Milan Gryndler

Organic and mineral fertilization, respectively, increase and decrease the development of external mycelium of arbuscular mycorrhizal fungi in a long-term field experiment

Effects of long-term mineral fertilization and manuring on the biomass of arbuscular mycorrhizal fungi (AMF) were studied in a field experiment. Mineral fertilization reduced the growth of AMF, as estimated using both measurements of hyphal length and the signature fatty acid 16:1ω5, whereas manuring alone increased the growth of AMF. The results of AMF root colonization followed the same pattern as AMF hyphal length in soil samples, but not AMF spore densities, which increased with increasing mineral and organic fertilization. AMF spore counts and concentration of 16:1ω5 in soil did not correlate positively, suggesting that a significant portion of spores found in soil samples was dead. AMF hyphal length was not correlated with whole cell fatty acid (WCFA) 18:2ω6,9 levels, a biomarker of saprotrophic fungi, indicating that visual measurements of the AMF mycelium were not distorted by erroneous involvement of hyphae of saprotrophs. Our observations indicate that the measurement of WCFAs in soil is a useful research tool for providing information in the characterization of soil microflora.

Interactions of arbuscular mycorrhizal fungi with other soil organisms

Interactions between arbuscular mycorrhizal (AM) fungi and other soil microorganisms (including bacteria, saprophytic microfungi, actinomycetes and some microarthropods) are described as well as the phenomena of the rhizosphere, mycorrhizosphere and mycosphere. Examples are given of how the growth of AM fungi and the formation of mycorrhizas are affected by the other soil organisms. All of these co-occuring positive and negative influences upon the interaction between AM fungi and plant roots are shown to underscore the complexity of the soil environment and difficulty in reaching general conclusions.

Bioaccumulation of silver in ectomycorrhizal and saprobic macrofungi from pristine and polluted areas

Macrofungi are effective accumulators of Ag. This study provides a comprehensive review of this phenomenon supported by original data on the Ag concentrations of macrofungi from pristine and Ag-polluted areas. In pristine areas, the median Ag concentrations of ectomycorrhizal (ECM) and saprobic (SAP) macrofungi were 0.79 and 2.94 mg kg(-1), respectively. In these areas, hyperaccumulation thresholds for Ag in ECM and SAP macrofungi are proposed as 100 and 300 mg kg(-1), respectively. In a Ag-polluted area, the Ag concentrations in macrofungi (ECM and SAP) were significantly elevated with the median value of 24.7 mg kg(-1) and the highest concentrations in Amanita spp. of the section Vaginatae (304-692 mg kg(-1)). The intracellular speciation of Ag in fruit-bodies of the Ag-accumulator Amanita submembranacea was inspected by size exclusion chromatography followed by sulfhydryl-specific fluorimetric assays of ligands using reverse phase high-performance liquid chromatography and improved polyacrylamide gel electrophoresis. Virtually all Ag was found to be intracellular and sequestered in the major 7 kDa and minor 3.3 kDa complexes. The lack of glutathione and phytochelatins and the presence of a single 3 kDa sulfhydryl-containing peptide in the isolated Ag-complexes suggest that detoxification of Ag in A. submembranacea may rely on metallothionein. Vertical distribution of Ag in a polluted forest soil profile has shown substantial enrichment in organic horizons; in polluted technosol, the highest Ag concentrations were found in surface layers. Standardized EDTA extraction of Ag in both the investigated soil profiles showed relatively low Ag extractibility, generally within the range of 2.2-7.7% of total Ag content.

Hyphal growth and mycorrhiza formation by the arbuscular mycorrhizal fungus Glomus claroideum BEG 23 is stimulated by humic substances

Effects of humic substances (humic acid or fulvic soil extract) or saprophytic microorganisms (Paecilomyces lilacinus and an unidentified actinomycete) on growth of mycelium and mycorrhiza formation by Glomus claroideum BEG23 were studied in a hydroponic system. Humic substances stimulated root colonization and production of extraradical mycelium by the mycorrhizal fungus. Both humic and fulvic acids tended to decrease populations of culturable bacteria and fungi in the cultivation system, indicating a moderately antibiotic activity. The addition of saprophytic microorganisms able to use humic substances to the cultivation system further stimulated the development of the mycorrhizal fungus. However, stimulation of G. claroideum was also observed when the saprophytic microorganisms were heat-killed, suggesting that their effect was not linked to a specific action on humic substances. The results indicate that humic substances may represent a stimulatory component of the soil environment with respect to arbuscular mycorrhizal fungi.

Treatment with culture fractions from Pseudomonas putida modifies the development of Glomus fistulosum mycorrhiza and the response of potato and maize plants to inoculation

Abstract Among different fractions of liquid Pseudomonas putida culture (living bacterial cells, crude cell extract and low molecular – MW 10 000 fractions) the low molecular fraction showed stimulatory effects on the growth of extraradical mycelium of arbuscular-mycorrhizal fungus Glomus fistulosum Skou and Jakobsen and its alkaline phosphatase activity. The effects were consistently observed in two experiments with potato (Solanum tuberosum L.) and maize (Zea mays L.) cultivated in vermiculite–sand mixture. Living cells of P. putida significantly stimulated growth and alkaline phosphatase activity of extraradical hyphae in the maize experiment only. Mycorrhizal colonization of potato roots was increased in the treatments inoculated with living cells of P. putida or with low-molecular fraction of bacterial culture homogenate added. The total leaf area of maize was significantly increased by inoculation with G. fistulosum alone or together with living cells of P. putida as compared to an uninoculated treatment. Shoot fresh weight, shoot/root ratio and plant height were decreased in treatments receiving low-molecular fraction of bacterial cell homogenate. Inoculation of potato plants with AM fungus increased the weight of the biggest tuber and the total weight of tubers per pot and thus the inoculation has potential use for post-vitro transplanted plants for microtuber production. Low molecular fraction of P. putida culture can be added along with mycorrhizal inoculation for enhancement of mycorrhization and activity of extraradical mycelium of plants.

