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  • RFLP  (201)
  • maize  (201)
  • Springer  (402)
  • American Physical Society
  • 1990-1994  (402)
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Entomologia experimentalis et applicata 67 (1993), S. 143-148 
    ISSN: 1570-7458
    Keywords: Nosema marucae ; microsporidium ; production ; biological control ; cereal stem borer ; Chilo partellus ; maize ; sorghum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In a study covering 3 years, experiments were carried out in order to determine the feasibility of producing a microsporidian pathogenNosema marucae in the spotted stalkborerChilo partellus. A maximum yield of 4.9×108 spores/larva (equivalent to 3.1×1010 spores/g fresh larval body weight) was obtained in 3rd instar larvae. It is considered that the production is inexpensive and can be readily adapted for small scale pathogen propagation systems in the tropics.
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  • 2
    ISSN: 1570-7458
    Keywords: Insecta ; Ostrina nubilalis ; pheromone trapping ; maize
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The response of male European corn borer, Ostrinia nubilalis (Hübner) to synthetic pheromone lures containing various isomeric blends of the sex pheromone 11-tetradecenyl acetate was measured in 13 counties in North Carolina. The blends consisted of either 3% Z (‘E strain’), 97% Z (‘Z strain’), or 35% Z (‘hybrid’) 11-tetradecenyl acetate. Response to E strain lures predominated in those counties located in the Coastal Plain (east) of the state, while response to the Z strain pheromone was dominant in the west. A zone of overlap of these broad strain distributions appears to occur in the eastern Piedmont. Within this zone there was substantial response to both E and Z blends. The proportion of these responses changed considerably between generations within years as well as between years. Significantly higher capture rates in hybrid baited traps in parts of the overlap zone may be indicative of increased rates of hybridization between the E and Z strains.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Entomologia experimentalis et applicata 65 (1992), S. 165-170 
    ISSN: 1570-7458
    Keywords: Lepidoptera ; Pyralidae ; stem borer ; suction trap ; behaviour ; maize ; dispersal ; pheromones ; activity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The diel flight periodicity of the nocturnal moth Chilo partellus (Swinhoe) (Lepidoptera;Pyralidae) was measured in the laboratory using an actograph and in the field with suction traps. Females showed almost no flight activity on the night of eclosion. Flight activity of mated females peaked before midnight, the period of peak oviposition activity. Male peak activity occurred after midnight coinciding with female eclosion. Presence or absence of females did not affect when or how long males were active. Data on flight activity and reproductive behaviour are discussed in relation to the use of pheromones to protect maize.
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  • 4
    Electronic Resource
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    Springer
    Entomologia experimentalis et applicata 64 (1992), S. 11-21 
    ISSN: 1570-7458
    Keywords: European corn borer ; Ostrinia nubilalis ; maize ; water ; drought ; stress ; development ; models ; microenvironment ; irrigation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract This study examined the impact of irrigation water on certain aspects of an insect-plant relationship in the field including the assessment of plant-mediated water effects on an herbivore's development, survival, and behavior, and plant damage parameters and host tissue water status. Maize (Zea mays L.) plants were arranged in a randomized complete block design in the field over two years in North Carolina (NC). Four blocks were subjected to three different irrigation treatments initiated ca. one week before anthesis: optimal, intermediate, deficit water supply. Each plant was infested with one (1986) or two (1987) black head stage, E-race European corn borer [Ostrinia nubilalis (Hübn.)] (ECB) egg masses at tasselling. ECB development, tunnelling site, and survival as well as plant tissue water status (tissue % water contents [θ] & leaf water potentials [Ψ]) were recorded through July. The irrigation effect on ECB parameters was slight and variable. Internal stalk temperatures of optimal plants were consistently cooler than their deficit counterparts (1 day-degree/day). With degree-days included as an explanatory variable in the analyses, there were no significant irrigation effects on the ECB parameters, except for total proportion of ECB's bored into maize plant parts. More ECB's bored into drier plants than in optimal plants; however, this trend was not significant in 1987. Plant water indices showed that though Ψ responded to irrigation, there were only minor changes in tissue θ, particularly in view of the larger diurnal tissue changes observed and the relatively high, sustained stalk θ levels seen over all treatments. Examination of ECB pupal θ confirmed that dietary water changes were minor or non-limiting to the insects' developmental physiology, because pupal θ was not sensitive to the irrigation treatments. Though water supply changes have drastic developmental and agronomic consequences for the maize plant, little or no changes were seen in the ECB feeding environment. Furthermore, a plant damage model was developed whereby the total % of ECB's tunnelled into maize was related to the mean larval age. The implications of this model on the understanding of ECB tunnelling behavior, damage potential, and pest management is noted.
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  • 5
    ISSN: 1570-7458
    Keywords: insecta ; Ostrinia nubilalis ; egg distribution ; maize
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The relationship between second generation European corn borer (Ostrinia nubilalis Hübner) egg mass numbers and subsequent field corn damage, as measured by stalk cavity numbers, was studied in 79 fields in northeastern North Carolina over three years. A mean of 0.028 egg masses per plant (645 egg masses/23400 plants) was found over the course of the study. Significant differences in oviposition rate were detected between fields and years. Ca. 85% of egg masses were deposited in a five leaf zone surrounding the primary ear; of these, 89% were found on the lower four leaves in this zone. Egg masses appeared to be distributed randomly within fields but at low rates of incidence, and oviposition was relatively uniform between sampling areas within individual fields. Under moderate to high oviposition pressure (mean number of egg masses per plant over the duration of the oviposition period 〉ca. 0.02), eggs laid during the early phases of the oviposition period account for more subsequent stalk damage than eggs laid during the later phases of the oviposition period. Variations in second generation egg mass numbers accounted for ca. 70% of variation in stalk cavity numbers.
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  • 6
    ISSN: 1420-9071
    Keywords: Mitochondrial DNA ; RFLP ; Leptinotarsa decemlineata ; Colorado potato beetle ; population genetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary This study demonstrates variability in restriction enzyme cleavage sites of mitochondrial DNA (mtDNA) among four popalations of Colorado potato beetle (CPB). A suite of three enzymes (EcoRI,HpaI,PstI) was sufficient to discriminate among the populations tested. Individuals heteroplasmic for restriction enzyme patterns were found in some populations. Variability in CPB mtDNA should prove useful in efforts to trace the origin and dispersal of the species in North America.
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  • 7
    Electronic Resource
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    Springer
    Entomologia experimentalis et applicata 55 (1990), S. 285-294 
    ISSN: 1570-7458
    Keywords: Graminella nigrifrons ; maize ; oats ; johnsongrass ; development ; fecundity ; host suitability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Description / Table of Contents: Résumé La dynamique des populations (durée de développement de l'œuf à l'adulte, poids et taille des adultes, fécondité) de G. nigrifrons Forbes (Homop. Cicadellidae) a été étudiée au laboratoire à 5 températures sur plantules de maïs (Zea mays L.), avoine (Avena sativa L.) et sorgho vivace (Shorgum halepense (L.) Pers.). Sur les 3 plantes, les mâles se développent en moyenne 1,2 j plus vite que les femelles. Les relations entre vitesse de développement et température ont été déterminées en utilisant à la fois un modèle linéaire et le modèle biophysique à 2 paramètres de Sharpe & DeMichele (1977). Les températures plus basses donnent des adultes des 2 sexes plus gros et plus lourds. Moins de G. nigrifrons se sont développés sur la graminée vivace que sur les 2 graminées annuelles à la température la plus élevée (30°C), tandis qu'à la température la plus basse (18°C) moins de cicadelles se sont développées sur les graminées annuelles. La température semble jouer un rôle significatif en déterminant l'adéquation des plantes comme hôtes convenant au développement de G. nigrifrons. Le potentiel de ponte de cette cicadelle avait été sous-estimé par les étudies précédentes.
    Notes: Abstract Population dynamics of the blackfaced leafhopper, Graminella nigrifrons (Forbes) (Homoptera: Cicadellidae), was studied at five temperatures (18, 21, 24, 27, & 30°C) in the laboratory on seedling maize (Zea mays L.), oats (Avena sativa L.), and the perennial johnsongrass (Sorghum halepense (L.) Pers.). Effects of temperature and host plant on egg to adult mean development time, adult size and weight, and fecundity were determined. Leafhoppers on all three hosts developed fastest at the highest temperature tested (21.3 days), and slowest at the lowest temperature tested (73.2 days). The duration from first to last adult eclosion was shortest at 30°C, (11.5 days) and longest at 18°C (43 days). The sex ratio of males to females did not differ from 1:1, but males developed an average of 1.2 days faster than females on all three hosts. Mean percent development/day ranged from 1.4% at 18°C to 4.7% at 30°C. The relationship of this development rate and temperature was determined using both a linear model and a variable parameter biophysical model. Based on these models, the developmental threshold is estimated at 12–15°C. The lowest temperature yielded larger and heavier adults (312 μg, dry weight) than did the highest temperature (225 μg). Fewer leafhoppers developed on the perennial than the annuals at 30°C and fewer on the annuals than the perennial at 18°C. Our results suggest that early in the season johnsongrass and perhaps other perennials are the superior developmental hosts for this leafhopper, whereas in midsummer when temperatures are highest, annuals are the better hosts.
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  • 8
    Electronic Resource
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    Springer
    Entomologia experimentalis et applicata 61 (1991), S. 169-177 
    ISSN: 1570-7458
    Keywords: African migratory locust ; crop environment ; oviposition behaviour ; oviposition sites ; maize
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Oviposition by the African migratory locust, Locusta migratoria migratorioides (Orthoptera: Acrididae), was studied in maize and wheat crops on the Orange Free State Highveld. Maize was shown to be the most important oviposition habitat with peak laying taking place in autumn and early winter when highest pod densities were recorded. Laying was mainly concentrated along the middle of the crop interrows in maize and within clearings in the wheat crop. Despite the uniform layout of these crops, the distribution of egg pods was found to be aggregated. Non-reproductive behaviour, such as locust aggregation, basking and feeding, as well as environmental factors appeared to influence the distribution of egg pods in these crops. Secondary selection for optinum soil moisture and compaction on the laying site enhanced the aggregation of pods.
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  • 9
    Electronic Resource
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    Springer
    Entomologia experimentalis et applicata 65 (1992), S. 57-64 
    ISSN: 1570-7458
    Keywords: Heliothis ; corn ; maize ; insecticide ; Aspergillus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The toxicity and anticholinesterase activity of tremorgenic fungal metabolites, territrems, to the corn earworm, Helioverpa zea (Boddie) (Lepidoptera, Noctuidae) were examined. In oral toxicity studies, territrem A significantly inhibited growth by 40% at 25 ppm and by 89% at 250 ppm. Territrem B and an epoxy-derivative significantly inhibited growth by ca. 45% at 250 ppm. Piperonyl butoxide administered orally synergized the toxicity of the territrems tested. In topical toxicity studies, the epoxy-derivative caused 26% mortality and 25% growth retardation at 10 mg/gm insects. Territrem A and B were not significantly lethal, but did reduce growth by 15–20% at 10 mg/gm insect. Paraoxon tested in the same way caused 100% mortality at 25 ppm orally and 10 mg/gm topically. However, all territrems tested in vitro as inhibitors of H. zea head acetylcholinesterase were at least comparable to or were more active than paraoxon. Topically administered epoxy-territrem B also inhibited H. zea head acetylcholinesterase. The potential for development of new insecticides from a territrem lead structure is discussed.
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  • 10
    ISSN: 1573-0832
    Keywords: Fusarium ; maize ; moniliformin ; mycotoxins ; trichothecenes ; zearalenone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Forty Fusarium isolates obtained from maize fields were screened for moniliformin production on maize kernels. Twelve isolates, including seven of F. subglutinans, were found to produce moniliformin at levels ranging from 0.4 to 64 ppm. Twenty six isolates were also screened for production of deoxynivalenol, diacetoxyscirpenol, T-2 toxin and zearalenone. Of these, 22, including all 11 isolates of F. graminearum, produced zearalenone at levels ranging from 0.1 to 96.0 ppm, while 13 produced T-2 toxin at low levels, (〈1.1 ppm). Deoxynivalenol and diacetoxyscirpenol were each produced by six isolates, also at low levels (〈1.0 ppm). Three isolates of F. graminearum and one of F. sambucinum produced four toxins simultaneously.
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  • 11
    Electronic Resource
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    Nutrient cycling in agroecosystems 23 (1990), S. 73-80 
    ISSN: 1573-0867
    Keywords: Nigerian savanna ; maize ; potassium ; zinc
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A three year field study was conducted at five locations in the Nigerian savanna to evaluate the response of early maturing maize variety to varying rates of K and Zn with a view to establishing the K and Zn requirements for maize production in this zone. Treatments consisted of 4 × 3 factorial combinations of 4 levels of K and 3 levels of Zn. Responses to K and Zn fertilization were sporadic and were obtained only in soils of the Southern Guinea savanna and in the soil formed on sedimentary sandstone. There seem to be no problem at present in soils of the Northern Guinea and Sudan savannas where leaching is less intense. It is inferred from this study that K and Zn deficiences are incipient in the high rainfall soils and in the sandstone derived soils. For these soils, 50 kg K/ha and 2–5 kg Zn/ha is suggested as adequate for an early maturing maize crop. Soil data showed that K and Zn responses can be expected when available K and Zn levels fall below 0.1 meq/100 g and 2 ppm respectively.
