<Emphasis Type="Italic">Rhizobium</Emphasis>,<Emphasis Type="Italic"> Bradyrhizobium</Emphasis> and<Emphasis Type="Italic"> Agrobacterium</Emphasis> strains isolated from cultivated legumes

The present study was conducted to isolate and characterize rhizobial strains from root nodules of cultivated legumes, i.e. chickpea, mungbean, pea and siratro. Preliminary characterization of these isolates was done on the basis of plant infectivity test, acetylene reduction assay, C-source utilization, phosphate solubilization, phytohormones and polysaccharide production. The plant infectivity test and acetylene reduction assay showed effective root nodule formation by all the isolates on their respective hosts, except for chickpea isolate Ca-18 that failed to infect its original host. All strains showed homology to a typical Rhizobium strain on the basis of growth pattern, C-source utilization and polysaccharide production. The strain Ca-18 was characterized by its phosphate solubilization and indole acetic acid (IAA) production. The genetic relationship of the six rhizobial strains was carried out by random amplified polymorphic DNA (RAPD) including a reference strain of Bradyrhizobium japonicum TAL-102. Analysis conducted with 60 primers discriminated between the strains of Rhizobium and Bradyrhizobium in two different clusters. One of the primers, OPB-5, yielded a unique RAPD pattern for the six strains and well discriminated the non-nodulating chickpea isolate Ca-18 from all the other nodulating rhizobial strains. Isolate Ca-18 showed the least homology of 15% and 18% with Rhizobium and Bradyrhizobium, respectively, and was probably not a (Brady)rhizobium strain. Partial 16S rRNA gene sequence analysis for MN-S, TAL-102 and Ca-18 strains showed 97% homology between MN-S and TAL-102 strains, supporting the view that they were strains of B. japonicum species. The non-infective isolate Ca-18 was 67% different from the other two strains and probably was an Agrobacterium strain.     

Salt tolerance in <Emphasis Type="Italic">Zea mays</Emphasis> (L). following inoculation with <Emphasis Type="Italic">Rhizobium</Emphasis> and <Emphasis Type="Italic">Pseudomonas</Emphasis>

This study aimed to investigate the effect of inoculation with plant growth-promoting Rhizobium and Pseudomonas species on NaCl-affected maize. Two cultivars of maize (cv. Agaiti 2002 and cv. Av 4001) selected on the basis of their yield potential were grown in pots outdoors under natural conditions during July. Microorganisms were applied at seedling stage and salt stress was induced 21 days after sowing and maintained up to 50% flowering after 120 days of stress. The salt treatment caused a detrimental effect on growth and development of plants. Co-inoculation resulted in some positive adaptative responses of maize plants under salinity. The salt tolerance from inoculation was generally mediated by decreases in electrolyte leakage and in osmotic potential, an increase in osmoregulant (proline) production, maintenance of relative water content of leaves, and selective uptake of K ions. Generally, the microbial strain acted synergistically. However, under unstressed conditions, Rhizobium was more effective than Pseudomonas but under salt stress the favorable effect was observed even if some exceptions were also observed. The maize cv. Agaiti 2002 appeared to be more responsive to inoculation and was relatively less tolerant to salt compared to that of cv. Av 4001.     

Impact of a soil feeding termite,<Emphasis Type="Italic"> Cubitermes niokoloensis</Emphasis>, on the symbiotic microflora associated with a fallow leguminous plant<Emphasis Type="Italic"> Crotalaria ochroleuca</Emphasis>

The influence of a soil-feeding termite nest (Cubitermes nikoloensis) on the development of a symbiotic microflora (rhizobia, arbuscular mycorrhizas) was tested in a pot experiment with a tropical legume (Crotalaria ochroleuca). Our results confirmed the role of soil-feeding termite nests as sites of high nutrient concentration, as a significantly higher content of available P and mineral-N was found in the mound wall. Arbuscular mycorrhizal spores increased in the soil near the termite mound. The mound soil itself almost totally depressed mycorrhizal establishment. The positive effect of the soils close to the mound was also evidenced by the number of nodules per root system as well as the nodule biomass per legume plant grown on this medium. Better growth of Crotalaria seedlings was observed in the soils from the mound wall; the shoot biomass increased by a factor of 9 and the root biomass by a factor of 6 as compared to the control soil (10 m away from the mound). Plant growth on soils from the immediate vicinity of the mound showed intermediate results but a higher N content per biomass unit. This probably reflected the association with arbuscular mycorrhiza and rhizobia. This work evidenced the linkage of plant nutrition to nutrient availability in mound material and the indirect mediating effect of the symbiotic microflora.     

