Induction of systemic resistance in rice against sheath blight disease by Pseudomonas fluorescens

Two Pseudomonas fluorescens strains viz., PF1 and FP7 which inhibited the mycelial growth of sheath blight fungus Rhizoctonia solani and increased the seedling vigour of rice plants in vitro were selected for assessing induced systemic resistance (ISR) against R. solani in rice. The Pseudomonas application as a bacterial suspension or a talc-based formulation through seed, root, soil and foliar application either alone or in combination (seed+root+soil+foliar) effectively reduced sheath blight disease incidence, promoted plant growth and ultimately increased yields under glasshouse or field conditions. Efficacy of Pseudomonas strains against R. solani was comparable to that of the fungicide carbendazim, which is normally used in the field to manage the disease. Pseudomonas treatment of rice cv IR50 led to induction of systemic resistance against R. solani, as a result of increase in chitinase and peroxidase activity. However, the extent of increase varied between treatments, Pseudomonas strains used and their duration. Though two chitinase isoforms (35 and 28 kDa) and five peroxidase isozymes (PO1–PO5) were found to be associated with the ISR, 35 kDa chitinase and three peroxidase isozymes (PO3–PO5) were established as the major determinants of ISR. Although a single application of a Pseudomonas strain resulted in ISR, the combined application through all of the four (seed, root, soil and foliar) methods increased the durability of ISR in rice plants. In addition, the Pseudomonas strains produced chitinase in the culture medium. It is presumed that the induced chitinase, peroxidase and bacterial chitinase may be either directly or indirectly involved in the reduction of sheath blight disease development in rice.     

Influence of soil type and indigenous pathogenic fungi on bean hypocotyl rot caused by Rhizoctonia solani AG4 HGI in Cuba

Calcisol, ferralsol and vertisol soils, representative of different bean production areas of Villa Clara province in Cuba, were selected to determine the impact of soil type on bean hypocotyl rot severity caused by Rhizoctonia solani AG4 HGI (isolate CuVC-Rs7). In inoculated autoclaved soil, hypocotyl rot was most severe in calcisol soil, followed by ferralsol soils and then vertisol soils. In inoculated natural soils, disease severity was lower in vertisol and calcisol soils and higher in ferralsol soil, indicating that biological factors are suppressing or stimulating the pathogenic efficiency of R. solani. Native binucleate Rhizoctonia AGF, Sclerotium rolfsii and R. solani AG 4 HGI were isolated from bean plants grown in natural calcisol, vertisol and ferralsol soils, respectively. Subsequent studies about the interaction between these fungi and R. solani indicated that they were involved in the variability of disease severity caused by R. solani. The addition of R. solani AG4 HGI (isolate CuVC-Rs7) into each autoclaved soil inoculated with binucleate Rhizoctonia or S. rolfsii resulted in a reduction of disease severity caused by this pathogen while in soils inoculated with native R. solani AG4 HGI, disease severity increased. Irrespective of fungal interactions, calcisol was always the most disease conducive soil and vertisol the most disease repressive soil. The mechanisms by which native pathogenic fungi could influence disease severity caused by R. solani are discussed.     

Screening of rhizobacteria isolated from maize (Zea mays L.) in Rio Grande do Sul State (South Brazil) and analysis of their potential to improve plant growth

Plant growth-promoting rhizobacteria (PGPR) are considered to have a beneficial effect on host plants and may facilitate plant growth by different mechanisms. In this work, the influence of different soil types on the bacterial diversity and the stimulatory effects of selected PGPR on two cultivars of maize were investigated. A set of 292 strains was isolated from the roots and rhizosphere soil of maize cultivated in five different areas of the Rio Grande do Sul State in Brazil. 16S rDNA-PCR-RFLP and 16S rDNA partial sequencing were used for identification, and the Shannon–Weaver index was used to evaluate bacterial diversity. We evaluated the ability of each isolate to produce indole acetic acid (IAA), siderophores and solubilize phosphates. On the basis of multiple PGP traits, six isolates were selected to test their potential as plant growth-promoting rhizobacteria on maize plants. In both the roots and the rhizospheric soil of maize, the dominant bacterial genera identified were Klebsiella and Burkholderia. IAA producers were distributed widely among isolates, regardless of the sampling site. Approximately 42% of the isolates exhibited at least two attributes, and 24% showed all three PGP traits. Three strains, identified as Achromobacter, Burkholderia, and Arthrobacter, were effective as PGPR in both of the cultivars evaluated.     