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  • 12
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    Nutrient cycling in agroecosystems 26 (1990), S. 249-252 
    ISSN: 1573-0867
    Keywords: Long-term manure trial ; residual effect ; model test ; nitrogen availability ; maize ; Italian ryegrass
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Sluijsmans and Kolenbrander developed a simple model to describe the availability of animal manure, assuming a readily available, an easily decomposable and a slowly decomposable N fraction. We tested this model on data from an experiment in which farmyard manure had been applied for eleven successive years to silage maize [Zea mays L.] grown on a light sandy soil. The residual effects of this FYM were then measured by growing Italian ryegrass [Lolium multiflorum Lamk.] in the 12th year. The measured uptake of N by the grass of the FYM residues was then compared with the computed values. The measured amounts of N taken up agreed fairly well with the calculated amounts for applications of 50 and 100 t FYM per ha per year. If the rates of manure application are adjusted to crop requirement, the model shows that the potential, long-term release of N from the residual N fraction of FYM will not exceed 20 kg N per ha. For cattle slurry with a smaller residual fraction, the release will be at most 10% of the total annual N application.
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  • 13
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    Nutrient cycling in agroecosystems 32 (1992), S. 209-222 
    ISSN: 1573-0867
    Keywords: Wheat ; maize ; barley ; rice ; foliar urea ; grain yield ; breadmaking quality
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract It has been suggested that there are several potential benefits of providing nitrogen to cereals via the foliage as urea solution. These include: reduced nitrogen losses through denitrification and leaching compared with nitrogen fertilizer applications to the soil; the ability to provide nitrogen when root activity is impaired e.g., in saline or dry conditions, and uptake late in the season to increase grain nitrogen concentration. Factors that influence the degree of foliar absorption in field conditions have not, however, been clearly defined and losses to the atmosphere and soil can occur. Foliar urea applications may also hinder crop productivity although the explanations for this vary, and include desiccation of leaf cells, aqueous ammonia and urea toxicity, biuret contamination and the disruption of carbohydrate metabolism. It has not yet been determined which one, or combinations, of these mechanisms are most important in field situations. When damage has not been severe, foliar urea applications have increased grain yield, particularly when applied before flag leaf emergence and when nitrogen availability is limiting. Increases in grain nitrogen content are often larger when applications of nitrogen fertilizers to the soil are reduced, and when the urea solution is sprayed either at anthesis or during the following two weeks. It is during this period that foliar urea sprays can be of greater benefit than soil applications with regard to nitrogen utilization by the crop. Increases in wheat grain nitrogen concentration following urea application can improve breadmaking quality. Responses in loaf quality may, however, be variable particularly when increases in grain nitrogen content have been large, and/or when the nitrogen: sulphur ratio in the grain is increased. These circumstances have lead to alterations in the proportions of the different protein fractions which influence breadmaking potential. To exploit the full potential benefits of foliar urea application to cereals, more needs to be known about the mechanisms, and thus how to prevent losses of nitrogen from the foliage, and to reduce the phytotoxic influences of sprays. More information is also required to exploit the reported effects that urea may have on limiting the development of cereal diseases.
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  • 14
    ISSN: 1573-0867
    Keywords: animal manure ; leaching ; maize ; nitrification inhibitor ; nitrogen recovery
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Field experiments with silage maize during eight years on a sandy soil in The Netherlands, showed that dicyandiamide (DCD) addition to autumn-applied cattle slurry retarded nitrification, thus reducing nitrate losses during winter. Spring-applied slurry without DCD, however, was on average associated with even lower losses and higher maize dry matter yields. Economically optimum supplies of mineral N in the upper 0.6 m soil layer in spring (EOSMN), amounted to 130–220 kg ha−1. Year to year variation of EOSMN could not be attributed to crop demand only. According to balance sheet calculations on control plots, apparent N mineralization between years varied from 0.36 to 0.94 kg ha−1 d−1. On average, forty percent of the soil mineral N (SMN) supply in spring, was lost during the growing season. Hence, the amounts of residual soil mineral N (RSMN) were lower than expected. Multiple regression with SMN in spring, N crop uptake and cumulative rainfall as explanatory variables, could account for 79 percent of the variation in RSMN. Postponement of slurry applications to spring and limiting N inputs to economically optimum rates, were insufficient measures to keep the nitrate concentration in groundwater below the EC level for drinking water.
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  • 15
    ISSN: 1572-9680
    Keywords: alley cropping ; maize ; soybean ; soil fertility ; Leucaena leucocephala ; Sesbania sesban ; Albizia falcataria ; Flemingia congesta ; Gliricidia sepium ; Cassia spectabilis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Theee trials to evaluat the potential of alley cropping in maize production on the low fertility, acidic soils in Northern Zambia are described. Leucaena leucocephala, Gliricidia sepium, Sesbania sesban, Albizia falcataria, Fleminga congesta, and Cassia spectabilis, were grown in alley crops with hybrid maize and soybean. All trials received recommended rates of P and K fertiliser; N fertiliser was applied at three rates as a subplot treatment. One trial received lime before establishment. Only in the limed trial was there a significant improvement in maize yields through alley cropping; when no N fertiliser was applied, incorporation of Leucaena leucocephala prunings resulted in an increase of up to 95% in yields, with a smaller improvement being produced by Flemingia congesta. There was a significant correlation between the quantity of prunings biomass applied and the proportional increase in maize yields over the control treatment. It is suggested that the lack of effect of most of the tree species on crop yields was due to low biomass production. An economic analysis showed that alley cropping with limed Leucaena was only profitable when fertiliser costs were high in relation to maize prices. However, lime is both expensive and difficult to obtain and transport for most small scale farmers in the region, and is therefore not a practical recommendation. It is suggested that future alley cropping research should focus on screening a wider range of tree species, including other species of Leucaena, for acid tolerance and higher biomass production.
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  • 16
    ISSN: 1572-9680
    Keywords: Erythrina ; Gliricidia ; alley cropping ; maize ; competition ; nitrogen availability ; Costa Rica
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Maize growing next toErythrina hedgerows had 44% lower biomass (p〈0.01) and 35% lower N content (p〈0.1) than maize growing in the middle of the alleys. Maize growing next toGliricidia hedgerows had the same biomass but 56% higher N content (p〈0.1) than maize growing in the middle of the alleys. However these differences did not develop until 2 months after sowing of the maize. Spatial variability in soil nitrogen mineralization and mulch nitrogen release did not explain any of the differences in growth or N uptake of the maize with respect to distance from the trees. It is hypothesized that the slower growth of the maize next to theErythrina trees after 2 months is due to increasing light and/or nutrient competition from the trees as the trees recover from pollarding. The apparent lack of competition fromGlirigidia may be due to different rates of regrowth or different shoot and root architecture. A theoretical model is described demonstrating that if a crop is to take advantage of the higher nutrient availability under alley cropping it must complete the major part of its growth before the trees recover significantly from pollarding, and start competing strongly with the crop.
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  • 17
    ISSN: 1572-9680
    Keywords: alley cropping ; maize ; nitrogen ; organic matter ; soil fertility ; Leucaena leucocephala ; Flemingia congesta
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A detailed study of the soil chemical and physical properties in seven-year-old alley cropping trial containingLeucaena leucocephala andFlemingia congesta in Northern Zambia is described. There was a strong correlation between the maize yield and the total amount of nitrogen applied, both from prunings and fertiliser, suggesting that a major reason for the observed benefit from alley cropping, particularly withLeucaena, was due to an improvement in nitrogen supply.Leucaena produced significantly more biomass, and its leaves had higher concentrations of nitrogen, phosphorous and potassium and lower C/N and C/P ratios than did those ofFlemingia. There was also evidence that the trees had a beneficial effect on other soil chemical properties; under the hedgerows, particularly those ofLeucaena, there were higher levels of organic carbon, Mg, K and ECEC, and pH values were also highest. It is suggested that higher levels of organic carbon in the alley crop treatments were responsible for the improvements observed in soil physical properties. Lower bulk density, lower penetration resistance, and a higher infiltration rate and pore volume fraction were measured in the alley crops, although there was no significant change in the soil water release parameters. A deteriorating effect of constant applications of nitrogen fertiliser on soil fertility was observed; as the level of urea application increased, there were significant decreases in Mg, K and pH, increases in Al and soil acidity, and higher penetrometer resistance. These results highlight the urgent need for further research on biological methods of maintaining soil fertility.
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  • 18
    ISSN: 1420-9098
    Keywords: Leptothorax acervorum ; mtDNA ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 27 nests ofLeptothorax acervorum were analysed for restriction fragment-length polymorphism (RFLP) in mitochondrial DNA (mtDNA), using four endonucleases. A substantial degree of variation was found between nests in the population (13 composite haplotypes). Intra-nest variation was detected in 15 % of the nests. The presence of occasional alien inseminated females indicates that polygyny in this species is caused by adoption of mated females. The occasional acceptance of alien females is difficult to explain, but interesting, since this behaviour could have given rise to inquilinism. Our results suggest that analysis of mtDNA RFLP is a method well suited for investigations of the population structure of ants.
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  • 19
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    Plant cell reports 12 (1993), S. 293-297 
    ISSN: 1432-203X
    Keywords: Lycopersicon esculentum ; Genetic marker ; Intraspecific genetic map ; DNA polymorphism ; Isozyme ; RFLP ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The existing molecular genetic maps of the tomato, Lycopersicon spp, are constructed based on isozyme and RFLP polymorphisms between tomato species. These maps are useful for certain applications but have few markers that exhibit sufficient polymorphisms for intraspecific analysis and manipulations within the cultivated tomato. The purpose of this study was to investigate the relative potential of RAPD technology, as compared to isozymes and RFLPs, to generate polymorphic DNA markers within cultivated tomatoes. Sixteen isozymes and 25 RFLP clones that were known to detect polymorphism between L. esculentum and L. pennellii, and 313 random oligonucleotide primers were examined. None of the isozymes and only four of the RFLP clones (i.e., 16%) revealed polymorphism between the cultivated varieties whereas up to 63% of the RAPD primers detected one or more polymorphic DNA fragments between these varieties. All RAPD primers detected polymorphism between L. esculentum and L. pennellii genotypes. These results clearly indicate that RAPD technology can generate sufficient genetic markers exploiting sequence differences within cultivated tomatoes to facilitate construction of intraspecific genetic maps.
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  • 20
    ISSN: 1432-203X
    Keywords: Protoplast fusion ; RFLP ; Mitochondrial DNA ; Chloroplast DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Somatic hybrid plants were recovered following fusion of leaf mesophyll protoplasts isolated from tomato (Lycopersicon esculentum) cultivar UC82 with protoplasts isolated from suspension cultured cells of L. chilense, LA 1959. Iodoacetate was used to select against the growth of unfused tomato protoplasts. Two somatic hybrids were recovered in a population of 16 regenerants. No tomato regenerants were recovered; all of the non-hybrid regenerants were L. chilense. The L. chilense protoplast regenerants were tetraploid. The hybrid nature of the plants was verified using species-specific restriction fragment length polymorphisms for the nuclear, chloroplast and mitochondrial genomes. The somatic hybrids had inherited the chloroplast DNA of the tomato parent, and portions of the mitochondrial DNA of the L. chilense parent. The somatic hybrids formed flowers and developed seedless fruit.
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  • 21
    ISSN: 1432-2145
    Keywords: Gametophytic self incompatibilityself-compatibility ; Lycopersicon peruvianum Lycopersicon hirsutum ; S-associated proteins ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Self-compatibility was investigated separately in two species of tomato, Lycopersicon peruvianum and L. hirsutum. The codominant expression of self-compatibility (SC)/self incompatibility (SI) was established using intraspecific hybrids of SC and SI hybrids. In SC L. peruvianum, a major stylar protein of approximately 29 kDa cosegregates with self-compatibility in the progeny of SC/SI hybrids. The SC/SI hybrids are self-fertile, but only partially so, since the SI allele present in the hybrids is capable of eliminating certain genotypes in the resultant progeny. In L. hirsutum, the majority of hybrids between one accession of SI L. hirsutum f. hirsutum and one of SC L. hirsutum f. glabratum are self-fertile. Analysis of the progeny revealed that the SC and SI alleles are codominant in this species as well. A protein product for the SC allele is not obvious in style extracts of L. hirsutum f. glabratum. Segregating progeny from SC/SI hybrids of L. hirsutum were used to map the S locus against five RFLP markers on chromosome 1, and estimated map distances are given. In addition, evidence is presented that indicates that one of the DNA markers, CD15, is duplicated in L. hirsutum f. glabratum, and the duplication is not linked to the S locus.
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  • 22
    ISSN: 1432-203X
    Keywords: Rice (Oryza sativa) ; Genetic Marker ; Genetic Map ; Integrated Linkage Map ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Application of genetic linkage maps in plant genetics and breeding can be greatly facilitated by integrating the available classical and molecular genetic linkage maps. In rice, Oryza sativa L., the classical linkage map includes about 300 genes which correspond to various important morphological, physiological, biochemical and agronomic characteristics. The molecular maps consist of more than 500 DNA markers which cover most of the genome within relatively short intervals. Little effort has been made to integrate these two genetic maps. In this paper we report preliminary results of an ongoing research project aimed at the complete integration and alignment of the two linkage maps of rice. Six different F2 populations segregating for various phenotypic and RFLP markers were used and a total of 12 morphological and physiological markers (Table 1) were mapped onto our recently constructed molecular map. Six linkage groups (i.e., chr. 1, 3, 7, 9, 11 and 12) on our RFLP map were aligned with the corresponding linkage groups on the classical map, and the previous alignment for chromosome 6 was further confirmed by RFLP mapping of an additional physiological marker on this chromosome. Results from this study, combined with our previous results, indicate that, for most chromosomes in rice, the RFLP map encompasses the classical map. The usefulness of an integrated genetic linkage map for rice genetics and breeding is discussed.
