Soil biological activity after a sixty-year fertilization practice in a wheat-maize crop rotation.

This study aimed to survey the long-term effects of fertilization practices on the functional diversity of the soil microbiota. A 60-year fertilization experiment with mineral fertilizers, farmyard manure and combined treatments was sampled in two consecutive years in maize (Zea mays L.) and wheat (Triticum aestivum L.). Soil chemical properties, plant growth and physiological parameters were measured. The MicroRespTM method was applied to assess the community level physiological profiles (CLPPs) of the rhizosphere soil, and the arbuscular mycorrhizal fungal (AMF) colonization of the roots was determined. Samples were taken in the early vegetative stages, at flowering, and at harvest in both years. The measured parameters were analysed using multifactorial ANOVA to determine treatment effects, crop-dependent differences, and seasonality. PCA analysis was performed on the data matrix to reveal more complex correspondences, and Pearson's product-moment correlation was used to confirm relationships between some of the measured soil and plant parameters. Fertilization treatments caused long-term changes in some biological parameters such as: MicroRespTM parameters, citrate utilization, total substrate-induced respiration value, and the ratio of utilization of amino acids and sugars. The rate of AMF colonization responded mainly to the plant nutrition status and the plant requirements, suggesting a plant-mediated effect in the case of mycorrhiza. Mineral nitrogen fertilization and soil acidification were found to be the main factors affecting the catabolic activity of soil microbiota, while AMF colonization responded to the balance of plant nutrition.

Electrical Capacitance versus Minirhizotron Technique: A Study of Root Dynamics in Wheat-Pea Intercrops.

This study evaluated the concurrent application and the results of the root electrical capacitance (CR) and minirhizotron (MR) methods in the same plant populations. The container experiment involved three winter wheat cultivars, grown as sole crops or intercropped with winter pea under well-watered or drought-stressed conditions. The wheat root activity (characterized by CR) and the MR-based root length (RL) and root surface area (RSA) were monitored during the vegetation period, the flag leaf chlorophyll content was measured at flowering, and the wheat shoot dry mass (SDM) and grain yield (GY) were determined at maturity. CR, RL and RSA exhibited similar seasonal patterns with peaks around the flowering. The presence of pea reduced the maximum CR, RL and RSA. Drought significantly decreased CR, but increased the MR-based root size. Both intercropping and drought reduced wheat chlorophyll content, SDM and GY. The relative decrease caused by pea or drought in the maximum CR was proportional to the rate of change in SDM or GY. Significant linear correlations (R2: 0.77-0.97) were found between CR and RSA, with significantly smaller specific root capacitance (per unit RSA) for the drought-stress treatments. CR measurements tend to predict root function and the accompanying effect on above-ground production and grain yield. The parallel application of the two in situ methods improves the evaluation of root dynamics and plant responses.

Density and Diversity of Microbial Symbionts under Organic and Conventional Agricultural Management.

The influence of organic and conventional farming and agroecology on the diversity and functioning of indigenous soil microbial communities was examined using a multifactorial analysis based on an extended minimum data set of classical status and functional tests. Main soil physicochemical properties and selected microbiological indicators, the quantity of heterotrophic or aerobic spore-forming bacteria, basal and substrate-induced respiration, catabolic activity with MicroResp™, and fluorescein diacetate enzyme activity were characterized. A pot experiment applying the most probable number method was designed with soil dilution series using Pisum sativum L. and Triticum spelta L. to assess the symbiotic infectivity and genetic diversity of key indicator groups of the plant microbiome, e.g. nitrogen-fixing bacteria (rhizobia) and arbuscular mycorrhizal fungi. Soil pH, humus content, CFU, enzyme activity, and soil respiration were significantly higher in organic soils. The activity of soil microorganisms was mainly related to clay, humus, calcium, and magnesium parameters. A redundancy analysis test of catabolic activities showed that samples were grouped according to different substrate utilization patterns and land uses were also clearly separated from each other. Farming practice influenced the abundance and diversity of microbial populations. Dark septate endophytic fungi were only found in conventional soils. In addition to confirming soil health improvements by organic management, our results highlight the importance of a complex evaluation including both classical status and functional parameters of soil microbiota, which may more reliably indicate a shift in the quality status of soils.

Symbiotic Effectivity of Dual and Tripartite Associations on Soybean (Glycine max L. Merr.) Cultivars Inoculated With Bradyrhizobium japonicum and AM Fungi.

