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31.
Lazos-Monterrosa FA C Orantes-García O Farrera-Sarmiento AG Verdugo-Valdez MS Sánchez-Cortés LE Ruíz-Meza 《Phyton》2015,84(1):138-143
The tempisque (Sideroxylon capiri) is a tree native to Mexico used by the rural population for housing construction, poles and hedges, as fuel (wood) and also for fodder and ornamental purposes, among others. It is considered an endangered species. In order to contribute to its preservation and sustainable management, it was considered important to determine the proportion of viable seeds, the loss of viability due to storage period and the germination process by applying pregerminative treatments. We found that freshly collected seeds showed 100% viability, which decreased to 0% after 5 months of storage. According to the cumulative germination significant differences between treatments (p≤0.01) were found. It was observed that seeds can accelerate their time of germination with the previous exposure of 24 h in water at room temperature. The soaking treatment in water for 24 h at room temperature obtained final germination of 55%, while with the control 39% was reached. Soaking in hydrogen peroxide and scarification were the treatments with lower germination percentage (33 and 23%, respectively). To get a higher percentage of germinated seeds in a short time, it is necessary to give a soaking treatment in water for 24 h before sowing. 相似文献
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33.
Cell-to-cell and phloem-mediated transport of potato virus X. The role of virions 总被引:13,自引:1,他引:12
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Movement-deficient potato virus X (PVX) mutants tagged with the green fluorescent protein were used to investigate the role of the coat protein (CP) and triple gene block (TGB) proteins in virus movement. Mutants lacking either a functional CP or TGB were restricted to single epidermal cells. Microinjection of dextran probes into cells infected with the mutants showed that an increase in the plasmodesmal size exclusion limit was dependent on one or more of the TGB proteins and was independent of CP. Fluorescently labeled CP that was injected into epidermal cells was confined to the injected cells, showing that the CP lacks an intrinsic transport function. In additional experiments, transgenic plants expressing the PVX CP were used as rootstocks and grafted with nontransformed scions. Inoculation of the PVX CP mutants to the transgenic rootstocks resulted in cell-to-cell and systemic movement within the transgenic tissue. Translocation of the CP mutants into sink leaves of the nontransgenic scions was also observed, but infection was restricted to cells close to major veins. These results indicate that the PVX CP is transported through the phloem, unloads into the vascular tissue, and subsequently is transported between cells during the course of infection. Evidence is presented that PVX uses a novel strategy for cell-to-cell movement involving the transport of filamentous virions through plasmodesmata. 相似文献
34.
A GFP-MAP4 reporter gene for visualizing cortical microtubule rearrangements in living epidermal cells 总被引:15,自引:7,他引:8
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J Marc CL Granger J Brincat DD Fisher Th Kao AG McCubbin RJ Cyr 《The Plant cell》1998,10(11):1927-1940
Microtubules influence morphogenesis by forming distinct geometrical arrays in the cell cortex, which in turn affect the deposition of cellulose microfibrils. Although many chemical and physical factors affect microtubule orientation, it is unclear how cortical microtubules in elongating cells maintain their ordered transverse arrays and how they reorganize into new geometries. To visualize these reorientations in living cells, we constructed a microtubule reporter gene by fusing the microtubule binding domain of the mammalian microtubule-associated protein 4 (MAP4) gene with the green fluorescent protein (GFP) gene, and transient expression of the recombinant protein in epidermal cells of fava bean was induced. The reporter protein decorates microtubules in vivo and binds to microtubules in vitro. Confocal microscopy and time-course analysis of labeled cortical arrays along the outer epidermal wall revealed the lengthening, shortening, and movement of microtubules; localized microtubule reorientations; and global microtubule reorganizations. The global microtubule orientation in some cells fluctuates about the transverse axis and may be a result of a cyclic self-correcting mechanism to maintain a net transverse orientation during cellular elongation. 相似文献
35.
36.
A. Yu. Skorokhodova A. A. Stasenko A. Yu. Gulevich V. G. Debabov 《Applied Biochemistry and Microbiology》2018,54(2):141-148
The effect of anaplerotic pathways activation on CO2-dependent anaerobic glucose utilization by Escherichia coli strains deficient in the main fermentation pathways and possessing a modified system of glucose transport and phosphorylation was studied. Intracellular CO2 generation in the strains was ensured resulting from oxidative decarboxylation of pyruvic acid by pyruvate dehydrogenase. Sodium bicarbonate dissolved in the medium was used as an external source of CO2. The genes of heterologous pyruvate carboxylase and native NADH-dependent malic enzyme were overexpressed in the strains to allow anaplerotic carboxylation of pyruvic acid to oxaloacetic or malic acid. The ability of the strains to reoxidize NADH utilizing carboxylation products was additionally increased due to enhanced expression of malate dehydrogenase gene. In the case of endogenous CO2 formation, the activation of anaplerotic pathways did not cause a notable increase in the anaerobic glucose consumption by the constructed strains. At the same time, the expression of pyruvate carboxylase led to a pronounced decrease in the secretion of pyruvic acid with the concomitant increase in the yield of four-carbon metabolites. Further enhancement of NADH-dependent malic enzyme expression provoked activation of a pyruvate–oxaloacetate–malate–pyruvate futile cycle in the strains. The availability in the medium of the external CO2 source sharply increased the anaerobic utilization of glucose by strains expressing pyruvate carboxylase. The activity of the futile cycle has raised with the increased malic enzyme expression and dropped upon enhancement of malate dehydrogenase expression. As a result, the efficiency of CO2-dependent anaerobic glucose utilization coupled to the formation of four-carbon carboxylation products increased in the studied strains resulting from the primary anaplerotic conversion of pyruvic acid into oxaloacetic acid followed by the involvement of the precursor formed in NADH-consuming biosynthetic reactions dominating over the reactions of the revealed futile cycle. 相似文献
37.