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  • 23
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    Current genetics 26 (1994), S. 456-460 
    ISSN: 1432-0983
    Keywords: Parasitella parasitica ; Zygomycetes ; RAPD ; PCR ; RFLP ; Electrophoretic karyotype ; Molecular taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In addition to conventional methods for the identification of fungi, molecular techniques at the DNA level are increasingly being employed. In order to check the validity of such experimental approaches, we have analyzed the well-defined species Parasitella parasitica, which belongs to the family Mucoraceae (Mucorales, Zygometes). The seven strains of this species, which are available from international strain collections, were analyzed by several molecular methods: restriction fragment length polymorphism analysis (RFLP), the random primer-dependent polymerase chain reaction (RAPD-PCR), and electrophoretic karyotyping. Unexpectedly, these strains are highly diverse at the molecular level. By these techniques they can be divided consistently into two different groups. Nevertheless, all seven strains belong to a single species. They show no morphological differences and sexual spores (zygospores) were found in all possible combinations either within or between the two groups. Southern-blot analysis of genomic DNA of all P. parasitica strains with RAPD-PCR-derived labelled probes shows the existence of repetitive elements characteristic for only one group of P. parasitica. In addition, chromosome sizes, which were separated by rotating-field electrophoresis, were highly divergent, and ranged from 3 to 6.5 Mb in one group and between 2 and 4.5 Mb in the other. The RAPD-PCR patterns also discriminate both groups of P. parasitica. However, they are very similar if strains of a single group are compared. Therefore, we propose that the determination of fungal species by molecular techniques should be vetted at least by morphological and physiological parameters and, whenever possible, by mating experiments.
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  • 24
    ISSN: 1432-0983
    Keywords: Cytoplasmic male sterility ; Antisense RNA ; RFLP ; Cybrids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Asymmetric cell-fusion of the japonica cultivar ofOryza sativa (rice) with cytoplasmic-male-sterile (CMS) plants bearing cytoplasm derived from Chinsurah Boro II, resulted in two classes of cytoplasmic hybrids (cybrids), fertile and CMS. Southern-blot analysis of the mitochondrial DNA (mtDNA) indicates recombination events around a number of genes; however, the appearance of the CMS character is tightly correlated to reorganization around theatp6 gene, suggesting recombination downstream from theatp6 gene is involved in CMS. The nucleotide sequence downstream fromatp6 contains a pseudogene which was probably created by recombination of the mitochondrial genome. Sense and antisense transcripts of the downstream region ofatp6 were found in CMS-and restored CMS (fertile)-lines, but not in the normal (fertile) line. In the CMS line, several antisense transcripts of theatp6 gene were also found. However, in the restored line which contains a nuclear-encoded gene,Rf-1, the levels of these transcripts were lower than in the CMS line. These results suggest abnormal transcripts of theatp6 gene produced in the antisense direction may be involved in CMS, and that products of the nuclear-encoded restorer gene may reduce abnormal transcription in this region of the mitochondrial genome.
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  • 25
    ISSN: 1432-0983
    Keywords: DNA fingerprinting of Trichoderma ; Trichoderma reesei ; RFLP ; Strain classification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have analyzed nine different species of the filamentous fungus Trichoderma and three strains of T. reesei for the presence of hypervariable loci in their genomes by hybridization with simple repeat oligonucleotides [(CT)8, (GTG)5, and (GACA)4]. On the basis of the DNA-fingerprints obtained, the Trichoderma aggregate is re-classified into five groups: I (T. reesei, T. todica), II (T. polysporum, T. longibrachiatum, T. koningii, and T. pseudokoningii), III (T. virgatum), IV (T. saturnisporum) and V (T. harzianum). These results contradict the claim that T. reesei is a subspecies of T. longibrachiatum. Furthermore, hybridization with (CA)8 allowed a subdivision of group II, wherein T. pseudokoningii formed a subgroup, IIb, which is highly homologous with, but distinct from subgroup IIa. The results show that RFLP analysis may be used to re-classify the Trichoderma aggregate.
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  • 26
    ISSN: 1573-4927
    Keywords: mutator ; transposable element ; alcohol dehydrogenase ; maize ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A secondary mutant, derived from an allele of maize alcohol dehydrogenase 1 (Adh1) carrying a Mutator transposable element (Mu1) in its first intron, was reported to exhibit a threefold decrease in ADH enzymatic activity and steady-state RNA levels compared to the original mutant. The original mutant,Adh1-S3034 (abbreviatedS3034), was previously characterized at the molecular level. The derivative, abbreviatedS3034b, has now been cloned; at the DNA sequence level the insertion and surroundingAdh1 sequences are indistinguishable fromS3034. Furthermore, in our lines there is no difference in relative ADH activities between products of the two putative alleles. A comparison of gene expression in heterozygotes obtained by crossing to different tester lines reveals a correlation between the measured decrease in levels of ADH polypeptide produced by the mutant allele and the background in which it is measured; this effect is distinct from any background-related variation in the expression of the progenitor allele. It does not appear to be attributable to alternative patterns of DNA modification. It appears to reflect a background-associated difference in the level of normalAdh1-RNA produced. Thus the previously reported distinction betweenS3034 andS3034b may be due to differences in the extent to which the mutant allele and a given genetic background interact to produce functionalAdh1-RNA.
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  • 27
    ISSN: 1573-1561
    Keywords: Leafhopper ; Dalbulus maidis ; Homoptera ; Cicadellidae ; hostfinding ; maize ; visual ; olfactory ; synergism ; pest
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Virtually nothing is known about the role plant volatiles play in host-finding by Homoptera in the Suborder Auchenorrhyncha. In laboratory bioassays, we examined the influence of plant volatiles on orientation and postcontact behaviors of the leafhopper,Dalbulus maidis, and determined the relationship between visual and olfactory stimuli during host-finding. When compared to the number of contacts made with reflected green light in the presence of a hexane control,D. maidis made more contacts when exposed to volatile extracts from its preferred host, maize; a similar number of contacts when exposed to volatiles from a marginal host, gamagrass; and fewer contacts when exposed to volatiles from a nonhost, sorghum. There was no difference between males and females in the number of contacts made with green light when exposed to maize volatiles compared to hexane alone. More contacts were made with green light than with white light of similar intensity, both in the presence and in the absence of olfactory stimuli; however, maize volatiles acted as a Synergist by increasing the number of contacts leafhoppers made with green light. After contacting the green light, exposure of maize volatiles significantly increased, relative to hexane, the amount of stationary time, but did not influence the amount of time spent moving, the distance traveled, or the speed while moving when within the boundaries of the green light. This study provides the first evidence for an interaction between visual and olfactory stimuli during host-finding for a leafhopper and also for olfactory mediation of postcontact behaviors not associated with feeding.
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  • 28
    ISSN: 1573-5028
    Keywords: carboxy-terminal processing ; glycosylation ; Hordeum vulgare L. ; Prx locus ; RFLP ; signal peptide ; targeting
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The major peroxidase of barley seed BP 1 was characterized. Previous studies showed a low carbohydrate content, low specific activity and tissue-specific expression, and suggested that this basic peroxidase could be particularly useful in the elucidation of the structure-function relationship and in the study of the biological roles of plant peroxidases (S.K. Rasmussen, K.G. Welinder and J. Hejgaard (1991) Plant Mol Biol 16: 317–327). A cDNA library was prepared from mRNA isolated from seeds 15 days after flowering. Full-length clones were obtained and showed 3′ end length variants, a G+C content of 69% in the translated region, a 90% G or C preference in the wobble position of the codons and a typical signal peptide sequence. N-terminal amino acid sequencing and sequence analysis of tryptic peptides verified 98% of the sequence of the mature BP 1 which contains 309 amino acid residues. BP 1 is the first characterized plant peroxidase which is not blocked by pyroglutamate. BP 1 polymorphism was observed. BP 1 is less than 50% identical to other plant peroxidases which, taken together with its developmentally dependent expression in the endosperm 15–20 days after flowering, suggests a unique biological role of this enzyme. The barley peroxidase is processed at the C-terminus and might be targeted to the vacuole. The single site of glycosylation is located near the C-terminus in the N-glycosylation sequon -Asn-Cys-Ser- in which Cys forms part of a disulphide bridge. The major glycan is a typical plant modified-type structure, Manα1-6(Xylβ1-2)Manβ1-4GlcNAcβ1-4(Fucα1-3)GlcNAc. The BP 1 gene was RFLP-mapped on barley chromosome 3, and we propose Prx5 as the name for this new peroxidase locus.
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  • 29
    ISSN: 1573-5028
    Keywords: cloning in YACs ; genome mapping ; maize ; PCR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have constructed a yeast artificial chromosome (YAC) library using high-molecular-weight DNA prepared from agarose-embedded leaf protoplasts of the maize inbred line UE95. This library contains 79 000 clones with an average insert size of 145 kb and should therefore represent approximately three haploid genome equivalents. The library is organised as an ordered array in duplicate microtitre plates. Forty-one pools of DNA from 1920 individual clones have been prepared for rapid screening of the library by the polymerase chain reaction (PCR). Using this approach, together with conventional colony hybridisation, we have been able to identify between one and eight positive clones for every probe used.
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  • 30
    ISSN: 1573-5028
    Keywords: CaMV 35S promoter ; leaf-specific DNA-binding protein ; maize ; PEP carboxylase gene promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract When gel shift assays were performed with maize nuclear extract and a DNA fragment containing the cauliflower mosaic virus (CaMV) 35S promoter, three DNA-protein complexes were observed. Analyses with nuclear extracts prepared from green leaves, etiolated leaves, stems and roots showed that the complexes resulted from the existence of at least two nuclear factors. One of them is presumably a constitutive nuclear factor found in all tissues tested, and another is a leaf-specific factor present both in green and etiolated leaves. This leaf-specific nuclear factor seemed to be identical to MNF1, previously identified as a factor interacting with the promoter of the maize gene for phosphoenolpyruvate carboxylase involved in the C4 photosynthesis. Deletion analysis revealed that MNF1 binds to the sequence from −281 to −235 relative to the transcription start site of the CaMV 35S promoter. MNF1-like nuclear protein was also found in tobacco nuclear extracts. The possibility that MNF1 participates as a positive trans-acting factor in the expression of genes in maize leaves is discussed.
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  • 31
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    Plant molecular biology 19 (1992), S. 563-575 
    ISSN: 1573-5028
    Keywords: maize ; ferritin ; iron stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The iron-storage protein ferritin has been purified to homogeneity from maize seeds, allowing to determine the sequence of the first 29 NH2-terminal amino acids of its subunit and to raise specific rabbit polyclonal antibodies. Addition of 500 μM Fe-EDTA/75 μM Fe-citrate to hydroponic culture solutions of maize plantlets, previously starved for iron, led to a significant increase of the iron concentration of roots and leaves, albeit root iron was mainly found associated with the apoplast. Immunodetection of ferritin by western blots indicated that this iron treatment induced ferritin protein accumulation in roots and leaves over a period of 3 days. In order to investigate this induction at the ferritin mRNA level, various ferritin cDNA clones were isolated from a cDNA library prepared from poly(A)+ mRNA isolated from roots 48 h after iron treatment. These cDNAs were classified into two groups called FM1 and FM2. Upstream of the sequence encoding the mature ferritin subunit, both of these cDNAs contained an in-frame coding sequence with the characteristics of a transit peptide for plastid targeting. Two members of the FM1 subfamily, both partial at their 5′ extremity, were characterized. They are identical, except in their 3′ untranslated region: FM1A extends 162 nucleotides beyond the 3′ terminus of FM1B. These two mRNAs could arise from the use of two different polyadenylation signals. FM2 is 96% identical to FM1 and contains 45 nucleotides of 5′ untranslated region. Northern analyses of root and leaf RNAs, at different times after iron treatment, revealed ferritin mRNA accumulation in response to iron. Ferritin mRNA accumulation was transient and particularly abundant in leaves, reaching a maximum at 24 h. The level of ferritin mRNA in roots was affected to a lesser extent than in leaves.
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  • 32
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    Plant molecular biology 19 (1992), S. 623-630 
    ISSN: 1573-5028
    Keywords: gene expression ; heat shock ; intron ; maize ; pollen ; RNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract While a heat shock treatment of 40 °C or 45 °C induced the vegetative tissues of maize to produce the typical heat shock proteins (HSPs), germinating maize pollen exposed to the same temperatures did not synthesize these characteristic HSPs. Comparison of RNA accumulation in shoot and tassel tissue showed that mRNAs for HSP70 and HSP18 increased several-fold, reaching high levels within 1 or 2 hours. At the higher temperature of 45 °C these vegetative tissues were blocked in removal of an intron from the HSP70 mRNA precursor, which accumulated to a high level in tassel tissue. In germinating pollen exposed to heat shock, mRNAs for these HSPs were induced but accumulated only to low levels. The stressed pollen maintained high levels of RNA for α-tubulin, a representative normal transcript. It is likely that the defective heat shock response of maize pollen is due to inefficient induction of heat shock gene transcription.
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  • 33
    ISSN: 1573-5028
    Keywords: gene duplication ; photosynthesis ; RFLP ; Southern blots
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A second locus (Lhb1B) encoding Photosystem II Type I chlorophyll a/b-binding (CAB) polypeptides was identified in Arabidopsis thaliana. This locus carries two genes in an inverted orientation. The predicted sequences of the polypeptides encoded by these two genes show substantial divergence in their amino termini relative to each other and to the proteins encoded by the three Lhb1 CAB genes previously characterized [10], but little divergence within the predicted primary structure of the mature protein. DNA probes derived from seven additional types of tomato CAB genes, encoding chlorophyll a/b-binding polypeptides of several antenna systems of the photosynthetic apparatus, were tested against A. thaliana. Each of these hybridized in Southern blots to unique DNA fragment(s), demonstrating the existence of each of these different types of CAB genes in the genome of A. thaliana. The number of genes encoding each CAB type in A. thaliana was estimated to be similar to that of tomato.
