A simple rapid procedure for obtaining axenic cultures from monoxenic cultures of myxomycete plasmodia

1999 ◽  
Vol 45 (10) ◽  
pp. 865-870 ◽  
Author(s):  
S Balaji ◽  
A Sujatha ◽  
I Kalyanasundaram
Keyword(s):  
Axenic Culture ◽  
Differential Staining ◽  
Complex Media ◽  
Associated Bacteria ◽  
Rapid Procedure ◽  
Substrate Preferences ◽  
Axenic Cultures ◽  
Time Required

Axenic culture of myxomycete plasmodia has been attempted from time to time by various authors, but with very little success. From over 500 known species of myxomycetes, fewer than 20 species have been reported in axenic culture to date, including axenic myxamoebal cultures. In these cultures, the plasmodia required either complex media, or a killed bacterial supplement for growth. Furthermore, the time required for attaining the axenic state varied from several months to years. In the present study, a simple, rapid procedure has been developed to render monoxenic plasmodial cultures axenic. This procedure is based on our discovery that plasmodia have certain unusual substrate preferences that are inhibitory to the associated bacteria using Physarella oblonga as a model. The presence or absence of the bacteria could be ascertained through incubation in four different bacteriological media and by the use of a differential staining technique.Key words: myxomycetes, axenic culture, hydrocarbon utilization, bacterial associates.

Colony initiation in flax rust in axenic culture: involvement of a volatile factor

1979 ◽  
Vol 57 (23) ◽  
pp. 2657-2662 ◽  
Author(s):  
Rosalinda Boasson ◽  
Michael Shaw
Keyword(s):  
Carbon Dioxide ◽  
Axenic Culture ◽  
Conclusive Evidence ◽  
Culture Flask ◽  
Melampsora Lini ◽  
Air Space ◽  
Lag Period ◽  
Axenic Cultures ◽  
Incubation Temperatures

In axenic cultures of flax rust (Melampsora lini) colonies are initiated after a lag period of 12–20 days, depending partly on incubation temperatures. Colony initiation is completely inhibited by removal of a volatile factor which is absorbed by KOH in the air space of the culture flask. The fungus remains sensitive to this inhibition for 8–10 days, i.e., until shortly before visible colonies would normally have developed. While in the presence of KOH, the fungus is not killed; cultures grow normally after removal of the KOH.Although conclusive evidence must await further work, the available data strongly suggest that carbon dioxide is responsible for this effect.

scholarly journals Effect of axenic culture and NAA in Vitro on masoyi (Cryptocarya massoy (Oken) Kosterm) seeds regeneration

2021 ◽  
Vol 914 (1) ◽  
pp. 012016
Author(s):  
Y Wibisono ◽  
A I Putri ◽  
Y Hadiyan ◽  
L Haryjanto ◽  
L Hakim ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Germination Rate ◽  
Axenic Culture ◽  
Culture Method ◽  
Ms Medium ◽  
Controlled Conditions ◽  
Axenic Cultures ◽  
One Year ◽  
The Impact

Abstract The high valuable endemic commodities in Papua, Masoyi’s (Cryptocarya massoy) population facing great threat due to unsustainable harvest system. Generative propagation faces significant challenges due to seed characteristics and habitat conditions. Controlled conditions and the role of hormones have an important effect on generative growth. This study aimed to determine the influence of axenic culture with sterilization treatments Isothiazolone Biocide (IB) and 1-Naphtaleaneacetic Acid (NAA) in Murashige and Skoog (MS) medium on seed regeneration and to observe the development of seedlings at the acclimatization stage. The tissue culture method was used. The highest percentage of axenic cultures (57%) was obtained with 5% of BI. The germination rate of masoyi seeds was achieved by 100%. Furthermore, it showed varied responses depending upon concentrations of NAA, the addition of 1 ml l−1 NAA in MS medium is recommended. Acclimatization has been successfully carried out in the greenhouse (67% survival rate) and excellent seedlings growth at nursery (52.35 + 0.6 cm in height after one year transferred). The impact of the controlled conditions and the addition of NAA to axenic cultures in vitro increased the germination of masoyi seeds. Axenic culture and hormones were also important requirements for mass propagation of masoyi by tissue culture.

