Soil Colonization Research Articles

Putting Plant Roots at Light: Temporal Imaging of Plant Roots and Soil Water with a Light Transmission Technique for Linking Water and Root Observations to Soil-Plant Models.

Estimating how the "hidden half" of plants, that is the roots, take up water or the influences of root system architecture or root physiological properties (such as root hydraulic conductance) on efficiency of water uptake is of prime importance for improving crops against water deficits. To unravel soil-root interactions for water, we describe a system that enables a dynamic imaging of the soil water content and of the root system, from the single root to the whole root system scales.This system uses plants grown in rhizotrons filled with sandy soil and is based on the variable attenuation of the intensity of light transmitted through the rhizotron with soil water content (the rhizotron is nearly translucent when saturated and becomes darker as soil water content decreases). Images of the transmitted light during plant water uptake (or exudation) phases are recorded with a camera, showing a qualitative pattern of water content variations. The gray levels of the image pixels are then quantitatively related to water content with a calibration.This system is affordable and can be easily implemented without specific equipment. It is scalable and quick to allow the phenotyping of a range of plant genotypes relative to their water uptake pattern. This pattern can be then related with root system properties (soil colonization, root architecture ) at different plant stages. In combination with modeling , imaging results help in obtaining physiological parameters such as root hydraulic conductivity, distributed root water uptake rates or root xylem water potential. Combination of modeling and experiment further helps in testing biological and physiological assumptions and in predicting the uptake behavior of plants in the field.

Terrestrial-type nitrogen-fixing symbiosis between seagrass and a marine bacterium

Symbiotic N2-fixing microorganisms have a crucial role in the assimilation of nitrogen by eukaryotes in nitrogen-limited environments1–3. Particularly among land plants, N2-fixing symbionts occur in a variety of distantly related plant lineages and often involve an intimate association between host and symbiont2,4. Descriptions of such intimate symbioses are lacking for seagrasses, which evolved around 100 million years ago from terrestrial flowering plants that migrated back to the sea5. Here we describe an N2-fixing symbiont, ‘Candidatus Celerinatantimonas neptuna’, that lives inside seagrass root tissue, where it provides ammonia and amino acids to its host in exchange for sugars. As such, this symbiosis is reminiscent of terrestrial N2-fixing plant symbioses. The symbiosis between Ca. C. neptuna and its host Posidonia oceanica enables highly productive seagrass meadows to thrive in the nitrogen-limited Mediterranean Sea. Relatives of Ca. C. neptuna occur worldwide in coastal ecosystems, in which they may form similar symbioses with other seagrasses and saltmarsh plants. Just like N2-fixing microorganisms might have aided the colonization of nitrogen-poor soils by early land plants6, the ancestors of Ca. C. neptuna and its relatives probably enabled flowering plants to invade nitrogen-poor marine habitats, where they formed extremely efficient blue carbon ecosystems7.

Land Use Modeling Predicts Divergent Patterns of Change Between Upper and Lower Elevations in a Subalpine Watershed of the Alps

The synergic influence of land use and climate change on future forest dynamics is hard to disentangle, especially in human-dominated forest ecosystems. Forest gain in mountain ecosystems often creates different spatial–temporal patterns between upper and lower elevation belts. We analyzed land cover dynamics over the past 50 years and predicted Business as Usual future changes on an inner subalpine watershed by using land cover maps, derived from five aerial images, and several topographic, ecological, and anthropogenic predictors. We analyzed historical landscape patterns through transition matrices and landscape metrics and predicted future forest ecosystem change by integrating multi-layer perceptron and Markov chain models for short-term (2050) and long-term (2100) timespans. Below the maximum timberline elevation of the year 1965, the dominant forest dynamic was a gap-filling process through secondary succession at the expense of open areas leading to an increase of landscape homogeneity. At upper elevations, the main observed dynamic was the colonization of unvegetated soil through primary succession and timberline upward shift, with an increasing speed over the last years. Future predictions suggest a saturation of open areas in the lower part of the watershed and stronger forest gain at upper elevations. Our research suggests an increasing role of climate change over the last years and on future forest dynamics at a landscape scale.

