Revival of saprotrophic and mycorrhizal basidiomycete cultures after 20 years in cold storage in sterile water

2008 ◽  
Vol 54 (8) ◽  
pp. 595-599 ◽  
Author(s):  
Dana L. Richter

Vegetatively colonized agar cores of 69 basidiomycete fungus isolates (48 species in 30 genera and 17 families) were stored at 5 °C in tubes of sterile distilled water without manipulation for 20 years. These were represented by 34 isolates of saprotrophic fungi (29 species in 19 genera) and 35 isolates of mycorrhizal fungi (19 species in 11 genera). Viability was evaluated based on revived growth on agar media at room temperature. Fifty-seven of the 69 isolates (82.6%) grew vigorously when revived after storage for 20 years; of the 34 saprotrophic fungus isolates, 30 revived (88.2%); of the 35 mycorrhizal fungus isolates, 27 revived (77.1%). Thirteen isolates of Laccaria were all viable after 20 years, indicating cold storage in sterile water to be a good method for maintaining this important genus of mycorrhizal fungi. In general, however, mycorrhizal fungus species demonstrated lower viability than saprotrophic fungi.

1989 ◽  
Vol 35 (11) ◽  
pp. 1055-1060 ◽  
Author(s):  
Dana L. Richter ◽  
Johann N. Bruhn

Vegetatively colonized agar cores of 135 basidiomycete isolates, representing 83 species in 38 genera, were stored at 5 °C in tubes of sterile distilled water. Viability, based on growth at room temperature, was determined after storage periods ranging from 3 to 48 months. Thirty-seven isolates of saprotrophic fungi represented 32 species of 22 genera; 98 isolates of mycorrhizal fungi represented 51 species of 16 genera. Most isolates of saprotrophic species were viable, even after the most extended periods of storage. Many isolates of mycorrhizal species demonstrated lower viability, especially over longer periods of storage. Among our isolates of mycorrhizal fungi, Laccaria bicolor (seven isolates) and Laccaria laccata (nine isolates) appeared well suited to this method of long-term storage, whereas Scleroderma species (22 isolates) did not.Key words: culture maintenance, culture viability, fungal preservation, long-term storage, vegetative cultures.


2016 ◽  
Vol 62 (11) ◽  
pp. 932-937 ◽  
Author(s):  
Dana L. Richter ◽  
Thomas G. Dixon ◽  
Jill K. Smith

Vegetatively colonized agar cores of 54 basidiomycete fungal isolates were stored at 5 °C in tubes of sterile distilled water without manipulation for 30 years. The cultures represented 28 isolates of saprotrophic fungi and 26 isolates of mycorrhizal fungi. These cultures came from a group of 57 fungal isolates that were determined to be viable after 20 years of cold-water storage. Overall, 47 of the 54 isolates (87%) grew vigorously when revived after storage for 30 years. Of the 28 saprotrophic fungal isolates, 26 revived (93%); of the 26 mycorrhizal fungal isolates, 21 revived (81%). Eight of 13 isolates (62%) of Laccaria were viable after 30 years, which was considerably less viable than what was found after 20 years for this genus of mycorrhizal fungi. However, a greater percentage of isolates of Laccaria bicolor (83%) were viable than isolates of Laccaria laccata (43%), suggesting that 30 years is approaching the maximum limit for storage in cold sterile water for certain species. Considering the original 135 fungal isolates that were stored in sterile cold water from which this set was derived, overall survival after 30 years of storage was 42%; however, saprotrophic fungi demonstrated considerably greater viability (70%) than mycorrhizal fungi (21%).


Author(s):  
K. Kumutha ◽  
R. Parimala Devi ◽  
P. Marimuthu ◽  
R. Krishnamoorthy

Background: Arbuscular Mycorrhizal Fungi (AMF) and Rhizobium are beneficial plant partners exhibiting mutual association with crop plants. Conventional carrier based formulation has lesser population and limited shelf life. Present study was aimed to evaluate and to assess the shelf life in new formulation as well as in coated seeds under storage.Methods: Three experiments are conducted in this study. Population of Rhizobium and AM fungi spore count were assessed in newer seed coat formulation under room temperature storage. Secondly survival of these organisms on the coated seeds of blackgram and greengram were evaluated. In third experiment the coated seeds stored for different duration were evaluated for germination, growth and vigour index.Result: Rhizobial population was maintained at 1011 g-1 and AM spore load was 900-1000 spores g-1 up to one year of storage. AM spore count was also found sufficient up to 9 months of storage in coated seeds of both crops. Growth and vigour index were significantly enhanced in inoculated plants over uninoculated control. From the study, it was evident that these formulations can be stored upto one year at room temperature without any loss in viable count and can be used for seed coating.


