sequences necessary for expression in pollen are present in a region from −100 to −54, while other sequences which amplify that expression reside between −260 and −100. The replacement of the normal terminator with a portion of the Zm13 3′ region containing the putative polyadenylation signal and site also increased GUS expression. While the −260 to −100 region contains sequences similar to other protein-binding domains reported for plants, the −100 to −54 region appears to contain no significant homology to other known promoter fragments which direct pollen-specific expression. The microprojectile bombardment of Tradescantia pollen appears to be a good test system for assaying maize and possibly other monocot promoter constructs for pollen expression.
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  • 59
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    The protein journal 12 (1993), S. 451-457 
    ISSN: 1573-4943
    Keywords: Carboxyl group ; Woodward's reagentK ; PEP carboxylase ; active site ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Maize leaf phosphoenolpyruvate carboxylase was completely and irreversibly inactivated by treatment with micromolar concentrations of Woodward's reagentK (WRK) for about 1 min. The inactivation followed pseudo-first-order reaction kinetics. The order of reaction with respect to WRK showed that the reagent causes formation of reversible enzyme inhibitor complex before resulting in irreversible inactivation. The loss of activity was correlated to the modification of a single carboxyl group per subunit, even though the reagent reacted with 2 carboxyl groups per protomer. Substrate PEP and PEP + Mg2+ offered substantial protection against inactivation by WRK. The modified enzyme showed a characteristic absorbance at 346 nm due to carboxyl group modification. The modified enzyme exhibited altered surface charge as seen from the elution profile on FPLC Mono Q anion exchange column. The modified enzyme was desensitized to positive and negative effectors like glucose-6-phosphate and malate. Pretreatment of PEP carboxylase with diethylpyrocarbonate prevented WRK incorporation into the enzyme, suggesting that both histidine and carboxyl groups may be closely physically related. The carboxyl groups might be involved in metal binding during catalysis by the enzyme.
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    Plant molecular biology 15 (1990), S. 121-125 
    ISSN: 1573-5028
    Keywords: Alcohol dehydrogenase ; DNA methylation ; Zea mays
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    Topics: Biology
    Notes: Abstract Using a battery of methylation-sensitive restriction enzymes, cytosine methylation at 23 sites in a 7.6 kb region surrounding the Alcohol dehydrogenase-1 (Adh1) gene was measured in DNA prepared from immature maize cobs. Both the 5′ upstream region and the entire coding region were hypomethylated in the two alleles examined. Methylation in Adh1 is independent of changes in Mutator transposable element methylation. The role of DNA methylation in Adh1 gene regulation is discussed.
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  • 61
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    Plant molecular biology 17 (1991), S. 265-268 
    ISSN: 1573-5028
    Keywords: cDNA sequence ; cytoplasmic ribosomes ; gene copy number ; ribosomal protein S11 ; steadystate mRNA level ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We isolated a Zea mays cDNA encoding the 40S subunit cytoplasmic ribosomal protein S11. The nucleotide sequence was determined and the derived amino acid sequence compared to the corresponding Arabidopsis thaliana protein showing an homology of 90%. This ribosomal protein is encoded by a small multigene family of at least two members. The mRNA steady-state level is about one order of magnitude higher in rapidly growing parts of the plant such as the roots and shoots of seedlings compared to fully expanded leaf tissue.
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  • 62
    ISSN: 1573-5028
    Keywords: chromatin structure ; DNA methylation ; DNA topoisomerase II ; plants ; Zea mays
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    Topics: Biology
    Notes: Abstract We have investigated the presence of higher-order chromatin structures in different maize tissues. Taking advantage of the pulsed-field gel electrophoresis technique to analyse large DNA fragments from intact nuclei and cells, we have determined the size distribution of the high-molecular-weight DNA fragments obtained from chromatin degradation by endogenous nucleases in isolated nuclei. Chromatin digestion leads to the appearance of stable DNA fragments of about 50 kb in all the tissues examined, suggesting the folding of DNA in higher-order chromatin domain structures. It has been reported that such chromatin domains are formed by loops of the 30 nm fibres anchored to the nuclear matrix by a complex set of proteins, including DNA topoisomerase II. Treatment of maize protoplasts with the calcium ionophore A23187 and the antitumour drug VM-26, which specifically inhibit the religation of the cleaved DNA in the topoisomerase II reaction, also produces the 50 kb structure. Analysis of the DNA contained in the 50 kb chromatin structure shows a higher degree of methylation than in bulk maize chromosomal DNA. The role of methylated DNA in the chromatin folding is discussed.
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  • 63
    ISSN: 1573-5028
    Keywords: chromatin ; high-mobility-group (HMG) proteins ; Vicia faba ; Zea mays
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    Notes: Abstract Chromosomal high-mobility-group (HMG) proteins occur ubiquitously in eukaryotes and their common structural and biochemical features indicate a critical role. In this context, we compared structural and functional aspects of HMG proteins from the monocotyledonous plant maize and the dicotyledonous plant Vicia faba. Besides biochemical similarities and immunological differences found between these proteins, the isolation and characterization of a cDNA encoding the V. faba homologue of the maize HMGa protein revealed great similarities between these two proteins, including the HMG-box DNA-binding motif and an acidic domain. Therefore, like the maize HMGa protein, the V. faba HMG protein belongs to the vertebrate HMG1 family, which consists of HMG proteins and transcription factors of various eukaryotes.
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  • 64
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    Plant molecular biology 23 (1993), S. 825-838 
    ISSN: 1573-5028
    Keywords: endosperm ; lysine ; opaque-2 ; non-zein proteins ; Zea mays ; zeins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The reduction of zein synthesis in the maize (Zea mays L.) opaque-2 mutant is associated with an increased percentage of lysine in the endosperm protein. When expressed on an endosperm basis, we found that W64A opaque-2 contains 490 μg of lysine compared with 350 μg in W64A normal. SDS-PAGE analysis of endosperm proteins indicated that several non-zein proteins are more abundant in the mutant than in normal genotype. To determine the subcellular origin of these proteins, we separated an endosperm homogenate from developing kernels by sucrose density gradient centrifugation and used marker enzyme assays and immunoblot analyses to identify cellular components. Amino acid analysis of proteins in the gradient fractions showed that the majority of the lysine occurs in soluble proteins at the top of the gradient. To identify these proteins, we prepared a complex antiserum against the entire soluble protein fraction and used it to immunoscreen an endosperm cDNA expression library. Sequence analysis of clones identified mRNAs involved in carbohydrate metabolism, amino acid biosynthesis, and protein synthesis. RNA dot blot hybridization analysis with these clones revealed significant variation in the levels of transcripts between normal and opaque-2 endosperm, but we identified several mRNAs that are elevated in opaque-2 and that may encode proteins responsible for the enhanced lysine content.