Soil fertility interactions with Sinorhizobium-legume symbiosis in a simulated Martian regolith; effects on nitrogen content and plant health.

Due to increasing population growth and declining arable land on Earth, astroagriculture will be vital to terraform Martian regolith for settlement. Nodulating plants and their N-fixing symbionts may play a role in increasing Martian soil fertility. On Earth, clover (Melilotus officinalis) forms a symbiotic relationship with the N-fixing bacteria Sinorhizobium meliloti; clover has been previously grown in simulated regolith yet without bacterial inoculation. In this study, we inoculated clover with S. meliloti grown in potting soil and regolith to test the hypothesis that plants grown in regolith can form the same symbiotic associations as in soils and to determine if greater plant biomass occurs in the presence of S. meliloti regardless of growth media. We also examined soil NH4 concentrations to evaluate soil augmentation properties of nodulating plants and symbionts. Greater biomass occurred in inoculated compared to uninoculated groups; the inoculated average biomass in potting mix and regolith (2.23 and 0.29 g, respectively) was greater than the uninoculated group (0.11 and 0.01 g, respectively). However, no significant differences existed in NH4 composition between potting mix and regolith simulant. Linear regression analysis results showed that: i) symbiotic plant-bacteria relationships differed between regolith and potting mix, with plant biomass positively correlated to regolith-bacteria interactions; and, ii) NH4 production was limited to plant uptake yet the relationships in regolith and potting mix were similar. It is promising that plant-legume symbiosis is a possibility for Martian soil colonization.

Assessing commercial and early-seral arbuscular mycorrhizal fungi inoculation to aid in restoring sagebrush steppe shrubs

Arbuscular mycorrhizal fungi (AMF) are drivers of plant communities as they influence plant competition by increasing plants’ root absorptive surface. As such, AMF could be an important tool for ecological restoration. However, a lack of research using AMF in sagebrush steppe communities implies a need for more assessments of its efficacy. Here, we assessed the ability of a commercial inoculum to colonize and influence growth of three native Artemisia spp. (sagebrush) shrubs and the exotic Taeniatherum caput-medusae (medusahead). We expected to increase colonization and biomass production with the addition of the inoculum. Before testing the inoculum, its viability was examined using Trifolium incarnatum (crimson clover) in autoclaved soil. Next, a factorial greenhouse experiment was conducted to evaluate the target species when grown in live or autoclaved field soil from a disturbed (early seral) site. Inoculation resulted in an average root colonization of 24.82% ± 4.86% for T. incarnatum but failed to colonize any of the sagebrush steppe species in autoclaved soil and had little to no influence on colonization in live soil. All sagebrush steppe species were colonized by AMF in live soil, and biomass production averaged 889 mg, 530 mg, 323 mg, and 1 852 mg, respectively; however, average biomass for all species was greater when grown in autoclaved soils (1 620 mg, 706 mg, 371 mg, and 3 902 mg, respectively). Our results reveal differential effects of a commercial AMF inoculum on root colonization of Artemisia spp. and T. incarnatum and highlight the need to assess the efficacy of AMF inoculum before field application.

Arbuscular mycorrhizal fungal community responses to drought and nitrogen fertilization in switchgrass stands

