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151.
以甘薯(1pomoeabatatas(L.)Lam.)品种栗子香的胚性悬浮细胞为受体材料,用根癌农杆菌介导法,获得了表达除草剂抗性基因bar基因的转HSl基因甘薯植株。共计380个遗传转化的胚性细胞团,在添加2mg/L2.4-D、100mg/L Carb和10mg/L Glu(glufosinate)的固体Ms培养基上选择培养9周后,得到了12个Glu抗性愈伤组织。将这些抗性愈伤组织转移到添加1mg/L ABA、100mg/L羧苄青霉素和10mg/L Glu的固体MS培养基上,其中的3个抗性愈伤组织再生出拟转基因植株。PCR鉴定它们为转基因植株。Southern blot分析表明,HS1基因已整合到基因组中。转基因植株具有稳定的除草剂抗性。结薯观察实验结果表明,转基因植株结薯正常。 相似文献
152.
1. Cultured dorsal root ganglion neurons of rat pups were depolarized by exposure to 50 mM K+ and the rise of [Ca2+]i was measured using fura-2 as an indicator.2. Lead in the extracellular solution reduced the rise of [Ca2+]i in a concentration-dependent manner, with a threshold concentration of 0.25 M. More than 80% of the calcium entry was prevented by 5 M lead. The IC50 and the Hill coefficient were 1.3 M and 1, respectively.3. This effect was considered to be due to a reduction of VACCCs, since applications of NMDA did not result in any rise of [Ca2+]i.4. Since Pb2+ itself changes the fura-2 signal in a typical and characteristic manner, fura-2 is also an indicator for Pb2+. No changes in fura-2 signals were detected when lead (5 M) was applied for several minutes in the absence of calcium, indicating that Pb2+ did not enter the cells.5. Thus it is concluded that lead prevents calcium entry by reducing VACCCs but does not cross the cell membrane itself. 相似文献
153.
Alfalfa (Medicago sativa L. cv. Canadian No. 1), tobacco (Nicotiana tabacum L. var. humilis) and wheat (triticum monococcum L.) cells were grown in a defined, liquid medium containing either ammonium sulfate, L-glutamine or potassium nitrate as the sole nitrogen source, and the effects of the tricarboxylic-acid (TCA) intermediates, citrate and -ketoglutarate (5, 10, 15 mM), on the growth (dry-weight increase) of these cells was observed. The three cell suspension cultures exhibited a different growth response to the TCA-cycle intermediate supplied, depending upon the concentration of the additive and the nitrogen source. Citrate (5 mM) greatly enhanced growth of alfalfa and wheat cells in an ammonium-based medium but was less effective at higher concentrations, and in the case of alfalfa cells markedly inhibited growth. Tobacco cell growth was inhibited by all citrate concentrations tested. In contrast, all concentrations of -ketoglutarate used stimulated the growth of all three cell cultures in an ammonium-based medium. Alfalfa and wheat cells grown in an L-glutamine-based medium were influenced by citrate in a manner similar to that in ammonium-based medium. The growth of tobacco cells was slightly enhanced by 5 mM citrate but inhibited by higher concentrations. -Ketoglutarate, at all concentrations tested, was stimulatory to the growth of the cells of all three species in a glutamine-based medium, except for alfalfa cells which were inhibited at 15 mM. Both TCA-cycle acids inhibited the growth of alfalfa and tobacco cells grown on a nitrate-based medium whereas the growth of wheat cells was almost unaffected. 相似文献
154.
The synthesis of cartilage collagen by rabbit and human chondrocytes in primary cell culture 总被引:2,自引:0,他引:2
Fred H. Schindler Marsha A. Ose Michael Solursh 《In vitro cellular & developmental biology. Plant》1976,12(1):44-47
Summary This report describes a method for preparing primary cell cultures of differentiated rabbit sternal and human vertebral cartilage
cells. These cell cultures were shown to synthesize primarily α1 chains, which is taken to mean that at least 82% of the collagen
produced is cartilage specific collagen (type II).
This work was supported in part by grant HD-05505 from NIH. 相似文献
155.
Adventitious shoot buds were induced from leaf and stem explants of Bacopa monnieri on Murashige and Skoog medium supplemented with benzyladenine or kinetin. The source of the explants as well as different
gelling agents in the medium were found to influence shoot induction and eventual shoot growth. The best response was obtained
in leaf explants taken from shoot cultures grown in medium supplemented with 2 μM benzyladenine and gelled with 0.2% gelrite. A transverse section of the leaf explant incubated in this medium showed several
shoot primordia emerging from the leaf surface. This system exhibited a potential for repeated harvesting of the shoots from
the original leaf explant as the latter continued to expand and regenerate new shoots, upon repeated periodical subculturing
onto fresh medium. However, the callusing response of the plant was very low. Qualitative TLC studies of the regenerated shoots
revealed a phytochemical profile similar to that of the field grown-plants.
Received: 20 March 1998 / Revision received: 1 December 1998 / Accepted: 12 December 1998 相似文献
156.