Axenic culture of flax rust isolated from cotyledons by cell-wall digestion

1972 ◽  
Vol 50 (12) ◽  
pp. 2601-2603 ◽  
Author(s):  
W. David Lane ◽  
Michael Shaw
Keyword(s):  
Cell Wall ◽  
Host Cell ◽  
Cell Walls ◽  
Hydrolytic Enzymes ◽  
Axenic Culture ◽  
Rust Fungi ◽  
Host Cells ◽  
Melampsora Lini ◽  
Large Numbers ◽  
Axenic Cultures

A technique is described for the isolation of colonies of flax rust (Melampsora lini (Ehrenb.) Lév., Race No. 3) from infected cotyledons. The technique depends on the digestion of the host cell walls with hydrolytic enzymes and washing of the liberated colonies. It is thus possible to collect large numbers of flax-rust colonies with only a few host cells adhering to them. Axenic cultures were established from colonies isolated in this way. This technique may be useful in establishing axenic cultures of other rust fungi.

scholarly journals Bacteria enhance the production of extracellular polymeric substances by the green dinoflagellate Lepidodinium chlorophorum

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Pauline Roux ◽  
Raffaele Siano ◽  
Karine Collin ◽  
Gwenael Bilien ◽  
Corinne Sinquin ◽  
...  
Keyword(s):  
Extracellular Polymeric Substances ◽  
Harmful Effect ◽  
Ne Atlantic ◽  
Associated Bacteria ◽  
Batch Cultures ◽  
Negative Effect ◽  
Axenic Cultures ◽  
High Concentrations ◽  
Axenic Conditions ◽  
Oyster Cultivation

AbstractHigh biomasses of the marine dinoflagellate Lepidodinium chlorophorum cause green seawater discolorations along Southern Brittany (NE Atlantic, France). The viscosity associated to these phenomena has been related to problems in oyster cultivation. The harmful effect of L. chlorophorum might originate from the secretion of Extracellular Polymeric Substances (EPS). To understand whether the EPS are produced by L. chlorophorum or its associated bacteria, or if they are a product of their interaction, batch cultures were performed under non-axenic and pseudo-axenic conditions for three strains. Maximum dinoflagellate cell abundances were observed in pseudo-axenic cultures. The non-sinking fraction of polymers (Soluble Extracellular Polymers, SEP), mainly composed of proteins and the exopolysaccharide sulphated galactan, slightly increased in pseudo-axenic cultures. The amount of Transparent Exopolymer Particles (TEP) per cell increased under non-axenic conditions. Despite the high concentrations of Particulate Organic Carbon (POC) measured, viscosity did not vary. These results suggest that the L. chlorophorum-bacteria interaction could have a detrimental consequence on the dinoflagellate, translating in a negative effect on L. chlorophorum growth, as well as EPS overproduction by the dinoflagellate, at concentrations that should not affect seawater viscosity.

scholarly journals Isolation and Culturing Axenic Microalgae: Mini–Review

2021 ◽  
Vol 15 (1) ◽  
pp. 111-119
Author(s):  
Saúl Fernandez-Valenzuela ◽  
Francisca Chávez-Ruvalcaba ◽  
Julio Cesar Beltran-Rocha ◽  
Pilar Morales San Claudio ◽  
Raúl Reyna-Martínez
Keyword(s):  
Axenic Culture ◽  
Biofuel Production ◽  
Simple Explanation ◽  
Biotechnological Applications ◽  
Isolation And Purification ◽  
Easy Task ◽  
Axenic Cultures ◽  
The Impact

Microalgae have several applications in nutraceuticals, pharmaceuticals, cosmetics, biofuel production, and bioremediation, among other fields. Isolation and purification are extremely important for obtaining axenic cultures of microalgae from different environments and crucial for their biotechnological applications, but it is not an easy task. In view of the above, it is fundamental to know the classical and advanced techniques and examples of how scientists from around the globe have applied such methods to isolate several genera and the impact of each step on successful algal purification. This review provides a brief and simple explanation of the methodology for sampling, growth, obtention of unialgal, and posterior axenic culture, which will facilitate the development of novel microalgae-related discoveries and applications for new researchers.

scholarly journals Metagenomic assembly of new (sub)arctic Cyanobacteria and their associated microbiome from non-axenic cultures

2018 ◽  
Author(s):  
Luc Cornet ◽  
Amandine R. Bertrand ◽  
Marc Hanikenne ◽  
Emmanuelle J. Javaux ◽  
Annick Wilmotte ◽  
...  
Keyword(s):  
Heterotrophic Bacteria ◽  
Point Of View ◽  
High Quality ◽  
Culture Collections ◽  
Associated Bacteria ◽  
Complete Genomes ◽  
Axenic Cultures ◽  
Metagenomic Assembly ◽  
Genome Assemblies ◽  
Public Collection