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  • 65
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    Plant molecular biology 25 (1994), S. 137-140 
    ISSN: 1573-5028
    Keywords: DNA acquisition ; retrotransposon ; retrovirus ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The recently identified maize retroelement Bs1 encodes domains of the plasma membrane H+-ATPase. This is the first example of host DNA captured by a plant retroelement and resembles the acquisition of oncogenes by vertebrate retroviruses. The ability to capture sequences from its host provides plant retroelements with a mechanism to alter gene structure which could be important for evolutionary adaptive change.
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  • 66
    ISSN: 1573-5028
    Keywords: intron ; maize ; splicing ; vectors ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The mechanisms of intron recognition and processing have been well-studied in mammals and yeast, but in plants the biochemistry of splicing is not known and the rules for intron recognition are not clearly defined. To increase understanding of intron processing in plants, we have constructed new pairs of vectors, pSuccess and pFail, to assess the efficiency of splicing in maize cultured cells. In the pFail series we use translation of pre-mRNA to monitor the amount of unspliced RNA. We inserted an ATG codon in the Bz2 (Bronze-2) intron in frame with luciferase: this construct will express luciferase activity only when splicing fails. In the pSuccess series the spliced message is monitored by inserting an ATG upstream of the Bz2 intron in frame with luciferase: this construct will express luciferase activity only when splicing succeeds. We show here, using both the wild-type Bz2 intron and the same intron with splice site mutations, that the efficiency of splicing can be estimated by the ratio between the luciferase activities of the vector pairs. We also show that mutations in the unique U-rich motif inside the intron can modulate splicing. In addition, a GC-rich insertion in the first exon increases the efficiency of splicing, suggesting that exons also play an important role in intron recognition and/or processing.
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  • 67
    ISSN: 1573-5028
    Keywords: Glutathione S-transferase ; herbicide safener ; inducible gene expression ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract Several GSTs have been characterised in maize. GST I is a homodimer of 29 kDa subunits, GST II a hetrodimer of 27 kDa and 29 kDa subunits and GST IV a homodimer of 27 kDa subunits. We report the isolation and characterization of a herbicide-safener inducible cDNA clone, GST-27. Based on partial amino acid sequence, GST-27 encodes the 27 kDa subunit present in both glutathione S-transferase isoforms GST II and IV. Northern blotting was used to compare the expression patterns of GST-27 with that of GST-29. Transcripts corresponding to GST-27 were found to be constitutively expressed in RNA isolated from the root, but no expression was detected in RNA isolated from aerial parts of the plant. The application of herbicide safener caused a dramatic increase in the expression of GST-27 in all aerial plant parts tested. GST-29 was found to be constitutively expressed in RNA isolated from a number of maize tissues. The basal level of GST-29 expression showed a minimal increase upon herbicide safener treatment. Although a range of hormonal, environmental and physiological stimuli failed to elevate GST-27 levels, some increase in GST-27 mRNA was observed in the late stages of leaf senescence and after treatments resulting in phytotoxic effects.
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    The journal of membrane biology 114 (1990), S. 133-142 
    ISSN: 1432-1424
    Keywords: Ca2+ transport ; plasma membrane ; Ca2+ pump ; pH gradient ; Ca2+/H+ antiporter ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Plasma membrane vesicles, which are mostly right side-out, were isolated from corn leaves by aqueous two-phase partitioning method. Characteristics of Ca2+ transport were investigated after preparing inside-out vesicles by Triton X-100 treatment.45Ca2+ transport was assayed by membrane filtration technique. Results showed that Ca2+ transport into the plasma membrane vesicles was Mg-ATP dependent. The active Ca2+ transport system had a high affinity for Ca2+(K m (Ca2+)=0.4 μm) and ATP(K m (ATP)=3.9 μm), and showed pH optimum at 7.5. ATP-dependent Ca2+ uptake in the plasma membrane vesicles was stimulated in the presence of Cl− or NO 3 − . Quenching of quinacrine fluorescence showed that these anions also induced H+ transport into the vesicles. The Ca2+ uptake stimulated by Cl− was dependent on the activity of H+ transport into the vesicles. However, carbonylcyanidem-chlorophenylhydrazone (CCCP) and VO 4 3− which is known to inhibit the H+ pump associated with the plasma membrane, canceled almost all of the Cl−-stimulated Ca2+ uptake. Furthermore, artificially imposed pH gradient (acid inside) caused Ca2+ uptake into the vesicles. These results suggest that the Cl−-stimulated Ca2+ uptake is caused by the efflux of H+ from the vesicles by the operation of Ca2+/H+ antiport system in the plasma membrane. In Cl−-free medium, H+ transport into the vesicles scarcely occurred and the addition of CCCP caused only a slight inhibition of the active Ca2+ uptake into the vesicles. These results suggest that two Ca2+ transport systems are operating in the plasma membrane from corn leaves, i.e., one is an ATP-dependent active Ca2+ transport system (Ca2+ pump) and the other is a Ca2+/H+ antiport system. Little difference in characteristics of Ca2+ transport was observed between the plasma membranes isolated from etiolated and green corn leaves.
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    The journal of membrane biology 119 (1991), S. 277-288 
    ISSN: 1432-1424
    Keywords: Ca2+-activated K+ current ; Ca2+ channel ; Zea mays ; 1,4-dihydropyridine ; phenylalkylamine ; patch clamp
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary The voltage- and time-dependent K+ current,I K + out , elicited by depolarization of corn protoplasts, was inhibited by the addition of calcium channel antagonists (nitrendipine, nifedipine, verapamil, methoxyverapamil, bepridil, but not La3+) to the extracellular medium. These results suggested that the influx of external Ca2+ was necessary for K+ current activation. The IC50, concentration of inhibitor that caused 50% reduction of the current, for nitrendipine was 1 μm at a test potential of +60 mV following a 20-min incubation period. In order to test whether intracellular Ca2+ actuated the K+ current, we altered either the Ca2+ buffering capacity or the free Ca2+ concentration of the intracellular medium (pipette filling solution). By these means,I K + out could be varied over a 10-fold range. Increasing the free Ca2+ concentration from 40 to 400nm also shifted the activation of the K+ current toward more negative potentials. Maintaining cytoplasmic Ca2+ at 500nm with 40nm EGTA resulted in a more rapid activation of the K+ current. Thus the normal rate of activation of this current may reflect changes in cytoplasmic Ca2+ on depolarization. Increasing intracellular Ca2+ to 500nm or 1 μm also led to inactivation of the K+ current within a few minutes. It is concluded thatI K + out is regulated by cytosolic Ca2+, which is in turn controlled by Ca2+ influx through dihydropyridine-, and phenylalkylamine-sensitive channels.