Conditions have been developed that induce maize (Zea mays L.) protoplasts to re-synthesize cell walls and to initiate cell divisions. Two types of embryogenic maize callus were used as a source of protoplasts: a heterogeneous callus (Type I) derived from immature embryos after three weeks in culture, and a friable, rapidly growing callus (Type II) selected from portions of the Type I callus. Many variables in the growth conditions of the donor tissue (type of medium, transfer schedule, age of callus), protoplast isolation solutions (pH, osmolarity, type and concentration of cell wall hydrolyzing enzymes, addition of polyamines) and conditions (amount of time in enzyme, amount of tissue per volume of enzyme incubation medium, agitation, preplasmolysis of source tissue, type of callus), and purification procedures (filtration and-or flotation), were found to affect both yield and viability of protoplasts (based upon fluorescein-diacetate staining). Our isolation procedure yielded high numbers of viable, uninucleated maize callus protoplasts which were densely cytoplasmic and varied in size from 20 to 50 m in diameter. Protoplasts plated in solid medium formed walls and divided several times. Of several gelling agents tested for protoplast propagation, only agarose resulted in protoplasts capable of sustained divisions leading to the formation of microcalli. Plating efficiency was established over a wide range of protoplast densities (103–107 protoplasts/ml). Highest plating efficiency (25%) was obtained at 1·106 protoplasts/ml). The resulting microcalli grew to be dense clusters of about 0.1–0.5 mm in diameter and then stopped growing. Nurse cultures of maize and carrot (Daucus carota L.), were used to establish that individual protoplasts (not contaminating cells or cell clusters) formed walls and divided. Nurse cultures also increased the efficiency of microcallus formation from protoplasts.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- MS
Murashige and Skoog (1962) salts
- MS 1D
Murashige and Skoog salts with 1 mg/l 2,4-D
- MS 2D
Murashige and Skoog salts with 2 mg/l 2,4-D
- N6
medium of Chu et al. (1975)
- NN67-mod
medium of Nitsch and Nitsch (1967) as modified in the present paper
- FDA
fluorescein diacetate
- LMP
low melting point 相似文献
157.
A soluble protein from Saccharomyces cerevisiae acts as a peroxidase but requires a NADPH-dependent thioredoxin system and was named thioredoxin peroxidase (TPx). The role of TPx in aging of stationary cultures of S. cerevisiae was investigated in a wild-type strain and a mutant yeast strain in which the tsa gene that encodes TPx was disrupted by homologous recombination. The occurrence of oxidative stress during aging of stationary cultures of the yeast has been proposed. Comparison of 5-day-old (young) stationary cultures of S. cerevisiae and of cultures aged for 3 months (old) revealed decreased viability, increased generation of reactive oxygen species, modulation of cellular redox status, and increased cellular oxidative damage reflected by increased protein carbonyl content and lipid peroxidation. The magnitude of this stress was augmented in yeast mutant lacking TPx. These results suggest that TPx may play a direct role in cellular defense against aging of stationary cultures presumably, functioning as an antioxidant enzyme. 相似文献
158.
Because of uniformity and small distances for transport, cell suspensions offer a system for rapid measurements of initial reactions of phytotoxic compounds. We had previously shown that a growth regulator, dikegulac (2,3:4,6 di-o-isopropylidine-2-keto-L-gulonate) inhibits amino acid incorporation into proteins. Using Solanum nigrum suspension cultures, it was found that dikegulac rapidly inhibits amino acid uptake into cells, before inhibiting incorporation, with time points starting at a few minutes, and kinetics that can be extrapolated back to time zero. With more rapid kinetics this compound induces leakage of a preloaded dye. The rate of leakage was less with stationary cells in suspension, reiterating that they are more resistant to the effects of this compound. It was thus concluded that at the concentrations used, the first effect of dikegulac (or one very close to the first effect) is on the cell membrane.Abbreviation FDA
fluorescein diacetate 相似文献
159.
Increase in anthocyanin yield from wild-carrot cell cultures by a selection system based on cell-aggregate size 总被引:2,自引:0,他引:2
Wild-carrot (Daucus carota L.) cell cultures were screened to yield small (less than 63 m) or large (greater than 170 m) cell aggregates which were then subcultured. Cultures of the small-size class had a higher, those of the large-size class a lower anthocyanin yield than the unscreened culture. This relationship became accentuated with an increasing number of passages with screening prior to subculture. At the end of six months (12 passages), the pigment yield of the small-size class was triple that of the unscreened cells. Following this selection period, the tendency of the small-size fraction to increase in clump size when subcultured without screening was much less than that of freshly isolated cell aggregates of the same size. These observations may be explainable on the basis of a differential distribution of cytokinin between aggregates of different sizes. High levels of cytokinin inhibit anthocyanin accumulation and inhibit cell separation; these effects result in large cell aggregates having low levels of anthocyanin. In support of this hypothesis, it is shown that addition of kinetin to cultures of small cell aggregates causes an increase in the size of cell aggregates and a parallel decrease in anthocyanin yield. 相似文献
160.
Johan U. Grobbelaar 《Journal of applied phycology》1989,1(4):333-340
It has been suggested that turbulence with the resultant light/dark cycle and light gradient through which phytoplankton move,
enhances their productivity. The stationary bottle incubation technique for estimating rates of primary productivity has mainly
been criticized because of bottle effects, the elimination of natural turbulence and the presence of photo-inhibition. In
a series of experiments where productivity was measured over static profiles and compared to the productivity in a mixed system,
no definite conclusion could be reached regarding the effect of varying light/dark cycles of medium frequency (seconds to
minutes). It appeared as though the ratio of the euphotic depth to mixing depth (Z
eu/Z
m) influenced productivity more than the duration of the light/dark cycle. The static bottle incubation method gave higher
integral productivities than the mixed samples at low ratio's ofZ
eu/Z
m. It is suggested that mixing has two separate, but synergistic effects i.e. it not only moves the phytoplankton cells through
a light/dark cycle, but also decreases the boundary layer, which increases the rate of exchange through the cell wall of nutrients
and metabolites. In doing so more nutrients are available and light could be utilized more efficiently and therefore, productivity
is increased. 相似文献