Soybean (Glycine max L. Merr.) is regarded worldwide as indisputably one of the most important crops for human food and animal feed. The presence of symbiotic bacteria and fungi is essential for soybean breeding, especially in low-input agricultural systems. Research on the cooperation between different microbial symbionts is a key to understanding how the health and productivity of the plant is supported. The symbiotic effectivity of dual and tripartite symbiotic agents was investigated in two pot experiments on different soybean cultivars with special regard to compatibility. In the Selection experiment, two out of sixteen soybean cultivars (Aliz, Emese) were chosen on the basis of their drought tolerance and used in all the other investigations. In the Compatibility experiment, the compatible coupling of symbiotic partners was selected based on the efficiency of single and co-inoculation with two Bradyrhizobium japonicum strains and two commercial arbuscular mycorrhizal fungal (AMF) products. Significant differences were found in the infectivity and effectivity of the microsymbionts. The rhizobial and AMF inoculation generally improved plant production, photosynthetic efficiency and root activity, but this effect depended on the type of symbiotic assotiation. Despite the low infectivity of AMF, inocula containing fungi were more beneficial than those containing only rhizobia. In the Drought Stress (DS) experiment, co-inoculated and control plants were grown in chernozem soil originating from organic farms. Emese was more resistant to drought stress than Aliz and produced a bigger root system. Under DS, the growth parameters of both microbially inoculated cultivars were better than that of control, proving that even drought tolerant genotypes can strengthen their endurance due to inoculation with AMF and nitrogen fixing bacteria. Root electrical capacitance (CR) showed a highly significant linear correlation with root and shoot dry mass and leaf area. The same root biomass was associated with higher CR in inoculated hosts. As CR method detects the absorptive surface increasing due to inoculation, it may be used to check the efficiency of the microbial treatment.

Stability of sunflower 2S albumins and LTP to physiologically relevant in vitro gastrointestinal digestion.

In order for a protein to elicit a systemic allergic response it must reach the circulatory system through the intestinal mucosa as a sufficiently large fragment with adequate structural integrity. Sunflower LTP and 2S albumins (SFA8 and three mixed fractions of Alb1 and Alb2) were digested in simulated gastric fluid (SGF) for 2h and the conditions were then changed to mimic the intestinal environment for a further 2h digestion. The effects of phosphatidylcholine (PC) and emulsification on the digestibility of the proteins were investigated. PC protected all of the proteins studied against both gastric and intestinal digestive enzymes but to different extents. Emulsification of SFA8 resulted in strong protection against digestion, which was further enhanced by the presence of PC in the SGF. These results highlight the importance of considering real food structures such as emulsified systems and also the gastrointestinal environment that proteins are exposed to once consumed when assessing allergenicity.

Transcription of two blue copper-binding protein isogenes is highly correlated with arbuscular mycorrhizal development in Medicago truncatula.

Expression profiling of two paralogous arbuscular mycorrhizal (AM)-specific blue copper-binding gene (MtBcp1a and MtBcp1b) isoforms was performed by real-time quantitative polymerase chain reaction in wild-type Medicago truncatula Jemalong 5 (J5) during the mycorrhizal development with Glomus intraradices for up to 7 weeks. Time-course analysis in J5 showed that expression of both MtBcp1 genes increased continuously and correlated strongly with the colonization intensity and arbuscule content. MtPT4, selected as a reference gene of the functional plant-fungus association, showed a weaker correlation to mycorrhizal development. In a second experiment, a range of mycorrhizal mutants of the wild-type J5 was assessed. Strictly AM-penetration-defective TRV25-C and TRV25-D (dmi3, Mtsym13), hypomycorrhizal TR25 and TR89 (dmi2, Mtsym2) mutants, and a hypermycorrhizal mutant TRV17 (sunn, Mtsym12) were compared with J5 3 and 7 weeks after inoculation. No MtBcp1 transcripts were detected in the mutants blocked at the appressoria stage. Conversely, TR25, TR89, and J5 showed a gradual increase of the expression of both MtBcp1 genes in 3- and 7-week-old plants, similar to the increase in colonization intensity and arbuscule abundance. The strong correlation between the expression level of AM-specific blue copper-binding protein-encoding genes and AM colonization may imply a basic role in symbiotic functioning for these genes, which may serve as new molecular markers of arbuscule development in M. truncatula.

Molecular diversity of arbuscular mycorrhizal fungi in onion roots from organic and conventional farming systems in the Netherlands.

Diversity and colonization levels of naturally occurring arbuscular mycorrhizal fungi (AMF) in onion roots were studied to compare organic and conventional farming systems in the Netherlands. In 2004, 20 onion fields were sampled in a balanced survey between farming systems and between two regions, namely, Zeeland and Flevoland. In 2005, nine conventional and ten organic fields were additionally surveyed in Flevoland. AMF phylotypes were identified by rDNA sequencing. All plants were colonized, with 60% for arbuscular colonization and 84% for hyphal colonization as grand means. In Zeeland, onion roots from organic fields had higher fractional colonization levels than those from conventional fields. Onion yields in conventional farming were positively correlated with colonization level. Overall, 14 AMF phylotypes were identified. The number of phylotypes per field ranged from one to six. Two phylotypes associated with the Glomus mosseae-coronatum and the G. caledonium-geosporum species complexes were the most abundant, whereas other phylotypes were infrequently found. Organic and conventional farming systems had similar number of phylotypes per field and Shannon diversity indices. A few organic and conventional fields had larger number of phylotypes, including phylotypes associated with the genera Glomus-B, Archaeospora, and Paraglomus. This suggests that farming systems as such did not influence AMF diversity, but rather specific environmental conditions or agricultural practices.