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  • 70
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    Mycorrhiza 5 (1994), S. 119-124 
    ISSN: 1432-1890
    Keywords: 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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  • 71
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    Theoretical and applied genetics 81 (1991), S. 635-640 
    ISSN: 1432-2242
    Keywords: Catalase null ; Zea mays ; Catalase transcript ; Transposable element
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The molecular basis for a shorter Cat2 transcript in maize lines null for the CAT-2 catalase iso/yme was investigated using cDNA libraries and genomic DNA blots. Sequence comparison of partial Cat2 cDNAs obtained from two CAT-2 null lines and a wild-type CAT-2 encoding cDNA showed that the Cat2 null transcripts diverged from the wild-type transcript but remained homologous to each other. Genomic DNA blots indicated that the missing portion of the transcript is present in the genomes of lines null for the CAT-2 isozyme. Differences in the hybridization patterns of normal and null lines were revealed when genomic DNA blots were probed with the full-length Cat2 cDNA, a Cat2 gene-specific probe, and a “null sequence” probe. Together, the DNA blotting results suggest that a rearrangement of the Cat2 gene has occurred in the CAT-2 null lines. The available data suggest that the CAT-2 null mutation in maize is due to a DNA insertion into the Cat2 gene.
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  • 72
    ISSN: 1432-2242
    Keywords: Sorghum ; Zea mays ; Phylogeny rDNA sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The phylogenetic relationships of the genus Sorghum and related genera were studied by sequencing the nuclear ribosomal DNA (rDNA) internal transcribed spacer region (ITS). DNA was extracted from 15 Sorghum accessions, including one accession from each of the sections Chaetosorghum and Heterosorghum, four accessions from Parasorghum, two accessions from Stiposorghum, and seven representatives from three species of the section Sorghum (one accession from each of S. propinquum and S. halepense, and five races of S. bicolor). The maize (Zea mays) line, H95, and an accession from Cleistachne sorghoides were also included in the study. Variable nucleotides were used to construct a strict consensus phylogenetic tree. The analyses indicate that S. propinquum, S. halepense and S. bicolor subsp. arundinaceum race aethiopicum may be the closest wild relatives of cultivated sorghum; Sorghum nitidum may be the closest 2n=10 relative to S. bicolor, the sections Chaetosorghum and Heterosorghum appear closely related to each other and more closely related to the section Sorghum than Parasorghum; and the section Parasorghum is not monophyletic. The results also indicate that the genus Sorghum is a very ancient and diverse group.
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  • 73
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    Theoretical and applied genetics 89 (1994), S. 259-264 
    ISSN: 1432-2242
    Keywords: RFLP ; Bootstrap ; Sampling variance ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Knowledge of genetic relationships among genotypes is useful in a plant breeding program because it permits the organization of germplasm and provides for more efficient sampling. The genetic distance (GD) among genotypes can be estimated using random restriction fragment length polymorphisms (RFLPs) as molecular markers. Knowledge of the sampling variance associated with RFLP markers is needed to determine how many markers are required for a given level of precision in the estimate of GD. The sampling variance for GD among all pairs of 37 maize (Z. mays L.) inbred lines was estimated from 1202 RFLPs. The 1202 polymorphisms were generated from 251 enzyme-probe combinations (EPC). The sampling variance was used to determine how large a sample of RFLPs was required to provide a given level of precision. The coefficient of variation (CV) associated with GD has a nearly linear relationship between its expected standard deviation and mean. The magnitude of the decrease in the mean CV for GD with increasing numbers of bands was dependent upon the sampling unit; e.g., individual polymorphic bands vs EPC, and the degree of relatedness among the inbreds compared. The rate of reduction in mean CV with increasing sample size was the same regardless of the restriction enzyme used, BamHI, EcoRI or HindIII, when the bootstrap sampling units were individual polymorphic bands. In constrast, although the rate of reduction (slopes) was the same, the intercepts of the mean CVs were different when EPCs were used as the bootstrap sampling unit. This difference was due to the higher number of bands per EPC in BamHI (4.94) compared with EcoRI (4.83) and HindIII (4.63).
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  • 74
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    Theoretical and applied genetics 81 (1991), S. 185-188 
    ISSN: 1432-2242
    Keywords: Zea mays ; Recombination ; A1 locus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We find that recombination between two alleles of the maize A1 locus that contain transposon insertions at known molecular positions can occur at 0.04–0.08 cM per kbp (centimorgan per kilobase pair), which is two orders of magnitude higher than the recombination rate for the whole maize genome. It is however, close to the rates found within the bronze locus, another maize structural gene for which both genetic and molecular data are available. This observation supports the idea that the genome consists of regions that are highly recombinogenic — in some cases, at least, structural genes — interspersed with regions that are less recombinogenic.
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  • 75
    ISSN: 1432-2242
    Keywords: Zea mays ; Chromosome doubling ; Amiprophos methyl ; Pronamide ; Oryzalin ; Trifluralin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Four antimicrotubule herbicides, amiprophosmethyl (APM), pronamide, oryzalin, and trifluralin, were evaluated for their ability to induce chromosome doubling in anther-derived, haploid maize callus. Effects of various herbicide treatments on the growth and regenerative capacity of callus along with the ploidy and seed set of regenerated plants were determined. Flow cytometric analysis was also used to measure changes in ploidy levels of callus cells following treatments. More than 50% of the cells were doubled in chromosome number after the haploid callus was treated with 5 or 10 μ M APM or 10 μ M pronamide for 3 days. A similar proportion of plants regenerated from the treated callus produced seed upon self-pollination. APM and pronamide did not inhibit callus growth at these concentrations and the treated callus retained a high plant regeneration capacity. Oryzalin very effectively induced chromosome doubling, but severely inhibited the growth of regenerable callus and plant regeneration. Trifluralin induced chromosome doubling in a small proportion of cells at lower concentrations (0.5 and 1 μ M), however, at a higher concentration (5 μM) it inhibited callus growth and plant regeneration. The results indicate that APM and pronamide may be useful agents for inducing chromosome doubling of anther-derived maize haploid callus at very low concentrations.
