Genetic diversity of rhizobia associated with alfalfa in Serbian soils

We have evaluated the genetic diversity and phylogeny of alfalfa rhizobia, originating from different types of soils in Serbia and their ability to establish an effective symbiosis with alfalfa (Medicago sativa L.). A collection of 65 strains isolated from root nodules of alfalfa were characterized by rep-PCR analysis, partial and complete 16S rDNA gene and recA gene sequencing, as well as atpD gene sequencing and DNA–DNA hybridizations. The results of the sequence analyses revealed that Sinorhizobium meliloti is the dominant species in alfalfa nodules. Only one strain was identified as Sinorhizobium medicae, two strains as Rhizobium tibeticum and one strain as Rhizobium sp. Despite the fact that the majority of strains were identified as S. meliloti, a high genetic diversity at strain level was detected. Almost all isolates shared the ability to nodulate and fix nitrogen with M. sativa, except 11 of them, which were incapable of fixing nitrogen with this species. About 50% of the isolates showed values of symbiotic effectiveness (SE) above 50%, while 10% of the strains were highly effective with SE values above 70%. Some of the strains which were highly effective in nitrogen fixation at the same time could intensively solubilize phosphates, offering a possibility for multipurpose inoculum development. This was the first genetic study of rhizobia isolated from this region and also the first report of natural presence of R. tibeticum in root nodules of M. sativa.     

Effects of long-term compost and fertilizer application on stability of aggregate-associated organic carbon in an intensively cultivated sandy loam soil

The study examined the influence of compost and mineral fertilizer application on the content and stability of soil organic carbon (SOC). Soil samples collected from a long-term field experiment were separated into macroaggregate, microaggregate, and silt + clay fractions by wet-sieving. The experiment involved seven treatments: compost, half-compost N plus half-fertilizer N, fertilizer NPK, fertilizer NP, fertilizer NK, fertilizer PK, and control. The 18-year application of compost increased SOC by 70.7–121.7%, and mineral fertilizer increased by 5.4–25.5%, with no significant difference between control soil and initial soil. The C mineralization rate (rate per unit dry mass) in microaggregates was 1.52–2.87 mg C kg⁻¹ day⁻¹, significantly lower than in macroaggregate and silt + clay fractions (P < 0.05). Specific C mineralization rate (rate per unit SOC) in silt + clay fraction amounted to 0.48–0.87 mg C g⁻¹ SOC day⁻¹ and was higher than in macroaggregates and microaggregates. Our data indicate that SOC in microaggregates is more stable than in macroaggregate and silt + clay fractions. Compost and mineral fertilizer application increased C mineralization rate in all aggregates compared with control. However, compost application significantly decreased specific C mineralization rate in microaggregate and silt + clay fractions by 2.6–28.2% and 21.9–25.0%, respectively (P < 0.05). By contrast, fertilizer NPK application did not affect specific C mineralization rate in microaggregates but significantly increased that in silt + clay fractions. Carbon sequestration in compost-amended soil was therefore due to improving SOC stability in microaggregate and silt + clay fractions. In contrast, fertilizer NPK application enhanced SOC with low stability in macroaggregate and silt + clay fractions.     

Are Primary Cultures of Trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) Ventricular Cardiomyocytes Metabolically Viable?

The objectives of this study were to establish a protocol for the isolation of metabolically viable ventricular cardiomyocytes from the rainbow trout and to determine which measures may best reflect viability 24 h after isolation. Cardiomyocytes were isolated by enzymatic digestion and maintained in cell suspension. Viability was assessed using Trypan blue dye exclusion, ATP content, oxygen consumption and lactate dehydrogenase (LDH) leakage into the medium. Viability, assessed by these measures did not significantly change over the time period of this study. ATP content did correlate significantly with oxygen consumption but not with Trypan blue exclusion. We conclude that primary cultured cardiomyocytes remain metabolically viable for at least 24 h after isolation. Also, it appears that ATP content and oxygen consumption most adequately reflect metabolic cell viability. To be confident with a culture, however, a combination of viability measures is necessary when isolating cardiomyocytes from fish.     

Solubility of kraft lignin-<Emphasis Type="Italic">g</Emphasis>-polyacrylonitrile copolymer in various ionic liquids and characterization of its solution