Comparison of nitrogen mineralisation patterns from root residues of<Emphasis Type="Italic"> Trifolium subterraneum</Emphasis> and<Emphasis Type="Italic"> Medicago sativa</Emphasis>

Crops grown in the first or second year after Medicago sativa L. (lucerne or alfalfa) in southern Australia have sometimes yielded less than crops grown after Trifolium subterraneum L. (subterranean clover), despite the greater annual accumulation of legume nitrogen (N) and enhanced N₂ fixation in lucerne-based pastures. To understand why, we studied the N mineralisation patterns of root residues collected from 16- or 20-week-old plants using incubation assays, in two separate experiments with contrasting soil types (a red loam and a grey clay loam). Fine roots of both species added to soil produced more mineral N than the control soil with no root residues added. In contrast, less mineral N accumulated in the presence of coarse roots than in the control soil. These patterns were not explained by differences in physical size or surface-area, but appeared to be more related to the carbon to nitrogen ratio (C:N) of fine and coarse roots. Fine roots of both species had a C:N of about 11, while the C:N of coarse roots ranged from 28 to 37. Subterranean clover had a high proportion of fine roots giving a weighted average C:N of 19 for the whole root system, and could be expected to result in a rapid net mineralisation of N. The root systems of lucerne on the other hand, consisted mainly of coarse roots giving an average C:N of 26, and would be likely to induce a transient net immobilisation of N and a delay in net mineralisation. The same general patterns of N mineralisation/immobilisation were observed in the second experiment. Tissue chemical analyses suggested that even where the total C:N of subterranean clover and lucerne residues were similar, the amounts of C and N in the soluble fraction of the residues and the C:N of that soluble fraction could largely account for the observed differences in N mineralisation/immobilisation.     

Interactions between arbuscular mycorrhizal fungi and phosphate-solubilizing fungus (<Emphasis Type="Italic">Mortierella</Emphasis> sp.) and their effects on <Emphasis Type="Italic">Kostelelzkya virginica</Emphasis> growth and enzyme activities of rhizosphere and bulk soils at different salinities

This study investigated the interactions between two arbuscular mycorrhizal fungi (AMF) (Glomus aggregatum and Glomus mosseae) and a P-solubilizing fungus (Mortierella sp.), with respect to their effects on growth of Kostelelzkya virginica and urease, invertase, neutral phosphatase, alkaline phosphatase, and catalase activities of rhizosphere and bulk soils at different salinity levels (i.e., 0, 100, 200, and 300 mM NaCl). Percentage of AMF colonization, Mortierella sp. populations, pH, electrical conductivity, and available P concentration in soil were also determined. Combined inoculation of AMF and Mortierella sp. increased the percentage of AMF colonization and Mortierella sp. populations under salt stress (i.e., 100, 200, and 300 mM NaCl). The dual inoculation of Mortierella sp. with AMF (G. aggregatum or G. mosseae) had significant effects on shoot and root dry weights and available P concentrations, pH values, and electrical conductivities of rhizosphere and bulk soils under salt stress. The inoculation of Mortierella sp. significantly enhanced the positive effects of AMF on some enzyme activities (i.e., neutral phosphatase, alkaline phosphatase, and catalase in bulk soil; neutral phosphatase and urease in rhizosphere soil); on the contrary, it produced negative effects on urease activities in bulk soil and invertase activities in bulk and rhizosphere soils. The results indicated that the most effective co-inoculation was the dual inoculation with Mortierella sp. and G. mosseae, which may help in alleviating the deleterious effects of salt on plants growth and soil enzyme activities.     