Assessing soil quality under intensive cultivation and tree orchards in Southern Italy

Concerns about groundwater contamination as well as pesticide residues in food and soil have fuelled vigorous debates about the sustainability of chemical-intensive agriculture. Search has been prompted for agronomic strategies with lower environmental hazards. In this multidisciplinary study we compared the characteristics of soils from 20 agricultural farms selected in five geographical areas of Southern Italy with different soil types. In each farm, fields with management regime classified as high-input (HIMR, intensive cultivation under plastic tunnels) or low-input (LIMR, tree orchards) were selected. Soil samples were analyzed for 31 parameters including physical and chemical properties (bulk density, water holding capacity, texture, pH, limestone, electrical conductivity, organic C to a depth of 0–20 and 20–40 cm, total N, P₂O₅, Ca²⁺, Mg²⁺, K⁺, Na⁺, cation exchange capacity), enzymatic activities (dehydrogenase, arylsulphatase, β-glucosidase, phosphatase and urease) and microbiological features (potential respiration, functional diversity of microbial populations by BIOLOG EcoPlates™, microbial biomass, fungal mycelium, culturable actinomycetes, bacteria and fungi, pseudomonads and bacterial species richness by 16S rDNA-DGGE). Finally, a soil bioassay was performed in order to evaluate the plant growth of a biotest plant (Lactuca sativa) and soil suppressiveness of the Rhizoctonia solani–L. sativa pathosystem.Results showed that many soil properties were influenced by management regime more than by the sampling area. Compared to LIMR, HIMR soils consistently had reduced soil organic C (−24%), enzymatic activities, microbial biomass and fungal mycelium (−40% and −18%, respectively), functional diversity (−18%) and bacterial species richness (−14%). On the contrary, the same soils showed a remarkable increase in the values of the parameters related to the mineral soil fraction (electrical conductivity +370%; P₂O₅ +72%; Na⁺ +86%). Management regime did not affect cation exchange capacity, pH, limestone and soil texture. The lettuce bioassay showed a higher plant growth (+17%) in the LIMR compared to HIMR soils, despite the lower content of mineral nutrients. Suppression of R. solani was not influenced by management regime, but significant differences were recorded among farms. Differences among the assessed soil parameters indicate a trend of soil quality deterioration under the high-input management regime.     

Effect of successive cauliflower plantings and Rhizoctonia solani AG 2-1 inoculations on disease suppressiveness of a suppressive and a conducive soil