AbstractCyanobacteria form one of the most diversified phylum of Bacteria. They are important ecologically as primary producers, for Earth evolution and biotechnological applications. Yet, Cyanobacteria are notably difficult to purify and grow axenically, and most strains in culture collections contain heterotrophic bacteria that were likely associated to Cyanobacteria in the environment. Obtaining cyanobacterial DNA without contaminant sequences is thus a challenging and time-consuming task. Here, we deploy a metagenomic pipeline that enables the easy recovery of high-quality genomes from non-axenic cultures. We tested this pipeline on 17 cyanobacterial cultures from the BCCM/ULC public collection and generated novel genome sequences for 15 arctic or subarctic strains, of which 14 early-branching organisms that will be useful for cyanobacterial phylogenomics. In parallel, we managed to assemble 31 co-cultivated bacteria from the same cultures and showed that they mostly belong to Bacteroidetes and Proteobacteria, some of them being very closely related in spite of geographically distant sampling sites.ImportanceComplete genomes of cold-adapted Cyanobacteria are underrepresented in databases, due to the difficulty to grow them axenically. In this work, we report the genome sequencing of 12 (sub)arctic and 3 temperate Cyanobacteria, along with 21 Proteobacteria and 5 Bacteroidetes recovered from their microbiome. Following the use of a state-of-the-art metagenomic pipeline, 12 of our new cyanobacterial genome assemblies are of high-quality, which indicates that even non-axenic cultures can yield complete genomes suitable for phylogenomics and comparative genomics. From a methodological point of view, we investigate the fate of SSU rRNA (16S) genes during metagenomic binning and observe that multi-copy rRNA operons are lost because of higher sequencing coverage and divergent tetranucleotide frequencies. Moreover, we devised a measure of genomic identity to compare metagenomic bins of different completeness, which allowed us to show that Cyanobacteria-associated bacteria can be highly related in spite of considerable distance between collection points.

Axenic culture of Cronartium fusiforme from three spore forms

1978 ◽  
Vol 56 (21) ◽  
pp. 2641-2647 ◽  
Author(s):  
Robert C. Hare
Keyword(s):  
Culture Medium ◽  
Rust Fungus ◽  
Synthetic Medium ◽  
Axenic Culture ◽  
Pinus Elliottii ◽  
Slash Pine ◽  
Fusiform Rust ◽  
Dikaryotic Mycelium ◽  
Axenic Cultures ◽  
First Time

Axenic cultures of the fusiform rust fungus (Cronartium fusiforme) were initiated from basidiospores, aeciospores, and uredospores. The morphology, cytology, and nuclear condition of the mycelium from the three spore stages are described and illustrated. Cultures derived from aeciospores and uredospores of C. fusiforme are reported for the first time. An improved culture medium is described. Inoculation of slash pine (Pinus elliottii var. elliottii) seedlings with suspensions of mycelium from basidiospores induced some typical galls which produced pycnial exudates and aeciospores. On synthetic medium, cultures from basidiospores were monokaryotic. Mostly dikaryotic mycelium was produced from aeciospores and uredospores, but some monokaryotic hyphae were observed.

Favored RLE for Axenic Culture of Neoaplectana glaseri

1961 ◽  
Vol 35 (S1) ◽  
pp. 169-174 ◽  
Author(s):  
Norman R. Stoll
Keyword(s):  
Growth And Development ◽  
Axenic Culture ◽  
Refrigerated Storage ◽  
Axenic Cultures ◽  
In Pregnancy

Favored RLE for use in promoting growth and development of Neoaplectana glaseri in axenic cultures include those made from the livers of rabbits sacrificed late in pregnancy or early postpartum, and used after subsequent refrigerated storage periods of 15–90 days.

scholarly journals Establish axenic cultures of armored and unarmored marine dinoflagellate species using density separation, antibacterial treatments and stepwise dilution selection

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Thomas Chun-Hung Lee ◽  
Ping-Lung Chan ◽  
Nora Fung-Yee Tam ◽  
Steven Jing-Liang Xu ◽  
Fred Wang-Fat Lee
Keyword(s):  
Density Gradient ◽  
Algal Blooms ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Algal Species ◽  
Axenic Culture ◽  
Serial Dilution ◽  
Epifluorescence Microscopy ◽  
Its Sequencing ◽  
Axenic Cultures

AbstractAcademic research on dinoflagellate, the primary causative agent of harmful algal blooms (HABs), is often hindered by the coexistence with bacteria in laboratory cultures. The development of axenic dinoflagellate cultures is challenging and no universally accepted method suit for different algal species. In this study, we demonstrated a promising approach combined density gradient centrifugation, antibiotic treatment, and serial dilution to generate axenic cultures of Karenia mikimotoi (KMHK). Density gradient centrifugation and antibiotic treatments reduced the bacterial population from 5.79 ± 0.22 log10 CFU/mL to 1.13 ± 0.07 log10 CFU/mL. The treated KMHK cells were rendered axenic through serial dilution, and algal cells in different dilutions with the absence of unculturable bacteria were isolated. Axenicity was verified through bacterial (16S) and fungal internal transcribed spacer (ITS) sequencing and DAPI epifluorescence microscopy. Axenic KMHK culture regrew from 1000 to 9408 cells/mL in 7 days, comparable with a normal culture. The established methodology was validated with other dinoflagellate, Alexandrium tamarense (AT6) and successfully obtained the axenic culture. The axenic status of both cultures was maintained more than 30 generations without antibiotics. This efficient, straightforward and inexpensive approach suits for both armored and unarmored dinoflagellate species.

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