Effects of ectomycorrhizal fungus <Emphasis Type="Italic">Boletus edulis</Emphasis> and mycorrhiza helper <Emphasis Type="Italic">Bacillus cereus</Emphasis> on the growth and nutrient uptake by <Emphasis Type="Italic">Pinus thunbergii</Emphasis>

The present greenhouse study was undertaken to evaluate the effects of co-inoculating the ectomycorrhizal (ECM) fungus Boletus edulis with the mycorrhiza helper bacterium Bacillus cereus HB12 or HB59 on the growth and nutrient uptake of Pinus thunbergii. The inoculation with mycorrhiza helper bacterium significantly (P?≤?0.05) increased the ectomycorrhizal colonization. Treatments with dual inoculum (the mycorrhiza helper bacterium plus mycorrhiza) significantly (P?≤?0.05) increased the P. thunbergii growth. Bacteria–mycorrhizae interactions resulted in a great utilization of phosphate and potassium. The single inoculation resulted in a higher root activity than the control while the co-inoculation led to the highest root activity. The 6-CFDA staining assay showed that B. cereus enhanced fungal activity in ectomycorrhizal symbiosis. The results conclusively suggest that B. cereus isolated from the rhizosphere of P. thunbergii can potentially be used as individual inoculant or co-inoculated with ECM fungi to increase the production in sustainable ecological systems. These results support the potential use of B. cereus (HB12 or HB59) and B. edulis as mixed inoculants stimulating growth of P. thunbergii.     

Benefits of inoculation of the common bean (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis>) crop with efficient and competitive<Emphasis Type="Italic"> Rhizobium tropici</Emphasis> strains

Cropping in low fertility soils, especially those poor in N, contributes greatly to the low common bean (Phaseolus vulgaris L.) yield, and therefore the benefits of biological nitrogen fixation must be intensively explored to increase yields at a low cost. Six field experiments were performed in oxisols of Paraná State, southern Brazil, with a high population of indigenous common bean rhizobia, estimated at a minimum of 10³ cells g^–1 soil. Despite the high population, inoculation allowed an increase in rhizobial population and in nodule occupancy, and further increases were obtained with reinoculation in the following seasons. Thus, considering the treatments inoculated with the most effective strains (H 12, H 20, PRF 81 and CIAT 899), nodule occupancy increased from an average of 28% in the first experiment to 56% after four inoculation procedures. The establishment of the selected strains increased nodulation, N₂ fixation rates (evaluated by total N and N-ureide) and on average for the six experiments the strains H 12 and H 20 showed increases of 437 and 465 kg ha^–1, respectively,in relation to the indigenous rhizobial population. A synergistic effect between low levels of N fertilizer and inoculation with superior strains was also observed, resulting in yield increases in two other experiments. The soil rhizobial population decreased 1 year after the last cropping, but remained high in the plots that had been inoculated. DGGE analysis of soil extracts showed that the massive inoculation apparently did not affect the composition of the bacterial community.     

Effect of the arbuscular mycorrhizal fungus <Emphasis Type="BoldItalic">Glomus intraradices</Emphasis> on the false root-knot nematode <Emphasis Type="BoldItalic">Nacobbus aberrans</Emphasis> in tomato plants

Information about the interaction between arbuscular mycorrhizal fungi (AMF) and the false root-knot nematode Nacobbus aberrans (Thorne, 1935) Thorne & Allen, 1944 is scarce. The effect of Glomus intraradices Schenk & Smith on tomato (Lycopersicon esculentum L.) cv. Platense inoculated with nematode juveniles from Lisandro Olmos (Argentina) was studied under greenhouse conditions. Six treatments with five replications were performed. After 80 days, nematode reproduction and percentage of AMF colonization in roots were estimated. Some plant growth parameters were also measured. In general, plants with AMF and AMF plus nematodes grew as well as the control without AMF and without nematodes. Furthermore, G. intraradices was beneficial in reducing nematode-induced damage in roots (lower number of galls) as well as in having a suppressive effect on parasite reproduction. This is the first study on the use of G. intraradices as a possible strategy in the control of N. aberrans in tomato.     

Characterization of rhizobia from <Emphasis Type="Italic">Sesbania</Emphasis> species native to seasonally wetland areas in Uruguay

Naturally growing Sesbania species with tolerance to unfavourable habitats are widely distributed in non-cultivated seasonally wetland areas in Uruguay. We investigated the relative abundance, diversity and symbiotic efficiency of Sesbania punicea and S. virgata rhizobia in three ecologically different undisturbed and water-logged sites in Uruguay. Numbers of native-soil rhizobia infective on S. punicea or S. virgata were low, with higher numbers associated with the presence of S. virgata. Plants of S. virgata inoculated with soil suspension showed aerial and nodule biomass greater than that obtained with S. punicea. The rhizobia nodulating Sesbania species in water-logged lands in different regions of Uruguay were diverse differing in growth rates, acid production, growth at 39°C and in LB medium, host range and symbiotic efficiency. Seventeen representative strains clustered into four groups on the basis of phenotypic characteristics, ARDRA and DNA fingerprinting (GTG₅-PCR). Partial sequence of 16S rRNA from eight of these strains classified them into at least two genera with four species: Azorhizobium doebereinerae, Rhizobium sp. related to R. etli and two different Rhizobium sp.-Agrobacterium. Our results confirm the presence of the specie Azorhizobium doebereinerae as microsymbionts of S. virgata in South America. No strain of Rhizobium etli has previously been reported as a microsymbiont of Sesbania, though R. etli like organisms have also been recovered from Dalea purpurea and Desmanthus illinoensis. Significant increases in dry matter production were obtained with S. virgata plants inoculated with selected rhizobial strains under growth chamber conditions.     