Disease suppressiveness against Rhizoctonia solani AG 2-1 in cauliflower was studied in two marine clay soils with a sandy loam texture. The soils had a different cropping history. One soil had a long-term (40 years) cauliflower history and was suppressive, the other soil was conducive and came from a pear orchard not having a cauliflower crop for at least 40 years. These two soils were subjected to five successive cropping cycles with cauliflower or remaining fallow in a greenhouse experiment. Soils were inoculated with R. solani AG 2-1 only once or before every crop. Disease decline occurred in all treatments cropped with cauliflower, either because of a decreased pathogen population or increased suppressiveness of the soil. Disease suppressiveness tests indicated that the conducive soil became suppressive after five subsequent cauliflower crops inoculated each cycle with R. solani AG 2-1. Suppressiveness of all treatments was measured in a seed germination test (pre-emergence damping-off) as well as by measuring the spread of R. solani symptoms in young plants (post-emergence damping-off). Results showed that suppressiveness was significantly stimulated by the successive R. solani inoculations; presence of the cauliflower crop had less effect. Suppressiveness was of biological origin, since it disappeared after sterilization of the soil. Moreover, suppressiveness could be translocated by adding 10% suppressive soil into sterilized soil. The suppressive soil contained higher numbers of culturable filamentous actinomycetes than the conducive soil, but treatments enhancing suppressiveness did not show an increased actinomycetes population. The suppressiveness of the soil samples did also not correlate with the number of pseudomonads. Moreover, no correlation was found with the presence of different mycoparasitic fungi, i.e. Volutella spp., Gliocladium roseum, Verticillium biguttatum and Trichoderma spp. The suppressive soil contained a high percentage of bacteria with a strong in vitro inhibition of R. solani. These bacteria were identified as Lysobacter (56%), Streptomyces (23%) and Pseudomonas (21%) spp. A potential role of Lysobacter in soil suppressiveness was confirmed by quantitative PCR detection (TaqMan), since a larger Lysobacter population was present in suppressive cauliflower soil than in conducive pear orchard soil. Our experiments showed that successive cauliflower plantings can cause a decline of the damage caused by R. solani AG 2-1, and that natural disease suppressiveness was most pronounced after subsequent inoculations with the pathogen. The mode of action of the decline is not yet understood, but antagonistic Lysobacter spp. are potential key organisms.     

Arsenic-contaminated soils genetically modified<Emphasis Type="Italic">Pseudomonas</Emphasis> spp. and their arsemc-phytoremediation potential

Sorghum was inoculated withPseudomonas bacteria, including strains harboring an As-resistance plasmid, pBS3031, to enhance As-extraction by the plants.Pseudomonas strains (P.fluorescens 38a, P.putida 53a, and P.aureofaciens BS1393) were chosen because they are antagonistic to a wide range of phyto-pathogenic fungi and bacteria, and they can stimulate plant growth. The resistance of natural rhizospheric pseudomonads to sodium arsenite was assessed. Genetically modifiedPseudomonas strains resistant to As(III)/As(V) were obtained via conjugation or transformation. The effects of the strains on the growth of sorghum on sodium-arsenite-containing soils were assessed. The conclusions from this study are: (1) It is possible to increase the survivability of sorghum growing in sodium-arsenite-containing soil by using rhizosphere pseudomonads. (2) The presence of pBS3031 offers the strains a certain selective advantage in arsenite-contaminated soil. (3) The presence of pBS3031 impairs plant growth, due to the As-resistance mechanism determined by this plasmid: the transformation of the less toxic arsenate into the more toxic, plant-root-available arsenite by arsenate reductase and the active removal of arsenite from bacterial cells. (4) Such a mechanism makes it possible to develop a bacteria-assisted phytoremediation technology for the cleanup of As-contaminated soils and is the only possible way of removing the soil-sorbed arsenates from the environment.     

Inoculation of wheat with Azospirillum brasilense and Pseudomonas fluorescens: Impact on the production and culturable rhizosphere microflora

Scientific evidence recognizes that the operation of a terrestrial ecosystem depends on soil microbial activity. Some Azospirillum strains produce plant growth regulators, increase the development of roots, and fix atmospheric nitrogen (N₂). Some Pseudomonas strains are capable of producing cytokinins and solubilizing organic phosphorus. A sustainability analysis requires a detailed knowledge of the interrelationships between the microorganisms added to the system and those present in the soil. This study examines the effect of three commercial inoculants Azospirillum brasilense Az1 and Az2 as well as Pseudomonas fluorescens Pf on biomass production, grain yield and rhizosphere colonization of wheat, combined with two levels of N-addition. Plate counts of rhizosphere soil showed that the inoculation and N-addition did not affect the number of P. fluorescens, whereas it significantly affected the number of Azospirillum. N-addition and inoculation did not change the communities of actinomycetes and bacteria but they changed the number of fungi at the rhizosphere of wheat plants. Community-level physiological profiles of carbon-source utilization of rhizosphere soil microbial communities were changed after inoculation with Az1, Az2 and Pf depending on the phenological stage of the crop. Although no significant responses were observed, in average, PGPB inoculation increased aerial biomass by 12%, root biomass by 40% and grain yield by 16%. These increases represent important earnings for the farmer and they may help to obtain a greater sustainability of the agroecosystems.     

