Emmanuel Frossard

Diversity and structure of AMF communities as affected by tillage in a temperate soil

Abstract. Arbuscular mycorrhizal fungi (AMF) were studied in differently tilled soils from a long-term field experiment in Switzerland. Diversity and structure of AMF communities were surveyed either directly on spores isolated from the field soil or on spores isolated from trap cultures, planted with different host plants. Single-spore cultures were established from the AMF spores obtained from trap cultures. Identification of the AMF was made by observation of spore morphology and confirmed by sequencing of ITS rDNA. At least 17 recognised AMF species were identified in samples from field and/or trap cultures, belonging to five genera of AMF – Glomus, Gigaspora, Scutellospora, Acaulospora, and Entrophospora. Tillage had a significant influence on the sporulation of some species and non-Glomus AMF tended to be more abundant in the no-tilled soil. The community structure of AMF in the field soil was significantly affected by tillage treatment. However, no significant differences in AMF diversity were detected among different soil tillage treatments. AMF community composition in trap cultures was affected much more by the species of the trap plant than by the original tillage treatment of the field soil. The use of trap cultures for fungal diversity estimation in comparison with direct observation of field samples is discussed. Electronic supplementary material to this paper can be obtained by using the Springer Link server located at http://dx.doi.org/10.1007/s00572-002-0163-z.

Potential for increasing the content and bioavailability of Fe, Zn and Ca in plants for human nutrition

This paper reviews the possibility and limits for increasing the content and bioavailability of iron (Fe), zinc (Zn) and calcium (Ca) in edible parts of staple crops, such as cereals, pulses, roots and tubers as a way to combat mineral deficiencies in human populations. Theoretically, this could be achieved by increasing the total level of Fe, Zn and Ca in the plant foods, while at the same time increasing the concentration of compounds which promote their uptake (ascorbic acid), and/or by decreasing the concentration of compounds which inhibit their absorption (phytic acid or phenolic compounds). The content of Zn and Ca in grains and fruits can in some cases be increased through soil and/or foliar applications of Zn and Ca fertilisers. Plant breeding and genetic engineering techniques, however, have the greatest potential to increase Fe and Zn content in grains, roots and tubers. The possibility of enhancing Ca and ascorbic acid content in plant foods by plant breeding and genetic engineering remained to be explored. The critical factor is to ensure that the extra minerals have an adequate bioavailability for man. Given the important role of phytic acid and polyphenols in plant physiology, reducing the levels of these compounds in the edible parts of plants does not appear to be wise although introduction of phytases which are active during digestion is an exciting possibility.

Organic phosphorus in the environment.

Separation, preconcentration and speciation Organic phosphorus speciation in natural waters by mass spectrometry Abiotic degradation of organic phosphorus compounds in the environment Enzymatic hydrolysis of organic phosphorus Abiotic stabilization of organic phosphorus Microbial tumover of phosphorus in soil Organic phosphorus dynamics in tropical agroecosystems Organic phosphorus transfer from terrestrial to aquatic environments Interactions in terrestrial ecosystems Organic phosphorus in the aquatic environment: speciation, transformations and interactions with nutrient cycles

SOIL TILLAGE AFFECTS THE COMMUNITY STRUCTURE OF MYCORRHIZAL FUNGI IN MAIZE ROOTS

In this study we tested whether communities of arbuscular mycorrhizal fungi (AMF) colonizing the roots of maize (Zea mays L.) were affected by soil tillage practices (plowing, chiseling, and no-till) in a long-term field experiment carried out in Tanikon (Switzerland). AMF were identified in the roots using specific polymerase chain reaction (PCR) markers that had been developed for the AMF previously isolated from the soils of the studied site. A nested PCR procedure with primers of increased specificity (eukaryotic, then fungal, then AMF species or species-group specific) was used. Sequencing of amplified DNA confirmed that the DNA obtained from the maize roots was of AMF origin. Presence of particular AMF species or species-group was scored as a presence of a DNA product after PCR with specific primers. We also used single-strand conformation polymorphism analysis (SSCP) of amplified DNA samples to check if the amplification of the DNA from maize roots matched the expected profile for a particular AMF ...