A. Yu. Skorokhodova A. A. Stasenko A. Yu. Gulevich V. G. Debabov 《Applied Biochemistry and Microbiology》2018,54(3):245-251
The effect of the introduction of a synthetic bypass, providing 2-ketoglutarate to succinate conversion via the intermediate succinate semialdehyde formation, on aerobic biosynthesis of succinic acid from glucose through the oxidative branch of the tricarboxylic acid cycle in recombinant Escherichia coli strains has been studied. The strain lacking the key pathways of acetic, lactic acid and ethanol formation from pyruvate and acetyl-CoA and possessing modified system of glucose transport and phosphorylation was used as a chassis for the construction of the target recombinants. The operation of the glyoxylate shunt in the strains was precluded resulting from the deletion of the aceA, aceB, and glcB genes encoding isocitrate lyase and malate synthases A and G. The constitutive activity of isocitrate dehydrogenase was ensured due to deletion of isocitrate dehydrogenase kinase/phosphatase gene, aceK. Upon further inactivation of succinate dehydrogenase, the corresponding strain synthesized succinic acid from glucose with a molar yield of 24.9%. Activation of the synthetic bypass by the induced expression of Mycobacterium tuberculosis 2-ketoglutarate decarboxylase gene notably increased the yield of succinic acid. Functional activity of the synthetic bypass in the strain with the inactivated glyoxylate shunt and opened tricarboxylic acid cycle led to 2.7-fold increase in succinate yield from glucose. As the result, the substrate to the target product conversion reached 67.2%. The respective approach could be useful for the construction of the efficient microbial succinic acid producers. 相似文献
38.
Peripheral alpha1,3-fucosylation of glycans occurs by the action of either
one of five different alpha1,3-fucosyltransferases (Fuc-Ts) cloned to date.
Fuc-TVI is one of the alpha1,3-fucosyltransferases which is capable to
synthesize selectin ligands. The major alpha1, 3- fucosyltransferase
activity in human plasma is encoded by the gene for fucosyltransferase VI,
which presumably originates from liver cells. While the sequence,
chromosomal localization, and kinetic properties of Fuc-TVI are known,
immunocytochemical localization and trafficking studies have been
impossible because of the lack of specific antibodies. Here we report on
the development and characterization of a peptide-specific polyclonal
antiserum monospecific to Fuc-TVI and an antiserum to purified soluble
recombinant Fuc-TVI crossreactive with Fuc-TIII and Fuc-TV. Both antisera
were applied for immunodetection in stably transfected CHO cells expressing
the full-length form of this enzyme (CHO clone 61/11). Fuc-TVI was found to
be a resident protein of the Golgi apparatus. In addition, more than 30% of
cell-associated and released enzyme activity was found in the medium.
Maturation and release of Fuc-TVI was analyzed in metabolically labeled CHO
61/11 cells followed by immunoprecipitation. Fuc-TVI occurred in two forms
of 47 kDa and 43 kDa bands, while the secreted form was detected as a 43
kDa. These two different intracellular forms arose by posttranslational
modification, as shown by pulse-chase experiments. Fuc-TVI was released to
the supernatant by proteolytic cleavage as a partially endo-H resistant
glycoform.
相似文献
39.
40.
L. A. Prasolova Yu. E. Gerbek R. G. Gulevich S. G. Shikhevich M. Yu. Konoshenko R. V. Kozhemyakina I. N. Oskina I. Z. Plyusnina 《Russian Journal of Genetics》2014,50(8):846-852
Two Norway rat (Rattus norvegicus) populations contrasting in behavior have been raised at the Institute of Cytology and Genetics, Novosibirsk, Russia with long-term selection for the absence or enhancement of an aggressive response towards humans. They are designated as tame and aggressive, respectively. In this work we investigated the effects of the selection on behavior, stress responsiveness, and function of testes in males of the 78th generation. It is shown that the difference between the strains in their response to humans remains the same as in previous generations. However, the differences in stress response and rate of maturation contradict earlier data. For the first time, we note a higher glucocorticoid-mediated response to restriction stress and retarded sexual maturation in tame rats compared to aggressive ones, according to morphometric indices of gonads and testosterone levels. It is conceivable that the change in selection effects is determined by the disjunction of the directions of selection for behavior and the modification of the stress response. This study is the first to characterize males recently (six or seven generations of propagation in captivity) caught in the wild with regard to the indices under consideration and used as a control group. Wild rats have the highest stress response and rate of sexual maturation as compared to those selected. 相似文献