Anthropogenic global change is increasing the severity and frequency of abiotic stresses such as drought that are likely to affect soil communities. Arbuscular mycorrhizal fungi (AMF) play important roles in many soil processes, so it is important to understand how drought affects AMF biodiversity. This is especially relevant in agricultural systems where crops rely on AMF associations for water and nutrient uptake, and where management decisions such as crop selection and fertilizer application may influence how the AMF community responds to drought. In this study, we examined the effects of reduced precipitation and nitrogen fertilization on AMF richness, community composition, and root and soil colonization in monocultures of two cultivars of switchgrass (Panicum virgatum) grown for bioenergy feedstock. We conducted a two-year field experiment using rain-out shelters to manipulate precipitation in mature stands of switchgrass growing in a long-term nitrogen fertilization (0 or 56 kg N ha−1) experiment at the W.K. Kellogg Biological Station Long-Term Ecological Research site in Michigan, USA. We expected that AMF richness and colonization would decrease due to drought, as predicted by the stress exclusion hypothesis. Contrary to our expectations, we found that drought stress increased AMF species richness in fertilized plots by 15%; there was no effect of drought on AMF richness in unfertilized plots. Drought also significantly altered AMF community composition, primarily due to increases in Rhizophagus taxa abundance, and reduced AMF root colonization in switchgrass by 6%. We also found variation in AMF richness and colonization across switchgrass cultivars as well as sampling dates. The changes in AMF richness and composition that we observed in this study may have implications for perennial bioenergy feedstock selection and management as changes in AMF communities may feedback to affect host plants.

Water-Soluble Humic Materials Modulating Metabolism and Triggering Stress Defense in Sinorhizobium fredii.

ABSTRACTBacteria have evolved a series of mechanisms to maintain their survival and reproduction in changeable and stressful environments. In-depth understanding of these mechanisms can allow for better developing and utilizing of bacteria with various biological functions. In this study, we found that water-soluble humic materials (WSHM), a well-known environment-friendly plant growth biostimulant, significantly promoted the free-living growth and survival of Sinorhizobium fredii CCBAU45436 in a bell-shaped, dose-dependent manner, along with more-efficient carbon source consumption and relief of medium acidification. By using RNA-Seq analysis, a total of 1,136 genes significantly up-/downregulated by external addition of WSHM were identified under test conditions. These differentially expressed genes (DEGs) were enriched in functional categories related to carbon/nitrogen metabolism, cellular stress response, and genetic information processing. Further protein-protein interaction (PPI) network analysis and reverse genetic engineering indicated that WSHM might reprogram the transcriptome through inhibiting the expression of key hub gene rsh, which encodes a bifunctional enzyme catalyzing synthesis and hydrolysis of the “magic spot” (p)ppGpp. In addition, the root colonization and viability in soil of S. fredii CCBAU45436 were increased by WSHM. These findings provide us with new insights into how WSHM benefit bacterial adaptations and demonstrate great application value to be a unique inoculant additive.IMPORTANCE Sinorhizobium fredii CCBAU45436 is a highly effective, fast-growing rhizobium that can establish symbiosis with multiple soybean cultivars. However, it is difficult to maintain the high-density effective viable cells in the rhizobial inoculant for the stressful conditions during production, storage, transport, and application. Here, we showed that WSHM greatly increased the viable cells of S. fredii CCBAU45436 in culture, modulating metabolism and triggering stress defense. The root colonization and viability in soil of S. fredii CCBAU45436 were also increased by WSHM. Our results shed new insights into the effects of WSHM on bacteria and the importance of metabolism and stress defense during the bacteria’s whole life. In addition, the functional mechanism of WSHM may provide candidate genes for improving environmental adaptability and application potential of bacteria through genetic engineering.

Herbivory and Soil Water Availability Induce Changes in Arbuscular Mycorrhizal Fungal Abundance and Composition.

We tested the prediction that abundance and composition of arbuscular mycorrhizal fungi (AMF) in Ipomopsis aggregata roots and soils are influenced by ungulate herbivory and drought conditions by examining the effects in a field setting over two years. We used a multi-metric approach to quantify AMF root colonization, AMF reproduction, and AMF community composition in roots and soils. We incorporated complimentary community characterization assays by morphologically identifying spores from trap cultures and the use of terminal restriction fragment length polymorphism (T-RFLP) fingerprinting. Herbivory caused a twofold increase in spore production, an increase in AMF taxa diversity in roots, and a shift in AMF species composition in rhizosphere soils. The impact of herbivory was dependent on water availability, which differed in the two contrasting years. This study demonstrates that both soil water availability and herbivory shape arbuscular mycorrhizal fungi communities. The changes to mycorrhizal communities may help in understanding mycorrhizal function in changing climates.