Evaluation and utilization of <Emphasis Type="Italic">Aegilops</Emphasis> and wild <Emphasis Type="Italic">Triticum</Emphasis> species for enhancing iron and zinc content in wheat

Grains of 80 accessions of nine species of wild Triticum and Aegilops along with 15 semi-dwarf cultivars of bread and durum wheat grown over 2 years at Indian Institute of Technology, Roorkee, were analyzed for grain iron and zinc content. The bread and durum cultivars had very low content and little variability for both of these micronutrients. The related non-progenitor wild species with S, U and M genomes showed up to 3–4 folds higher iron and zinc content in their grains as compared to bread and durum wheat. For confirmation, two Ae. kotschyi Boiss. accessions were analyzed after ashing and were found to have more than 30% higher grain ash content than the wheat cultivars containing more than 75% higher iron and 60% higher zinc than that of wheat. There were highly significant differences for iron and zinc contents among various cultivars and wild relatives over both the years with very high broad sense heritability. There was a significantly high positive correlation between flag leaf iron and grain iron (r = 0.82) and flag leaf zinc and grain zinc (r = 0.92) content of the selected donors suggesting that the leaf analysis could be used for early selection for high iron and zinc content. ‘Chinese Spring’ (Ph ^I ) was used for inducing homoeologous chromosome pairing between Aegilops and wheat genomes and transferring these useful traits from the wild species to the elite wheat cultivars. A majority of the interspecific hybrids had higher leaf iron and zinc content than their wheat parents and equivalent or higher content than their Aegilops parents suggesting that the parental Aegilops donors possess a more efficient system for uptake and translocation of the micronutrients which could ultimately be utilized for wheat grain biofortification. Partially fertile to sterile BC₁ derivatives with variable chromosomes of Aegilops species had also higher leaf iron and zinc content confirming the possibility of transfer of required variability. Some of the fertile BC₁F₃ and BC₂F₂ derivatives had as high grain ash and grain ash iron and zinc content as that of the donor Aegilops parent. Further work on backcrossing, selfing, selection of fertile derivatives, leaf and grain analyses for iron and zinc for developing biofortified bread and durum wheat cultivars is in progress. Nidhi Rawat, Vijay K. Tiwari, and Neelam Singh have contributed equally to the work.     

Changes in soil properties and their effects on maize productivity following<Emphasis Type="Italic"> Sesbania sesban</Emphasis> and<Emphasis Type="Italic"> Cajanus cajan</Emphasis> improved fallow systems in eastern Zambia

Improved fallows with leguminous trees have been developed in Southern Africa as a viable alternative to inorganic fertilizers but the changes in soil properties that are responsible for crop productivity improvement and implications of mixing litter and fresh leaves from the same tree species on soil fertility are not fully understood. Our objectives were to quantify (1) some changes in soil properties that are responsible for crop production improvement under improved fallow systems; (2) the N mineralization patterns of mixtures of litter and fresh leaves from the same tree species. The treatments used in the study were 2-year planted Sesbania sesban (sesbania) and Cajanus cajan (cajanus) and controls of natural fallow, continuous fertilized and unfertilized maize. At fallow clearing sesbania contributed 56 kg N ha^–1 through litter and fresh leaves. Sesbania (fresh leaves + litter) showed high N mineralization after 10 weeks compared to the mixture of cajanus fresh leaves with litter. Maize yields were significantly correlated with preseason NO₃^–-N and total inorganic-N content of the top 20-cm soil layer. Soil penetrometer resistance at 4 weeks after planting was lowest in the sesbania land-use system (2.2 Mpa), whereas the highest percentage of water-stable aggregates at fallow clearing and crop harvest was in sesbania (83%) and cajanus (77%), respectively. The improved soil conditions and N contribution of sesbania and cajanus fallows to the subsequent maize crop was evidenced by increased maize yields of between 170–200% over maize without fertilizer.     