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  • 76
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    Theoretical and applied genetics 84 (1992), S. 798-802 
    ISSN: 1432-2242
    Keywords: Zea mays ; C-banding ; Cell size ; Chloroplast number ; Cold tolerance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary C-band number, guard cell length, and chloroplast number per guard cell were determined for eight maize populations. These populations consisted of maize selected for cold tolerance at the University of Nebraska as well as the original unselected populations. The genome size of these populations had previously been determined. C-band number fluctuated concertedly with the changes in genome size indicating that deletions and additions of constitutive heterochromatin occurred during selection, resulting in altered genome sizes. Guard cell size of all the cold tolerant populations was greater than the cell size of the respective nonselected populations. Chloroplast number per guard cell was also higher in all the cold tolerant populations than in their parental populations, but the increases were not statistically significant. The results indicate that changes in genome size that occurred during selection for cold tolerance are the result of changes in amounts of C-band heterochromatin and that the selection process results in an increase in cell size in the cold tolerant populations.
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  • 77
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    Theoretical and applied genetics 86 (1993), S. 975-984 
    ISSN: 1432-2242
    Keywords: Chemiluminescence ; Genotyping costs ; RAPD ; RFLP ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three molecular marker protocols, chemiluminescent restriction fragment length polymorphisms (c-RFLPs), radioactivity-based restriction fragment length polymorphisms (r-RFLPs), and randomly amplified DNA polymorphisms (RAPDs) were compared in terms of cost and time efficiency. Estimates of cost of supplies and time requirements were obtained from simulations of maize (Zea mays L.) genotyping experiments utilizing protocols currently in use. The increase in total cost with increasing numbers of individuals genotyped and markers analyzed is higher for RAPDs than for RFLPs. RAPDs were generally found to be more cost and time efficient for studies involving small sample sizes, while RFLPs have the advantage for larger sample sizes. Because of the shorter exposure times involved, c-RFLPs require less time than r-RFLPs to obtain a given amount of information. Variations in the protocols, such as number of re-uses of Southern blots or cost of Taq DNA polymerase per reaction of amplification, also affect the relative merits of RAPDs and RFLPs. Two examples were analyzed where molecular markers are used: a germ plasm survey and quantitative trait loci (QTL) mapping in a segregating population. No protocol was found to be the most cost and time efficient over the entire range of sample sizes and number of marker loci studied.
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  • 78
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    Theoretical and applied genetics 88 (1994), S. 780-784 
    ISSN: 1432-2242
    Keywords: Acetochlor tolerance ; Gene expression Pollen selection ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The objectives of this research were to determine if genes controlling the reaction to the herbicide acetochlor in maize (Zea mays L.) are active during both the haploid and the diploid phases of the life cycle and if pollen selection can be utilized for improving sporophytic resistance. Pollen of eight inbred lines, previously characterized through sporophytic analysis for the level of tolerance to acetochlor, showed a differential reaction to the herbicide forin vitro tube length; moreover, such pollen reactions proved to be significantly correlated (r =0.786*,df=6) with those of the sporophytes producing the pollen. Pollen analysis of two inbred lines (i.e. Mo17, tolerant, and B79, susceptible) and their single cross showed that thein vitro pollen-tube length reaction of the hybrid was intermediate between those of two parents. An experiment on pollen selection was then performed by growing tassels of Mo17xB79 in the presence of the herbicide. Pollen obtained from treated tassels showed a greater tolerance to acetochlor, assessed asin vitro tube length reaction, than pollen obtained from control tassels. Moreover, the backcross [B79 (Mo17xB79)] sporophytic population obtained using pollen from the treated tassels was more tolerant (as indicated by the fresh weight of plants grown in the presence of the herbicide) than was the control backcross population. The two populations did not differ when grown without the herbicide. These findings indicate that genes controlling the reaction to acetochlor in maize have haplodiploid expression; consequently, pollen selection can be applied for improving plant tolerance.
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  • 79
    ISSN: 1432-2242
    Keywords: Denaturing gradient gel electrophoresis ; DNA polymorphism ; Restriction fragment length polymorphism ; Zea mays ; B-A translocation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have used denaturing gradient gel electrophoresis (DGGE) to identify genomic DNA polymorphism in maize (Zea mays L.). DGGE probes detect polymorphism in maize at a frequency comparable to the incidence of restriction fragment length polymorphism (RFLP). Probes identifying polymorphism were mapped to maize chromosome arms by utilizing DGGE and maize lines carrying B-A chromosomal translocations. The methods for library construction, probe screening, and genome analysis, described here for maize, can also be applied to the genomic analysis of other organisms.
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  • 80
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    Theoretical and applied genetics 80 (1990), S. 705-711 
    ISSN: 1432-2242
    Keywords: Zea mays ; Catalase ; Gene expression ; Pollen ; superoxide dismutase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The expression of the Cat and Sod genes encoding the multiple catalases and superoxide dismutases in maize has been studied in maize pollen at anther dehiscence. Pollen from three catalase activity variant inbred lines was examined. Zymorgram analysis, immunoassays, and RNA blots show that, in the lines used, only the CAT-1 catalase isozyme is expressed in mature pollen; the CAT-2 and CAT-3 isozymes are not. The data presented further demonstrate that the Cat1 gene is transcribed and translated after tetrad formation. The relative protein levels of the various superoxide dismutase (SOD) isozymes appear to be similar in pollen and scutella, and correspond to the relatively low levels observed for the different Sod transcripts in these tissues. The presence of “double” transcripts for Sod3 and Sod4A is discussed.
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  • 81
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    Theoretical and applied genetics 81 (1991), S. 162-165 
    ISSN: 1432-2242
    Keywords: Genotype x environment interaction ; Stability ; Maize ; Zea mays ; Heterogeneity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Genotype x environment (GE) interaction encountered in experiments complicates genotype selection and varietal recommendation. The integration of yield and stability of genotypes into a single parameter may make selection and recommendation easier. Kang developed a rank-sum method that allows selection for both yield and the stability variance statistics (σ i 2 or s i 2 ) of Shukla. The objective of this research was to compare the rank-sum selection method to selection based on yield alone in five international maize (Zea mays L.) yield trials. Ranks were assigned for yield (the highest mean yield received a rank of 1) and for σ i 2 and s i 2 (the lowest value received a rank of 1). The yield and σ i 2 ranks and/or the yield and s i 2 ranks for each genotype were summed. Each trial contained two reference entries (REs). Yield rank or rank-sum of each genotype was compared to yield rank or rank-sum of the best RE (BRE). GE interaction was significant for all trials. Heterogeneity in the GE interaction due to the linear effect of a covariate (differences in fertility and/or cultural practices) was significant in Trials 1, 2, and 5. Overall, in all trials, 29 genotypes were selected on the basis of yield alone. On the basis of σ i 2 and yield rank-sum, 32 genotypes were identified, with 11 being lower yielding than the 29 yield-based selections. On the basis of s i 2 and yield rank-sum, 31 genotypes were selected, with 11 being lower yielding than the yield-bases selections. Obviously, yield is sacrificed when the rank-sum method is used in the selection process. However, selection based on yield alone may not be adequate when GE interaction is significant because of testing in diverse environments.