Site-Dependent Relationships Between Fungal Community Composition, Plant Genotypic Diversity and Environmental Drivers in a Salix Biomass System.

Soil fungi are strongly affected by plant species or genotypes since plants modify their surrounding environment, but the effects of plant genotype diversity on fungal diversity and function have not been extensively studied. The interactive responses of fungal community composition to plant genotypic diversity and environmental drivers were investigated in Salix biomass systems, posing questions about: (1) How fungal diversity varies as a function of plant genotype diversity; (2) If plant genotype identity is a strong driver of fungal community composition also in plant mixtures; (3) How the fungal communities change through time (seasonally and interannually)?; and (4) Will the proportion of ECM fungi increase over the rotation? Soil samples were collected over 4 years, starting preplanting from two Salix field trials, including four genotypes with contrasting phenology and functional traits, and genotypes were grown in all possible combinations (four genotypes in Uppsala, Sweden, two in Rostock, Germany). Fungal communities were identified, using Pacific Biosciences sequencing of fungal ITS2 amplicons. We found some site-dependent relationships between fungal community composition and genotype or diversity level, and site accounted for the largest part of the variation in fungal community composition. Rostock had a more homogenous community structure, with significant effects of genotype, diversity level, and the presence of one genotype ("Loden") on fungal community composition. Soil properties and plant and litter traits contributed to explaining the variation in fungal species composition. The within-season variation in composition was of a similar magnitude to the year-to-year variation. The proportion of ECM fungi increased over time irrespective of plant genotype diversity, and, in Uppsala, the 4-mixture showed a weaker response than other combinations. Species richness was generally higher in Uppsala compared with that in Rostock and increased over time, but did not increase with plant genotype diversity. This significant site-specificity underlines the need for consideration of diverse sites to draw general conclusions of temporal variations and functioning of fungal communities. A significant increase in ECM colonization of soil under the pioneer tree Salix on agricultural soils was evident and points to changed litter decomposition and soil carbon dynamics during Salix growth.

Manipulation of the soil microbiome regulates the colonization of plants by arbuscular mycorrhizal fungi.

Arbuscular mycorrhizal fungi (AMF) are important symbionts of many plant species, facilitating the acquisition of soil nutrients by roots. We hypothesized that AMF root colonization is strongly influenced by the composition of the soil microbiome. Here, we evaluated mycorrhizal colonization of two plants, the grass Urochloa brizantha (Brachiaria) and the legume Crotalaria juncea (Crotalaria). These were cultivated in the same soil but hosting eight distinct microbiomes: natural soil (i); soil exposed to heat treatments for 1h at 50 ºC (ii), 80 ºC (iii), or 100 ºC (iv); sterilized soil by autoclaving (AS) followed by re-inoculation of dilutions of the natural soil community at 10-1 (v), 10-3 (vi), and 10-6 (vii); and AS without re-inoculation (viii). Microbial diversity (bacteria and fungi) was assessed through 16S rDNA and ITS1 metabarcoding, respectively, and the soil acid phosphatase activity (APASE) was measured. Sequencing results showed the formation of distinct microbial communities according to the soil manipulations, which also correlated with the decline of APASE. Subsequently, seedlings of Brachiaria and Crotalaria were grown in those soils inoculated separately with three AMF (Acaulospora colombiana, Rhizophagus clarus, and Dentiscutata heterogama) which were compared to an AMF-free control treatment. Brachiaria showed higher colonization in natural soil when compared to the microbial community manipulations, regardless of the AMF species inoculated. In contrast, two mycorrhiza species were able to colonize Crotalaria under modified microbial communities at similar rates to natural soil. Furthermore, Brachiaria showed a possible inverse relationship between APASE and mycorrhization, but this trend was absent for Crotalaria. We conclude that mycorrhizal root colonization and soil acid phosphatase activity were associated with the structure of the soil microbiome, depending on the plant species evaluated.

