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71.
The organization of the microtubule (Mt) cytoskeleton during mitosis and cytokinesis of the generative cell (GC) in Ornithogalum virens L. (bicellular pollen type, chromosome number, n = 3) from prophase to telophase/sperm formation was investigated by localization of -tubulin immunofluorescence using a conventional fluorescence microscope and a confocal laser scanning microscope. Chromosomes were visualized with DNA-binding fluorochrome dyes (ethidium bromide and 46-diamino-2-phenyl-indole). The GC of O. virens is characterized by G2/M transition within the pollen grain and not in the pollen tube as occurs in the majority of species with bicellular pollen. It was found that prophase in the GC starts before anthesis and prometaphase takes place after 10 min of pollen germination. The prophase Mts are organized into three prominent bundles, located near the generative nucleus. The number of these Mt bundles is the same as the number of GC chromosomes, a relation which has not previously been considered in other species. The most evident feature in the prophase/ prometaphase transition of O. virens GC is a direct rapid rearrangement of Mt bundles into a network which appears to interact with kinetochores and form a typical prometaphase Mt organization. The metaphase chromosomes are arranged into a conventional equatorial plate, and not in tandem as is thought to be characteristic of GC metaphase. The metaphase spindle consists of kinetochore fibres and a few interzonal fibres which form dispersed poles. Anaphase is characterized by a significant elongation of the mitotic spindle concomitant with the extension of the distance between the opposite poles. At anaphase the diffuse poles converge. Cytokinesis is realized by cell plate formation in the equatorial plane of the GC. The phragmoplast Mts between two future sperm nuclei appear after Mts of the mitotic spindle have disappeared.Abbreviations DAPI
46-diamino-2-phenyl-indole
- GC
generative cell
- GN
generative nucleus
- Mt
microtubule
This research was made possible in part due to TEMPUS Programme and Global Network for Cell and Molecular Biology UNESCO grants to Magorzata Bana. The experimental part of the work was done in Siena University. M. Banas is very grateful to Prof. Mauro Cresti and his group for scientific interest, offering the excellent laboratory facilities, and kind reception. 相似文献
72.
P. N. Joshi B. Biswal G. Kulandaivelu U. C. Biswal 《Radiation and environmental biophysics》1994,33(2):167-176
Response of senescing leaves of wheat seedlings to ultraviolet A (UVA) radiation (365 nm) has been examined. The results indicate that senescence-induced disorganization of thylakoid membrane, decline in carotenoid-to-chlorophyll energy transfer, and enhancement of lipid peroxidation are furthered by radiation. The senescence-induced decline in photochemical activity of photosystem II further declines on irradiation. UVA does not specifically alter any site other than those damaged by senescence. 相似文献
73.
Summary Production of microspore-derived embryos from cultured anthers is now a well established technique for the isolation of homozygous lines in many crop plants. We describe here a culture method for embryo induction and plant regeneration from anthers of four sunflower genotypes. For preliminary experiments, anthers of uninucleate microspores were cultured on four types of basal media viz., Murashige and Skoog's MS, Gamborg's B5, Nitsch and Nitsch, and White's W, supplemented with 1.0 mg/l 2,4 dichlorophenoxy acetic acid and 0.5 mg/l 6-benzylaminopurine and 40 g/l sucrose. MS basal medium, being more responsive for embryo induction, was used for further experimentation. To optimise the culture requirement MS basal medium was supplemented with 0.2–2.0 mg/l 2,4 dichlorophenoxy acetic acid and 0.5 and 1.0 mg/l 6-benzylaminopurine. The effect of cold pretreatment, hormone regime and sucrose concentration were tested for embryogenic efficiency. Genotype had a significant effect on the capacity of embryo induction. Addition of silver nitrate (2.5 mg/l), an ethylene inhibitor, stimulated embryo germination. Plantlets were obtained (10–15%) from embryos of only one genotype.Abbreviations 2,4-D
2,4 dichlorophenoxy acetic acid
- NAA
-naphthalene acetic acid
- IAA
indole-3-aceticacid
- BAP
6-benzylaminopurine
- KN
Kinetin
- ABA
abscisic acid
- GA3
gibberellic acid 相似文献
74.
75.
Sushama Chaphalkar Rajdeep Dongre Deepa Joshi Sabita Dey 《Biotechnic & histochemistry》1993,68(3):166-168
A synthetic aromatic polymer has been used for preparing replicas of different microorganisms. This method of preparing highly concentrated (9.6 k) microbiological samples for scanning electron microscopy was compared with a standard method. The micrographs of the replicated samples are satisfactory. This method is rapid, cost effective and produces good results, especially in the case of spore-forming mycelial microorganisms. 相似文献
76.
The probit analysis model is generally used for the deterininationof lethal dosages in bioassay applications. However, the logitmodels, which use the logistic curve instead of the integratednonnal curve, could also be effectively used for the determinationof lethal dosages and regression coefficients. In this paper,two lypes of logit models, namely minimum logit chi square andmaximum likelihood, have been described in detail for the estimationof the parameters. The results of these two models are alsocompared with those of the probit model. 相似文献
77.