Mycorrhizal hyphae as ecological niche for highly specialized hypersymbionts - or just soil free-riders?

Mycorrhizal fungi interconnect two different kinds of environments, namely the plant roots with the surrounding soil. This widespread coexistence of plants and fungi has important consequences for plant mineral nutrition, water acquisition, carbon allocation, tolerance to abiotic and biotic stresses and interplant competition. Yet some current research indicates a number of important roles to be played by hyphae-associated microbes, in addition to the hyphae themselves, in foraging for and acquisition of soil resources and in transformation of organic carbon in the soil-plant systems. We critically review the available scientific evidence for the theory that the surface of mycorrhizal hyphae in soil is colonized by highly specialized microbial communities, and that these fulfill important functions in the ecology of mycorrhizal fungal hyphae such as accessing recalcitrant forms of mineral nutrients, and production of signaling and other compounds in the vicinity of the hyphae. The validity of another hypothesis will then be addressed, namely that the specific associative microbes are rewarded with exclusive access to fungal carbon, which would qualify them as hypersymbionts (i.e., symbionts of symbiotic mycorrhizal fungi). Thereafter, we ask whether recruitment of functionally different microbial assemblages by the hyphae is required under different soil conditions (questioning what evidence is available for such an effect), and we identify knowledge gaps requiring further attention.

Long-term tracing of Rhizophagus irregularis isolate BEG140 inoculated on Phalaris arundinacea in a coal mine spoil bank, using mitochondrial large subunit rDNA markers

During the last decade, the application of arbuscular mycorrhizal fungi (AMF) as bioenhancers has increased significantly. However, until now, it has been difficult to verify the inoculation success in terms of fungal symbiont establishment in roots of inoculated plants because specific fungal strains could not be detected within colonized roots. Using mitochondrial large subunit ribosomal DNA, we show that Rhizophagus irregularis (formerly known as Glomus intraradices) isolate BEG140 consists of two different haplotypes. We developed nested PCR assays to specifically trace each of the two haplotypes in the roots of Phalaris arundinacea from a field experiment in a spoil bank of a former coal mine, where BEG140 was used as inoculant. We revealed that despite the relatively high diversity of native R. irregularis strains, R. irregularis BEG140 survived and proliferated successfully in the field experiment and was found significantly more often in the inoculated than control plots. This work is the first one to show tracing of an inoculated AMF isolate in the roots of target plants and to verify its survival and propagation in the field. These results will have implications for basic research on the ecology of AMF at the intraspecific level as well as for commercial users of mycorrhizal inoculation.

Trichloroacetic acid in Norway spruce/soil-system I. Biodegradation in soil

Trichloroacetic acid (TCA) as a phytotoxic substance affects health status of coniferous trees. It is known as a secondary air pollutant (formed by photooxidation of tetrachloroethene and 1,1,1-trichloroethane) and as a product of chlorination of humic substances in soil. Its break-down in soil, however, influences considerably the TCA level, i.e. the extent of TCA uptake by spruce roots. In connection with our investigations of TCA effects on Norway spruce, microbial processes in soil were studied using 14C-labeling. It was shown that TCA degradation in soil is a fast process depending on TCA concentration, soil properties, humidity and temperature. As a result, the TCA level in soil is determined by a steady state between uptake from the atmosphere, formation in soil, leaching and degradation. The process of TCA degradation in soil thus participates significantly in the chlorine cycle in forest ecosystems.

Chitin stimulates development and sporulation of arbuscular mycorrhizal fungi

Abstract Chitin added to sand-soil based cultivation substrates stimulated the root colonization, growth of extraradical mycelium and production of spores of arbuscular mycorrhizal fungi (AMF) in three experiments with Allium amppelloprasum , Plantago lanceolata and Lactuca sativa as host plants. Stimulation of AMF sporulation was also observed when autoclaved mycelium of Fusarium oxysporum was used instead of chitin. Increased numbers of actinomycetes in the substrate as a result of chitin treatment were recorded.

Effect of soil bacteria on hyphal growth of the arbuscular mycorrhizal fungus Glomus claroideum.

The effect of 46 bacterial strains isolated from tilled and non-tilled soils collected at 3 localities on the growth of intraradical hyphae of the arbuscular mycorrhizal (AM) fungusGlomus claroideum was demonstrated. A larger number of stimulatory bacterial isolates was obtained from tilled soils, but the bacteria showing the strongest stimulation of hyphal growth were isolated from a soil that had not been cultivated. Isolates obtained from hyphae of AM fungi showed no substantial stimulatory effects, but produced more uniform effects on hyphal growth than the isolates of bacteria obtained from soil. Bacterial cenoses present in 3 different soils differ significantly in their effects on AM fungi.