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  • 1
    ISSN: 1432-203X
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract An embryogenic suspension culture of orchardgrass (Dactylis glomerata L.) consisting of small, embryogenic cell clusters was obtained from callus formed on basal sections of young leaves through a process of selective enrichment. These suspensions were used as a source of protoplasts. The isolated protoplasts divided at a frequency of 0.5–10% when plated in an agarose solidified culture medium. Conditioned medium, in which embryogenic Dactylis suspension cultures had been grown, was found to increase the rate of cell colony formation. Protoplast-derived colonies grew rapidly in a bead-type culture system of floating agarose slabs in liquid medium. New suspension cultures formed as the colonies grew out of the agarose. These cultures were embryogenic and formed green plantlets when plated on a solid medium lacking auxin. The plantlets were established in soil and grown to mature plants.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 2
    ISSN: 1617-4623
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary Protoplasts isolated from a 5-methyltryptophan resistant nonregenerable Nicotiana tabacum (L.) Xanthi suspension culture were fused via the dextran method (Kameya et al. 1981) with protoplasts isolated from N. glutinosa L. leaf mesophyll cells. Prospective somatic hybrids were selected by their ability to produce chlorophyll and to grow on medium containing 5-methyltryptophan and eventually to regenerate into complete plants. Twenty-eight plants regenerated from selected colonies were classified into three groups: (1) Four plants had intermediate morphology, were sterile, had hybrid isozyme patterns for four different enzymes, and contained fraction 1 protein large subunit patterns indicative of a N. glutinosa chloroplast origin. (2) One plant had N. tabacum morphology, was fertile, had two hybrid isozyme patterns, and contained fraction 1 protein large subunits indicative of the N. tabacum chloroplast type. Although this hybrid contained both parental small subunit patterns of fraction 1 protein, two-dimensional gel electrophoresis showed the N. glutinosa-type small subunit to consist of several molecular weights suggestive of errors in the processing steps. Progeny of the Type 2 hybrid (selfed) contained no N. glutinosa characteristics. (3) Twenty-three plants were N. glutinosa escapes. Chromosome numbers ranged from 60 (some chromosome loss) in two hybrids to 34–35 (extensive chromosome loss) in three hybrids. These results suggest that amino acid analog resistance and regeneration capability can be utilized as complementing markers for selecting and identifying somatic hybrids after protoplast fusion. Mesophyll protoplasts from N. glutinosa can also be regenerated into morphologically normal fertile plants.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 3
    Digitale Medien
    Digitale Medien
    Springer
    Plant cell, tissue and organ culture 39 (1994), S. 239-244 
    ISSN: 1573-5044
    Schlagwort(e): carbon ; elevated CO2 ; nitrogen ; suspension culture
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract A photoautotrophic soybean suspension culture (SB-P) was used to study CO2 assimilation while exposed to elevated or ambient CO2 levels. These studies showed that under elevated CO2 (5% v/v) malate is the dominant fixation product, strongly suggesting that phosphoenolpyruvate carboxylase (PEPCase) is the primary enzyme involved in carbon fixation in these cells under their normal growth conditions. Citrate and [aspartate + glutamate] were also significant fixation products during fifteen minutes of exposure to 14CO2. During the ten minute unlabeled CO2 chase however, 14C-malate continued to increase while citrate and [aspartate + glutamate] declined. Fixation of 14CO2 under ambient CO2 levels (0.037%) showed a very different product pattern as 3-phosphoglycerate was very high in the first one to two minutes followed by increases in [serine + glycine] and [aspartate + glutamate]. Hexose phosphates were also quite high initially but then declined relatively rapidly. Thus, the carbon fixation pattern at ambient CO2 levels resembles somewhat that seen in C3 leaf cells while that seen at elevated CO2 levels more closely resembles that of a C4 plant. The initial fixation product of C3 plants, 3-PGA, was never detectable under high CO2 conditions. These data suggest that an in vitro photoautotrophic system would be suitable for studying carbon fixation physiology during photosynthetic and non-photosynthetic growth.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 4