Nutrient composition and short-term release from <Emphasis Type="Italic">Croton macrostachyus</Emphasis> Del. and <Emphasis Type="Italic">Millettia ferruginea</Emphasis> (Hochst.) Baker leaves

A field study was conducted to investigate the nutrient content of green and abscised leaves of Croton macrostachyus Del. and Millettia ferruginea (Hochst.) Baker and their decomposition to return these nutrients to the soil in the short-term. Green and abscised leaves were collected from trees of comparable size in Wondo Genet, Ethiopia, and were incorporated into litterbags to decompose for a period of 12 weeks. Green leaves of C. macrostachyus had significantly higher nutrient concentrations than those of M. ferruginea. In both species, green leaves had significantly higher (P<0.05) C, N and P and significantly lower (P<0.05) lignin and polyphenol concentrations than abscised leaves. Fifty percent of the biomass applied was lost during the first 7 weeks in C. macrostachyus, which was about 3 times faster than that of M. ferruginea. The half-lives of N and P were 5–8 weeks and 4–6 weeks, respectively, in C. macrostachyus; the corresponding values for M. ferruginea were 6–8 and about 22 weeks, respectively. Only 15.7% and 26.8% of green and abscised leaves of C. macrostachyus, respectively, were recovered after the 12 weeks of decomposition. The corresponding values were about 3.5-fold and 2.5-fold higher for M. ferruginea. Generally, leaves with higher initial lignin, polyphenol, lignin:N and C:N ratios had lower decomposition and mineralization rates. In both species, removal of leaf biomass before abscission (e.g. by pruning) could enhance both the quantity and rate of nutrient return to the soil.     

Diversity of <Emphasis Type="Italic">Citrus</Emphasis><Emphasis Type="Italic">depressa</Emphasis> Hayata (Shiikuwasha) revealed by DNA analysis

Citrus depressa Hayata is an indigenous mandarin species on the Ryukyu Islands located in the subtropical region of Japan. We deduced its phylogenetic relationships by evaluating accessions grown on various Ryukyu Islands via cleaved amplified polymorphic sequence analysis of cpDNA and sequence-related amplified polymorphism (SRAP). The cpDNA results indicated that C. depressa could be classified into two types. SRAP revealed patterns of diversity within C. depressa consistent with our cpDNA results. These results indicate that maternal origin may influence or is correlated with the constitution of the nuclear genome of C. depressa. Another Japanese mandarin species, Citrus tachibana (Makino) Tanaka was distinguished from C. depressa by SRAP markers. Moreover, both C. depressa and C. tachibana could be distinguished from other Citrus species. Our results suggest that Japanese mandarin possesses a characteristic genome with the genus Citrus.     

A new mycorrhizal helper bacterium, <Emphasis Type="Italic">Ralstonia</Emphasis> species,in the ectomycorrhizal symbiosis between <Emphasis Type="Italic">Pinus thunbergii</Emphasis> and <Emphasis Type="Italic">Suillus granulatus</Emphasis>

In a Robinia-pseudoacacia-dominated coastal forest in Tottori prefecture Japan, the growth and survival of Pinus thunbergii seedlings and the natural regeneration of P. thunbergii was disturbed by R. pseudoacacia. In order to improve the growth of P. thunbergii seedling in the Tottori sand dune, we tried to find a mycorrhiza helper bacteria (MHB) from P. thunbergii mycorrhizosphere in a Tottori sand dune. Two MHB, Ralstonia sp. and Bacillus subtilis, were selected from the nine bacterial species isolated from the mycorrhizosphere of P. thunbergii. The bacterial effect on the ectomycorrhizal fungus Suillus granulatus was investigated by confrontation assay and a microcosm experiment. The confrontation assay showed that Ralstonia sp. promoted the hyphal growth of S. granulatus. Moreover, the S. granulatus–P. thunbergii symbiosis was significantly stimulated by Ralstonia sp. and B. subtilis. Ralstonia sp. and B. subtilis were regarded as MHB associated with P. thunbergii. This is the first report of Ralstonia sp. as an MHB.     