The earthworm (Aporrectodea caliginosa) primes the release of mobile and available micronutrients in soil

The objectives of our study were to quantify the impact of endogeic earthworms Aporrectodea caliginosa (Savigny) on iron (Fe), manganese (Mn) and zinc (Zn) mobility and availability in soil. Dried rye straw (Cecale cereale L.), clover aboveground parts (Trifolium pratense L.) or calcium carbonate were added to determine the effects on soil micronutrient mobility. To test the importance of soil–water saturation mediated by earthworms, soil samples were modified to 60% (control) and 100% (as in casts) water holding capacity (WHC). To assess availability of micronutrients, a cucumber plant (Cucumis sativus L.) bioassay were used. Earthworm casts had generally higher amounts of water-soluble micronutrients compared with bulk soils regardless of their moisture contents. The increased micronutrient mobility was more pronounced in casts from soil samples amended with plant residues (especially with straw) and was significantly higher than mobility in control soil for at least 1 week after the casts were deposited. Pre-incubation of soils amended with clover or straw with living earthworms for 4 weeks produced an increase in both shoot biomass and translocation rate of micronutrients (Mn, Zn) into xylem sap of cucumber compared to soils not worked by earthworms. The earthworm-mediated plant performances were determined 4 weeks after the earthworms were removed. The results demonstrated that earthworms can significantly impact the formation of mobile and available micronutrients in a soil. The relationship between micronutrient availability to cucumber plants and earthworm contribution to nitrogen (N) mineralization and micronutrient mobility are discussed.     

Interactive effects of plant growth–promoting rhizobacteria and organic fertilization on P nutrition of Zea mays L. and Brassica napus L.

The organic matter supply can promote the dispersal and activity of applied plant growth–promoting rhizobacteria (PGPR), but the complementary effect of organic fertilization and PGPR application on the turnover of P is scarcely known. The effects of the application of two PGPR strains (Pseudomonas fluorescens strain DR54 and Enterobacter radicincitans sp. nov. strain DSM 16656) alone and in combination with organic fertilization (cattle manure and biowaste compost) on growth and P uptake of maize (Zea mays L.) and oilseed rape (Brassica napus L.) were investigated under semi–field conditions. Furthermore, P pools and phosphatase activities in soil and the arbuscular mycorrhizal colonization of maize were examined. The organic‐fertilizer amendments increased the growth and P uptake of both plant species and the soil P pools. The application of the E. radicincitans strain increased P uptake of oilseed rape when no organic fertilizer was added. Furthermore, the application of both bacterial strains increased the activities of phosphatases under both plant species. Here, the effect of the PGPR application even exceeded the effect of organic fertilization. The magnitude of this effect varied between the different fertilizing treatments and between the two bacterial strains. Phosphatase activities were increased to the greatest extent after application of P. fluorescens in the unfertilized soil. Under rape increases of 52% for acid phosphatase activities (ACP), 103% for alkaline phosphatase activities (ALP), and 133% for phosphodiesterase (PDE) were observed therewith. In the unfertilized soil, the application of P. fluorescens also resulted in a strong increase of the arbuscular mycorrhizal colonization of maize. We conclude that application of PGPR can promote the P mobilization and supply of crops in P‐deficient soils, however, in combination with organic fertilization these effects might be masked by a general improved P supply of the crops. Interactive effects of applied bacterial strains and organic fertilization depend on the sort of organic fertilizer and crop species used.     