2019 ◽  
Vol 54 (1) ◽  
Author(s):  
Agnieszka Jamiołkowska ◽  
Ali Hamood Thanoon ◽  
Elżbieta Patkowska ◽  
Jarosław Grządziel

Mycorrhizal fungi influence the development and activity of communities of soil microorganisms. The purpose of this study was to estimate the effect of arbuscular mycorrhizal fungus <em>Claroideoglomus etunicatum</em> (W. N. Becker &amp; Gerd.) C. Walker &amp; Schüβler on the population structure of fungal colonies in the rhizosphere of tomatoes grown in a plastic tunnel. The field experiment was conducted from 2015 to 2017 at an ecological farm in Grądy, central eastern Poland. The object of study were the three tomato cultivars: ‘Antalya F<sub>1</sub>’, ‘Esmira F<sub>1</sub>’, and ‘Pelikan F<sub>1</sub>’. Tomato seedlings were inoculated with <em>C. etunicatum</em>; spores were introduced about 5 cm deep in the rhizosphere of the studied plants (25–30 spores of <em>C. etunicatum</em> for each plant). Each year, mycological analysis of the tomato rhizosphere was conducted using Warcup’s method; structure of fungal communities of the tomato rhizosphere varied depending on the AMF applied. Saprotrophic fungi such as <em>Trichoderma</em> ssp., <em>Mucor</em> spp., and <em>Penicillium</em> spp. were often more isolated from the rhizosphere of plants inoculated with <em>C. etunicatum</em> than that of the control samples. It can be concluded that AMF directly impacted the development of fungal biodiversity in the tomato rhizosphere, particularly regarding the number of saprotrophs in the soil.


Author(s):  
P. A. Madden ◽  
W. R. Anderson

The intestinal roundworm of swine is pinkish in color and about the diameter of a lead pencil. Adult worms, taken from parasitized swine, frequently were observed with macroscopic lesions on their cuticule. Those possessing such lesions were rinsed in distilled water, and cylindrical segments of the affected areas were removed. Some of the segments were fixed in buffered formalin before freeze-drying; others were freeze-dried immediately. Initially, specimens were quenched in liquid freon followed by immersion in liquid nitrogen. They were then placed in ampuoles in a freezer at −45C and sublimated by vacuum until dry. After the specimens appeared dry, the freezer was allowed to come to room temperature slowly while the vacuum was maintained. The dried specimens were attached to metal pegs with conductive silver paint and placed in a vacuum evaporator on a rotating tilting stage. They were then coated by evaporating an alloy of 20% palladium and 80% gold to a thickness of approximately 300 A°. The specimens were examined by secondary electron emmission in a scanning electron microscope.


Author(s):  
Jane Payne ◽  
Philip Coudron

This transmission electron microscopy (TEM) procedure was designed to examine a gram positive spore-forming bacillus in colony on various solid agar media with minimal artifact. Cellular morphology and organization of colonies embedded in Poly/Bed 812 resin (P/B) were studied. It is a modification of procedures used for undecalcified rat bone and Stomatococcus mucilaginosus.Cultures were fixed and processed at room temperature (RT) under a fume hood. Solutions were added with a Pasteur pipet and removed by gentle vacuum aspiration. Other equipment used is shown in Figure 3. Cultures were fixed for 17-18 h in 10-20 ml of RT 2% phosphate buffered glutaraldehyde (422 mosm/KgH2O) within 5 m after removal from the incubator. After 3 (30 m) changes in 0.15 M phosphate buffer (PB = 209-213 mosm/KgH2O, pH 7.39-7.41), colony cut-outs (CCO) were made with a scalpel.


2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Anurag Chaturvedi ◽  
Joaquim Cruz Corella ◽  
Chanz Robbins ◽  
Anita Loha ◽  
Laure Menin ◽  
...  