Coinoculaton of endophytic diazotroph with PGPR and AM fungi for enhancing sugarcane production

The field experiment was conducted at Sugarcane Research Station, Cuddalore during 2010 – 2012, to evaluate the response of sugarcane variety CoC 24 to the application of bioinoculants viz., Gluconoacetobacter diazotrophicus, AM fungi and Azophos (Azospirillum and phosphobacteria), under different levels of N, P2O5 and K2O inorganic fertilizer in plant and ratoon crop. The results revealed that the application of mycorrhizae, G. diazotrophicus, Azospirillum and phosphobacteria significantly produced higher cane yield in plant crop. The application of Gluconoacetobacter diazotrophicus @10 kg + AM fungi + Azophos @10 kg + 75 % NPK recorded the maximum germination and tiller population and also maximum mean millable cane population of 1.32 lakhs /ha, cane yield (137.45 t/ha) and sugar yield (16.96 t/ha). Similar results were recorded with the ratoon crop. The population of Azospirillum and Pseudomonas are higher compared to that of Glucanoacetobacter diazotrophicus and phosphobacteria. The application of bioinoculants improves soil microbial biomass and their by enhancing soil organic matter content. The applied bacterial sources helps in nitrogen fixation and also in continuous mobilizing and solubilisation of nutrients and their persistence and colonization in soil is an added advantage and also enhances the soil fertility. The usage of these bioinoculants inturn reduces the inorganic fertilizer input and thereby reduces the cost of cultivation.

Effect of oil pollution on the ecological condition of soils and bottom sediments of the arctic region (Yakutia)

Oil and petroleum products are known to be among the most widespread soil pollutants. The risk of emergencies is sure to increase greatly in conditions of abnormally low temperatures. Oil and oil products are not only toxic to the environment, but can also have a negative impact on the state of the permafrost zone, accelerating the processes of permafrost degradation. The goal of the research was to study the soils and bottom sediments for oil pollution in the Arctic region of Yakutia. The research was carried out with using the complex of geochemical and microbiological methods of analysis. It had shown that at present oil pollution was mainly concentrated on the objects bearing a high technogenic load. However, some migration of hydrocarbons was observed with melt, seasonal melt and rainwaters, as a result of which the natural background of the nearby territories became technogenic character. In the Arctic conditions for the first time according to the obtained data on geochemical and microbiological studies oxidative destruction of oil pollutants in soil occurred mainly under the influence of physic and chemical environmental factors, not by microbial oxidation. Sluggish processes of mineralization of organic residues and the transformation of oil pollutants by the type of putrefaction led to the colonization of oil-polluted soils of the Arctic with putrefying and pathogenic microorganisms. The purpose of further research will be studying the possibility of intensification of soil remediation processes of technologically disturbed soils at abnormally low temperatures.

Colonization Characteristics of Poplar Fungal Disease Biocontrol Bacteria N6-34 and the Inhibitory Effect on Pathogenic Fungi by Real-Time Fluorescence Quantitative PCR Detection.