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    Theoretical and applied genetics 81 (1991), S. 227-232 
    ISSN: 1432-2242
    Keywords: Callus cultures ; RFLP ; Zea mays ; Oryza sativa ; Regeneration ; Somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Tissue culture of the Zea mays inbred line A188 resulted in the regeneration of plants having a high level of phenotypic variation compared to seed-grown control plants. To determine how such variation was induced and whether this could be related to specific in vitro culture methods, callus cultures were established and maintained on different, commonly used culture media. Plants were regenerated and the genomic DNA of callus cultures and regenerants analysed for RFLP differences. The results show that regardless of the gene probe used, callus formation resulted in significant deviations from the DNA pattern normally found in seed-grown control plants. Alterations in gene copy number also occurred. As differentiation and organogenesis began, the level of DNA variation fell, and most of the regenerated plants showed a genetic similarity to the controls; those with RFLP differences were the somaclonal variants.
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  • 83
    ISSN: 1432-2242
    Keywords: Zea mays ; Parentage determination ; Varietal identification ; Genetic polymorphisms ; Genetic markers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Using a novel procedure based on the polymerase chain reaction, we have developed a rapid, efficient, and economical method for identifying plant genotypes. The arbitrarily primed polymerase chain reaction (AP-PCR) generates reproducible fingerprints from any organism, without the need for DNA sequence information. These fingerprints include DNA fragment polymorphisms that can be (1) used for varietal identification and parentage determination, (2) followed in segregating populations produced by crosses, (3) used as markers for the construction of genetic maps, and (4) used to generate dendograms of phylogenetic relationships, especially at the intraspecific level. AP-PCR requires only minute quantities of DNA (10–25 ng per reaction) and therefore can be used in situations in which DNA is limiting. We demonstrate the use of AP-PCR to identify inbred parents of hybrid maize plants in double-blind experiments.
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  • 84
    ISSN: 1432-2242
    Keywords: Zea mays ; Restriction fragment length polymorphisms (RFLPs)-Phosphorus (P) stress-quantitative trait loci (QTLs)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary An understanding of the genetic nature underlying tolerance to low-phosphorus (low-P) stress could aid in the efficient development of tolerant plant strains. The objective of this study was to identify the number of loci in a maize (Zea mays L.) population segregating for tolerance to low-P stress, their approximate location, and the magnitude of their effect. Seventy-seven restriction fragment length polymorphisms (RFLPs) were identified and scored in a maize F2 population derived from a cross between line NY821 and line H99. The F2 individuals were self-pollinated to produce F3 families. Ninety F3 families were grown in a sand-alumina system, which simulated diffusion-limited, low-P soil conditions. The F3 families were evaluated for vegetative growth in a controlled-environment experiment. To identify quantitative trait loci (QTLs) underlying tolerance to low-P stress, the mean phenotypic performances of the F3 families were contrasted based on genotypic classification at each of 77 RFLP marker loci. Six RFLP marker loci were significantly associated with performance under low-P stress (P〈0.01). One marker locus accounted for 25% of the total phenotypic variation. Additive gene action was predominant for all of the QTLs identified. Significant marker loci were located on four separate chromosomes representing five unlinked genomic regions. Two marker loci were associated with an additive by additive epistatic interaction. A multiple regression model including three marker loci and the significant epistatic interaction accounted for 46% of the total phenotypic variation. Heterozygosity per se was not predictive of phenotypic performance.
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  • 85
    ISSN: 1432-2242
    Keywords: Zea mays ; Aspartate kinase Threonine-overproducing mutants ; Lysine ; Methionine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The aspartate-derived amino-acid pathway leads to the production of the essential amino-acids lysine, methionine, threonine and isoleucine. Aspartate kinase (AK) is the first enzyme in this pathway and exists in isoforms that are feedback inhibited by lysine and threonine. Two maize (Zea mays L.) threonine-overproducing, lysine-insensitive AK mutants (Ask1-LT19 and Ask2-LT20) were previously isolated. The present study was conducted to determine the map location of Ask2 and to examine the amino-acid profiles of the Ask mutants. The threonine-overproducing trait conferred by Ask2-LT20 was mapped to the long arm of chromosome 2. Both mutants exhibited increased free threonine concentrations (nmol/mg dry weight) over wild-type. The percent free threonine increased from approximately 2% in wild-type kernels to 37–54% of the total free amino-acid pool in homozygous mutant kernels. Free methionine concentrations also increased significantly in homozygous mutants. Free lysine concentrations were increased but to a much lesser extent than threonine or methionine. In contrast to previous studies, free aspartate concentrations were observed to decrease, indicating a possible limiting factor in threonine synthesis. Total (free plus protein-bound) amino-acid analyses demonstrated a consistent, significant increase in threonine, methionine and lysine concentrations in the homozygous mutants. Significant increases in protein-bound (total minus free) threonine, methionine and lysine were observed in the Ask mutants, indicating adequate protein sinks to incorporate the increased free amino-acid concentrations. Total amino-acid contents (nmol/kernel) were approximately the same for mutant and wild-type kernels. In five inbred lines both Ask mutations conferred the threonine-overproducing phenotype, indicating high expressivity in different genetic backgrounds. These analyses are discussed in the context of the regulation of the aspartate-derived amino-acid pathway.
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  • 86
    ISSN: 1432-2242
    Keywords: Zea mays ; Opaque-2 ; RFLPs Marker-assisted selection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Quality Protein Maize (QPM) is a name given to genetically modified opaque-2 maize with hard endosperm. The opaque-2 mutation conditions a reduction in the amount of zein seed storage protein; zeins are deficient in the essential amino acids lysine and tryptophan, and mutant seed have a higher nutritional value. To utilize the potential of opaque-2 maize, elite inbreds can be converted to o2/o2 forms and subsequently to hard endosperm opaque-2. Since opaque-2 is recessive and endosperm specific, conventional backcross procedures to convert elite inbreds to opaque-2 forms are inefficient. To alleviate this problem, a marker-assisted selection procedure was developed for the Texas A&M University Quality Protein Maize breeding program. Hybridization of an O2 cDNA probe to blots of DNA from plants carrying O2 and o2 alleles showed that restriction fragment length polymorphisms (RFLPs) exist between the W64A o2 allele and O2 alleles of Mo17 and TX5855 inbred lines. To identify the opaque2 genotypes in segregating populations, an RFLP marker assay combining the O2 cDNA probe and HindIII-digestion of genomic DNA was developed. The effectiveness of the O2 RFLP marker assay was tested under field conditions using F2 and backcross populations of several hard endosperm opaque-2 lines. A comparison of the genotypes identified by RFLP analysis with the seed phenotypes of the next generation indicated that this procedure is accurate and can be used for identifying O2/O2, O2/o2, and o2/o2 genotypes of individual juvenile plants in breeding populations.