A significant proportion of indigenous rhizobia from India associated with soybean (<Emphasis Type="Italic">Glycine max</Emphasis> L.) distinctly belong to <Emphasis Type="Italic">Bradyrhizobium</Emphasis> and <Emphasis Type="Italic">Ensifer</Emphasis> genera

The diversity among 269 rhizobia isolated from naturally occurring root nodules of soybean collected from two different agro-ecological regions of India, based on RFLP and sequences of the intergenic spacer (IGS) between the 16S and 23S rRNA genes, growth rate, and indole acetic acid production, revealed their significant, site-dependent genomic diversity. Among these bacteria, nine IGS genotypes were identified with two endonucleases. They were distributed into five divergent lineages by sequence analysis of each IGS representative strain, i.e., (1) comprising IGS genotypes I, II, III, and reference Bradyrhizobium yuanmingense; (2) with genotype IV and strains of unclassified bradyrhizobia genomic species; (3) including genotypes V, VI, and Bradyrhizobium liaoningense; (4) with IGS genotype VII and Bradyrhizobium elkanii strains; and (5) comprising IGS genotypes VIII, IX, and different Ensifer genus bacteria. Host-specificity test revealed that all rhizobia-nodulated soybean and cowpea and only part of them formed nodules on Arachis hypogeae and Cajanus cajan. The great diversity of soybean nodulators observed in this study emphasises that Indian soil is an important reservoir of nitrogen-fixing rhizobia.     

<Emphasis Type="Italic">Trichoderma</Emphasis> improves the growth of <Emphasis Type="Italic">Leymus chinensis</Emphasis>

Bio-fertilizer application has been proposed as a strategy for enhancing soil fertility, regulating soil microflora composition, and improving crop yields, and it has been widely applied in the agricultural yields. However, the application of bio-fertilizer in grassland has been poorly studied. We conducted in situ and pot experiments to investigate the practical effects of different fertilization regimes on Leymus chinensis growth, with a focus on the potential microecological mechanisms underlying the responses of soil microbial composition. L. chinensis biomass was significantly (P?<?0.05) increased by treatment with 6000 kg ha^?1 of Trichoderma bio-fertilizer compared with other treatments. We found a positive (R² =?0.6274, P <?0.001) correlation between bacterial alpha diversity and L. chinensis biomass. Hierarchical cluster analysis and nonmetric multidimensional scaling (NMDS) revealed that soil bacterial and fungal community compositions were all separated according to the fertilization regime used. The relative abundance of the most beneficial genera in bio-fertilizer (BOF) (6000 kg ha^?1Trichoderma bio-fertilizer) was significantly higher than in organic fertilizer (OF) (6000 kg ha^?1 organic fertilizer) or in CK (non-amend fertilizer), there the potential pathogenic genera were reduced. There were significant negative (P?<?0.05) correlations between L. chinensis biomass and the relative abundance of several potential pathogenic genera. However, the relative abundance of most beneficial genera were significantly (P?<?0.05) positively correlated with L. chinensis biomass. Soil properties had different effects on these beneficial and on these pathogenic genera, further influencing L. chinensis biomass.     

Karyotypic variation in <Emphasis Type="Italic">Nicotiana</Emphasis> section Suaveolentes

Nicotiana section Suaveolentes (Solanaceae) currently includes 28 species and subspecies that are endemic to Australasia and the South Pacific and one African species, N. africana. The section is monophyletic and of allotetraploid origin, but relationships among the species in it and its diploid progenitors are poorly understood. Here we report chromosome numbers for 20 of the 29 taxa from the Suaveolentes, including a count for one recently proposed species for which no number has previously been available. Many of the published chromosome numbers for the Suaveolentes are confirmed in this study. However, six counts were different from the published numbers including n = 15 for N. maritima and N. suaveolens, which is a new chromosome number for the genus. Nicotiana goodspeedii and N. rotundifolia were n = 16, and the same number was found in the suggested species N. sp. ‘Corunna’. Nicotiana suaveolens contains polyploid races of n = 32 and here we report the probable existence of an n = 31 race as well. Karyotypic variation within species and within the section is apparently much greater than previously thought and further investigation is warranted.     