Isolation and characterization of phosphate-solubilizing bacteria from betel nut (Areca catechu) and their effects on plant growth and phosphorus mobilization in tropical soils

Phosphate-solubilizing bacteria (PSB) were isolated and characterized from the rhizosphere and bulk soils of Areca catechu plants. A long history of phosphate fertilizer use has elicited a direct effect on the incidence of soil PSB. Their abundance and ability to solubilize insoluble phosphate were significantly greater (P?<?0.0001) in soils with low available phosphorus (P) content than in other soil types. Three efficient PSB strains, namely, ASL12, ASG34, and ADH302, were identified as Acinetobacter pittii, Escherichia coli, and Enterobacter cloacae by characterizing 16S rRNA sequences and biochemical characteristics; they produced gluconic acid at concentrations of 7862.4, 4306.5, and 2663.8 mg L^?1, respectively. The highest amount of solubilized P was determined in Pikovskaya (PVK) medium for the bacterial strain ASL12. The secretion of gluconic acid was related to the available P of rhizosphere soils and P solubilization. Under shaded conditions, the application of these three strains significantly improved plant height, shoot and root dry weight, and nutrient uptake of A. catechu seedlings. A further increase in P solubilization was observed by co-inoculating the three strains and also applying tricalcium phosphate (TCP) or aluminum phosphate (AP). A significant (P?<?0.05) correlation was also observed between P-solubilization activity and A. catechu plant growth in pot experiments. Thus, the three strains can be potentially applied as inoculants in tropical and aluminum-rich soils.     

Improvement of Brassica napus growth under cadmium stress by cadmium-resistant rhizobacteria

This study focuses on the characterization of four bacterial isolates from heavy metal-polluted rhizosphere in order to examine their plant growth promoting (PGP) activity. The PGP activity on the canola (Brassica napus) of the strains which showed cadmium resistance and multiple PGP traits was assessed in the presence and in the absence of Cd²⁺. The strains, Pseudomonas tolaasii ACC23, Pseudomonas fluorescens ACC9, Alcaligenes sp. ZN4 and Mycobacterium sp. ACC14 showed 1-aminocyclopropane-1-carboxylate deaminase (ACCD) activity. They also synthesized ACCD enzyme in vitro when 0.4 mM Cd²⁺ was added to the growth medium. The presence of the metal, however, reduced the ACCD activity in Alcaligenes sp. ZN4 and Mycobacterium sp. ACC14, while it did not affect the ACCD activity of P. tolaasii ACC23 and P. fluorescens ACC9. ACC9 and ACC23 produced indole acetic acid (IAA) and siderophores, while ACC14 produced only IAA. IAA and siderophores were produced more actively under Cd-stress.Root elongation assays conducted on B. napus under gnotobiotic conditions demonstrated increases (from 34% up to 97%) in root elongation of inoculated canola seedlings compared to the control plants. Subsequently, the effect of inoculation with these strains on growth and uptake of Cd²⁺ in roots and shoots of canola was studied in pot experiments using Cd-free and Cd-treated (15 μg Cd²⁺ g^?1 dw) soil. Inoculation with P. tolaasii ACC23, P. fluorescens ACC9 and Mycobacterium sp. ACC14 promoted the growth of plants at concentrations of 0 and 15 μg Cd²⁺ g^?1 soil. The maximum growth was observed in the plants inoculated with P. tolaasii ACC23. The strains did not influence the specific accumulation of cadmium in the root and shoot systems, but all increased the plant biomass and consequently the total cadmium accumulation.The present observations showed that the bacterial strains used in this study protect the plants against the inhibitory effects of cadmium, probably due to the production of IAA, siderophores and ACCD activity.     