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  • 87
    ISSN: 1573-9368
    Keywords: transient gene expression ; β-glucuronidase ; luciferase ; Hordeum vulgare ; Zea mays ; Nicotiana tabacum ; quantitation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The particle gun approach was used for the quantification of promoter efficiency in a test system for transient gene expression. β-Glucuronidase was used as reporter gene for determining promotote strength. The variability inherent in this gene transfer system was considerably reduced by calculating a transformation efficiency factor given by the expression of a cotransferred second reporter gene (firefly luciferase). The calibration of β-glucuronidase activity by the transformation efficiency factor caused a lower statistical variance of the values and allowed reliable results to be obtained with a smaller set of repetitions. The CaMV 35S promoter (as a control) and the monocot-specific promoters for maize polyubiquitin1, rice actin 1 and the maize-derivedEmu were characterized and compared with respect to expression strength, as tested under identical conditions in suspension cell cultures of maize, barley and tobacco. Compared to the 35S promoter, the monocot-specific promoters show up to 15-fold higher expression in maize and barley but give only weak expression in tobacco. No expression was found for the rice actin 1 promoter in tobacco. The level of reporter gene expression is influenced by the osmotic potential in the agar medium. For theEmu promoter, the calibrated β-glucuronidase activities remained mearly constant at low sucrose concentrations. Above 8% sucrose, the calibrated activities increased steadily with increasing osmotic conditions, reaching a three-to four-fold higher level at the highest sucrose concentration (32%) as compared to the standard concentration (4% sucrose) in the medium.
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  • 88
    ISSN: 1615-6102
    Keywords: Cell fractionation ; Glucan synthase ; Glycosyl linkage ; Golgi apparatus ; Xylan synthase ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Membranes from etiolated maize seedlings were isolated using sucrose gradients for in vitro studies of polysaccharide synthesis. Following downward centrifugation, flotation centrifugation improved the purity of membrane fractions, in particular the Golgi apparatus. Based on naphthylphthalamic acid binding to plasma membrane and inosine-5′-diphosphatase activity in Golgi apparatus, flotation centrifugation removed about 70% of the plasma membrane which cosedimented with the Golgi apparatus in downward centrifugation. The addition of chelators during flotation centrifugation allowed separation of the Golgi apparatus from endoplasmic reticulum, as indicated by NADH cytochromec reductase activity. Glucan and xylan synthase activities were measured as the radioactivity incorporated from either UDP-14C-glucose or UDP-14C-xylose into 80% ethanol insoluble materials. Glucan synthase activity at a substrate concentration of 1 mM UDP-glucose without CaCl2 was greatest in fractions enriched in Golgi apparatus, but in the presence of 3 mM CaCl2 the activity was greatest in fractions enriched in plasma membrane. Glucan synthase activity at a substrate concentration of 10μM UDP-glucose in the presence of 3 mM MnCl2 was greatest in fractions enriched in plasma membrane, but was also high in fractions enriched in Golgi apparatus. Xylan synthase activity, at a substrate concentration of 1 μM UDP-xylose in the presence of 3 mM MnCl2, was greatest in fractions enriched in Golgi apparatus. To further characterize these synthase reactions, the glycosyl linkages of the products formed were analyzed with a gas chromatograph coupled to a radiogas proportional counter. With the substrate, UDP-14C-glucose, and fractions enriched in Golgi apparatus, both (1→3)- and (1→4)-radioactive glucosyl linkages were formed, whereas the main linkage formed by fractions enriched in plasma membrane was (1→3)-glucosyl. With the substrate, UDP-14C-xylose, mostly (1→4)-xylosyl and some terminal-xylosyl linkages were formed by fractions enriched in Golgi apparatus. Only xylan synthase activity copurified with Golgi apparatus and, because plasma membrane lacked this activity, xylan synthase may be used as a reasonable indicator of Golgi apparatus.
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    Protoplasma 169 (1992), S. 49-56 
    ISSN: 1615-6102
    Keywords: Lectin ; Plasma membrane ; Protoplast isolation ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary An improved method for the mechanical release of protoplasts from plant tissues is described. The historically-low yield of mechanically-released protoplasts is greatly increased by use of a simple electrically-driven tissue sheer and by optimization of various other steps in the procedure. As counted by light microscopy of a purified preparation, the number of mechanically-released protoplasts obtained is about 6×104 per gram fresh weight of cortical tissue from the primary root of maize (Zea mays L. WF9×Mo 17) seedlings. Nuclear staining of the preparation, however, shows that about half of these protoplasts lack a nucleus and thus are actually subprotoplasts. Comparison of lectin binding to the plasma membranes of mechanically-and enzymatically-released protoplasts shows that both types contain binding sites forRicinus communis agglutinin. Binding sites for peanut (Arachis hypogaea) agglutinin are not naturally present on mechanically-released protoplasts but are generated by exposure to a mixture of Cellulysin and Pectolyase Y-23, the cell wall-degrading enzymes used to prepare enzymatically-released protoplasts.
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  • 90
    ISSN: 1615-6102
    Keywords: Auxin ; Blue-light photoreceptor ; Microtubule orientation ; Phytochrome ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The effects of red and blue light on the orientation of cortical microtubules (MTs) underneath the outer epidermal wall of maize (Zea mays L.) coleoptiles were investigated with immunofluorescent techniques. The epidermal cells of dark-grown coleoptiles demonstrated an irregular pattern of regions of parallel MTs with a random distribution of orientations. This pattern could be changed into a uniformly transverse MT alignment with respect to the long cell axis by 1 h of irradiation with red light. This response was transient as the MTs spontaneously shifted into a longitudinal orientation after 1–2 h of continued irradiation. Induction/reversion experiments with short red and far-red light pulses demonstrated the involvement of phytochrome in this response. In contrast to red light, irradiation with blue light induced a stable longitudinal MT alignment which was established within 10 min. The blue-light response could not be affected by subsequent irradiations with red or far-red light indicating the involvement of a separate blue-light photoreceptor which antagonizes the effect of phytochrome. In mixed light treatments with red and blue light, the blue-light photoreceptor always dominated over phytochrome which exhibited an apparently less stable influence on MT orientation. Long-term irradiations with red or blue light up to 6 h did not reveal any rhythmic changes of MT orientation that could be related to the rhythmicity of helicoidal cell-wall structure. Subapical segments isolated from dark-grown coleoptiles maintained a longitudinal MT arrangement even in red light indicating that the responsiveness to phytochrome was lost upon isolation. Conversely auxin induced a transverse MT arrangement in isolated segments even in blue light, indicating that the responsiveness to blue-light photoreceptor was eliminated by the hormone. These complex interactions are discussed in the context of current hypotheses on the functional significance of MT reorientations for cell development.
