Behavior in tissue culture of nitrogen-fixing root nodules of Ceanothus integerrimus

1980 ◽  
Vol 58 (10) ◽  
pp. 1121-1128 ◽  
Author(s):  
George S. Ellmore ◽  
Ruth Strand ◽  
W. M. Laetsch
Keyword(s):  
Tissue Culture ◽  
Root Nodule ◽  
Root Nodules ◽  
Tissue Growth ◽  
Cultivated Plants ◽  
In Vitro Growth ◽  
Cell Cytoplasm ◽  
Host Cell Cytoplasm ◽  
Agar Media

This study documents the in vitro growth of N2-fixing root nodules from the dicotyledonous shrub Ceanothus integerrimus (Rhamnaceae). Root nodule lobes were removed from cultivated plants, surface sterilized, and cultured on agar media containing varied amounts of nitrogen (N). Uninfected cells inside the cultured nodules divide and form a callus which is visible after 7 days of growth. Cells infected by the N2-fixing actinomycete do not divide, but rather degenerate. Host cell cytoplasm disappears and organelles and the plasma membrane are no longer seen under the electron microscope. Endophyte structure also deteriorates. Nucleoid areas within the terminal vesicles become diffuse. Cross walls separating vesicles from the hyphae disappear and cytoplasm of the hyphae withdraws from the actinomycete wall. Ceanothus nodules readily form callus in tissue culture. Callus proliferates from uninfected cortical parenchyma outside the infection zone. The callus is free of living endophyte and is incapable of fixing N2 as measured by acetylene reduction. Nodule tissue growth in vitro is more vigorous on media with high N than on N-deficient media.

Virulence of tissue culture-propagated canine distemper virus

1980 ◽  
Vol 29 (3) ◽  
pp. 940-944 ◽  
Author(s):  
A E Metzler ◽  
R J Higgins ◽  
S Krakowka ◽  
A Koestner
Keyword(s):  
Tissue Culture ◽  
Neurological Disease ◽  
Canine Distemper Virus ◽  
In Vitro Growth ◽  
Canine Distemper ◽  
Passage Level ◽  
Pulmonary Macrophage

Virulence of canine distemper virus (CDV) adapted to in vitro growth in Vero or bovine cells was determined by inoculation into CDV-susceptible neonatal gnotobiotic dogs. When compared with dogs given virulent R252-CDV, Vero R252-CDV was attenuated at passage level 14. In contrast, dogs inoculated with bovine R252-CDV at the same passage level experienced rapid fatal neurological disease. Virulence was not linked to ability to infect or replicate in canine pulmonary macrophage cultures. Retention of virulence by bovine R252-CDV is unique and worthy of further study.

Eimeria canadensis (Protozoa, Sporozoea): ultrastructure of the host–parasite interface during development of first-generation schizonts in vitro

1980 ◽  
Vol 58 (11) ◽  
pp. 2018-2025 ◽  
Author(s):  
Bodo E. G. Mueller
Keyword(s):  
Host Cell ◽  
Parasitophorous Vacuole ◽  
Eimeria Species ◽  
Outer Zone ◽  
Ultrastructural Observation ◽  
Cell Cytoplasm ◽  
Cell Organelles ◽  
Host Cell Cytoplasm ◽  
Host Parasite

Eimeria canadensis sporozoites were inoculated into monolayer cultures of Madin–Darby bovine kidney and primary bovine embryonic kidney cells. Sporozoites retained their shape for at least 9 days. At that time, the nucleus was enlarged and contained a prominent nucleolus, and amylopectin granules were no longer apparent. The width of the parasitophorous vacuole (pv) between host cell cytoplasm and parasite pellicle widened during transformation of sporozoites into multinucleate schizonts. Areas of altered host cell cytoplasm immediately adjacent to the pv membrane increased in size and became confluent, resulting in the formation of two distinct layers of cytoplasm. The outer zone contained the host cell nucleus, mitochondria, Golgi stacks, and ER, whereas the inner layer appeared granular and was void of all cell organelles except structures resembling ribosomes. Microfilaments were abundant at the border between inner and outer zone. In the most advanced stages observed, host cell organelles persisted only in the perinuclear region. The remaining, attenuated cytoplasm resembled the former inner zone.The novel ultrastructural observation of a bilayered cytoplasm of cells harbouring E. canadensis schizonts is compared with light microscope reports of similar effects caused by other Eimeria species of ruminants and with electron microscope findings of altered intestinal and abomasal cells of sheep harbouring "globidial" schizonts.

scholarly journals Trafficking of malarial proteins to the host cell cytoplasm and erythrocyte surface membrane involves multiple pathways.

