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1.
2.
Summary Maize (Zea mays) leaf protoplasts were isolated from various leaves of two-week (4-leaf) seedlings and from sections of the third leaf blades. Microtubules (MTs) were visualized using immunofluorescence microscopy. Only freshly isolated protoplasts from the third and fourth leaf blades contained MTs, with protoplasts from the fourth leaf containing the most i.e. 13% of fourth-leaf protoplasts contained MTs. In general, protoplasts with fewer and smaller chloroplasts had more MTs. Initially 90–95% of protoplasts from basal portions of leaves had MTs but the percentage decreased slightly during culture particularly after 10 days. The antioxidant n-propyl gallate was beneficial in maintaining MT content. Few protoplasts from older sections intitially contained MTs but in all sections at least some protoplasts regained a significant MT content during culture (e.g., 10% of protoplast from the tip section possessed microtubules after 7 days of culture). Far fewer MTs were observed in individual leaf protoplasts than those isolated from suspension culture.Abbreviations BMS
Black Mexican Sweet
- MT
microtubule
- MtSB
microtubule stabilizing buffer
- PBS
phosphate buffered saline 相似文献
3.
DNA replication in maize leaf protoplasts 总被引:1,自引:0,他引:1
H. Wang Adrian J. Cutler M. Saleem Larry C. Fowke 《Plant Cell, Tissue and Organ Culture》1989,18(1):33-46
Maize leaf protoplasts were investigated for their metabolic competence and capacity to synthesize DNA. When protoplasts were incubated at elevated temperatures, they exhibited a heat shock response with specific proteins being preferentially synthesized. This indicated that the protoplasts were fully metabolically functional and capable of responding to environmental stimuli. Significant DNA synthesis was observed in these protoplasts after incorporation of 3H-thymidine into chromatin by trichloroacetic acid precipitation and by incorporation of 5-bromo-2-deoxyuridine (BrdU), an analog of thymidine, detected by immunofluorescence. The immunocytochemical method revealed that about 50% of nuclei in the maize leaf protoplasts were labelled after 3 days of culture and that most of these nuclei were labelled as intensely as normal mitotic cells. Aphidicolin, an inhibitor of DNA polymerase-, decreased the percentage of labelled nuclei, demonstrating that the labelling was substantially due to replicative DNA synthesis. However, chromosome condensation was not observed. It is proposed that these protoplasts are capable of DNA synthesis, but incapable of nuclear division. Effects of media additives on the number of nuclei entering S phase in these protoplasts were also assessed by the immunocytochemical method. Inclusion of 80mM Ca2+ in the enzyme solution increased protoplast yield and also appeared beneficial to DNA synthesis. The antioxidant, n-propyl gallate, which was used to stabilize the protoplasts, delayed the onset of DNA synthesis. Arginine and spermidine produced a slight increase in DNA synthesis.Abbreviations BrdU
5-bromo-2-deoxyuridine
- DMSO
dimethyl sulfoxide
- n-PG
n-propyl gallate
- PBS
phosphate-buffered saline
Dedicated to Dr. Friedrich Constabel on the occasion of his 60th birthday 相似文献
4.
We have isolated the delta-globin gene of the New-World spider monkey,
Ateles geoffroyi, and compared its nucleotide sequence with those of other
primate delta- and beta-globin genes. Among primate delta-globin genes, the
rate of nonsynonymous substitutions is much less than the rate of
synonymous substitutions. This suggests that primate delta- globin genes
may remain under evolutionary conservation, perhaps because hemoglobin A2
has an as yet unknown physiological importance.
相似文献
5.
The neem tree (Azadirachta indica) contains promising pest-control substances found effective against many economically important pests. These materials are easy to process by village-level industries and easy to use by limited-resource farmers, thereby offering potential for crop protection and off-farm income generation. The tree’s numerous pharmacological and other complementary uses make it doubly attractive for incorporation in large-scale rural development efforts. 相似文献
6.
Biochemical pathways in prokaryotes can be traced backward through evolutionary time 总被引:10,自引:0,他引:10
For the first time, a credible prokaryotic phylogenetic tree is being
assembled by Woese and others using quantitative sequence analysis of
oligonucleotides in the highly conservative rRNA. This provides an
evolutionary scale against which the evolutionary steps that led to the
arrangement and regulation of contemporary biochemical pathways can be
measured. This paper presents an emerging evolutionary picture of aromatic
amino acid biosynthesis within a large superfamily assemblage of
prokaryotes that is sufficiently developed to illustrate a new perspective
that will be applicable to many other biochemical pathways.
相似文献
7.