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  • 91
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    Protoplasma 179 (1994), S. 131-141 
    ISSN: 1615-6102
    Keywords: Inosine diphosphatase ; Golgi membranes ; Zea mays ; Roots
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Light microsomes of corn roots, enriched in endoplasmic reticulum and Golgi membranes, have an IDPase activity which is stimulated by Triton X-100 and by cold storage. In the native state, the enzyme activity does not follow Michaelis-Menten kinetics. It hydrolyses IDP with K0.5 of about 900 μM and Vmax of 300–400 nmol Pi/mg protein/min. In the presence of Triton X-100, the enzyme is maximally stimulated and it renders to a Michaelis-Menten behavior with a Km of about 500 μM and a Vmax of 800–1200 nmol Pi/mg protein/min. The maximal effect of the detergent occurs at about 1 mM IDP (270%), being reduced (190%) at high IDP concentrations (〉2 mM) which, per se, have a slight stimulatory effect on the enzyme. On the other hand, we observed that ATP (〉2 mM) and adenosine inhibit the IDPase. The effects of the nucleotides and of the adenosine are abolished in the presence of Triton X-100, which makes the enzyme fully active. Furthermore, we observed that detergent treatment of the membranes reduces the change in the activation energy which occurs at 10 °C and eliminates cooperative effects, as revealed by the Arrhenius analysis and the Hill analysis, respectively. We also observed that IDPase inhibition by ATP is maximal at low IDP concentrations (1 mM), whereas it decreases at high concentrations of IDP (4 mM), which promote maximal velocities in the native enzyme. Conversely, the inhibitory effect of adenosine is not reduced at high IDP concentrations. Pyrophosphate also inhibits the IDPase, but the effect is non-competitive and it is cumulative with that of ATP. We also observed that the latent activity of the IDPase (Triton-stimulated IDPase) is reduced by pre-treatment of the membranes with glutaraldehyde. The results indicate that Golgi IDPase is an allosteric enzyme which is positively modulated by IDP and negatively modulated by ATP and adenosine. Pyrophosphate inhibits the IDPase, but it seems to act at the catalytic site, whereas the other modulators appear to interact with a distinct regulatory site.
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  • 92
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    Protoplasma 165 (1991), S. 11-26 
    ISSN: 1615-6102
    Keywords: Zea mays ; Caffeine ; Microtubules ; Wall thickening ; Stomatal pore opening ; Microtubule organizing centres
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Treatment ofZea mays seedlings with a 5 mM caffeine solution inhibits cytokinesis in guard cell mother cells (GMCs), producing unicellular, binucleate aberrant stomata (a-stomata). Ventral wall (VW) strips of limited length, which usually meet the wall portions of GMCs adjoining the cortical zone of the preprophase microtubule band (PMB), are laid down in many a-stomata. In a-stomata with or without VW-strips, the periclinal walls are lined by numerous microtubules (Mts) converging on their mid-region, where local wall thickenings are deposited. When the VW-strips reach the mid-region of the periclinal walls, thickenings lined by numerous Mts rise at their free margins. In certain a-stomata an anticlinal wall column, surrounded by a dense Mt bundle, grows centripetally from either or both of the periclinal wall thickenings. In wall thickenings, the cellulose microfibrils are co-aligned with the adjacent Mts. Pore formation is initiated in all a-stomata. Deposition of an electron dense intra-wall material followed by lysis precedes “pore” opening. This process is closely related to the a-stornata morphogenesis. These observations show that the primary morphogenetic phenomenon in a-stomata is the establishment of an intense and stable polarity in the cytoplasm abutting on the mid-region of the periclinal walls and/or the adjacent plasmalemma area. Prime morphogenetic factor(s), including microtubule organizing centres (MTOCs), seem to function in these sites. Morphogenesis in a-stomata is a Mt-dependent process that is carried out as in normal stomata but in the absence of a VW.
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  • 93
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    Protoplasma 167 (1992), S. 33-42 
    ISSN: 1615-6102
    Keywords: Daucus carota ; Zea mays ; Vigna radiata ; Helicoidal cell walls ; Polylamellate cell walls ; Rapid-freeze deep-etch
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary This paper reports on preliminary investigations into the structure of cell walls of varying complexity as revealed by the rapidfreeze deep-etch technique. Three cell types from different species were examined in order to compare the three-dimensional arrangement of random, polylamellate and helicoidal walls. Each cell type displayed a distinctive level of organisation with respect to the cellulose microfibrils and the matrix material. In polylamellated walls, the microfibrils within each layer were linked to each other by 16–20 nm long side chains regularly spaced along the length of the microfibril. In helicoidal walls, the shifting of the microfibrils could cleary be seen, yet no recognisable structures were observed which could mediate this movement.
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  • 94
    ISSN: 1615-6102
    Keywords: Auxin ; Mechanical stress ; Mechanosensor ; Microtubule orientation ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Plants respond to mechanical stress by adaptive changes in growth. Although this phenomenon is well established, the mechanism of the perception of mechanical forces by plant cells is not yet known. We provide evidence that the cortical microtubules sub-adjacent to the growth-controlling outer epidermal cell wall of maize coleoptiles respond to mechanical extension and compression by rapidly reorientating perpendicular to the direction of the effective force change. These findings shed new light on many seemingly unrelated observations on microtubule reorientation by growth factors such as light or phytohormones. Moreover, our results suggest that microtubules associated with the plasma membrane are causally involved in sensing vectorial forces and provide vectorial information to the cell that can be utilized in the orientation of plant organ expansion.
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  • 95
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    Protoplasma 162 (1991), S. 12-19 
    ISSN: 1615-6102
    Keywords: Zea mays ; Chromium ; Roots ; Cell cycle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Four inbred lines ofZea mays (33.16, B 68, N 7B, B 77) were grown in nutrient solution to which K2Cr2O7 was added to give final concentration of 5 mg/l Cr (VI). The most evident differences in metal tolerance were observed between the B 68 and 33.16 line: in fact, even though the level of Cr (VI) was almost the same in the root tissues of both lines after 6 d of treatment, in the B 68 line, Cr induced marked alterations of nuclear structure and a progressive arrest of the cell cycle in G 1. In the 33.16 line, on the contrary, the integrity of the nuclei was well preserved and the progression of the cell cycle was only barely affected.