AbstractEarly-diverging fungi (EDF) are distinct from Dikarya and other eukaryotes, exhibiting high N6-methyldeoxyadenine (6mA) contents, rather than 5-methylcytosine (5mC). As plants transitioned to land the EDF sub-phylum, arbuscular mycorrhizal fungi (AMF; Glomeromycotina) evolved a symbiotic lifestyle with 80% of plant species worldwide. Here we show that these fungi exhibit 5mC and 6mA methylation characteristics that jointly set them apart from other fungi. The model AMF, R. irregularis, evolved very high levels of 5mC and greatly reduced levels of 6mA. However, unlike the Dikarya, 6mA in AMF occurs at symmetrical ApT motifs in genes and is associated with their transcription. 6mA is heterogeneously distributed among nuclei in these coenocytic fungi suggesting functional differences among nuclei. While far fewer genes are regulated by 6mA in the AMF genome than in EDF, most strikingly, 6mA methylation has been specifically retained in genes implicated in components of phosphate regulation; the quintessential hallmark defining this globally important symbiosis.


Author(s):  
Vincenza Cozzolino ◽  
Hiarhi Monda ◽  
Davide Savy ◽  
Vincenzo Di Meo ◽  
Giovanni Vinci ◽  
...  

Abstract Background Increasing the presence of beneficial soil microorganisms is a promising sustainable alternative to support conventional and organic fertilization and may help to improve crop health and productivity. If the application of single bioeffectors has shown satisfactory results, further improvements may arise by combining multiple beneficial soil microorganisms with natural bioactive molecules. Methods In the present work, we investigated in a pot experiment under greenhouse conditions whether inoculation of two phosphate-solubilizing bacteria, Pseudomonas spp. (B2) and Bacillus amyloliquefaciens (B3), alone or in combination with a humic acids (HA) extracted from green compost and/or a commercial inoculum (M) of arbuscular mycorrhizal fungi (AMF), may affect maize growth and soil microbial community. Phospholipid fatty acid (PLFA) and denaturing gradient gel electrophoresis (DGGE) fingerprinting analysis were performed to detect changes in the microbial community composition. Results Plant growth, N and P uptake, and mycorrhizal root colonization were found to be larger in all inoculated treatments than in the uninoculated control. The greatest P uptake was found when B. amyloliquefaciens was applied in combination with both HA and arbuscular mycorrhizal fungi (B3HAM), and when Pseudomonas was combined with HA (B2HA). The PLFA-based community profile revealed that inoculation changed the microbial community composition. Gram+/Gram− bacteria, AMF/saprotrophic fungi and bacteria/fungi ratios increased in all inoculated treatments. The greatest values for the AMF PLFA marker (C16:1ω5) and AMF/saprotrophic fungi ratio were found for the B3HAM treatment. Permutation test based on DGGE data confirmed a similar trend, with most significant variations in both bacterial and fungal community structures induced by inoculation of B2 or B3 in combination with HA and M, especially in B3HAM. Conclusions The two community-based datasets indicated changes in the soil microbiome of maize induced by inoculation of B2 or B3 alone or when combined with humic acids and mycorrhizal inoculum, leading to positive effects on plant growth and improved nutrient uptake. Our study implies that appropriate and innovative agricultural management, enhancing the potential contribution of beneficial soil microorganisms as AMF, may result in an improved nutrient use efficiency in plants.


1985 ◽  
Vol 25 (06) ◽  
pp. 945-953 ◽  
Author(s):  
Mark A. Miller ◽  
H.J. Ramey