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  • 34
    ISSN: 1573-5028
    Keywords: bark storage protein ; gene clustering ; RFLP ; systemic response
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two wound-inducible cDNAs from poplar leaves show sequence identity to vegetative storage proteins (VSP) that accumulate seasonally in poplar bark tissues. We have compared the genomic organization, cDNA sequences and expression of the genes encoding the wound-inducible cDNAs (win4) with that of a bark VSP (called bark storage protein, or BSP). There appear to be several win4 genes in the poplar genome which segregate as a single locus and are therefore likely to be clustered. The same is true of the BSP genes. The win4 locus is linked (map distance of 5 cM) to the BSP locus, consistent with a common evolutionary origin of the genes. A near full-length win4 cDNA shows 75% sequence identity to BSP cDNAs. Both win4 and BSP are systemically wound-inducible; win4 transcripts accumulate in leaves and stems, whereas BSP transcripts accumulate almost exclusively in stems. A phloem transport-dependent signaling mechanism appears to be involved in systemic win4 expression after wounding. In contrast to BSP gene expression, win4 genes are not expressed in response to short day conditions. The data indicate win4 and BSP genes are differentially regulated, and their products may play important roles in the storage and reallocation of nitrogen in perennial plants.
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  • 35
    ISSN: 1573-5028
    Keywords: cell wall ; ethylene ; genetic transformation ; HRGP ; maize ; wounding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The expression of the maize gene coding for a hydroxyproline-rich glycoprotein (HRGP) has been studied by measuring the mRNA accumulation after wounding or ethylene treatment. RNA blot and in situ hybridization techniques have been used. The temporal and tissue-specific expression has been observed: the cells related to the vascular system show the more intense HRGP mRNA accumulation. Transcriptional constructions of the maize HRGP promoter have been tested on different maize tissues by microbombarding. A 582 bp promoter is able to direct the expression of the gus gene on calli and young leaves. Constructions having shorter promoter sequences lose this ability. The 582 bp construction retains the general specificity of expression observed for the HRGP gene.
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  • 36
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    Plant molecular biology 20 (1992), S. 581-588 
    ISSN: 1573-5028
    Keywords: histone variants ; cDNA ; expression ; maize
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The complete amino acid sequences of two variants of histone H2B of maize were deduced from the cDNAs isolated from a maize cDNA library. The two encoded proteins are 150 (H2B(1)) and 149 (H2B(2)) amino acids long and shows the classical organization of H2B histones. The hydrophobic C-terminal region is highly conserved as compared to that of the animal counterparts with only 21 changes (13 conservative) among the 90 residues. Between the N-terminal part and the C-terminal region we note the presence of a basic cluster (9 residues) characteristic of histones H2B. The N-terminal third is extended as compared to the animal consensus H2B and has the same size as the H2B histone of wheat. Up to 9 acidic residues and a five time repeated pentapeptide PA/KXE/KK are present in this region. Southern-blot hybrization showed that the H2B histones are encoded by a multigenic family like the other core histones (H3 and H4) of plants. The general expression pattern of these genes was not significantly different from that of the H3 and H4 genes neither in germinating seeds nor in different tissues of adult maize.
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  • 37
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    Plant molecular biology 20 (1992), S. 857-867 
    ISSN: 1573-5028
    Keywords: cDNA ; differential screening ; genomic cloning ; lignin ; maize ; O-methyltransferase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The isolation and characterization of cDNA and homologous genomic clones encoding the lignin O-methyltransferase (OMT) from maize is reported. The cDNA clone has been isolated by differential screening of maize root cDNA library. Southern analysis indicates that a single gene codes for this protein. The genomic sequence contains a single 916 bp intron. The deduced protein sequence from DNA shares significant homology with the recently reported lignin-bispecific caffeic acid/5-hydroxyferulic OMTs from alfalfa and aspen. It also shares homology with OMTs from bovine pineal glands and a purple non-sulfur photosynthetic bacterium. The mRNA of this gene is present at different levels in distinct organs of the plant with the highest accumulation detected in the elongation zone of roots. Bacterial extracts from clones containing the maize OMT cDNA show an activity in methylation of caffeic acid to ferulic acid comparable to that existing in the plant extracts. These results indicate that the described gene encodes the caffeic acid 3-O-methyltransferase (COMT) involved in the lignin biosynthesis of maize.
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  • 38
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    Plant molecular biology 21 (1993), S. 99-108 
    ISSN: 1573-5028
    Keywords: fusion protein ; Oenothera ; plastome ; RFLP ; unidentified open reading frame
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The chloroplast DNA encodes genes for components of photosynthesis and the transcription-translation machinery; a number of unidentified open reading frames (ORFs) are also present. To determine whether a large ORF in the inverted repeat of chloroplast DNA of tobacco (ORF2280) encodes a chloroplast protein, a conserved region of the ORF was expressed in Escherichia coli. An antibody against the ORF protein was prepared using the purified fusion protein as an antigen. When incubated with proteins from the soluble fraction of tobacco, spinach and Oenothera chloroplasts, the antiserum detects relatively labile polypeptides, which have apparent molecular weights of 170 to 180 kDa. The ORF in tobacco and spinach is large enough to encode a protein of 240–250 kDa, thus it is possible that post-transcriptional or post-translational processing reduces the size of the expression product. Analysis of Oenothera chloroplasts representing four different plastome types revealed endonuclease restriction fragment length polymorphisms in chloroplast DNA indicative of insertion/deletion events in a region of the chloroplast DNA that shared significant sequence similarity with ORF2280. The ORF2280 antiserum was used to demonstrate that there are qualitative differences in the ORF proteins from different Oenothera plastome types.
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  • 39
    ISSN: 1573-5028
    Keywords: transposable element ; Activator ; gene dosage ; maize ; tobacco
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of Ac copy number on the frequency and timing of germinal transposition in tobacco was investigated using the streptomycin phosphotransferase gene (SPT) as an excision marker. The activity of one and two copies of the element was compared by selecting heterozygous and homozygous progeny of transformants carrying single SPT::Ac inserts. It was observed that increasing gene copy not only increases the transposition frequency, but also occasionally alters the timing of transposition such that earlier events are obtained. The result is that some homozygous plants generate multiple streptomycin resistant progeny carrying the same transposed Ac (trAc) element. We have also investigated the effect of modification of the sequence in the region around 82 bp downstream of the polyadenylation site and 177 bp from the 3′ end of the element on germinal excision frequencies. Alteration of three bases to create a BglII site at this location caused a minor decrease in germinal excision events, but insertion of four bases to create a Cla I site caused a 10-fold decrease in the transposition activity of the Ac element.
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  • 40
    ISSN: 1573-5028
    Keywords: ABA-responsive element ; maize ; tissue-specific factors ; rab genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The maize gene rab28 has been identified as ABA-inducible in embryos and vegetative tissues. It is also induced by water stress in young leaves. The proximal promoter region contains the conserved cis-acting element CCACGTGG (ABRE) reported for ABA induction in other plant genes. Transient expression assays in rice protoplasts indicate that a 134 bp fragment (-194 to -60 containing the ABRE) fused to a truncated cauliflower mosaic virus promoter (35S) is sufficient to confer ABA-responsiveness upon the GUS reporter gene. Gel retardation experiments indicate that nuclear proteins from tissues in which the rab28 gene is expressed can interact specifically with this 134 bp DNA fragment. Nuclear protein extracts from embryo and water-stressed leaves generate specific complexes of different electrophoretic mobility which are stable in the presence of detergent and high salt. However, by DMS footprinting the same guanine-specific contacts with the ABRE in both the embryo and leaf binding activities were detected. These results indicate that the rab28 promoter sequence CCACGTGG is a functional ABA-responsive element, and suggest that distinct regulatory factors with apparent similar affinity for the ABRE sequence may be involved in the hormone action during embryo development and in vegetative tissues subjected to osmotic stress.
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  • 41
    ISSN: 1573-5028
    Keywords: β-tubulin ; microtubules ; maize ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Four different β-tubulin coding sequences were isolated from a cDNA library prepared from RNA from maize seedling shoots. The four genes (designated tub4, tub6, tub7 and tub8) represented by these cDNA clones together with the tub1 and tub2 genes reported previously encode six β-tubulin isotypes with 90–97.5% amino acid sequence identity. Results from phylogenetic analysis of 17 β-tubulin genes from monocot and dicot plant species indicated that multiple extant lines of β-tubulin genes diverged from a single precursor after the appearance of the two major subfamilies of α-tubulin genes described previously. Hybridization probes from the 3′ non-coding regions of six β-tubulin clones were used to quantify the levels of corresponding tubulin transcripts in different maize tissues including developing anthers and pollen. The results from these dot blot hybridization experiments showed that all of the β-tubulin genes were expressed in most tissues examined, although each gene showed a unique pattern of differential transcript accumulation. The tub1 gene showed a high level of transcript accumulation in meristematic tissues and almost no accumulation in the late stages of anther development and in pollen. In contrast, the level of tub4 transcripts was very low during early stages of male flower development but increased markedly (more than 100 times) during the development of anthers and in pollen. Results from RNAse protection assays showed that this increased hybridization signal resulted from expression of transcripts from one or two genes closely related to tub4. The tub4-related transcripts were not present in shoot tissue. Transcripts from the tub2 gene accumulated to very low levels in all tissues examined, but reached the highest levels in young anthers containing microspore mother cells. RNAse protection assays were used to measure the absolute levels of α- and β-tubulin transcripts in seedling shoot and in pollen. The α-tubulin gene subfamily I genes (tua1, tua2, tua4) contributed the great majority of α-tubulin transcripts in both shoot and pollen. Transcripts from the β-tubulin genes tub4, tub6, tub7, and tub8 were predominant in shoot, but were much less significant than the tub4-related transcripts in pollen.
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  • 42
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    Plant molecular biology 24 (1994), S. 449-463 
    ISSN: 1573-5028
    Keywords: AU-rich ; intron ; maize ; splicing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Plant introns are generally short (〈200 nt) and AU-rich, and an elevated AU content is necessary for efficient splicing. Further, an intron in some plant genes enhances gene expression by a post-transcriptional mechanism that results in an increase of cytoplasmic mRNA. The specific intron features responsible for efficient splicing and enhancement are not well characterized in plants. Internal deletions of up to 80% of two maize introns, Adh1 intron 1 and maize actin intron 3, indicate that large segments of these introns are dispensable for normal function. However, extensive deletion (〉75%) of Adh1 intron 1 diminishes both intron enhancement and splicing efficiency. This finding suggests that there are internal sequence motifs required for intron function, and that these motifs are redundant. We attempted to repair a deletion-impaired Adh1 intron 1 variant by adding back either oligomers of defined sequence content or fragments of maize internal intron sequence. The addition of AU-rich oligomers improved splicing efficiency and in one example, a U-rich oligomer activated a cryptic 3′ splice acceptor. We also found that replacing the region proximal to the Adh1 intron 1 3′ acceptor with U-rich sequence improved splicing. We found that adding G- and C-rich oligomers did not improve intron function, but a C-rich oligomer activated a cryptic 3′ acceptor. The addition of internal intron sequence to an impaired intron improved splicing, and in one case, resulted in the activation of a cryptic 3′ acceptor. We present evidence that U-rich sequence immediately upstream of the 3′ splice junction increases splicing efficiency and contributes to, but does not uniquely specify, 3′ acceptor AG choice.
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  • 43
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    Biochemical genetics 30 (1992), S. 233-246 
    ISSN: 1573-4927
    Keywords: phosphogluconate dehydrogenase ; isozymes ; maize ; gene dosage ; tissue-specific expression ; null alleles
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Maize (Zea mays L.) cytosolic 6-phosphogluconate dehydrogenase isozymes (EC 1.1.1.44; 6-PGD) are encoded by unlinked lociPgd1 andPgd2. Two families from a Robertson's Mutator line were isolated which have no detectable expression ofPgd2. ThesePgd2-null mutants and aPgd1-null line were used to generate plants homozygous for null alleles at both cytosolic 6-PGD loci. The specific activity of 6-PGD in the double-null mutant was between 20 and 30% of wild-type levels in root extracts. The double-null mutant was reproductively viable in a moderate environment, suggesting that wild-type levels of cytosolic 6-PGD activity are not essential for growth. Isozyme dimer ratios in roots, leaves, and scutellum were binomial and reflected the wild-type gene copy number. 6-PGD isozymes showed tissue- and cell type-specific expression.
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  • 44
    ISSN: 1573-4927
    Keywords: alcohol dehydrogenase ; maize ; Zea mays ; mutation ; cDNA
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    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
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    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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    Biochemical genetics 30 (1992), S. 233-246 
    ISSN: 1573-4927
    Keywords: phosphogluconate dehydrogenase ; isozymes ; maize ; gene dosage ; tissue-specific expression ; null alleles
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Maize (Zea mays L.) cytosolic 6-phosphogluconate dehydrogenase isozymes (EC 1.1.1.44; 6-PGD) are encoded by unlinked lociPgd1 andPgd2. Two families from a Robertson's Mutator line were isolated which have no detectable expression ofPgd2. ThesePgd2-null mutants and aPgd1-null line were used to generate plants homozygous for null alleles at both cytosolic 6-PGD loci. The specific activity of 6-PGD in the double-null mutant was between 20 and 30% of wild-type levels in root extracts. The double-null mutant was reproductively viable in a moderate environment, suggesting that wild-type levels of cytosolic 6-PGD activity are not essential for growth. Isozyme dimer ratios in roots, leaves, and scutellum were binomial and reflected the wild-type gene copy number. 6-PGD isozymes showed tissue- and cell type-specific expression.
