Effect of Soil Type and In Vitro Proliferation Conditions on Acclimation and Growth of Willow Shoots Micropropagated in Continuous Immersion Bioreactors.

Salix viminalis L. is a species with high capacity for micropropagation and acclimation and could therefore be used to evaluate emergent techniques in the field of plant propagation. The aims of this study were to propagate willow in liquid medium with a continuous immersion system, to explore the application of photoautotrophic conditions and to investigate the adaptation of willow plantlets to different soils that could be used as alternatives to commercial peat. For proliferation, we used 3% sucrose or sugar-free medium, and as substrates, we used commercial peat, a soil from an oak forest with high organic matter content and a crop soil with low organic matter content. The effect of sugar supplementation during proliferation and the soil characteristics during acclimation and growth were evaluated on the basis of aerial and root growth and the hydrolytic and dehydrogenase enzymatic activities of the soils. The results indicate that under photoautotrophic conditions, the supplementation of sucrose during micropropagation did not affect the subsequent growth of the plantlets. All plants acclimated without loss, but the type of soil influenced the height and vigor. Plants produced the highest shoots in peat, whereas the most root development occurred in crop soil. Soil enzyme activities were more influenced by the type of soil than by the presence of plants.

Environmental and ecological factors influencing soil functionality of biologically crusted soils by different lichen species in drylands.

Biocrusts are an essential soil surface cover at drylands where ecosystems are especially fragile to soil degradation processes due to climatic peculiarities. In the present work, (micro)biological and physicochemical properties indicative of soil functionality were studied in two different biocrust types dominated by Dipolschistes diacapsis and Lepraria isidiata and in underlying soil at two different depths (SL1, soil layer right below the biocrusts, and SL2, soil layer underlying SL1) at the Tabernas desert (southeast Spain). The influence of climatic factors (rainfall and temperature) and general soil properties on the (micro)biological properties were also analyzed in different environmental (climatic) conditions over a period of two years. PERMANOVA analyses showed significant statistical differences (Pseudo-F = 63.9; P (perm) = 0.001) among biocrust and soil layers. Throughout the study period, enzyme activities involved in C, N, and P cycles; microbial biomass-C; basal respiration; and several properties directly related to ecosystem productivity (total organic carbon, total nitrogen, concentration of ammonium and nitrate) were higher in both biocrust types than in the underlying soil layers, showing that biocrusts improved soil functions related to nutrient cycling. These properties progressively diminished in successive soil layers under the biocrusts (biocrusts > SL1 > SL2). Biocrusts showed greater similarity to each other and to SL1 than to SL2 in (micro)biological properties. A distance-based linear model analysis showed that total organic carbon, rainfall, pH, mineralized N-NH4+, and total nitrogen were the most important variables for predicting (micro)biological soil properties in biocrusts. Different biochemical behavior between the biocrusts and successive underlying soil layers has been found in wet periods. After rainfall periods, the biocrusts showed important peaks in basal soil respiration and in enzyme activities involved in C and P cycles. Nevertheless, soil biochemical properties hardly showed any peak in SL1 and did not change in SL2 despite soil moisture being higher in the soil layers below the biocrusts. Correlation analyses corroborated the existence of different relationships between soil moisture and enzymatic activities. In biocrusts, soil moisture showed a greater number of significant positive correlations with enzymes such as ß-glucosidase, invertase, and phosphomonoesterase among others, whereas in SL1 it was only correlated with cellulase and in SL2 with dehydrogenase. A change in rainfall regime, as predicted by models based on climate change in arid and semiarid zones, could affect the activity of soil enzymes in the biocrusts and underlying layers, thus aggravating the degradation of these fragile dryland ecosystems.

Use of soil enzyme activities to assess the recovery of soil functions in abandoned coppice forest systems.

Coppicing consists of periodically cutting back tree stems to ground level to stimulate the growth of multiple stems from the stool. In Central Europe, many coppiced forests were abandoned at the beginning of the last century owing to a decline in the demand for charcoal and wood. This was assumed to enable the forests to recover and the properties to become similar to those of unmanaged forest (high forest). Most studies on abandoned coppiced forest have focused on forest recovery, while soil recovery has generally been overlooked. With the aim of filling this gap, this study investigated the effect of coppicing abandonment on soil recovery by analysing the changes in soil enzyme activities (dehydrogenase, ß-glucosidase, invertase, urease, acid phosphatase and arylsulphatase). Two differently managed sessile oak (Quercus petraea) forests were selected for study: a former coppice forest, abandoned >90 years ago, and an undisturbed forest. The analytical data were compared to assess the degree of recovery of the soil in the abandoned coppice forest. The soil organic matter content was two times lower in the abandoned coppice than in the high forest, suggesting that organic matter depletion due the past coppicing is a long-term effect. All of the absolute enzyme activities were also two times lower in the abandoned coppice forest soil than in the high forest soil. However, the specific enzyme activities were similar in both types of soil. This indicates that metabolic activity is similar in both soil types, suggesting that it either recovers faster than organic matter and soil enzyme activity or that, despite the depletion in organic matter and enzyme activities, metabolic activity was sustained in coppiced forest soil. However, in the latter case this would imply that organic matter and soil enzymes were lost in exactly the same proportion, which is highly improbable.