Rhizosphere colonization by Serratia marcescens for the control of Sclerotium rolfsii

Two-hundred and three different strains of bacteria were isolated from the rhizosphere of bean, peanut and chickpea plants grown in Sclerotium rolfsii infested soil. A bacterium, identified as Serratia marcescens, was found to be the best biocontrol agent of the pathogen, under greenhouse conditions (up to 75% disease reduction). Populations of 10⁵ or 10⁶ CFU g⁻¹ soil were the most effective in disease control. The drench and drip application of S. marcescens suspension were more effective in controlling S. rolfsii than spraying, mixing in soil or seed coating. This bacterium significantly reduced damping-off incidence of bean, caused by Rhizoctonia solani, by 50%, but was not effective against Pythium aphanidematum in cucumber. A natural mutant of S. marcescens, resistant to the antibiotic rifampicin, was isolated. The mutant, effective as the wild type, was used to study rhizosphere colonization. The highest population density of the bacteria was found on the proximal portion of the root, decreasing significantly until the tips, where they increased again.     

Effects of soil,grass and legume root extracts on heterotrophic bacteria,nitrogen mineralization and nitrification in soil

The effects of root extracts of four grasses and two legumes and extracts of soils supporting these plants on the growth of five strains of heterotrophic soil bacteria, and on the rate of nitrogen mineralization and nitrification were measured in culture and in soil. All the root extracts inhibited the growth in culture of the five bacteria by 9–98 per cent. The legume-soil and one of the grass-soil extracts did not inhibit bacterial growth. Only two of the grass-soils, Andropogon lectorum and Pennisetum purpureum markedly inhibited the five bacteria. Incubation of soils with extracts of grass roots or grass-soil increased the rates of nitrogen mineralization and nitrification and incubation with legume root and soil extracts increased the rates of nitrogen mineralization and nitrification even further.     

Effect of single and dual phosphate-solubilizing bacterial strain inoculations on overall growth of mung bean plants

Available phosphorus is limiting in most cultivable soils in several parts of India, including Rajasthan. Four phosphate-solubilizing bacterial strains viz. Pseudomonas fluorescens BAM-4, Burkholderia cepacia BAM-6, B. cepacia BAM-12 and Aeromonas vaga BAM-77 were isolated from the rhizosphere of pearl millet (Pennisetum glaucum, cv. Raj 171), mung bean (Phaseolus aureus, cv. RMG 492) and sesame (Sesamum indicum, cv. RT 46). To the best of our knowledge, this is the first report on phosphate solubilization by Aeromonas vaga. Seed inoculation of mung bean with or without tricalcium phosphate (TCP) was performed to study the effect of single and dual bacterial inoculations in pot trials having sterilized sandy loam soil, and was found to enhance the growth and yield of plants. The results were on a par with chemical fertilizer, single superphosphate (SSP) and commercial biofertilizers, PSB (Bacillus polymyxa) and MC (Pseudomonas striata), used as standard reference. Addition of TCP to soil gave better results and dual inoculation was more effective than single inoculation of bacteria. Among the four strains studied, A. vaga and P. fluorescens were found to be more valuable as single inoculants in terms of plant growth, whereas in combination treatments, P. fluorescens along with B. cepacia and A. vaga performed very well both in the presence and absence of TCP.     

Characterization of functional traits of two fluorescent pseudomonads isolated from basidiomes of ectomycorrhizal fungi

Some functional traits of Pseudomonas fluorescens 92 and BBc6, two strains isolated, respectively, from the basidiome of the ectomycorrhizal fungi Suillus grevillei and Laccaria laccata, were evaluated. A rifampicin-resistant mutant of P. fluorescens 92 (P. fluorescens 92R1) showed a significant in vivo plant growth promotion effect on cucumber plants. Quantitative analysis of enzymatic and physiological activities on different substrates showed that P. fluorescens 92 produced about a three times higher level of avicelase than BBc6, while equivalent amounts of β-glucosidase were produced by both strains. Satisfactory levels of neutral phosphomonoesterase and medium levels of acid phosphomonoesterase were produced by both. Only P. fluorescens BBc6 produced a very low amount of phosphodiesterase. Both strains produced high amounts of IAA and siderophores. Both strains showed on an iron deficient medium a very high antagonistic activity against the phytopathogenic fungus Heterobasidion annosum. Purification of fluorescent siderophores by copper-chelate chromatography showed that P. fluorescens 92 produced one pyoverdin (Pf92) and BBc6 two pyoverdins (PfBI and PfBII). A good inhibitory activity against mycelial growth of H. annosum was also observed when using the pyoverdines purified by affinity chromatography. Further purification by reverse phase high pressure liquid chromatography produced multiple fractionation of the three pyoverdins. Analysis of the reverse-phase purified pyoverdines by electronspray ionization mass spectrometry gave for pyoverdin Pf92 the mass value of 1213.8 and for both PfBI and PfBII the mass value of 1305.7. The presence of iron-chelating forms and sodium adducts were also evidenced.     