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  • 47
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    Plant molecular biology 14 (1990), S. 333-347 
    ISSN: 1573-5028
    Keywords: intergenic spacer ; maize ; methylation ; rDNA ; teosinte
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The restriction endonucleases Hpa II and Msp I were used to examine cytosine methylation in the ribosomal RNA genes (rDNA) of inbred lines of maize and species of teosinte. In all of the rDNAs examined, Msp I (not sensitive to mCpG) digestion yielded a distribution of lower molecular weight fragments indicative of multiple recognition sites. The majority of the rDNA arrays in an individual were inaccessible to Hpa II (sensitive to mCpG) cleavage, but a significant fraction (10–25%) was cleaved at least once by Hpa II into repeat unit length fragments (9.1 kbp). In some maize inbred lines, one or two additional fragment populations (less than 9.1 kbp in length) were also produced by Hpa II digestion. All of the unmethylated Hpa II sites mapped to the intergenic spacer (IGS), and the major unmethylated site was located approximately 800 bp 5′ to the start of the 18S RNA coding sequence. An Eco RI polymorphism, present in the 26S gene of certain inbred lines and hybrids, was utilized to investigate the organization of unmethylated repeat units in the rDNA array. In double digest experiments with Hpa II/Eco RI, the fragments from repeat units with two Eco RI sites were sensitive to Hpa II digestion, whereas, the fragments from repeat units with a single Eco RI site were almost completely resistant to Hpa II digestion. Similar digestion patterns were also observed in Eco RII (sensitive to mCNG)/Eco RI digests. These results suggest that unmethylated and Eco RI polymorphic sites occur in the same repeat units.
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  • 48
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    Keywords: retrotransposon ; Pisum ; segregation ; PCR ; RFLP ; mapping
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    Notes: Abstract A DNA sequence between two legumin genes in Pisum is a member of the copia-like class of retrotransposons and represents one member of a polymorphic and heterogeneous dispersed repeated sequence family in Pisum. This sequence can be exploited in genetic studies either by RFLP analysis where several markers can be scored together, or the segregation of individual elements can be followed after PCR amplification of specific members.
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  • 49
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    Plant molecular biology 15 (1990), S. 783-785 
    ISSN: 1573-5028
    Keywords: sorghum ; maize ; glycine-rich proteins ; RNA-binding
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  • 50
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    Plant molecular biology 16 (1991), S. 919-923 
    ISSN: 1573-5028
    Keywords: Zea mays ; maize ; embryogenesis ; abscisic acid ; seed maturation
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    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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  • 51
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    Plant molecular biology 16 (1991), S. 935-936 
    ISSN: 1573-5028
    Keywords: maize ; mitochondrial DNA ; repeated sequence
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  • 52
    ISSN: 1573-5028
    Keywords: hygromycin ; inheritance ; maize ; tissue culture ; transformation
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    Topics: Biology
    Notes: Abstract Embryogenic maize (Zea mays L.) callus cultures were transformed by microprojectile bombardment with a chimeric hygromycin phosphotransferase (HPT) gene and three transformed lines were obtained by selecting for hygromycin resistance. All lines contained one or a few copies of the intact HPT coding sequence. Fertile, transgenic plants were regenerated and the transmission of the chimeric gene was demonstrated through two complete generations. One line inherited the gene in the manner expected for a single, dominant locus, whereas two did not.
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  • 53
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    Plant molecular biology 18 (1992), S. 201-210 
    ISSN: 1573-5028
    Keywords: maize ; transformation ; inheritance ; phosphinothricin acetyltransferase ; cotransformation ; microprojectile bombardment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Progeny recovered from backcrossed transgenic maize tissue culture regenerants (R0) were analyzed to determine the segregation, expression, and stability of the introduced genes. Transgenic A188×B73 R0 plants (regenerated from embryogenic suspension culture cells transformed by microprojectile bombardment; see [9]) were pollinated with nontransformed B73 pollen. Inheritance of a selectable marker gene, bar, and a nonselectable marker gene, uidA, was analyzed in progeny (R1) representing four independent transformation events. Activity of the bar gene product, phosphinothricin acetyltransferase (PAT), was assessed in plants comprising the four R1 populations. The number of R1 plants containing PAT activity per total number of R1 plants recovered for each population was 2/7, 19/34, 3/14 and 73/73. Molecular analysis confirmed the segregation of bar in three R1 populations and the lack of segregation in one R1 population. Cosegregation analysis indicated genetic linkage of bar and uidA in all four R1 populations. Analysis of numerous R2 plants derived from crossing transformed R1 plants with nontransformed inbreds revealed 1:1 segregation of PAT activity in three of four lines, including the line that failed to segregate in the R1 generation. Integrated copies of bar in one line appeared to be unstable or poorly transmitted.
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  • 54
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    Keywords: maize ; protease inhibitor ; trypsin ; activated Hageman factor ; cDNA
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  • 55
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    Plant molecular biology 17 (1991), S. 1259-1261 
    ISSN: 1573-5028
    Keywords: anaerobic genes ; maize ; pyruvate decarboxylase ; DNA sequence
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  • 56
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    Plant molecular biology 18 (1992), S. 1181-1184 
    ISSN: 1573-5028
    Keywords: alcohol dehydrogenase-1 ; maize ; plant ; promoter ; Z-DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Z-DNA is a left-handed helix which can form within tracts of alternating purines and pyrimidines. Tracts of potential Z-DNA identified by sequence inspection are often noted within regulatory portions of genes, but evidence that these tracts of sequence actually exist as Z-DNA is very limited, and not available for any plant gene. In this study, the chemical probes osmium tetroxide, diethylpyrocarbonate and hydroxylamine were used to show that a tract of alternating purines and pyrimidines in the Adh1 promoter (from -311 to -325) actually assumes a Z-DNA conformation under superhelical stress in vitro.
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  • 57
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    Plant molecular biology 21 (1993), S. 885-893 
    ISSN: 1573-5028
    Keywords: cell wall ; deglycosylation ; extensin ; hydroxyproline-rich ; immunoprecipitation ; maize
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract This study concerned the molecular basis for the protein size heterogeneity of extensin from two maize (Zea mays L.) varieties. We studied the physical properties of extensin, a hydroxyproline-rich glycoprotein (HRGP), from the silk and pericarp of Golden X Bantam (GXB) sweet corn and Japanese Hulless (JHL) popcorn. Extensin from GXB has a molecular mass of 66 kDa whereas extensins from JHL have molecular masses of 76 and 66 kDa. Treatment with anhydrous hydrogen fluoride to deglycosylate proteins reduced the size of all extensins by 5 kDa. Probing with a 500 bp fragment from a genomic clone of maize extensin identified two transcripts (1.9 and 1.5 kb) on northern blots. JHL contained both transcripts and GXB contained only the 1.5 kb transcript. The probe also hybridized to two larger transcripts (6.2 and 4.5 kb) that were found in both varieties. We immunoprecipitated two proteins (66 and 56 kDa) from translated RNA isolated from JHL and one protein (56 kDa) from GXB. These results demonstrate that these extensins differ in the size of their peptide moiety and not in their extent of glycosylation.
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  • 58
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    Plant molecular biology 19 (1992), S. 533-536 
    ISSN: 1573-5028
    Keywords: chlorophyll a/b-binding protein ; CAB gene ; nucleotide sequence ; maize
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  • 59
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    Plant molecular biology 22 (1993), S. 783-792 
    ISSN: 1573-5028
    Keywords: proteinase inhibitor ; wound induction ; wound-induced gene ; maize
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have cloned and sequenced a wound-inducible cDNA clone designated WIP1 (for wound-induced protein) from maize coleoptiles. It was isolated by differential screening of a cDNA library prepared from excised maize coleoptile segments. The deduced amino acid sequence predicts a secretory, cysteine-rich protein of 102 residues with a calculated molecular mass of 11 kDa and a typical N-terminal signal sequence. The protein has about 30% identity with various Bowman-Birk type proteinase inhibitors. Most interestingly, it is novel in that it is double-headed with exclusive specificity for chymotrypsin. WIP1 is strongly wound-induced in contrast to other members of the Bowman-Birk proteinase inhibitor family, which occur in seeds and are regulated during development. The response is fast, similar to defenceinduced genes, and measurable as early as 30 min after wounding. Induction can also be evoked in the intact coleoptiles and the signal is systemically transmitted in the coleoptile to adjacent regions of the wounded area. Isolation and analysis of the corresponding genomic clone reveals that WIP1 contains an intron of 90 nucleotides.
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  • 60
    ISSN: 1573-5028
    Keywords: C4 photosynthesis ; maize ; plastid gene expression ; Sorghum
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    Notes: Abstract The transcription of plastome-encoded genes in mesophyll and bundle-sheath chloroplasts of the monocotyledonous NADP-malic enzyme-type C4 species Zea mays L. (maize) and Sorghum bicolor (L.) Moench. was investigated. RNA accumulation and transcription were assayed starting from isolated mesophyll and bundle-sheath chloroplasts and using quantitative northern and run-on transcription analysis. Determination of the mesophyll to bundle-sheath ratios of transcript abundance in maize and Sorghum chloroplasts showed that the mRNAs of the plastome-encoded photosystem II genes analysed (psbA, psbB, psbD, psbH and psbE/F) varied from 2.5- to 4.0-fold (maize) and 3.1- to 5.2-fold (Sorghum), respectively. The rbcL transcript, in contrast, was more abundant in bundle-sheath chloroplasts of both species, about 3-fold in maize and more than 10-fold in Sorghum. On the other hand, transcripts of genes encoding the 16S ribosomal RNA (r16) and subunits of photosystem I (psaA) and the cytochrome b/f complex (petB, petA) accumulated to similar levels in both types of chloroplasts. Determination of absolute transcript levels for rbcL and psbA in chloroplasts from maize and Sorghum demonstrated that for both genes, the mesophyll to bundle-sheath differences in transcript abundance were more pronounced in Sorghum. Measurements of the transcriptional activities of rbcL and psbA showed that the transcription rate of rbcL is higher in bundle-sheath chloroplasts while psbA is more actively transcribed in mesophyll chloroplasts. The differences in the transcription rates between the two chloroplast types were again more pronounced in Sorghum, thus reflecting the differences between maize and Sorghum in the relative levels of the rbcL and psbA transcripts. However, although transcription rate and mRNA abundance are correlated, they did not exactly match one another. This indicates additional regulation of transcript abundance at the level of RNA stability.
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  • 61
    ISSN: 1573-5028
    Keywords: Cell biology ; epigenetics ; maize ; transformation ; transgenes
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    Topics: Biology
    Notes: Abstract Zea mays transformants produced by particle bombardment of embryogenic suspension culture cells of the genotype A188 × B73 and selected on kanamycin or bialaphos were characterized with respect to transgene integration, expression, and inheritance. Selection on bialaphos, mediated by thebar orpat genes, was more efficient than selection on kanamycin, mediated by thenptII gene. Most transformants contained multicopy, single locus, transgene insertion events. A transgene expression cassette was more likely to be rearranged if expression of that gene was not selected for during callus growth. Not all plants regenerated from calli representing single transformation events expressed the transgenes, and a non-selectable gene (uidA) was expressed in fewer plants than was the selectable transgene. Mendelian inheritance of transgenes consistent with transgene insertion at a single locus was observed for approximately two thirds of the transformants assessed. Transgene expression was typically, but not always, predictable in progeny plants-transgene silencing, as well as poor transgene transmission to progeny, was observed in some plant lines in which the parent plants had expressed the transgene.
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  • 62
    ISSN: 1573-5028
    Keywords: cDNA clones ; EST sequencing ; maize ; RFLP mapping
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    Notes: Abstract As one component of a maize genome project, we report the analysis of a number of randomly selected cDNAs, by a combination of measuring mRNA expression, ‘single-pass’ sequencing (SPS), and genome mapping. Etiolated seedling (490) and membrane-free polysomal endosperm cDNA clones (576) were evaluated for their transcription levels by hybridizing with a probe prepared from total mRNA and categorized as corresponding to abundantly or rarely expressed mRNAs and as either constitutive or tissue-specific. A total of 313 clones from the two libraries were submitted to ‘single-pass’ sequencing from the presumed 5′ end of the mRNA and the nucleotide sequence compared with the GenBank database. About 61% of the clones showed no significant similarities within GenBank, 14% of the clones exhibited a high degree of similarity, while the remaining 25% exhibited a lesser degree of similarity. The chromosomal location of more than 300 clones was determined by RFLP mapping using standard populations. The results demonstrate that a combination of analyses provides synergistic information in eventually deducing the actual function of these types of clones.
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  • 63
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    Plant molecular biology 26 (1994), S. 1065-1071 
    ISSN: 1573-5028
    Keywords: Brassica ; polyploid ; gene expression ; RT-PCR ; RFLP
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    Topics: Biology
    Notes: Abstract One of the essential issues regarding evolution of polyploid species is how duplicate genes are expressed. Most studies on gene expression in polyploids have been based on isozyme analyses; RNA analysis has not been widely used partially due to difficulties in distinguishing homologous transcripts which usually have the same length and similar or almost identical sequences. In this study, a method combining RT-PCR with RFLP was used to analyze transcripts of homologous genes in natural and synthetic Brassica amphidiploids. Sequences coding for several known genes were selected and used to synthesize gene-specific primers. Total RNAs were used as templates for RT-PCR to amplify homologous transcripts in three diploid parental species, three cultivated amphidiploid species and six synthetic amphidiploids. For each gene, initial PCR products amplified in all species had identical length; however, homologous transcripts in the diploid and amphidiploid species could be distinguished after digesting the PCR products with restriction enzymes. Preliminary results based on three genes indicated that both transcripts from the diploid parents were expressed in the synthetic and natural amphidiploids. This study represents the first application of RT-PCR and RFLP analysis to investigate expression of homologous genes in higher plants. The technique is a sensitive, simple and efficient method for distinguishing homologous transcripts in a mixed RNA population and can be applied to many types of studies on expression of homologous genes.