In Drosophila pseudoobscura, the amylase (Amy) multigene family is
contained within a series of inversions, or gene arrangements, on the third
chromosome. The Standard (ST), Santa Cruz (SC), and Tree Line (TL)
inversions are central to the phylogeny of arrangements, and have clusters
of other arrangements derived from them. The gene arrangements belonging to
each of these three clusters have a characteristic number of Amy genes,
ranging from three in ST to two in SC to one in TL. This distribution
pattern can reflect a history of either duplications or deletions, although
the data available in the past did not permit a decision between these
alternatives. We provide unambiguous evidence that three Amy genes were
present before the divergence of the ST, SC, and TL arrangements. Thus, the
current status of the Amy multigene family is the result of deletions in
the TL and SC arrangements, which created three new pseudogenes: TL
Amy2-psi, TL Amy3-psi, and SC Amy3- psi. Analysis of pseudogene sequences
revealed that, in the SC and ST arrangements, pseudogene evolution has been
retarded, most likely due to the homogenization effect of gene conversion.
Finally, by determining the original copy number, we have reconstructed the
evolutionary history of the Amy multigene family and linked it with the
evolution of the central gene arrangements.
相似文献
8.
Raman Kapur Mohammed Saleem Bryan L. Harvey Adrian J. Cutler 《In vitro cellular & developmental biology. Plant》1993,29(4):200-206
Summary Barley leaf blade protoplasts accumulate malonaldehyde, a product of lipid peroxidation, during culture. In addition, glutathione
levels fall after protoplast isolation and the proportion of glutathione in the oxidized state rises. These data indicate
oxidative stress after protoplast isolation and during culture. The cause of this phenomenon is revealed by data showing that
the activities of enzymes associated with antioxidative processes including glutathione reductase and ascorbate peroxidase
decrease after barley protoplast isolation. In contrast, protoplasts isolated from suspension cultured cells of bromegrass
and soybean exhibit little evidence for oxidative stress and increased activities of glutathione reductase and ascorbate peroxidase.
We suggest that an antioxidative response is associated with mitosis and colony formation from protoplasts, as exhibited by
bromegrass and soybean. Conversely, failure of an antioxidative response is associated with low viability and absence of mitosis,
as in barley. Increased viability of barley leaf protoplasts cultured on feeder layer cells is correlated with increased glutathione
content and higher glutathione reductase activity. 相似文献
9.
The application of sensory methodology for measuring deodorizing effect of an air conditioner equipped with electric plasma was introduced. Deodorizing effect was measured using chemical and sensory methods at different time (0, 30 and 60 min) and mode (control, blowing and cooling) of an air conditioner. Smoke from a roll of cigarette in a closed room was used as a source of odor and the concentrations of acetic acid and ammonia were measured as odorous chemical components. As one of the sensory methods triangle test was used and as a first step to obtain deodorizing effects by triangle test, the threshold of each panelist was obtained as the log dilution ratio of odor concentration at which the difference from odorless air was detected. The odor concentration at each time and mode was calculated using the threshold of the panel and the deodorizing effect was obtained on the basis of the odor concentration. In addition to a triangle test, scaling methods such as category scaling or magnitude estimation were used to measure deodorizing effect of an air conditioner. Deodorizing effects by scaling methods were calculated based on odor intensity with time at each mode. The regression analysis was done between the efficacy of deodorizing effect by sensory test and those by acetic acid and ammonia, the R2 values of the regression equations for triangle test, category scale, and magnitude estimation were 0.84, 0.72 and 0.69, respectively. Deodorizing effect by triangle test explained the decrease of acetic acid and ammonia better than those by category scaling or magnitude estimation while high cost and time consuming labor involved in triangle tests reduced the merit. The results of this study demonstrated that various sensory methods could be used to measure deodorizing effect of air conditioners and further researches on fast and reliable methods are needed to establish the official procedures. 相似文献
10.
Sequence requirements for the termination of rolling-circle replication of plasmid pT181 总被引:3,自引:0,他引:3
Most small multicopy plasmids of Gram-positive bacteria and many in Gram-negative bacteria replicate by a rolling-circle (RC) mechanism. The replication initiator proteins encoded by the RC plasmids and single-stranded bacteriophages of Escherichia coli have origin-specific nicking-closing activities that are required for the initiation and termination of RC replication. We have investigated the sequence requirements for termination of RC replication of plasmid pT181. The initiator nick site is located in the loop of a hairpin region (IRII) within the pT181 origin of replication. By mutational analysis, we have found that several nucleotides within the stem of IRII which are critical for the initiation activity are dispensable for termination of replication. We also demonstrate that nucleotides in the right arm of IRII, but not the left arm, are absolutely required for termination of RC replication. We have also identified specific nucleotides in IRII that are critical for its termination activity. The sequence of the right arm of the hairpin must be located downstream of the initiator nick site for termination, suggesting that termination requires a specific orientation of the initiator protein at the origin. 相似文献