Influence of growth-promoting bacteria on the growth of wheat in different soils and temperatures

Plant-growth-promoting bacteria isolated from the rhizosphere, phyllosphere and soil of the root zone in different climatic regions of Germany and Uzbekistan were analysed for plant-growth-promoting effects and nutrient uptake on winter wheat on different soils and under different temperature regimes. The investigations were carried out in pot experiments using loamy sand and sandy loam soils from Müncheberg, Germany and Calcisol soil from Tashkent, Uzbekistan. The temperature and soil types were found to influence growth-promoting effects. Inoculation with bacterial strains Pseudomonas fluorescens PsIA12, Pantoea agglomerans 050309 and Mycobacterium sp. 44 isolated from Müncheberg (semi-continental climate) was found to significantly increase the root and shoot growth of winter wheat at 16 °C compared to 26 °C in loamy sand. Mycobacterium phlei MbP18 and Mycoplana bullata MpB46 isolated from Tashkent (semi-arid climate) were found to significantly increase the root and shoot growth of winter wheat in nutrient-poor Calcisol at 38 °C as well as in nutrient-rich loamy sand at 16 °C. Bacterial inoculation also resulted in significantly higher N, P, and K contents of plant components. The bacteria isolates were able to survive in the rhizosphere and in the soil of winter wheat after root and shoot inoculation.     

The invasive plant Solidago canadensis L. suppresses local soil pathogens through allelopathy

Recent studies suggest that invasive plants pose a significant effect on local soil pathogens, which in turn affects on the plant invasion. However, the mechanisms by which invasive plants affect soil pathogens were less well known. We conducted four experiments to test the hypothesis that the invasive plant species Solidago canadensis L. may affect soilborne pathogens through exudation of allelochemicals. Two common soilborne pathogens Pythium ultimum and Rhizoctonia solani were used in the study. Tomato (Lycopersicon esculentum Mill) variety Qianhong No.1 which is sensitive to soil pathogens P. ultimum and R. solani was used to indicate pathogenic activity (in terms of seedling mortality and damping-off). Extracts from root and rhizome of S. canadensis significantly suppressed the growth and pathogenic activity of both pathogens under Petri dish culture and sand culture (experiments 1 and 2), providing direct evidence that S. canadensis exerts allelopathic effects on these pathogens. Subsequently, a pathogen inoculation experiment under sand culture showed that pathogenic activity of both P. ultimum and R. solani was lower under the soil with S. canadensis compared to that under the soil with a common native plant Kummerowia striata (Thunb.) Schindl (experiment 3), implying that invasive S. canadensis had but native K. striata did not have allelopathic effects on soil pathogens through root and rhizome exudation. Finally, results from field soil tests showed that mortality and damping-off rate of tomato seedlings were significantly lower under the soils collected from the fields dominated by S. canadensis than that dominated by native plants at both sampling sites, suggesting that suppression of pathogens also occurs in the field. From the present experimental results we suggest that invasive S. canadensis may acquire spreading advantage in non-native habitat by using “novel weapons” to inhibit not only local plants but also soilborne pathogens.     