Morphological characteristics of some accessions of grass pea (<Emphasis Type="Italic">Lathyrus sativus</Emphasis> L.) grown in Europe and nutritional traits of their seeds

Thirty-one European accessions of grass pea (Lathyrus sativus L.) originating from Italy, Spain, Northern France, Germany and Poland grown in the same soil-climatic conditions in Poland were investigated. The morphological traits as well as the seed morphology and nutrient contents were studied. Substantial differentiation in some morphological characteristics was noticed. The plants could be assigned to two groups: one group comprising the lines from the Mediterranean basin (Italy and Spain) and the other group containing the accessions from West-central Europe (Northern France, Germany and Poland). The grass pea plants produced from the seeds obtained from West-central Europe appeared to be about 10 cm taller than the Mediterranean accessions and had predominantly white flowers. Their seeds were more than two times smaller than seeds of the accessions of Italian or Spanish origin. There were not noted any considerable differences in nutrient contents or antinutritional factors in seeds between these two groups of Lathyrus accessions. A negative correlation between β-ODAP and tannin contents (r = −0.379, P ≤ 0.05) and a positive correlation (r = 0.843, P ≤ 0.01) between a tannin content and flower color intensity were found. A white or crème colour was associated with a low tannin level in the seeds, while the colourful blooming accessions had higher tannin contents.     

Dominant Characteristics Between <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis> and <Emphasis Type="Italic">Cyclotella</Emphasis> Sp. Accompanying Dilution Process in Eutrophic Lake

Although dilution of lake water has been used for improvement of water quality and algal blooms control, it has not necessarily succeeded to suppress the blooms. We hypothesized that the disappearance of algal blooms by dilution could be explained by flow regime, nutrient concentrations, and their interaction. This study investigated the effects of daily renewal rate (d), nitrogen (N) and phosphorus (P) concentration, and their interaction on the domination between Microcystis aeruginosa and Cyclotella sp. through a monoxenic culture experiment. The simulation model as functions of the N:P mass ratio and dilution rate (D) (calculated from d) was constructed, and the dominant characteristics of both species were predicted based on the model using parameters obtained in a monoculture experiment and our previous study. Results of monoxenic culture experiment revealed that M. aeruginosa dominated in all conditions (d = 5 or 15%; N = 1.0 or 2.5 or 5.0 mg-N L^?1; P = 0.1 or 0.5 mg-P L^?1) and the predicted cell densities were substantially correspondent to experimental data. Under various N:P ratios and D values, characteristics of domination for each species were predicted, indicating that Cyclotella sp. tended to be dominant under high P concentrations (P ≥ 0.36 mg-P L^?1) when the N:P ratio was less than 7.0, and M. aeruginosa could not form algal blooms at the N:P ratio ≤ 7.0 (N ≤ 0.7 mg-N L^?1). It was also suggested that the dilution rate leading to the Cyclotella sp. domination required 0.20 day^?1 or higher regardless of the N:P ratios.

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Graphical Abstract ? M. aeruginosa and Cyclotella sp. could be a superior competitor in nutrient-limited and nutrient-rich conditions, respectively. ? The simulation model in this study indicated that the predicted cell density and nutrient concentration were substantially correspondent to experimental data. ? The model predicted that Cyclotella sp. tended to be dominant at the P ≥ 0.36 mg-P L^?1 when the N:P ratio was less than 7.0, and M. aeruginosa could not form algal blooms at the N:P ratio ≤ 7.0 (N ≤ 0.7 mg-N L^?1).

     

<Emphasis Type="Italic">Achromobacter insolitus</Emphasis> and <Emphasis Type="Italic">Zoogloea ramigera</Emphasis> associated with wheat plants (<Emphasis Type="Italic">Triticum aestivum</Emphasis>)

This study reports for the first time the presence of diazotrophic bacteria belonging to the genera Achromobacter and Zoogloea associated with wheat plants. These bacterial strains were identified by the analysis of 16S rDNA sequences. The bacterium IAC-AT-8 was identified as Azospirillum brasiliense, whereas isolates IAC-HT-11 and IAC-HT-12 were identified as Achromobacter insolitus and Zoogloea ramigera, respectively. A greenhouse experiment involving a non-sterilized soil was carried out with the aim to study the endophytic feature of these strains. After 40 days from inoculation, all the strains were in the inner of roots, but they were not detected in soil. In order to assess the location inside wheat plants, an experiment was conducted under axenic conditions. Fifteen days after inoculation, preparations of inoculated plants were observed by the scanning electron microscope, using the cryofracture technique, and by the transmission electron microscope. It was observed that all isolates were present on the external part of the roots and in the inner part at the elongation region, in cortex cells, but not in the endodermis or in the vascular bundle region. No colonizing bacterial cells were observed in wheat leaves.