Extraction and quantification of chlorophenolate molecules in soils spiked with 2,4-dichlorophenol and 2,4,5-trichlorophenol.

The compounds 2,4-dichlorophenol (2,4-DCP) and 2,4,5-trichlorophenol (2,4,5-TCP) are classified as priority pollutants, with potentially hazardous impacts on the environment. In soil, dissociation of the phenol group occurs, resulting in the simultaneous presence of neutral phenol and anionic phenolate. Although the toxicity of 2,4-DCP and 2,4,5-TCP to soil microbiota has been suggested to be mainly due to the phenolate anion, this hypothesis cannot be tested due to the lack of appropriate methods of extracting and quantifying the anionic form of these compounds (unlike the neutral form, which can be easily quantified). In this study, we developed a method that enables extraction and quantification of phenolate ions. The method could therefore be used to elucidate the processes that regulate the behaviour of chlorophenolic molecules in soil and to clarify the distribution and toxicity of these compounds in the edaphic environment. The proposed method uses saline solutions (CaCl2, KCl and K2SO4) of low ionic strength to extract the chlorophenolate anion from soil, followed by sequential transformation of the anion from the aqueous solution to an organic solvent that enables subsequent identification and quantification of the molecule by gas chromatography. Two soils of contrasting pH were used to test whether the proposed method was practicable. The method enabled analysis of the distribution of the neutral and anionic forms of the chlorophenols in both types of soil considered and revealed the influence of soil pH in this distribution.

Relationships among bulk soil physicochemical, biochemical, and microbiological parameters in an organic alfalfa-rice rotation system.

The microbial communities of bulk soil of rice paddy fields under an ancient organic agriculture regimen, consisting on an alfalfa-rice rotation system, were characterized. The drained soil of two adjacent paddies at different stages of the rotation was compared before rice seeding and after harvesting. The relationships among the soil microbial, physicochemical, and biochemical parameters were investigated using multivariate analyses. In the first year of rice cropping, aerobic cultivable heterotrophic populations correlated with lineages of presumably aerobic bacteria (e.g., Sphingobacteriales, Sphingomonadales). In the second year of rice cropping, the total C content correlated with presumable anaerobic bacteria (e.g., Anaerolineae). Independently of the year of rice cropping, before rice seeding, proteolytic activity correlated positively with the cultivable aerobic heterotrophic and ammonifier populations, the soil catabolic profile and with presumable aerobes (e.g., Sphingobacteriales, Rhizobiales) and anaerobes (e.g., Bacteroidales, Anaerolineae). After harvesting, strongest correlations were observed between cultivable diazotrophic populations and bacterial groups described as comprising N2 fixing members (e.g., Chloroflexi-Ellin6529, Betaproteobacteria, Alphaproteobacteria). It was demonstrated that chemical parameters and microbial functions were correlated with variations on the total bacterial community composition and structure occurring during rice cropping. A better understanding of these correlations and of their implications on soil productivity may be valid contributors for sustainable agriculture practices, based on ancient processes.

Biochemical properties of vineyard soils in Galicia, Spain.

In the present study we investigated changes in soil biochemical activity in vineyard soils. With this aim, soil samples (0-10 cm) from 15 vineyard soils developed on diverse parent materials were collected during winter. All soil samples were analysed for a large number of both general and specific biochemical properties. The values of all of the biochemical parameters analysed were extremely low, between 3 and 39% of the usual values obtained for native Galician soils under climax vegetation. To estimate the level of biochemical quality of vineyard soils, while avoiding the problem of comparing soils with totally different organic matter contents, a biochemical equilibrium equation was used. The results showed that vineyard soils are highly degraded soils, for which in the most extreme cases, the value of the biochemical equilibrium index was 13%, i.e. almost 8 times lower that in high quality soils such as climax soils.

Suitability of the OCDE tests to estimate contamination with 2,4-dichlorophenol of soils from Galicia (NW Spain).

The objective of the present study was to verify whether the generic reference levels (GRL) for soils contaminated with 2,4-dichlorophenol (2,4 DCP), established by Spanish legislation and published in the Real Decreto 9/2005, are accurate for Galician soils. For this, the surface horizons of seven soils under different types of land use were experimentally contaminated with different doses (between 0 and 10,000 times the GRL) of 2,4 DCP, and were then were subject to OECD toxicity test numbers 208 (root emergence and elongation) and 216 (soil nitrogen mineralization). The results obtained for the nitrogen mineralization test were difficult to interpret because they varied among soils, whereas the results of the root germination and elongation test were more coherent -- the values decreased with increasing doses of contaminant added to the soil. The results suggest that the root elongation test reflects the effect of this contaminant more clearly than the soil nitrogen mineralization test. Nevertheless, considering that in the lowest quality soils (i.e. agricultural soils containing a very low level of organic matter) contaminant doses of up to 1000 times the GRL did not affect root germination and elongation, it is clear that the GRL indicated in the relevant legislation are very low for the soils under study and that the threshold should be established taking into account the soil characteristics.