Botryosphaeria dothidea is one of the most important diseases which can cause poplar canker. In our previous study, the endophytic Bacillus subtilis N6-34 screened from poplar tissue was found to be an antagonistic strain against B. dothidea. In order to ascertain the colonization rule of B. subtilis N6-34 in poplar plants, colonization of B. subtilis N6-34 labeled with a green fluorescent protein (GFP) was investigated in poplar plants and the rhizosphere soil. To confirm the inhibitory effect of the strain N6-34 on pathogenic fungi, real-time fluorescent quantitative PCR experiment with Fusarium oxysporum as the target strain was carried out. Firstly, a plasmid (pHT01-P43GFPmut3a) containing gfp gene was successfully transformed into wild B. subtilis N6-34, which has the similar characteristics with the strain N6-34 in cell growth and antifungal activity. The poplar pot experiments were carried out to examine the colonization rules and colonization quantity in poplar plants and rhizosphere soil. Observation with a confocal laser scanning microscope showed that GFP-labeled B. subtilis N6-34 (N6-34-GFP) could colonize in primary root, lateral root and adventitious root. With the extension of inoculation time, the colonization quantity of N6-34-GFP in the rhizosphere soil and poplar plants showed a trend of first increasing, then stabilizing for a period of time and then decreasing. The real-time fluorescent quantitative PCR result showed a gradual decrease in the number of F. oxysporum with increasing inoculation time. Therefore, N6-34-GFP exhibited colonization in the rhizosphere soil and different parts of poplar plants. In addition, the strain N6-34 could inhibit the growth of pathogenic fungi. The ability of B. subtilis N6-34 to colonize in the rhizosphere soil and poplar plants and to inhibit fungal growth in vitro suggest a potential application of this strain as a biological control agent.

Polyketide Synthase-Terpenoid Synthase Hybrid Pathway Regulation of Trap Formation through Ammonia Metabolism Controls Soil Colonization of Predominant Nematode-Trapping Fungus.

Polyketide synthase-terpenoid synthase (PKS-TPS) hybrid pathways for biosynthesis of unique sesquiterpenyl epoxy-cyclohexenoids (SECs) have been found to be widely distributed in plant pathogenic fungi. However, the natural and ecological functions of these pathways and their metabolites still remain cryptic. In this study, the whole PKS-TPS hybrid pathway in the predominant nematode-trapping fungus Arthrobotrys oligospora was first proposed according to all the intermediates and their derivatives from all the A. oligospora mutants with a deficiency in each gene involved in SEC biosynthesis. Most mutants displayed significantly increased trap formation which was correlated with alteration of the ammonia level. Further analysis revealed that the main metabolites involved in ammonia metabolism were largely increased in most mutants. However, significantly retarded colonization in soil were observed in most mutants compared to the wild-type strain due to significantly decreased antibacterial activities. Our results suggested that A. oligospora used the PKS-TPS hybrid pathway for fungal soil colonization via decreasing fungal nematode-capturing ability. This also provided solid evidence that boosting fungal colonization in soil was the secondary metabolite whose biosynthesis depended on a PKS-TPS hybrid pathway.

The mineralosphere\u2014interactive zone of microbial colonization and carbon use in grassland soils

To improve our understanding of early microbial colonization of pristine minerals and their group-specific C utilization, we exposed minerals (illite/goethite/quartz) amended with artificial root exudates (ARE, glucose, and citric acid) in grassland soils for a period of 24 weeks. FTIR spectra indicated that mineral-associated ARE were used within the first 2 weeks of exposure and were replaced by other carbohydrates derived from living or dead cells as well as soil-borne C sources transported into the mineralosphere after heavy rain events. Fungi and Gram-positive bacteria incorporated ARE-derived C more rapidly than Gram-negative bacteria. Gram-negative bacteria presumably profited indirectly from the ARE by cross-feeding on mineral-associated necromass of fungi and Gram-positive bacteria. The Gram-negative bacterial phyla Verrucomicrobia, Planctomycetes, Gemmatimonadetes, Armatimonadetes, and Chloroflexi showed a positive correlation with Gram-negative PLFA abundances. After 24 weeks of exposure in the grassland soils, abundances of soil microorganisms in the mineralosphere reached only 3.1% of the population density in soil. In conclusion, both bacteria and fungi slowly colonize new surfaces such as pristine minerals, but quickly assimilate artificial root exudates, creating an active microbial community in the mineralosphere.