Phosphorus transformations in an oxisol under contrasting land-use systems: The role of the soil microbial biomass

It is generally assumed that phosphorus (P) availability for plant growth on highly weathered and P-deficient tropical soils may depend more on biologically mediated organic P (Po) turnover processes than on the release of adsorbed inorganic P (Pi). However, experimental evidence showing the linkages between Po, microbial activity, P cycling and soil P availability is scarce. To test whether land-use systems with higher soil Po are characterized by greater soil biological activity and increased P mineralization, we analyzed the partitioning of P among various organic and inorganic P fractions in soils of contrasting agricultural land-use systems and related it to biological soil properties. Isotopic labeling was used to obtain information on the turnover of P held in the microbial biomass. Soil samples were taken from grass–legume pasture (GL), continuous rice (CR) and native savanna (SAV) which served as reference. In agreement with estimated P budgets (+277, +70 and 0 kg P ha−1 for CR, GL and SAV, respectively), available P estimated using Bray-2 and resin extraction declined in the order CR > GL > SAV. Increases in Bray-2 and resin Pi were greater in CR than GL relative to total soil P increase. Organic P fractions were significantly less affected by P inputs than inorganic fractions, but were a more important sink in GL than CR soils. Extractable microbial P (Pchl) was slightly higher in GL (6.6 mg P kg−1) than SAV soils (5.4 mg P kg−1), and significantly lowest in CR (2.6 mg P kg−1). Two days after labeling the soil with carrier free 33P, 25, 10 and 2% of the added 33P were found in Pchl in GL, SAV and CR soils, respectively, suggesting a high and rapid microbial P turnover that was highest in GL soils. Indicators of P mineralization were higher in GL than CR soils, suggesting a greater transformation potential to render Po available. Legume-based pastures (GL) can be considered as an important land-use option as they stimulate P cycling. However, it remains to be investigated whether crops planted in pasture–crop rotations could benefit from the enhanced Po cycling in grass–legume soils. Furthermore, there is need to develop and test a direct method to quantify Po mineralization in these systems.

Low-P tolerance by maize (Zea mays L.) genotypes: Significance of root growth, and organic acids and acid phosphatase root exudation

We investigated some mechanisms, which allow maize genotypes to adapt to soils which are low in available P. Dry matter production, root/shoot-ratio, root length and root exudation of organic acids and acid phosphatase were investigated in four maize genotypes grown under P-deficient and P-sufficient conditions in sterile hydroponic culture. A low-P tolerant, an acid-tolerant and a low-P susceptible genotype of maize were compared with a Swiss commercial cultivar. The study found increased root development and increased exudation of acid phosphatase under P-deficient conditions in all maize genotypes, except for the Swiss cultivar. Effects on root formation and acid phosphatase were greater for the low-P tolerant than for the low-P susceptible, and the acid soil tolerant genotypes. Organic acid contents in root tissues were increased under P deficiency and related to increased PEPC activity. However, the increase in contents was associated with an increase in exudation for the low-P tolerant genotype only. The low-P susceptible genotype was characterized by high organic acid content in roots and low organic acid exudation. The organic acids content in the phloem exudates of shoots was related to root exudation under different P supply, to the difference between lines in organic acids root content, but not to the low-P tolerance or susceptibility of maize genotypes.

Release of phenols from Lupinus albus L. roots exposed to Cu and their possible role in Cu detoxification

The mechanisms enabling plants to tolerate high concentrations of available Cu in their rhizosphere are still poorly understood. To better understand the mechanisms involved, Lupinus albus L. (white lupin) was grown over 40 days in a hydroponic system compelling roots to develop under sterile conditions in the presence of a nutrient solution containing 0.5, 20 or 62 μM Cu. The following parameters were investigated in detail: low molecular weight phenols in nutrient solution (colorimetric assay), high molecular weight phenols in roots and in solution (HPLC-MS, HPLC-UV), pH, redox potential in solution (electrochemistry) and Cu distribution in the plant (AAS) as well as in apical root sections (EDX microanalysis). Finally, in vitro adsorption studies using voltammetry were conducted to evaluate the Cu adsorption behaviour of different phenolic compounds. When exposed to 62 μM Cu, biomass production of white lupin was strongly reduced. Plants grown in the presence of 20 μM Cu had a similar dry matter production compared to the control plants grown in a 0.5 μM Cu solution. However, an increased release of soluble and high molecular weight phenols into the solution was observed. The concentration of polyphenolic compounds in the roots (particularly isoflavonoids like genistein and genistein-(malonyl)-glucoside) was significantly higher for lupins grown in a 20 μM Cu solution compared to the control plants. As shown by an in vitro adsorption study, these phenolic compounds can bind Cu ions. In addition, plants exposed to 20 and 62 μM Cu cumulated high Cu amounts in root cell walls whereas only low amounts reached the symplasm. Therefore, it is proposed that the complexation of Cu2+ ions in the rhizosphere and in the roots apoplasm by phenolic compounds could alleviate Cu-mediated toxicity.