1992 ◽  
Vol 119 (6) ◽  
pp. 1481-1495 ◽  
Author(s):  
J A Gormley ◽  
R J Howard ◽  
T F Taraschi
Keyword(s):  
Host Cell ◽  
Protein Trafficking ◽  
Surface Membrane ◽  
Lipid Vesicle ◽  
Cell Cytoplasm ◽  
Host Cell Membrane ◽  
Trophozoite Stage ◽  
Host Cell Cytoplasm ◽  
Texas Red

During the asexual stage of malaria infection, the intracellular parasite exports membranes into the erythrocyte cytoplasm and lipids and proteins to the host cell membrane, essentially "transforming" the erythrocyte. To investigate lipid and protein trafficking pathways within Plasmodium falciparum-infected erythrocytes, synchronous cultures are temporally analyzed by confocal fluorescence imaging microscopy for the production, location and morphology of exported membranes (vesicles) and parasite proteins. Highly mobile vesicles are observed as early as 4 h postinvasion in the erythrocyte cytoplasm of infected erythrocytes incubated in vitro with C6-NBD-labeled phospholipids. These vesicles are most prevalent in the trophozoite stage. An immunofluorescence technique is developed to simultaneously determine the morphology and distribution of the fluorescent membranes and a number of parasite proteins within a single parasitized erythrocyte. Parasite proteins are visualized with FITC- or Texas red-labeled monoclonal antibodies. Double-label immunofluorescence reveals that of the five parasite antigens examined, only one was predominantly associated with membranes in the erythrocyte cytoplasm. Two other parasite antigens localized only in part to these vesicles, with the majority of the exported antigens present in lipid-free aggregates in the host cell cytoplasm. Another parasite antigen transported into the erythrocyte cytoplasm is localized exclusively in lipid-free aggregates. A parasite plasma membrane (PPM) and/or parasitophorous vacuolar membrane (PVM) antigen which is not exported always colocalizes with fluorescent lipids in the PPM/PVM. Visualization of two parasite proteins simultaneously using FITC- and Texas red-labeled 2 degrees antibodies reveals that some parasite proteins are constitutively transported in the same vesicles, whereas other are segregated before export. Of the four exported antigens, only one appears to cross the barriers of the PPM and PVM through membrane-mediated events, whereas the others are exported across the PPM/PVM to the host cell cytoplasm and surface membrane through lipid (vesicle)-independent pathways.

Toxin action b. Novyi (b. Oedematiens Weinberg-Sequin) on tissue growth in vitro and on cell metabolism. To the study of anaerobes and their toxin using the explantation method

1935 ◽  
Vol 31 (2) ◽  
pp. 271-272
Author(s):  
G. Barg
Keyword(s):  
Tissue Culture ◽  
Cell Metabolism ◽  
Anaerobic Bacteria ◽  
Culture Method ◽  
Tissue Growth ◽  
Tissue Culture Method ◽  
Series Of Experiments

The author has shown in a series of experiments that the tissue culture method is quite applicable for the study of anaerobic bacteria and their toxins, especially those of them that are significantly weakened by filtration.

scholarly journals Cooperation, Competition, and Specialized Metabolism in a Simplified Root Nodule Microbiome

mBio ◽  
2020 ◽  
Vol 11 (4) ◽  
Author(s):  
Bridget L. Hansen ◽  
Rita de Cassia Pessotti ◽  
Monika S. Fischer ◽  
Alyssa Collins ◽  
Laila El-Hifnawi ◽  
...  
Keyword(s):  
Potential Role ◽  
Root Nodule ◽  
Root Nodules ◽  
Imaging Mass Spectrometry ◽  
In Planta ◽  
Content Type ◽  
Specialized Metabolites ◽  
Brevibacillus Brevis