Abstract Over the past 20 years, a number of studies have reported temperature effects on two-phase relative permeabilities in porous media. Some of the reported results, however, have been contradictory. Also, observed effects have not been explained in terms of fundamental properties known to govern two-phase flow. The purpose of this study was to attempt to isolate the fundamental properties affecting two-phase relative permeabilities at elevated temperatures. Laboratory dynamic-displacement relative permeability measurements were made on unconsolidated and consolidated sand cores with water and a refined white mineral oil. Experiments were run on 2-in. [5.1-cm] -diameter, 20-in. [52.-cm] -long cores from room temperature to 300F [149C]. Unlike previous researchers, we observed essentially no changes with temperature in either residual saturations or relative permeability relationships. We concluded that previous results may have been affected by viscous previous results may have been affected by viscous instabilities, capillary end effects, and/or difficulties in maintaining material balances. Introduction Interest in measuring relative permeabilities at elevated temperatures began in the 1960's with petroleum industry interest in thermal oil recovery. Early thermal oil recovery field operations (well heaters, steam injection, in-situ combustion) indicated oil flow rate increases far in excess of what was predicted by viscosity reductions resulting from heating. This suggested that temperature affects relative permeabilities. One of the early studies of temperature effects on relative permeabilities was presented by Edmondson, who performed dynamic displacement measurements with crude performed dynamic displacement measurements with crude and white oils and distilled water in Berea sandstone cores. Edmondson reported that residual oil saturations (ROS's) (at the end of 10 PV's of water injected) decreased with increasing temperature. Relative permeability ratios decreased with temperature at high water saturations but increased with temperature at low water saturations. A series of elevated-temperature, dynamic-displacement relative permeability measurements on clean quartz and "natural" unconsolidated sands were reported by Poston et al. Like Edmondson, Poston et al. reported a decrease in the "practical" ROS (at less than 1 % oil cut) as temperature increased. Poston et al. also reported an increase in irreducible water saturation. Although irreducible water saturations decreased with decreasing temperature, they did not revert to the original room temperature values. It was assumed that the cores became increasingly water-wet with an increase in both temperature and time; measured changes of the IFT and the contact angle with temperature increase, however, were not sufficient to explain observed effects. Davidson measured dynamic-displacement relative permeability ratios on a coarse sand and gravel core with permeability ratios on a coarse sand and gravel core with white oil displaced by distilled water, nitrogen, and superheated steam at temperatures up to 540F [282C]. Starting from irreducible water saturation, relative permeability ratio curves were similar to Edmondson's. permeability ratio curves were similar to Edmondson's. Starting from 100% oil saturation, however, the curves changed significantly only at low water saturations. A troublesome aspect of Davidson's work was that he used a hydrocarbon solvent to clean the core between experiments. No mention was made of any consideration of wettability changes, which could explain large increases in irreducible water saturations observed in some runs. Sinnokrot et al. followed Poston et al.'s suggestion of increasing water-wetness and performed water/oil capillary pressure measurements on consolidated sandstone and limestone cores from room temperature up to 325F [163C]. Sinnokrot et al confirmed that, for sandstones, irreducible water saturation appeared to increase with temperature. Capillary pressures increased with temperature, and the hysteresis between drainage and imbibition curves reduced to essentially zero at 300F [149C]. With limestone cores, however, irreducible water saturations remained constant with increase in temperature, as did capillary pressure curves. Weinbrandt et al. performed dynamic displacement experiments on small (0.24 to 0.49 cu in. [4 to 8 cm3] PV) consolidated Boise sandstone cores to 175F [75C] PV) consolidated Boise sandstone cores to 175F [75C] with distilled water and white oil. Oil relative permeabilities shifted toward high water saturations with permeabilities shifted toward high water saturations with increasing temperature, while water relative permeabilities exhibited little change. Weinbrandt et al. confirmed the findings of previous studies that irreducible water saturation increases and ROS decreases with increasing temperature. SPEJ P. 945


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Sabaiporn Nacoon ◽  
Sanun Jogloy ◽  
Nuntavun Riddech ◽  
Wiyada Mongkolthanaruk ◽  
Jindarat Ekprasert ◽  
...  

AbstractIn this work, the effects of co-inoculation between an arbuscular mycorrhizal fungus (AMF) and a phosphate solubilizing bacteria (PSB) to promote the growth and production of sunchoke under field condition were investigated during 2016 and 2017. Four treatments were set up as follows: plants without inoculation, with AMF inoculation, with PSB inoculation and with co-inoculation of PSB and AMF. The results showed the presence of PSB and AMF colonization at the harvest stage in both years. This suggested the survival of PSB and successful AMF colonization throughout the experiments. According to correlation analysis, PSB positively affected AMF spore density and colonization rate. Also, both AMF and PSB positively correlated with growth and production of sunchoke. Co-inoculation could enhance various plant parameters. However, better results in 2016 were found in co-inoculation treatment, while AMF inoculation performed the best in 2017. All of these results suggested that our AMF and PSB could effectively promote growth and production of sunchoke under field conditions. Such effects were varied due to different environmental conditions each year. Note that this is the first study showing successful co-inoculation of AMF and PSB for promoting growth and yield of sunchoke in the real cultivation fields.


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