Phosphorus budget and phosphorus availability in soils under organic and conventional farming

The aim of this work was to assess to which extent organic farming practices would affect the accumulation of total and available phosphorus (P) in a cropped soil in comparison to conventional practices. In order to achieve this, soil samples were taken from a long-term field trial comparing a non-fertilised control (NON), two conventionally cultivated treatments (MIN, CON), and two organically cultivated treatments (ORG, DYN). Soil samples were taken from each treatment at two depths (0-20 and 30-50 cm) before starting the field trial (1977) and at the end of every three crop rotations (1984, 1991 and 1998). They were then analysed for total P (Pt), total inorganic P (Pi), total organic P (Po) and isotopically exchangeable Pi. After 21 years, the average P input-output budget reached -20.9 kg P ha−1 a−1 for NON, -7.8 for DYN, -5.7 for ORG, -5.0 for MIN and +3.8 for CON. Total P, Pi as well as the amount of Pi isotopically exchangeable within 1 minute (E1) were positively correlated to the P budget. Comparison between P budget and Pt in the top- and subsoils of the fertilised treatments suggested a net transfer of P from the 0–20 to the 30–50 cm layers between 13 and 26 kg P ha−1 a−1during the first rotation and between 3 and 12 kg P ha−1 a−1during the second rotation. During the third rotation a net upward movement of P from the subsurface to the topsoil ranging between 3.7 and 10.5 kg P ha−1 a−1was estimated. In the topsoil, E1decreased from an initial value of 12 mg P kg−1 to 11 in CON, 8 in MIN, 6 in ORG, 5 in DYN and 2 in NON after 21 years. In the subsoil, E1 increased from an initial value of 2 mg P kg−1 to 4 in MIN, ORG, DYN and NON and to 6 in CON. These results show that, with the exception of NON, all treatments had still an adequate level of available P after 21 years of trial and that, in this low to moderately P sorbing soil, an equilibrated input-output budget allows to maintain P availability at a constant level. In the organic systems, yields have so far partly been attained at the expense of soil reserves or residual P from earlier fertiliser applications.

Aluminum resistance in two cultivars of Zea mays L.: Root exudation of organic acids and influence of phosphorus nutrition

Root exudation of organic acids as Al-chelating compounds and P nutrition have been suggested to play a major role in Al-resistance in higher plants. Effects of Al exposure on maize plant growth, and organic acid root content and root exudation under various levels of P nutrition were examined. Sikuani, a Colombian maize cultivar tolerant to acid soils with high Al saturation, and Corso, a Swiss cultivar, were grown in sterile hydroponic conditions for 21 days. Al-caused inhibition of root growth was lower in Sikuani than in Corso. Al effect on plant growth was decreased with increasing P content in roots. Al content in roots increased with increasing P content and was higher in Sikuani than in Corso. When exposed to Al, the contents in root apices as well as the root exudation of citric and malic acids in Corso and citric, malic and succinic acids in Sikuani increased, and were higher in Sikuani than in Corso. Increased PEP carboxylase (PEPC) activity in root apices after Al exposure partially explained the variations of organic acid content in the roots. These Al-induced changes in PEPC activity, organic acid content and exudation were reduced in plants supplied with higher P concentrations during the 21 days prior to treatment. Increased secretion of organic acids after exposure to Al appeared to be specific to Al and was not totally explained by increased root content in organic acids.

Glomus intraradices dominates arbuscular mycorrhizal communities in a heavy textured agricultural soil

Arbuscular mycorrhizal fungal (AMF) spore communities were surveyed in a long-term field fertilization experiment in Switzerland, where different amounts of phosphorus (P) were applied to soil. Plots receiving no P as well as plots systematically fertilized in excess to plant needs for 31 years were used to test the hypothesis that application of P fertilizer changes the composition and diversity of AMF communities. AMF spores were isolated from the field soil, identified, and counted so as to quantify the effect of P fertilization on AMF spore density, composition, and diversity. Trap cultures were established from field soil with four host plants (sunflower, leek, maize, and Crotalaria grahamiana), and the spore communities were then analyzed in substrate samples from the pots. Altogether, nine AMF species were detected in the soil. No evidence has been acquired for effect of P fertilization on spore density, composition, and diversity of AMF in both the field soil and in trap cultures. On the other hand, we observed strong effect of crop plant species on spore densities in the soil, the values being lowest under rapeseed and highest under Phacelia tanacetifolia covercrop. The identity of plant species in trap pots also significantly affected composition and diversity of associated AMF communities, probably due to preferential establishment of symbiosis between certain plant and AMF species. AMF spore communities under mycorrhizal host plants (wheat and Phacelia in the fields and four host plant species in trap pots) were dominated by a single AMF species, Glomus intraradices. This resulted in exceptionally low AMF spore diversity that seems to be linked to high clay content of the soil.