Microenvironmental Conditions Drive the Differential Cyanobacterial Community Composition of Biocrusts from the Sahara Desert

The Sahara Desert is characterized by extreme environmental conditions, which are a unique challenge for life. Cyanobacteria are key players in the colonization of bare soils and form assemblages with other microorganisms in the top millimetres, establishing biological soil crusts (biocrusts) that cover most soil surfaces in deserts, which have important roles in the functioning of drylands. However, knowledge of biocrusts from these extreme environments is limited. Therefore, to study cyanobacterial community composition in biocrusts from the Sahara Desert, we utilized a combination of methodologies in which taxonomic assignation, for next-generation sequencing of soil samples, was based on phylogenetic analysis (16S rRNA gene) in parallel with morphological identification of cyanobacteria in natural samples and isolates from certain locations. Two close locations that differed in microenvironmental conditions were analysed. One was a dry salt lake (a “chott”), and the other was an extension of sandy, slightly saline soil. Differences in cyanobacterial composition between the sites were found, with a clear dominance of Microcoleus spp. in the less saline site, while the chott presented a high abundance of heterocystous cyanobacteria as well as the filamentous non-heterocystous Pseudophormidium sp. and the unicellular cf. Acaryochloris. The cyanobacteria found in our study area, such as Microcoleus steenstrupii, Microcoleus vaginatus, Scytonema hyalinum, Tolypothrix distorta, and Calothrix sp., are also widely distributed in other geographic locations around the world, where the conditions are less severe. Our results, therefore, indicated that some cyanobacteria can cope with polyextreme conditions, as confirmed by bioassays, and can be considered extremotolerant, being able to live in a wide range of conditions.

Cyanobacteria in early soil development of deglaciated forefields: Dominance of non-heterocytous filamentous cyanobacteria and phosphorus limitation of N-fixing Nostocales

Cyanobacteria are integral to soil development in the earliest stages of primary succession by fixing carbon (C) and nitrogen (N) essential for organismal growth. In this study, we examined soil cyanobacterial communities at the earliest stage of succession (<5 years) at two disparate glacial forefields to reveal cyanobacterial patterns central to ecosystem development. Despite vast differences in climate and the relative dominance of heterotrophic versus autotrophic bacteria in these forefields, we found common patterns in cyanobacterial community composition in early soils. This work adds to limited research suggesting the importance of non-heterocytous filamentous cyanobacteria in initial soil colonization and development of primary succession of glacier forefields. Further, a phosphorus (P) addition experiment suggests P-limitation as a mechanism underlying the restricted presence of ecologically important, putatively N-fixing, Nostocales in these emerging landscapes.

EVALUATION OF EFFICACY OF PASTEURIA PENETRANS ALONE AND IN COMBINATION WITH VERTICILLIUM CHLAMYDOSPORIUM AGAINST MELOIDOGYNE JAVANICA

The potential control of Meloidogyne javanica using Pasteuria penetrans (Pp) alone and in combination with Verticillium chlamydosporium (Vc) was tested in earthen pots following a cropping sequence "tomato-tomato-tomato” over three crop cycles. After the final harvesting, analysis of variance showed significant effect of treatments (P0.01) regarding number of eggmasses, galls and nematode female populations. Similarly, significant effect of treatments (P0.01) was also recorded in case of infected nematode females with Pasteuria and number of eggs/eggmass while no significant effect (P 0.0 5) was observed in case of endospore production. Higher numbers of eggmasses (360) and root galling (6.2) was observed where biocontrol agents were absent (control). The treatments showed 46.58, 58.85 and 33.13 percent reduction in number of galls, eggmasses and nematodes in Pp alone and 43.34, 55.21 and 30.09 percent reductions in Vc+Pp treatments respectively. Numbers of females infected with the endospores of P. penetrans were recorded higher in Pp-3 alone treatment (13.2) followed by Vc+Pp combined treatment (13.0) and maize rotated treatment (10.4) respectively. Significantly lesser number of eggmasses, galls and nematodes were recorded in pots where tomato was rotated with maize (treatment 3) compared with control. Thus rotation prevented the buildup of nematode population and resulted in a 72% decrease in numbers of eggmasses, 38% in root galling and 46% regarding female populations over the control after the final harvest. Maximum colony forming units of V. chlamydosporium per gram of soil were recorded after its addition to the soil. The fungus established in the soil during the first crop and soil colonization of the fungus was also observed after final crop.