ABSTRACT Microbiomes associated with various plant structures often contain members with the potential to make specialized metabolites, e.g., molecules with antibacterial, antifungal, or siderophore activities. However, when and where microbes associated with plants produce specialized metabolites, and the potential role of these molecules in mediating intramicrobiome interactions, is not well understood. Root nodules of legume plants are organs devoted to hosting symbiotic bacteria that fix atmospheric nitrogen and have recently been shown to harbor a relatively simple accessory microbiome containing members with the ability to produce specialized metabolites in vitro. On the basis of these observations, we sought to develop a model nodule microbiome system for evaluating specialized microbial metabolism in planta. Starting with an inoculum derived from field-grown Medicago sativa nodules, serial passaging through gnotobiotic nodules yielded a simplified accessory community composed of four members: Brevibacillus brevis, Paenibacillus sp., Pantoea agglomerans, and Pseudomonas sp. Some members of this community exhibited clear cooperation in planta, while others were antagonistic and capable of disrupting cooperation between other partners. Using matrix-assisted laser desorption ionization–imaging mass spectrometry, we found that metabolites associated with individual taxa had unique distributions, indicating that some members of the nodule community were spatially segregated. Finally, we identified two families of molecules produced by B. brevis in planta as the antibacterial tyrocidines and a novel set of gramicidin-type molecules, which we term the britacidins. Collectively, these results indicate that in addition to nitrogen fixation, legume root nodules are likely also sites of active antimicrobial production.

scholarly journals The effect of hypoxanthine on the growth of an avian tissue in vitro

1944 ◽  
Vol 132 (868) ◽  
pp. 253-257 ◽  
Keyword(s):  
Tissue Culture ◽  
Tissue Growth ◽  
The Other ◽  
Purine Base ◽  
Other Hand

The experiments described in this paper have shown that the purine base hypoxanthine, added in suitable concentration to a medium commonly used for tissue culture, is capable of markedly increasing both the rate and duration of the tissue growth. On the other hand, the related base adenine acts unfavourably on the growth. The nature of the influence of the former base is perhaps not clear, but certain possibilities are discussed.

Toxoplasma gondii reorganizes the host cell architecture during spontaneous cyst formation in vitro

Parasitology ◽  
2017 ◽  
Vol 145 (8) ◽  
pp. 1027-1038 ◽  
Author(s):  
T. C. Paredes-Santos ◽  
E. S. Martins-Duarte ◽  
W. de Souza ◽  
M. Attias ◽  
R. C. Vommaro
Keyword(s):  
Toxoplasma Gondii ◽  
Host Cell ◽  
Cyst Wall ◽  
Cyst Formation ◽  
Acute Stage ◽  
Host Cells ◽  
Cell Cytoplasm ◽  
Chronic Stage ◽  
Host Cell Cytoplasm

AbstractToxoplasma gondii is an intracellular protozoan parasite that causes toxoplasmosis, a prevalent infection related to abortion, ocular diseases and encephalitis in immuno-compromised individuals. In the untreatable (and life-long) chronic stage of toxoplasmosis, parasitophorous vacuoles (PVs, containing T. gondii tachyzoites) transform into tissue cysts, containing slow-dividing bradyzoite forms. While acute-stage infection with tachyzoites involves global rearrangement of the host cell cytoplasm, focused on favouring tachyzoite replication, the cytoplasmic architecture of cells infected with cysts had not been described. Here, we characterized (by fluorescence and electron microscopy) the redistribution of host cell structures around T. gondii cysts, using a T. gondii strain (EGS) with high rates of spontaneous cystogenesis in vitro. Microtubules and intermediate filaments (but not actin microfilaments) formed a ‘cage’ around the cyst, and treatment with taxol (to inhibit microtubule dynamics) favoured cystogenesis. Mitochondria, which appeared adhered to the PV membrane, were less closely associated with the cyst wall. Endoplasmic reticulum (ER) profiles were intimately associated with folds in the cyst wall membrane. However, the Golgi complex was not preferentially localized relative to the cyst, and treatment with tunicamycin or brefeldin A (to disrupt Golgi or ER function, respectively) had no significant effect on cystogenesis. Lysosomes accumulated around cysts, while early and late endosomes were more evenly distributed in the cytoplasm. The endocytosis tracer HRP (but not BSA or transferrin) reached bradyzoites after uptake by infected host cells. These results suggest that T. gondii cysts reorganize the host cell cytoplasm, which may fulfil specific requirements of the chronic stage of infection.