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    Plant molecular biology 14 (1990), S. 1-15 
    ISSN: 1573-5028
    Keywords: cDNA ; differential screening ; genomic cloning ; maize ; tandem genes ; α-tubulin
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    Topics: Biology
    Notes: Abstract The identification of a cDNA (MR19) corresponding to a maize α-tubulin and homologous genomic clones (MG19/6 and MG19/14) is described. The cDNA has been isolated by differential screening of a cDNA maize root library. We have found two α-tubulin genes in a tandem arrangement in the genomic clones, separated by approximately 1.5 kbp. One of the genes (gene I) contains an identical nucleotide sequence which corresponds to the cDNA clone. The two deduced proteins from DNA sequences are very similar (only two conservative replacements in 451 amino acids) and they share a high homology as compared with the published α-tubulin sequences from other systems and in particular with the Arabidopsis thaliana and Chlamydomonas reinhardtii sequences reported. The structure of both genes is also very similar; it includes two introns, of 1.7 kbp and 0.8 kbp respectively, in each gene and only one intron placed at a homologous position in relation to Arabidopsis thaliana genes. By using specific 3′ probes it appears that both genes are preferentially expressed in the radicular system of the plant. The α-tubulin gene family of Zea mays seems to be represented by at least 3 or 4 members.
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  • 65
    ISSN: 1573-5028
    Keywords: maize ; Coix ; zein ; coixin ; phylogenetic relationship
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    Notes: Abstract Zeins from Zea mays L cv. Maya and coixins from Coix lacryma-jobi L. cv. Adlay were fractionated to obtain α-, β-, and γ-zein and α-, β-, and γ-coixin. The α-coixins were composed of 4 polypeptide classes of 27 kDa (C1), 25 kDa (C2), 17 kDa (C4) and 15 kDa (C5) with solubility properties very similar to those of the 22 kDa and 19 kDa α-zeins. Like the α-zeins, the C1 and C2 α-coixins corresponded to 80% of total Coix prolamins. The fraction corresponding to γ-coixin contained only one protein band of 22 kDa (C3). This coixin fraction has solubility properties similar to those of γ-zein and represents 15% of the total coixin. The β-zein fraction was composed of a major 17 kDa protein band, while the β-coixin fraction consisted of a mixture of α- and γ-coixins. Polyclonal antibodies raised against C1 recognized C1 and C2 and cross-reacted strongly with the 22 kDa α-zein, as did C4 and C5 antisera. The antiserum against γ-coixin showed strong cross-reaction with γ-zein. The homology between coixins and zeins was further investigated by using Southern hybridization analyses. The genomic DNA of maize and Coix were digested with several restriction enzymes and probed with cDNA clones representing 19 and 22 kDa α-zeins as well as the 28 and 16 kDa γ-zeins. The Coix genome showed complex cross-hybridization sequences with the 22 kDa α-zein cDNA, while no cross-hybridization was observed with the 19 kDa cDNA clone. The cDNA clone representing the 28 kDa γ-zein cross-hybridized with only one band of Coix genomic DNA, in contrast to the three bands observed in maize. This same Coix sequence also cross-hybridized with the cDNA clone representing the 16 kDa γ-zein. The relevance of these findings are discussed in the context of the origin of zein and coixin genes.
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    Plant molecular biology 16 (1991), S. 831-839 
    ISSN: 1573-5028
    Keywords: RFLP ; Oryza ; rice ; genome evolution ; allotetraploid
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    Topics: Biology
    Notes: Abstract Restriction fragment length polymorphisms (RFLPs) were studied in fourteen accessions of CCDD genome allotetraploid wild rice species (Oryza latifolia, O. alta and O. grandiglumis). Fourteen nuclear RFLP markers previously mapped in AA genome-cultivated rice were used as probes. A phylogenetic tree, constructed by parsimony analysis based on RFLPs, grouped the accessions according to their geographic origin from Central or South America. Oryza alta, O. grandiglumis and one accession of O. latifolia grouped together as a subgroup, and our results suggested that the three taxa should be considered as populations of a single complex species. Duplicate loci, representing the two constituent genomes of the allotetraploid, were observed for most RFLP markers. By comparing RFLPs from the allotetraploids with those from a CC genome diploid wild species (O. officinalis), it was possible to detect RFLPs specific for both the CC and DD genomes of the allotetraploid. In inter-accession F2 populations, independent segregation of RFLP markers for CC and DD genomes was observed.
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  • 67
    ISSN: 1573-5028
    Keywords: anaerobic stress ; enolase ; gene regulation ; maize
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    Notes: Abstract A cDNA encoding maize enolase (2-phospho-D-glycerate hydrolase) was purified by functional genetic complementation using an enolase deficient mutant of Escherichia coli, DF261. This cDNA, pZM245, was characterized by restriction mapping and DNA sequence analysis. The cDNA contained an open reading frame encoding a protein of 446 amino acids with a high degree of similarity to enolase sequences from other organisms (72% identity to yeast enolase and 82% identity to human enolase). The pZM245 contains a correctly positioned consensus prokaryotic translation initiation sequence. The specific activity of enolase in maize increases to about twice its initial level after 48 hours of anaerobiosis. Northern-blot analysis showed a five-fold anaerobic induction in enolase mRNA, while heat shock or cold shock increased enolase mRNA levels only slightly. Southern-blot analysis of maize genomic DNA indicated that there is one copy of the pZM245 hybridizing sequence per haploid genome in maize.
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  • 68
    ISSN: 1573-5028
    Keywords: chromosome assignment ; dihaploid progenies ; heterozygosity ; potato ; rDNA ; RFLP
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    Notes: Abstract Restriction map and organization of rDNA was inferred from analysis of dihaploid progenies of two tetraploid genotypes of cultivated potato. Each tetraploid genotype was characterized by a specific distribution of different types of rDNA repetition units on their four homologous chromosomesII. The genotypes were heterozygous and differed by the kind of units carried by each chromosomeII. Models for the generation of the observed organization are discussed and supported by first cloning studies.
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  • 69
    ISSN: 1573-5028
    Keywords: cDNA family ; maize ; pollen-expressed genes ; polygalacturonase ; signal peptide
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    Notes: Abstract A full-length cDNA clone, named PG1, abundantly expressed in late stages of pollen development, has been isolated from a cDNA library using a differential screening method with cDNA probes representative of microspores at early or late developmental stages. The encoded 410 amino acid polypeptide has significant homology with various polygalacturonases (PG) described elsewhere. Two polypeptides, of 49 and 53 kDa respectively, have been identified in the active PG fraction, isolated from mature pollen by immuno-cross-reaction with tomato PG antibodies. According to their N-terminal sequence, they can be identified as being mature peptides encoded by the PG1 cDNA clone. We propose that these two proteins derive from a unique precursor through several post-translational events, including the excision of a 22 amino-terminal signal peptide and glycosylation. PG-encoding genes form a small genomic family. Sequence analysis of three PG cDNA clones shows that they are closely related. The divergence of nucleotides between these three cDNA clones is 1%. They encode the same product.
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  • 70
    ISSN: 1573-5028
    Keywords: electroporation ; heat shock ; maize ; promoter ; splicing ; ubiquitin
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    Notes: Abstract Two genomic clones (λUbi-1 and λUbi-2) encoding the highly conserved 76 amino acid protein ubiquitin have been isolated from maize. Sequence analysis shows that both genes contain seven contiguous direct repeats of the protein coding region in a polyprotein conformation. The deduced amino acid sequence of all 14 repeats is identical and is the same as for other plant ubiquitins. The use of transcript-specific oligonucleotide probes shows that Ubi-1 and Ubi-2 are expressed constitutively at 25°C but are inducible to higher levels at elevated temperatures in maize seedlings. Both genes contain an intron in the 5′ untranslated region which is inefficiently processed following a brief, severe heat shock. The transcription start site of Ubi-1 has been determined and a transcriptional fusion of 0.9 kb of the 5′ flanking region and the entire 5′ untranslated sequence of Ubi-1 with the coding sequence of the gene encoding the reporter molecule chloramphenicol acetyl transferase (CAT) has been constructed (pUBI-CAT). CAT assays of extracts of protoplasts electroporated with this construct show that the ubiquitin gene fragment confers a high level of CAT expression in maize and other monocot protoplasts but not in protoplasts of the dicot tobacco. Expression from the Ubi-1 promoter of pUBI-CAT yields more than a 10-fold higher level of CAT activity in maize protoplasts than expression from the widely used cauliflower mosaic virus 35S promoter of a 35S-CAT construct. Conversely, in tobacco protoplasts CAT activity from transcription of pUBI-CAT is less than one tenth of the level from p35S-CAT.
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  • 71
    ISSN: 1573-5028
    Keywords: expression ; germination ; histone genes ; maize ; subfamilies ; tissues
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    Notes: Abstract The steady-state levels of H3 and H4 mRNAs transcribed from three H3 and two H4 multigene subfamilies were studied during germination and in different organs of maize. During germination the five subfamilies are expressed in parallel to DNA synthesis, but a 5-fold difference in the quantity of mRNAs transcribed per gene copy was found from our subfamily to another. In adult plants H3 and H4 mRNA levels are highest in organs containing meristematic tissues but also high in non-proliferating tissues. No strict tissue specificity expression could be detected but some subfamilies show preferential expression in some tissues.
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  • 72
    ISSN: 1573-5028
    Keywords: cell cycle ; maize ; non-systemic expression ; polyubiquitin gene ; stress response
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    Notes: Abstract We have used the promoter, 1st exon and 1st intron of the maize polyubiquitin gene (Ubi-1) for rice transformation experiments and revealed the characteristic expression of Ubi-1 gene: (1) Ubi-1 gene is not regulated systemically but rather individual cells respond independently to the heat or physical stress; (2) Ubi-1 gene changes its tissue-specific expression in response to stress treatment; (3) the expression of Ubi-1 gene is dependent on cell cycle.
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  • 73
    ISSN: 1573-5028
    Keywords: cDNA ; cyanogenesis ; β-glucosidase ; RFLP ; Trifolium repens
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    Topics: Biology
    Notes: Abstract The Li locus in white clover controls the presence of cyanogenic β-glucosidase (linamarase) activity in leaf tissue, such that plants homozygous for the ‘null’ allele (li) have no linamarase activity in this tissue. The isolation of a cDNA clone from linamarase mRNA is described. The cDNA clone is used to further characterise alleles of the Li locus. Northern blot analysis shows that plants homozygous for the ‘null’ allele (li li) produce very reduced levels of mRNA which hybridises to the cDNA. Heterozygous plants (Li li), which have intermediate levels of enzyme activity, produce intermediate levels of mRNA. Southern blot analysis of Hind III digested genomic DNA shows that the white clover genome contains three genes with homology to the linamarase cDNA and that at least two of these genes segregate independently. Analysis of the cosegregation of linamarase activity and the presence of genomic restriction fragments identifies the genomic sequence specifying linamarase structure and indicates either a structural or cis acting control function of the Li locus.
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  • 74
    ISSN: 1573-5028
    Keywords: maize ; ABA-induced gene ; protein phosphorylation
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    Topics: Biology
    Notes: Abstract The ABA-induced MA12 cDNA from maize, which encodes a set of highly phosphorylated embryo proteins, was used to isolate the corresponding genomic clone. This gene, called RAB-17 (responsive to ABA), encodes a basic, glycine-rich protein (mol. wt. 17 164) containing a cluster of 8 serine residues, seven of them contiguous. It is a homologue of the rice RAB-21 gene (Mundy J, Chua NH, EMBO J 7; 2279–2286, 1988). Phosphoamino acid analysis of the isolated protein indicates that only the serine residues are phosphorylated and a putative casein-type kinase phosphorylatable sequence was identified in the protein. The pattern of expression and in vivo phosphorylation of the RAB-17 protein was studied during maize embryo germination and in calli of both meristematic or embryonic origin. ABA treatment induced the synthesis of RAB-17 mRNA and protein in calli, however, the RAB-17 proteins were found to be highly phosphorylated only in embryos.
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  • 75
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    Plant molecular biology 15 (1990), S. 913-920 
    ISSN: 1573-5028
    Keywords: Adh1 ; chloramphenicol acetyl transferase ; enhancer ; intron ; maize ; transient expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Chimeric genes containing the coding sequence for bacterial chloramphenicol acetyl transferase (CAT) have been introduced by electroporation into maize protoplasts (Black Mexican Sweet) and transient expression monitored by enzyme assays. Levels of CAT expression were enhanced 12-fold and 20-fold respectively by the inclusion of maize alcohol dehydrogenase-1 introns 2 and 6 in the chimeric construct. This enhancement was seen when the intron was placed within the 5′ translated region but not when it was located upstream of the promoter or within the 3′ untranslated region. Deletion of exon sequences adjacent to intron 2 abolished its ability to mediate enhancement of CAT gene expression. Northern analysis of protoplasts electroporated with intron constructs revealed elevated levels of CAT mRNA. However, this elevation was insufficient to account for the increased enzyme activity. One explanation of these results is that splicing affects both the quantity of mRNA.