Influence of grass species and soil type on rhizosphere microbial community structure in grassland soils

A number of studies have reported species specific selection of microbial communities in the rhizosphere by plants. It is hypothesised that plants influence microbial community structure in the rhizosphere through rhizodeposition. We examined to what extent the structure of bacterial and fungal communities in the rhizosphere of grasses is determined by the plant species and different soil types. Three grass species were planted in soil from one site, to identify plant-specific influences on rhizosphere microbial communities. To quantify the soil-specific effects on rhizosphere microbial community structure, we planted one grass species (Lolium perenne L.) into soils from three contrasting sites. Rhizosphere, non-rhizosphere (bulk) and control (non-planted) soil samples were collected at regular intervals, to examine the temporal changes in soil microbial communities. Rhizosphere soil samples were collected from both root bases and root tips, to investigate root associated spatial influences. Both fungal and bacterial communities were analysed by terminal restriction fragment length polymorphism (TRFLP). Both bacterial and fungal communities were influenced by the plant growth but there was no evidence for plant species selection of the soil microbial communities in the rhizosphere of the different grass species. For both fungal and bacterial communities, the major determinant of community structure in rhizospheres was soil type. This observation was confirmed by cloning and sequencing analysis of bacterial communities. In control soils, bacterial composition was dominated by Firmicutes and Actinobacteria but in the rhizosphere samples, the majority of bacteria belonged to Proteobacteria and Acidobacteria. Bacterial community compositions of rhizosphere soils from different plants were similar, indicating only a weak influence of plant species on rhizosphere microbial community structure.     

Evaluation of pinewood biochar as a carrier of bacterial strain Enterobacter cloacae UW5 for soil inoculation

The large-scale production of biochar for carbon sequestration provides an opportunity for using these materials as inoculum carriers to deliver plant growth-promoting rhizobacteria (PGPR) into agricultural soils. Here, we evaluated the suitability of a biochar produced from pinewood pyrolyzed at 300 °C as a carrier for a well-studied PGPR strain, Enterobacter cloacae UW5. This strain was genetically modified to produce a green fluorescent protein marker that enabled tracking of the inoculum. Results from selective plate count assays and quantitative PCR (qPCR) confirmed that cell survival was slightly improved by addition of bacteria to soil using biochar as a carrier for the inoculant, as compared to soil directly inoculated. Total 16S rRNA genes were quantified using qPCR and DNA templates from the same soil treatments to distinguish the impact of biochar on total bacterial abundance from its influence on inoculum survival. Here total bacterial abundance was not influenced by biochar. All treatments resulted in bacterial colonization of roots at population densities of approximately 10⁵ CFU g⁻¹ root mass. Cucumber plants grown in the biochar amended soils had significantly greater biomass and root development than those planted in un-amended soil, regardless of the presence of inoculum. The ability of bacteria to colonize the plant roots and produce a plant growth hormone was not affected by biochar. However, UW5 inoculum did not promote root development in cucumber in any of the soils tested here. Overall, these experiments suggest that the 300 °C pine biochar is effective for evenly distributing inoculum into soil and promotes cucumber development in sandy loams.     

Competitive ability of selected Cyclopia Vent. rhizobia under glasshouse and field conditions

This study tested the competitive ability of three locally isolated Cyclopia rhizobia and strain PPRICI3, the strain currently recommended for the cultivation of Cyclopia, a tea-producing legume. Under sterile glasshouse conditions, the three locally isolated strains were equally competitive with strain PPRICI3. In field soils, the inoculant strains were largely outcompeted by native rhizobia present in the soil, although nodule occupancy was higher in nodules growing close to the root crown (the original inoculation area). In glasshouse experiments using field soil, the test strains again performed poorly, gaining less than 6% nodule occupancy in the one soil type. The presence of Cyclopia-compatible rhizobia in field soils, together with the poor competitive ability of inoculant strains, resulted in inoculation having no effect on Cyclopia yield, nodule number or nodule mass. The native rhizobial population did not only effectively nodulate uninoculated control plants, they also out-competed introduced strains for nodule occupancy in inoculated plants. Nonetheless, the Cyclopia produced high crop yields, possibly due to an adequate supply of soil N.