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  • 96
    ISSN: 1615-6102
    Keywords: Zea mays ; (1→3), (1→4)-β-D-glucan ; Glucan synthase ; Golgi apparatus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The synthesis of (1→3), (1→4)-β-D-glucan (MG) is associated with the Golgi apparatus of maize. Identification of in vitro reaction products by enzymic hydrolysis and separation of diagnostic oligosaccharides by HPLC was used as a specific assay for MG synthase activity. Large quantities of highly enriched membrane are needed to study the enzyme components of MG synthesis. We directly obtained highly enriched Golgi apparatus in a single flotation centrifugation, without the necessity of an initial downward centrifugation. This new procedure has improved the yield of Golgi apparatus, and has improved recovery of MG synthase activity. The substrate in glucan synthase reactions is UDP-Glc, but UDP-Glc is also a substrate in many other reactions, including the production of simple glucosides. In addition, much of the labeled Glc from UDP-Glc is broken down to Glc-1-P and Glc under MG synthase reaction conditions. We have explored some inhibitors of phosphatase, phosphorylase, phosphodiesterase, and glucosidase activities in order to minimize these competing reactions and increase the activity of MG synthase.
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  • 97
    ISSN: 1615-6102
    Keywords: Carbohydrates ; Proteins ; Root cap mucilage ; Root rhizodermal mucilage ; Root surface ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A strategy to obtain fractions enriched in mucilages secreted by root caps or produced by the rhizodermis of axenicallygrown maize seedlings is proposed. It involves a two-step procedure allowing the successive collection of root exudates and surface extracts from the same set of intact, sterile maize plants. Cytological controls were performed at each phase of collection. Whereas root cap mucilage is easily collected in water after one day's extraction, under conditions favouring secretory activity, rhizodermal mucilage remains tightly adherent to the root surface. It can be better extracted using neutral saline buffer assisted by gentle shaking at low temperature. Acidic saline buffer is unsuitable as it induces cell lysis and release of cell wall components. Biochemical analyses confirm that fractions enriched in root cap mucilage contain very high levels of fucose and galactose, high levels of arabinose, xylose and glucose and trace amounts of mannose. Fractions enriched in rhizodermal mucilage contain large amounts of glucose, moderate amounts of arabinose, xylose, mannose and galactose and trace levels of fucose. Isoelectric focusing and SDS-PAGE indicate that there are numerous similarities in the protein composition of materials enriched in root cap mucilages from root exudates or aqueous root surface extracts. However, specific protein bands that could be characteristic of rhizodermal mucilage are obtained using neutral saline buffer extracts. According to these biochemical data, the two-step procedure used in the present study appears to be useful for further biochemical characterization of both types of mucilages.
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  • 98
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    Protoplasma 171 (1992), S. 123-133 
    ISSN: 1615-6102
    Keywords: Endosperm ; Floury-2 ; Immunocytochemistry ; Protein bodies ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The seed storage proteins of maize (Zea mays L.) are synthesized during endosperm development on membrane-bound polyribosomes. These proteins, collectively called zeins, are translocated into the lumen of the rough endoplasmic reticulum, where they assemble into protein bodies. Protein body formation in normal genotypes occurs via an ordered deposition of the various types of zeins, and leads to the formation of spherical structures with a diameter of about 1 μm. These structures consist of a central core that contains predominantly α-zein; this central region is surrounded by a peripheral layer of β- and γ-zeins, and the entire structure is bounded by rough endoplasmic reticulum. In the endosperm mutant floury-2 the levels of all classes of zeins are reduced; these kernels exhibit an opaque phenotype instead of the vitreous phenotype observed in normal genotypes. In contrast to the discrete, spherical protein bodies which are formed in normal maize endosperm, the protein bodies within floury-2 endosperm are irregular and the zeins are disorganized; patches of β- and γ-zeins occur within irregularly lobed clusters of α-zein within the lumen of the rough endoplasmic reticulum. The implications of this aberrant distribution are discussed, both with respect to protein body development and kernel characteristics.
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  • 99
    ISSN: 1615-6102
    Keywords: Cytoplasmic calcium ; Fura ; Indo ; Protoplasts ; Gravitropism ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Calcium has been implicated as a key component in the signal transduction process of root gravitropism. We measured cytoplasmic free calcium in protoplasts isolated from the elongation zone and cap of primary roots of light-grown, vertically oriented seedlings ofZea mays L. Protoplasts were loaded with the pentapotassium salts of fura-2 and indo-1 by incubation in acidic solutions of these calcium indicators. Loading increased with decreasing pH but the pH dependence was stronger for indo-1 than for fura-2. In the case of fura-2, loading was enhanced only at the lowest pH (4.5) tested. Dyes loaded in this manner were distributed predominantly in the cytoplasm as indicated by fluorescence patterns. As an alternative method of loading, protoplasts were incubated with the acetoxymethylesters of fura-2 and indo-1. Protoplasts loaded by this method exhibited fluorescence both in the cytoplasm and in association with various organelles. Cytoplasmic calcium levels measured using spectrofluorometry, were found to be 160±40 nM and 257±27 nM, respectively, in populations of protoplasts from the root cap and elongation zone. Cytoplasmic free calcium did not increase upon addition of calcium to the incubation medium, indicating that the passive permeability to calcium was low.
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  • 100
    ISSN: 1615-6102
    Keywords: Endosperm ; Zea mays ; Immunolocalization ; Rough endoplasmic reticulum ; Binding protein ; Protein bodies
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A maize 75 kDa protein recently has been identified as a plant homolog of the mammalian binding protein (BiP). To better understand the function of BiP in protein body formation in maize endosperm, immunomicroscopy studies were conducted on three maize endosperm mutants, floury-2, Mucronate, and Defective endosperm-B 30, in which the level of BiP is highly elevated. Our results showed that protein body morphology in all three mutants was altered. In addition, BiP was localized in both the ER and peripheral regions of the abnormal protein bodies. The degree to which protein body morphology differed from normal was positively correlated with increased amounts of BiP. In addition, the accumulation of BiP in abnormal protein bodies increased with protein body maturation. In the three endosperm mutants, the arrangement of zeins within protein bodies had been perturbed, yet none of the specific zein subclasses exhibited the staining pattern found for BiP. The association of BiP with abnormal packaging of proteins in protein bodies may reflect a biological function to mediate protein folding and assembly in maize endosperm.
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