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  • 76
    ISSN: 1573-5028
    Keywords: germination ; fungi ; maize ; pathogenesis-related ; seed
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    Topics: Biology
    Notes: Abstract Pathogenesis-related proteins (PRs) are plant proteins produced in leaves in response to infection by pathogens including viruses, viroids, fungi and bacteria. Information on the presence and/or expression of PRs in monocotyledonous plants is scarce. Here we report the identification of cDNA and genomic clones coding for a basic form of a protein from germinating maize seeds having a high homology with the group of PR-1 from tobacco. A cDNA library enriched in aleurone-specific sequences was prepared from maize seeds two days after germination. One clone was found to contain an open reading frame encoding a protein homologous to PR proteins from tomato (p14) and tobacco (PR-1 group). Sequence analysis of the corresponding genomic clone revealed that it was encoded by a single exon. Besides, DNA blot hybridization indicates that this PR-like protein is encoded by a single-copy gene in maize. The accumulation of its mRNA increases after rehydration of desiccated seeds. Furthermore, a relationship was found between its expression and infection by a natural pathogen of maize, the fungus Fusarium moniliforme. The possible role of this protein as a response mechanism following fungal infection in cereal seeds is discussed.
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  • 77
    ISSN: 1573-5028
    Keywords: heat shock ; heat shock cDNAs ; maize ; small heat shock proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Heat-shocked maize seedlings (cv. Oh43) synthesize a characteristic set of heat-shock proteins (hsps) which include an 18 kDa family containing at least six major isoelectric variants. A cDNA library was constructed from poly(A)+ RNAs isolated from the radicles of heat-shocked maize seedlings and screened with a DNA fragment from the theoretical open reading frame of a putative Black Mexican Sweet maize hsp 18 genomic clone. Two clones, cMHSP18-3 and cMHSP18-9, were isolated, and the RNA transcripts generated from them were translated into proteins which immunoreact with antibodies directed against the maize 18 kDa hsps and exhibit the same electrophoretic characteristics as two different members of the 18 kDa hsp family. Nucleotide sequence analyses of the cDNAs in these clones reveal that their 5′ and 3′ untranslated regions exhibit 33–34% identity and that their protein encoding regions share 93% identity. The deduced amino acid sequences of these clones show 90% identity, and the apparent molecular masses and isoelectric points of these proteins agree with those established for two different 18 kDa hsps, numbered 3 and 6. This report substantiates that at least two of the 18 kDa hsps in maize are products of different but related genes. Moreover, it establishes that transcripts for these proteins accumulate during heat shock and that both their nucleotide and deduced amino acid sequences share extensive similarities with the class VI small hsps in soybean and with transcripts expressed during meiosis in Lilium.
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  • 78
    ISSN: 1573-5028
    Keywords: enhanced gene expression ; exon ; intron ; maize ; rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Both exon 1 and intron 1 of the maize Shrunken-1 (Sh1) gene individually stimulate expression of reporter genes in transient gene expression experiments if present within the transcription unit. The Sh1 exon 1 mediates a 10-fold increase in activity when inserted at the 5′ end of the bacterial chloramphenicol transacetylase (CAT) marker gene in both monocot and dicot protoplasts. The Sh1 intron 1 enhances chimeric gene expression in rice and maize protoplasts approximately 100-fold but inhibits CAT expression in tobacco protoplasts. In combination, the stimulatory effects of Sh1 exon 1 and intron 1 are multiplicative in monocot protoplasts resulting in a final enhancement of up to 1000-fold compared to the unmodified CAT or luciferase marker genes.
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  • 79
    ISSN: 1573-5028
    Keywords: anthocyanin ; B-Peru ; maize ; transcription activator
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  • 80
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    Plant molecular biology 17 (1991), S. 305-307 
    ISSN: 1573-5028
    Keywords: adenine nucleotide translocator ; ADP/ATP translocase ; mitochondria ; maize ; inner mitochondrial membrane
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  • 81
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    Plant molecular biology 17 (1991), S. 361-369 
    ISSN: 1573-5028
    Keywords: chloroplast genome ; cloning in YACs ; maize ; stability analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The 139 kb circular maize chloroplast genome was cloned in a linear form in a single yeast artificial chromosome (YAC), and several such clones were then analyzed to study the stability of large DNA sequences in YACs. The YAC clones were isolated from a partial maize DNA YAC library. The library was constructed byBal-31 nuclease digestion of high-molecular-weight total maize genomic DNA followed by blunt-end ligation into a modified YAC vector, pYAC11. All chloroplast genome-containing YACs (cpYACs) were characterized by comparing their restriction enzyme digestion patterns with that of purified maize chloroplast DNA (MCD). Comparison of the restriction patterns of four cpYACs, MY503, MY504, MY518 and MY526, with that of MCD did not show any change in the size of the restriction fragments except at the chloroplast DNA-vector junction fragments. These altered size fragments were confirmed as junction fragments by hybridization of the pYAC11 border fragments. The end-specific T3 and T7 transcripts synthesized from MY503 also identified the same junction fragments as did the pYAC11 border fragments. Most cpYACs were completely stable through the analyzed one hundred generations.
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  • 82
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    Plant molecular biology 17 (1991), S. 679-690 
    ISSN: 1573-5028
    Keywords: herbicide safener ; maize ; inducible plant genes ; differential gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A search of compounds capable of inducing specific gene expression in plants without affecting growth and development led to the examination of changes in the pattern of gene expression in corn after treatment with substituted benzenesulfonamide herbicide safeners. Following hydroponic treatment of corn with the safener N-(aminocarbonyl)-2-chlorobenzenesulfonamide (2-CBSU), the specific induction of new translatable mRNA species was observed. Replicate copies of a cDNA library made using RNA from 2-CBSU-treated corn roots were differentially screened with cDNA probes made from either the same mRNA fraction used for library construction or mRNA isolated from roots treated with 2-chlorobenzenesulfonamide (2-CBSA), an inactive analog of the safener. Colonies showing hybridization only with the probe made using mRNA from 2-CBSU-treated roots were further characterized to assess the specificity of the induction and decay of the corresponding induced RNA species. RNA blot analyses showed two clones, designated In2-1 and In2-2, contained plasmids that hybridized to RNAs that were induced from an undetectable background in corn roots within 30 minutes after treatment with 2-CBSU. Leaf and meristem tissues showed similar inductions of the In2-1 and In2-2 RNA species after a delay of several hours. In addition, both RNA species were induced in corn by foliar application of 2-CBSU. In contrast, neither RNA species was induced following stress treatments of plants. These results indicate a substituted benzenesulfonamide safener might be used with the promoters from the In2-1 and In2-2 genes to develop a new inducible gene expression system for plants.
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  • 83
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    Plant molecular biology 21 (1993), S. 805-821 
    ISSN: 1573-5028
    Keywords: DNaseI sensitivity ; maize ; methylation ; nucleolar dominance ; rDNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An Eco RI polymorphism, present in the 26S ribosomal RNA gene (rDNA) of the maize hybrid Sx19 (B73×Mo17), was utilized to correlate DNaseI sensitivity, undermethylation and expression in rDNA. We had previously shown that in double digest experiments with methylation-sensitive restriction enzymes and Eco RI, Sx19 rDNA fragments originating from repeat units with two Eco RI sites (8.0 kb) are undermethylated, whereas the fragments originating from repeat units with a single Eco RI site (9.1 kb) are completely methylated. In the present study, Sx19 rDNA chromatin structure was examined by purifying intact nuclei and digesting them briefly with increasing amounts of DNaseI. Analysis of this DNA with Eco RI showed that the 8.0 kb rDNA fragments are extremely sensitive to DNaseI digestion, while the 9.1 kb rDNA fragments are relatively resistant to digestion even at high levels of DNasel. Specific sites hypersensitive to DNaseI cleavage were mapped to a region in the intergenic spacer (IGS) near the major undermethylated site. Analysis of polymerase chain reaction (PCR) products synthesized using Sx19, B73, and Mo17 DNAs as templates indicated that the Eco RI polymorphism is due to a base change in the recognition site. Direct rRNA sequencing identified a single-base change in Mo17 rRNA relative to B73 rRNA. Allele-specific oligonucleotide probes containing the region surrounding and including the Eco RI polymorphic site were utilized to detect a nucleolar dominance effect by quantitating levels of rRNA transcripts in Sx19 and the reciprocal cross. Results from these single-base-pair mismatch hybridization experiments indicate that the majority of the rRNA transcripts in Sx19 orginate from the DNaseI-sensitive, undermethylated, Eco RI-polymorphic rDNA repeat units.
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  • 84
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    Plant molecular biology 22 (1993), S. 323-336 
    ISSN: 1573-5028
    Keywords: gene cluster ; maize ; multigene family ; transcriptional regulation ; zein ; early in-frame stop codon
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cluster of five α-zein subfamily 4 (α-zein SF4) genes are present in a 56 kb region of the maize W22 genome. Two types of α-zein SF4 genes are in the cluster. One of the genes, termed a type 1 (T1) α-zein SF4 gene, contains no early in-frame stop codons. Four of the genes, termed type 2 (T2) α-zein SF4 genes, contain one or two early in-frame stop codons. The base sequence of the T1 α-zein SF4 gene is similar (〉90%) to the sequences of any of the four T2 α-zein SF4 genes. However, their sequences differ markedly at distances greater than -875 bp upstream from the translation initiation codon of the α-zein coding region. This region of dissimilarity is well inside the functional 5′-flanking region for the genes since a 1.8 kb transcript is initiated in this region and the sequences of the T2 α-zein SF4 genes are similar in this region. Two sizes of mRNA transcripts, 1.8 kb and 0.9 kb, were detected in a gene specific manner for 4 of the 5 genes in this α-zein SF4 gene cluster. One of the T2 α-zein SF4 genes had only the 0.9 kb transcript. The RNA level for the 0.9 kb transcript of the T1 α-zein SF4 gene was 5- to 10-fold higher than the transcript levels of any of the T2 α-zein SF4 genes. In each case, the amount of the 0.9 kb transcript detected was at least 5-fold higher than the amount of the 1.8 kb transcript. A cDNA clone with a sequence identical to a T2 α-zein SF4 gene was isolated, providing the first direct evidence for the transcription of T2 α-zein genes containing early in-frame stop codon(s) in maize endosperm.
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  • 85
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    Plant molecular biology 22 (1993), S. 437-446 
    ISSN: 1573-5028
    Keywords: RFLP ; genome ; PFGE ; multicopy markers ; symbiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Pulsed-field gel electrophoresis (PFGE) was used to study a cluster of molecular markers in the soybean genome. There were 550 kb per centimorgan (cM) in the cluster, which is close to the calculated average for the whole genome. The analysis was complicated by the presence of duplicated sequences, and some ambiguities arising from this were resolved by using second-dimension conventional electrophoresis to relate physical maps to the RFLP map of soybean. The results show that there is a high degree of conservation of ‘rare cutter’ sites between homoeologous regions. Finally, PFGE can confirm physical linkage of monomorphic copies of markers, which can aid in the study and comparison of homoeologous regions that are invisible to RFLP analysis.
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  • 86
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    Plant molecular biology 19 (1992), S. 693-697 
    ISSN: 1573-5028
    Keywords: maize ; hypoxia ; lactate dehydrogenase ; anaerobic regulatory element
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In cereal root tissue, hypoxia induces the enzyme lactate dehydrogenase (LDH); (S)-lactate:NADH oxidoreductase, EC 1.1.1.27). In barley, both biochemical and genetic data indicate that five isozymes are induced under hypoxia. These isozymes are tetramers and arise from the random association of the products of two Ldh genes. The induction of LDH activity in root tissue has been shown to be correlated to an increase in LDH protein and Ldh mRNA. In order to more fully characterize the hypoxic induction of LDH, we have isolated a maize Ldh genemic clone which has strong homology at both the amino acid and nucleotide level to the barley LDH cDNA clones. The Ldh1 gene consists of two exons separated by a 296 bp intron, has the expected eukaryotic regulatory signals and a sequence that has strong homology to the maize anaerobic regulatory element.
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  • 87
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    Plant molecular biology 22 (1993), S. 1031-1038 
    ISSN: 1573-5028
    Keywords: maize ; catalase ; genomic clone ; sequence ; gene structure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A maize genomic DNA library (Zea mays L. W64A) was constructed using the lambda GEM-11 vector. A gene-specificCat3 cDNA probe was used to screen the library by plaque hybridization. Two independent clones, λ24C and gl27C, which hybridized to theCat3 cDNA probe, were isolated. Restriction maps were created for the two clones (λ24C and λ27C) encoding the maizeCat3 catalase gene. ASal I restriction fragment from each clone was subcloned into pBluescript KS+. Subsequent subcloning steps were performed to obtain plasmids suitable for sequencing. Both strands of the λ24C clone were sequenced. Computer analysis of the putativeCat3 promoter region has not revealed any known transcription factor binding motifs. Analysis of the deduced C-terminal amino acids encoded byCat3 shows that CAT-3 lacks the SRL peroxisomal targeting sequence contained in the CAT-1 and CAT-2 isozymes. The DNA sequence and physical structure of theCat3 gene presented here will be used in characterizingcis- andtrans-acting factors affectingCat3 gene expression.
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  • 88
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    Plant molecular biology 22 (1993), S. 1135-1143 
    ISSN: 1573-5028
    Keywords: maize ; Adh1 gene ; nuclear matrix ; MAR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nuclear matrices were isolated from maize leaves by the two conventional methods usually employed for the preparation of the corresponding structures of animal origin. It is demonstrated that functionally competent matrices, recognizing and specifically binding the MAR-containing DNA of the mousek-immunoglobulin gene may be prepared by both 2 M NaCl and LIS extractions of maize nuclei. A DNA region with a high affinity for the nuclear matrix was identified at the 5′ end of the maizeAdh1-S gene, distal to the promoter region. The presence of sites of reported altered chromatin structure in this particular region is discussed. While the proximity and the cohabitation of MARs with different regulatory elements is a common feature of matrix association regions in animal systems, this is the first plant MAR identified in a region of known significance for gene regulation.