scholarly journals Effect of synthetic auxins on in vitro and ex vitro bromeliad rooting

2013 ◽  
Vol 43 (2) ◽  
pp. 138-146 ◽  
Author(s):  
João Paulo Rodrigues Martins ◽  
Edilson Romais Schimildt ◽  
Rodrigo Sobreira Alexandre ◽  
Breno Régis Santos ◽  
Gizele Cristina Magevski
Keyword(s):  
Tissue Culture ◽  
Acetic Acid ◽  
Naphthalene Acetic Acid ◽  
Root Induction ◽  
In Vitro Growth ◽  
Free Medium ◽  
Ex Vitro ◽  
Synthetic Auxins ◽  
Parameters Analysis

The tissue culture can contribute to the propagation of several economic species, such as the bromeliads. This research aimed at evaluating the auxins type and concentration in the in vitro and ex vitro rhizogenesis of Neoregelia concentrica bromeliad. N. concentrica shoots were induced in a growth medium with 15.0 µM of 6-benzylaminopurine, for 80 days, followed by sub-cultivation in phytoregulator-free medium, for 45 days. In the in vitro rhizogenesis, the shoots grew in a medium supplemented with indole-3-butyric acid (IBA) or naphthalene-acetic acid (NAA), at the concentrations of 0.0 µM, 1.0 µM, 2.0 µM, 3.0 µM and 4.0 µM. In the ex vitro rhizogenesis, the bases of shoots were immersed, for 60 minutes, in IBA or NAA solutions, at the concentrations of 0.0 µM, 5.0 µM, 10.0 µM and 15.0 µM. After immersion, the shoots were planted in plastic trays with vermiculite. At the end of each rhizogenesis method, the phytotechnical parameters analysis was carried out. For the in vitro rhizogenesis, a higher number of roots were observed when the shoots were cultivated in concentrations higher than 1.0 µM of NAA, when compared to the IBA. However, the rooting rate differed only at 30 days after the in vitro growth, with a higher root induction in the shoots grown with NAA. At 60 days, the rooting rate was higher than 90% and statistically similar in all treatments. In the ex vitro rhizogenesis, a better formation of the rooting system was observed when 5.0 µM of IBA was applied, with higher rooting averages and number of roots.

scholarly journals Photoautotrophic System: A Review and Potential Application for Plant Propagation In Vitro

2018 ◽  
Vol 5 (2) ◽  
pp. 73-78
Author(s):  
Krisantini Krisantini ◽  
Ni Made Armini Wiendi
Keyword(s):  
Tissue Culture ◽  
Culture Media ◽  
Co2 Enrichment ◽  
Advantages And Disadvantages ◽  
System A ◽  
Leaf Chlorophyll ◽  
Micro Propagation ◽  
Agar Media ◽  
Propagation Methods

AbstractThe standard method of in vitro plant micro propagation uses of tightly closed culture bottles using agar media containing macro and micro nutrients and sucrose as a source of carbon for the explants. The closed bottle culture is usually kept in a temperature and light controlled environment which is lower and of different quality from the natural sunlight, resulting in high relative humidity and no air exchange inside the bottles.  Explants produced in vitro have malfunctioned stomata, undeveloped cuticles and lower leaf chlorophyll levels, and hyper hydration of the plantlets. Photoautotrophic tissue culture is micro propagation without or with a reduced sugar level in the culture media, so the growth or accumulation of carbohydrates of the explants is dependent fully upon photosynthesis and inorganic nutrient uptake. This method is usually combined with ventilation or CO2 enrichment, and recently, with incorporating porous materials such as vermiculite, gum or paper pulp to the agar media to promote better root system of the explants. This article discuss the advantages and disadvantages of the photoautotrophic micro propagation compared to the standard micro propagation methods, and provided the results of the photo autotrophic micro propagation studies conducted at Laboratory of Tissue Culture II of the Department of Agronomy and Horticulture, Bogor Agricultural University, Indonesia.

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