    Digitale Medien
    Digitale Medien
    Springer
    Plant cell, tissue and organ culture 39 (1994), S. 245-250 
    ISSN: 1573-5044
    Schlagwort(e): asparagine ; cell culture ; Glycine max L. ; nitrogen metabolism
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract A photoautotrophic soybean suspension culture was used to study free amino acid pools during a subculture cycle. Free amino acid analysis showed that the intracellular concentrations of asparagine, serine, glutamine, and alanine reached peaks of 200, 10, 9 and 7 mM, respectively, at specific times in the 14-day subculture cycle. Asparagine and serine levels peaked at day 14 but glutamine level rose quickly after subculture, peaking at day three and then declined gradually. Roughly similar patterns were found in the conditioned culture medium although the levels were 1000-fold lower than those found in cells. Photoautotrophic (SB-P) and photomixotrophic (SB-M) cultures were quantitatively similar with regard to free asparagine and serine but not glutamine or free ammonia. Heterotrophic (SB-H) cells had 81–85% less free asparagine on day seven than did SB-M or SB-P cells. Hence, similar to the phloem sap of a soybean plant, asparagine, glutamine, alanine and serine were the predominant amino acids in photoautotrophic soybean cell cultures. Varying the amount of total nitrogen in culture medium for two subcultures at 10, 25, 50, and 100% Of normal levels showed that growth was inhibited only at the 10 and 25% levels but that growth on medium containing 50% of the normal nitrogen was as good as that on 100% nitrogen. Moreover, cellular chlorophyll content correlated exceptionally well with initial nitrogen content of the medium. Thus, the photosynthesis of SB-P cells was not limited by chlorophyll content. SB-P cells grown for two subcultures on 10% nitrogen contained very low free amino acid levels and only 1% of the free ammonia levels found in cells growing on a full nitrogen complement.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 5
    Digitale Medien
    Digitale Medien
    Springer
    Plant cell, tissue and organ culture 30 (1992), S. 85-91 
    ISSN: 1573-5044
    Schlagwort(e): cell culture ; photoautotrophic ; soybean
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract We have attempted to optimize the conditions under which a photoautotrophic soybean suspension culture line (SB-P; Horn et al. 1983) is grown. Magnesium, phosphate, and calcium concentrations were varied individually from one-tenth to five times the normal level found in the Murashige and Skoog (1962) recipe. After two subcultures, only phosphate at one-tenth the normal level caused the cells to show a substantial reduction in fresh and dry weight increase and chlorophyll level. Nitrate and ammonium levels were inversely varied in 20 millimolar increments of potassium nitrate and ammonium chloride. Neither N-source alone could support growth through two subcultures. A ratio of 40 millimolar potassium nitrate to 20 millimolar ammonium gave significantly better fresh and dry weight increases than did a ratio of 20:40 or 30:30 but the chlorophyll level was unchanged. The minor salts as a group resulted in a small improvement in growth when provided at twice the normal level. Indole-3-acetic acid at five milligrams per liter resulted in significantly better fresh and dry weight increases than did α-naphthaleneacetic acid at any level but the final chlorophyll level was not changed. There was no correlation between growth and kinetin level and this resulted in the discovery that SB-P cells are cytokinin-autotrophic, as are heterotrophic SB cells, with regard to both growth and greening ability. Growing SB-P cells under a 14 h:10 h day:night photoperiod resulted in a slow but inevitable death. Increasing the carbon dioxide level to 10% for four weeks gave no increase in SB-P cell growth or chlorophyll level, but SB-P cells would not grow with carbon dioxide levels below 0.4%. The results clearly show that SB-P cells, despite their tenuous existence, are capable of adapting to a wide range of culture conditions. A simplified and improved culture medium for photoautotrophic cultures is given.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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