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  • 89
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    Plant molecular biology 23 (1993), S. 685-695 
    ISSN: 1573-5028
    Keywords: extensin ; gene expression ; hydroxyproline ; maize ; silk ; vegetative tissues
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract This study concerned the developmental regulation of wall-localized, hydroxyproline-containing proteins in maize tissues and organs. Silk and pericarp cell walls contained more peptidyl hydroxyproline than did walls of any vegetative tissue, although all tissues and organs accumulated these proteins as they matured. In many tissues, hydroxyproline-rich proteins are first associated with the wall in a soluble form before being insolubilized through covalent attachment to the matrix. Because hydroxyproline was more soluble earlier than later in development, it appears that insolubilization was occurring in maize tissues and organs as well. Tissue prints reacted with an anti-extensin antibody gave positive results, indicating the presence of a soluble form of this common hydroxyproline-rich glycoprotein (HRGP). Silk and pericarp cells actively synthesized this extensin from abundant transcripts. In vegetative tissues, extensin transcripts were somewhat more abundant in seedlings than in pre-anthesis or mature plants, but levels were much lower than in silk and pericarp. Southern blots of maize genomic DNA indicated that these extensin transcripts are encoded by a small multigene family. Potential roles for extensin in reproductive/protective tissues versus the embryo or vegetative tissues are suggested.
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  • 90
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    Plant molecular biology 20 (1992), S. 343-345 
    ISSN: 1573-5028
    Keywords: maize ; microsporogenesis ; pollen ; polygalacturonase
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  • 91
    ISSN: 1573-5028
    Keywords: maize ; 1-acyl-sn-glycerol-3-phosphate acyltransferase ; complementation cloning
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    Notes: Abstract We selected cDNA plasmid clones that corrected the temperature-sensitive phenotype of Escherichia coli strain JC201, which is deficient in 1-acyl-sn-glycerol-3-phosphate acyltransferase activity. A plasmid-based maize endosperm cDNA library was used for complementation and a plasmid that enabled the cells to grow at 44°C on ampicillin was isolated. Addition of this plasmid (pMAT1) to JC201 restored 1-acyl-sn-glycerol-3-phosphate acyltransferase activity to the cells. Total phospholipid labelling showed that the substrate for the enzyme, lysophosphatidic acid, accumulated in JC201 and was further metabolised to phosphatidylethanolamine in complemented cells. Membranes isolated from such cells were able to convert lysophosphatidic acid to phosphatidic acid in acyltransferase assays. The cDNA insert of pMAT1 contains one long open reading frame of 374 amino acids which encodes a protein of relative molecular weight 42 543. The sequence of this protein is most similar to SLC1, which is thought to be able to acylate glycerol at the sn-2 position during synthesis of inositol-containing lipids. Homologies between the SLC1 protein, the 1-acyl-sn-glycerol-3-phosphate acyltransferase of E. coli (PlsC) and the maize ORF were found with blocks of conserved amino acids, whose spacing was conserved between the three proteins, identifiable.
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  • 92
    ISSN: 1573-5028
    Keywords: intron ; maize ; splicing ; vectors ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
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    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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    The journal of membrane biology 121 (1991), S. 11-22 
    ISSN: 1432-1424
    Keywords: channel ; protoplast ; K+ current ; patch clamp ; corn ; maize
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Whole-cell sealed-on pipettes have been used to measure electrical properties of the plasmalemma surrounding protoplasts isolated from Black Mexican sweet corn shoot cells from suspension culture. In these protoplasts the membrane resting potential (V m ) was found to be −59±23 mV (n=23) in 1mm K o − . The meanV m became more negative as [K−] o decreased, but was more positive than the K+ equilibrium potential. There was no evidence of electrogenic pump activity. We describe four features of the current-voltage characteristic of the plasmalemma of these protoplasts which show voltagegated channel activity. Depolarization of the whole-cell membrane from the resting potential activates time- and voltage-dependent outward current through K+-selective channels. A local minimum in the outward current-voltage curve nearV m =150 mV suggests that these currents are mediated by two populations of K+-selective channels. The absence of this minimum in the presence of verapamil suggests that the activation of one channel population depends on the influx of Ca2+ into the cytoplasm. We identify unitary currents from two K+-selective channel populations (40 and 125 pS) which open when the membrane is depolarized; it is possible that these mediate the outward whole-cell current. Hyperpolarization of the membrane from the resting potential produces time- and voltage-dependent inward whole-cell current. Current activation is fast and follows an exponential time course. The current saturates and in some cases decreases at membrane potentials more negative than −175 mV. This current is conducted by poorly selective K+ channels, whereP Cl/P K=0.43±0.15. We describe a low conductance (20 pS) channel population of unknown selectivity which opens when the membrane is hyperpolarized. It is possible that these channels mediate inward whole-cell current. When the membrane is hyperpolarized to potentials more negative than −250 mV large, irregular inward current is activated. A third type of inward whole-cell current is briefly described. This activates slowly and with a U-shaped current-voltage curve over the range of membrane potentials −90〈V m 〈0 mV.
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    Theoretical and applied genetics 79 (1990), S. 577-582 
    ISSN: 1432-2242
    Keywords: Genetic marker ; RFLP ; Quantitative traits
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Digestion of genomic DNA with the restriction endonuclease Avail disclosed a probable insertion deletion of approximately 200 base pairs (bp) near the prolactin gene. Two alleles were apparent as three distinct hybridization patterns. These alleles were statistically associated with quantitative trait loci among sons of one elite Holstein sire family. The favorable genotype was correlated with the presence of a 1.15-kb hybridization band inherited from the sire when genomic DNA was probed with a full-length cDNA for prolactin. Pedigree estimates of genetic merit among genotypes were similar, differing by only 19.3 kg for milk in ancestor merit. Comparisons of genetic estimates for quantitative yield traits in offspring of this heterozygous sire showed significant (P〈0.05) differences between homozygous genotypes for predicted difference milk (PDM), predicted difference dollars (PD$), cheese yield dollars, and protein dollars. The estimated differences between homozygous genotypes for USDA Transmitting Abilities of PDM, PD$, Cheese Yield $ and Protein $ were 282.93 kg, $74.35, $48.58 and $53.67, respectively. However, the estimated breeding values from progeny ranged over 900 kg in transmitting ability for milk. Frequency of the favorable marker allele was estimated to be 0.231 in the elite cow population used as dams of sons. These results demonstrate the potential of molecular biological techniques to discriminate between individuals within a family and to predict breeding values for selection schemes.
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  • 95
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    Theoretical and applied genetics 80 (1990), S. 673-679 
    ISSN: 1432-2242
    Keywords: Rice ; Tissue culture ; Somaclonal variation ; RFLP ; Methylation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Regenerants of rice were examined by RFLP analysis to determine the occurrence and extent of somaclonal variation. DNA polymorphisms were observed both among plants regenerated from different callus cultures as well as among sibling plants derived from a single callus. Regardless of the basal medium, a higher degree of genetic instability was found among plants regenerated from callus cultures maintained for longer incubation periods (67 days) than among those from shorter incubation periods (28 days). Detailed analysis showed that in several regenerants, there was a close correlation among those plants exhibiting DNA rearrangements and those with apparent methylation changes. Such alterations were observed with both structural and housekeeping genes.
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  • 96
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    Theoretical and applied genetics 80 (1990), S. 680-686 
    ISSN: 1432-2242
    Keywords: Retrotransposon ; Zea ; RFLP ; Bs1 ; Transposable element
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Thirty-eight accessions from Zea and 20 accessions from related genera were probed for the presence of Bs1, a retrotransposon originally found in maize. All maize and teosinte plants tested show the presence of Bs1 in one to five densely hybridizing bands. The mean copy numbers of Bs1 elements among the maize and teosinte accessions were similar: 2.92 and 3.25, respectively, with no large differences between any subgroups. Most exotic maize samples exhibited two common bands of 7.8 kb and 4.7 kb. Section Zea teosintes (but not teosintes of section Luxuriantes) also show the presence of a common band of the same size as the smaller common band in maize. At reduced stringency, Tripsacum dactyloides exhibited a single hybridizing band at 6.9 kb. Results argue for the evolution of maize from a mexicana or parviglumis teosinte, and the evolution of the Bs1 element within the tribe Andropogoneae. Additionally, recombinant inbred lines were probed for the presence of Bs1, in order to map the chromosomal locations of Bs1 elements in four different maize lines. Two of the recombinant inbred parental lines had an element (Bs1-F) on chromosome 5, while the other two lines had an element (Bs1-S) on chromosome 8. Restriction site polymorphisms have apparently arisen in the vicinity of Bs1-S since its insertion. Segregation analysis of other lines was also performed; the data indicate that Bs1 has the distribution expected of a transposable element, different locations in different lines, and not that of a fixed gene locus. However, the common bands in the Zea mays lines and the recombinant inbred data imply that Bs1 is not highly mobile.
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  • 97
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    Theoretical and applied genetics 82 (1991), S. 393-398 
    ISSN: 1432-2242
    Keywords: Maize ; Helminthosporium turcicum race1 ; RFLP ; NILs ; Mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have identified tight linkage of an RFLP marker to theHt1 gene of maize that confers resistance to the fungal pathogenHelminthosporium turcicum race 1. This was accomplished by the use of four pairs of near isogenic lines (NILs; B73, A619, W153R, and CM105), each differing by the presence or the absence of the geneHt1. SinceHt1 maps to chromosome 2, 26 clones already mapped to this chromosome were labeled and probed against Southern blots of these NILs DNA digested with three restriction enzymes:EcoRI,BamHI, andHindIII. Six markers exhibited an RFLP for at least one pair of NILs. Presumptive linkage was further tested by analyzing the segregation of five of the six markers (one was monomorphic in the cross studied) and resistance toH. turcicum race 1 on 95 F2 individuals from the cross DF20 × LH146Ht. The results indicate a tight linkage between one of the DNA markers,UMC150B, and theHt1 gene.
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  • 98
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    Theoretical and applied genetics 82 (1991), S. 697-703 
    ISSN: 1432-2242
    Keywords: RFLP ; Bamboo ; Phyllostachys ; Chloroplast DNA ; Germplasm screening
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Bamboo species are difficult to identify because flowering material is seldom available and taxonomy is of necessity based on vegetative characters. To evaluate the utility of restriction fragment length polymorphism (RFLP) analysis in bamboo systematics and germplasm screening, a library of random genomic probes from a Phyllostachys nigra PstI library was constructed. Probes from the library were used to screen bamboo germplasm consisting mostly of temperate bamboos of the genus Phyllostachys. RFLP variation was abundant, and species-specific patterns were readily obtained. Chloroplast DNA showed little variation among the bamboo accessions analyzed.
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  • 99
    ISSN: 1432-2242
    Keywords: RFLP ; PCR ; Rice ; Inheritance ; Nonradioactive
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Thirty mapped Indica rice genomic (RG) clones were partially sequenced from each end. From such sequence data, pairs of oligonucleotides were synthesized to act as primers for polymerase chain reaction (PCR) amplification of the corresponding loci in crude total DNA preparations. The PCR products from DNA of Indica varieties were of the sizes expected from the sizes of the corresponding RG clones. However, size polymorphisms were seen between PCR products from Indica and Japonica varieties, and among wildOryza species. Restriction fragment length polymorphism (RFLP) was observed between PCR products of Indica varieties simply by electrophoretic analysis of restricted products, without the need for Southern hybridization or radiolabelling. The RFLPs noted between varieties ARC6650 and Phalguna were inherited in recombinant inbred lines derived from a cross between them. The RFLPs were detectable in PCR products amplified from DNA extracted by a simple procedure from single seedlings or leaves, and revealed genetic heterogeneity in cultivated lines. An approach is described that is relevant to the acceleration of classical plant breeding through molecular techniques.
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  • 100
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    Theoretical and applied genetics 83 (1991), S. 89-96 
    ISSN: 1432-2242
    Keywords: RFLP ; Alfalfa ; Genetic diversity ; Phylogenetic tree ; Gene duplication
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Alfalfa (Medicago sativa L.) is a major forage crop throughout the world. Although alfalfa has many desirable traits, continued breeding is required to incorporate pest resistances and other traits. We conducted this study to determine the amount of restriction fragment length polymorphism (RFLP) variability present within and between diploid and tetraploid alfalfa populations, and whether or not this variability is sufficient for construction of an RFLP map. Diploid plants from M. sativa ssp. falcata, ssp. coerulea, and ssp. sativa and tetraploid spp. sativa cultivars ‘Apollo,’ ‘Florida 77,’ and ‘Spredor 2’ were included. A total of 19 cDNA clones was probed onto genomic Southern blots containing DNA digested by EcoRI, HindIII, or BamHI. Phylogenetic trees were produced, based on parsimony analysis of shared restriction fragments. Evidence for extensive gene duplication was found; most probes detected complex patterns of restriction fragments. Large amounts of variation are present within all diploid subspecies. M. sativa ssp. falcata plants formed clusters distinct from ssp. sativa or ssp. coerulea plants, which were not distinctly clustered. Some M. sativa ssp. falcata plants were more similar to the other groups than to other plants within ssp. falcata. Variation among tetraploid cultivars showed that Florida 77 and Apollo had more similarities than either showed with Spredor 2. All three cultivars showed large within-population variation, with Apollo being the most diverse and Spredor 2 the least. Based on these results, development of an RFLP map at the diploid level appears possible. Also, differentiation of cultivars, particularly ones of divergent origin, seems possible based on RFLP patterns.
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