Quantification of Pseudomonas protegens FD6 and Bacillus subtilis NCD-2 in soil and the wheat rhizosphere and suppression of root pathogenic Rhizoctonia solani AG-8

Bacillus subtilis strain NCD-2 and Pseudomonas protegens strain FD6 produce a range of antimicrobial metabolites known to be effective against soil-borne plant pathogens. Quantitative qPCR assays were designed from sequence characterized amplified fragment length polymorphisms (AFLP) diagnostic for each inoculant genotype. The qPCR minimum detection thresholds (MDT) for NCD-2 and FD6 ranged between 5.2 × 103 (log 3.71) and 1.5 × 106 (log 6.18) genome (cell) equivalents g−1 of soil or wheat root tissue and were independent of soil type. Wheat rhizosphere populations of strains FD6 and NCD-2 significantly declined during the first 14 days postinoculation and then remained stable for the next 14 days. Colonization of wheat roots, rhizosphere soil and bulk (non-rhizosphere) soil by FD6 was significantly greater than that of NCD-2. Strains FD6 and NCD-2 significantly decreased in vitro growth of wheat root pathogenic Rhizoctonia solani AG-8, Pythium irregulare and Gaeumannomyces graminis var. tritici, fungal antibiosis by FD6 greater than NCD-2. Wheat rhizosphere colonization by NCD-2 and FD6 significantly decreased abundance of R. solani AG-8 in the rhizosphere by 3- and 13-fold respectively, pathogen suppression increasing (21%–29%) foliar biomass of wheat seedlings to levels not significantly different to the non-diseased control. The qPCR assays developed in this study were used to monitor soil and rhizosphere populations of P. protegens FD6 and B. subtilis NCD-2 and their suppressive efficacies against R. solani AG-8 in wheat. These assays can be applied to monitor the dynamics of these inoculants in suppressing Rhizoctonia root rot and other crop diseases.

Overcoming field barriers to restore dryland soils by cyanobacteria inoculation

Cyanobacteria inoculation to promote biocrust formation and improve soil properties has shown positive results in indoor studies, but limited success when applied under field conditions. Successful results of application of this technology in the field have been only found in desert sand dunes in China. Therefore, further development of this technique is necessary to extend its applicability to other soil types and regions. In this study, we inoculated a consortium of three N-fixing native cyanobacteria (Nostoc commune, Scytonema hyalinum and Tolypothrix distorta) on degraded soils from three semiarid study sites in south-eastern Spain with differing soil properties and soil development. After two years, chlorophyll a spectral absorption and albedo in inoculated and control plots were similar. Consequently, a second experiment was conducted to test the effectiveness of progressive cyanobacterial preacclimation before inoculation as well as the use of habitat amelioration techniques, consisting of covering the inoculated soils with a vegetal fiber mesh or a plastic grid, to improve cyanobacteria performance. Our results showed that: 1) hardening these cyanobacterial strains did not enhance their colonization capability, and 2) covering inoculated soils with a vegetal mesh did promote soil colonization by cyanobacteria, as shown by higher chlorophyll a soil content and Chla spectral absorption and lower albedo than in the uncovered plots. Moreover, it promoted the presence of more condensed, tightly-bound exopolysaccharides (EPS) and higher molecular weight molecules in the more soluble loosely-bound EPS fraction, both of which could be involved in the improvement of soil aggregation. Finally, higher abundance of xylose and galactose was also found in this treatment, likely indicating a greater development of the induced biocrusts. The results of this study show that direct soil inoculation with cyanobacteria, whether subjected to desiccation hardening or not, did not promote the artificial formation of biocrusts in the field. However, more positive results were found when the inoculated soils were covered with a vegetal mesh to help cyanobacteria cope with abiotic stress and soil erosion. Therefore, developing diversified efficient habitat amelioration strategies might be key in the successful application of this restoration technique in the field.