Michael Vreones Maja Mustapic Ruin Moaddel Krishna A. Pucha Jacqueline Lovett Douglas R. Seals Dimitrios Kapogiannis Christopher R. Martens 《Aging cell》2023,22(1):e13754
Declining nicotinamide adenine dinucleotide (NAD+) concentration in the brain during aging contributes to metabolic and cellular dysfunction and is implicated in the pathogenesis of aging-associated neurological disorders. Experimental therapies aimed at boosting brain NAD+ levels normalize several neurodegenerative phenotypes in animal models, motivating their clinical translation. Dietary intake of NAD+ precursors, such as nicotinamide riboside (NR), is a safe and effective avenue for augmenting NAD+ levels in peripheral tissues in humans, yet evidence supporting their ability to raise NAD+ levels in the brain or engage neurodegenerative disease pathways is lacking. Here, we studied biomarkers in plasma extracellular vesicles enriched for neuronal origin (NEVs) from 22 healthy older adults who participated in a randomized, placebo-controlled crossover trial (NCT02921659) of oral NR supplementation (500 mg, 2x /day, 6 weeks). We demonstrate that oral NR supplementation increases NAD+ levels in NEVs and decreases NEV levels of Aβ42, pJNK, and pERK1/2 (kinases involved in insulin resistance and neuroinflammatory pathways). In addition, changes in NAD(H) correlated with changes in canonical insulin–Akt signaling proteins and changes in pERK1/2 and pJNK. These findings support the ability of orally administered NR to augment neuronal NAD+ levels and modify biomarkers related to neurodegenerative pathology in humans. Furthermore, NEVs offer a new blood-based window into monitoring the physiologic response of NR in the brain. 相似文献
78.
R. Ashton Lavoie Jeffrey T. Zugates Andrew T. Cheeseman Matt A. Teten Srivatsan Ramesh Julia M. Freeman Summer Swango Jeremy Fitzpatrick Amod Joshi Bradley Hollers Zufan Debebe Tyler K. Lindgren Amber N. Kozak Vinay K. Kondeti Mary K. Bright Eric J. Yearley Alexander Tracy Jacob A. Irwin Michael Guerrero 《Biotechnology and bioengineering》2023,120(10):2953-2968
Adeno-associated virus-based gene therapies have demonstrated substantial therapeutic benefit for the treatment of genetic disorders. In manufacturing processes, viral capsids are produced with and without the encapsidated gene of interest. Capsids devoid of the gene of interest, or “empty” capsids, represent a product-related impurity. As a result, a robust and scalable method to enrich full capsids is crucial to provide patients with as much potentially active product as possible. Anion exchange chromatography has emerged as a highly utilized method for full capsid enrichment across many serotypes due to its ease of use, robustness, and scalability. However, achieving sufficient resolution between the full and empty capsids is not trivial. In this work, anion exchange chromatography was used to achieve empty and full capsid resolution for adeno-associated virus serotype 5. A salt gradient screen of multiple salts with varied valency and Hofmeister series properties was performed to determine optimal peak resolution and aggregate reduction. Dual salt effects were evaluated on the same product and process attributes to identify any synergies with the use of mixed ion gradients. The modified process provided as high as ≥75% AAV5 full capsids (≥3-fold enrichment based on the percent full in the feed stream) with near baseline separation of empty capsids and achieved an overall vector genome step yield of >65%. 相似文献
79.
Larry J. Heilmann Patrick M. Trewitt A. Krishna Kumaran 《Archives of insect biochemistry and physiology》1993,23(3):125-134
The soluble proteins of the eggs of the coleopteran insect Anthonomus grandis Boheman, the cotton boll weevil, consist almost entirely of two vitellin types with Mrs of 160,000 and 47,000. We sequenced their N-terminal ends and one internal cyanogen bromide fragment of the large vitellin and compared these sequences with the deduced amino acid sequence from the vitellogenin gene. The results suggest that both the boll weevil vitellin proteins are products of the proteolytic cleavage of a single precursor protein. The smaller 47,000 M vitellin protein is derived from the N-terminal portion of the precursor adjacent to an 18 amino acid signal peptide. The cleavage site between the large and small vitellins at amino acid 362 is adjacent to a pentapeptide sequence containing two pairs of arginine residues. Comparison of the boll weevil sequences with limited known sequences from the single 180,000 Mr honey bee protein show that the honey bee vitellin N-terminal exhibits sequence homology to the N-terminal of the 47,000 Mr boll weevil vitellin. Treatment of the vitellins with an N-glycosidase results in a decrease in molecular weight of both proteins, from 47,000 to 39,000 and from 160,000 to 145,000, indicating that about 10–15% of the molecular weight of each vitellin consists of N-linked carbohydrate. The molecular weight of the deglycosylated large vitellin is smaller than that predicted from the gene sequence, indicating possible further proteolytic processing at the C-terminal of that protein. © 1993 Wiley-Liss, Inc. 1 This article is a US Government work and, as such, is in the public domain in the United States of America. 相似文献
80.
In animal cells, microtubule assembly is usually initiated at one specialized structure, the centrosome. By contrast, in plant cells, microtubule assembly begins at a variety of locations within the cell. A member of the tubulin gene family, gamma-tubulin, is localized to the centrosome in animal cells and is important in the assembly of microtubules in vivo. Recent reports have identified gamma-tubulin genes in plants and have described the complex intracellular distribution of the encoded polypeptides. Here, Harish Joshi and Barry Palevitz comment upon how this information may help elucidate the organizing principles of the complex arrays of microtubules in plant cells. 相似文献