Lignin is a potential precursor for low-cost carbon fiber production, but it is difficult to spin and spool lignin because of its complex and interconnected molecular structure. This disadvantage can be overcome by introducing g-polyacrylonitrile (PAN) to lignin. However, the resulting copolymer is insoluble in common organic solvents. In this study, kraft lignin (KL)-g-polyacrylonitrile copolymers with different KL/PAN proportions were prepared via atom transfer radical polymerization (ATRP) method and their solubility in ionic liquids (ILs) was investigated at different temperatures. 1-Ethyl-3-methylimidazolium acetate ([EMIM]Ac), 1,3-dimethylimidazolium methyl sulfate ([MMIM]MeSO4), 1-butyl-3-methylimidazolium chloride ([BMIM]Cl), and 1-butyl-3-methylimidazolium bromide ([BMIM]Br) were used as the ILs. At all investigated temperatures, the highest solubility of KL-g-PAN was observed in [EMIM]Ac, with the order of [MMIM]MeSO4 > BMIM]Br > [BMIM]Cl. The solubility in BMIM]Br and [BMIM]Cl was remarkably low, reaching values of less than 4 g/Kg. The viscosity and surface tension of the KL-g-PAN/[EMIM]Ac solution increased and decreased, respectively, with increasing amounts of PAN and further by the addition of dimethylformamide (DMF) to the solution. FTIR spectra of KL-g-PAN copolymers before and after dissolution suggested that PAN was partially depolymerized from the copolymer during the dissolution process.     

Factors influencing stump sprouting by pondcypress (Taxodium distichum var. nutans (Ait.) Sweet)

Sprouting can be an important mechanism of regeneration in harvested cypress (Taxodium sp.) wetlands; however, limited information exists on factors influencing sprouting. We examined the influence of harvesting practices, tree size, and site conditions on stump sprouting in eight pondcypress (Taxodium distichum var. nutans (Ait.) Sweet) wetlands in Florida, USA. Stumps were cut at three heights: 20 cm below high water level, at high water, and 40 cm above high water. Half of each site was harvested using a sawhead feller and the other half using shearhead. We monitored sprouting for 2 years post-harvest. Stump height affected first year sprouting, but the effect disappeared during year 2. Felling equipment did not affect sprouting; however, the shearhead caused greater stump damage. Sprouting declined with increasing parent tree size; percentage of stumps with sprouts after 2 years was 68% for trees 10–14 cm dbh and 28% for trees 20–24 cm dbh. Stump sprouting ranged among sites from 23 to 54% after 2 years. Results indicate that sprouting may be an important, but inadequate, form of regeneration. At sites with short hydroperiods, dense understories, and/or predominance of trees greater than 20 cm dbh, reduced sprouting may occur, and planting seedlings may be beneficial.     

Manufacture and mechanical properties of binderless boards from kenaf core

Binderless boards were prepared from finely ground powders of kenaf (Hibiscus cannabinus L.) core under varying manufacturing conditions. This research was designed to investigate their mechanical properties and evaluate the various manufacturing conditions: pressing temperature and time, pressing pressure, board density, board thickness, grain size of raw materials, and addition of furfural. The mechanical properties (i.e., modulus of rupture and elasticity, internal bonding strength) of boards increased with increasing board density and met the requirement for 15 type medium-density fiberboard (MDF) by JIS A 5905-1994. Thickness swelling and water absorption of boards exceeded the maximum permitted levels for 15 type MDF and S20 grade hardboard by JIS A 5905-1994, which indicates the low water-resistant property of binderless boards. In contrast to that in usual wood-based materials, internal bonding strength showed significant correlations with other board properties: modulus of rupture and elasticity, thickness swelling, and water absorption. We confirmed experimentally that the best manufacturing conditions proved to be as follows: pressing temperature 180°C, time 10min; pressing pressure 5.3MPa; board thickness 5mm; board density 1.0g/cm³; average grain size 53µm; and powder with no furfural content.Part of this paper was presented at the 52nd Annual Meeting of the Japan Wood Research Society, Gifu, April 2002     

Application of a modified rate equation to the acetylation of wood

A modified rate equation, in which the diffusion of reagent was taken into consideration, was proposed and applied to results reported previously for the acetylation of wood. The equation was fitted more successfully to the prolonged as well as the early stage of the reaction than a common first-order rate equation. Although the conclusions were virtually unchanged, the reliability was enhanced because reaction parameters such as the rate constant, ultimate extent of the reaction, and activation energy calculated for the modified equation were based on much more data than those reported previously. The equation proposed here may be applicable not only to acetylation but to other chemical modifications of cellulosic materials in heterogeneous systems.     

Dynamic excitation and static loading tests of glulam lattice floor

A wooden lattice floor with high stiffness and damping capacity has been developed to solve noise problems in wooden apartment houses. The lattice floor consisted of Douglas fir glulam beams with inserted steel plate joints and drift pins. To examine the structural performance of the floor, dynamic excitation and static loading tests were conducted on the full size floor. The first and second order resonance frequencies of the floor were 13.5Hz and 27.0Hz, respectively. These frequencies are similar to the peak frequency of a conventional wooden floor and the combined floor fabricated from glued laminated timber and iron. The maximum static load of the floor was 127kN. The apparent flexural rigidity was less than half the value of several floors studied in the past. However, it is considered that the stiffness is improved by constructing panels and this floor has almost equivalent performance. Relative deflection was not affected by the loading history.Part of this study was presented at the International Wood Engineering Conference, New Orleans, October 1996.     

Leaf-litter decomposition of 15 tree species in a lowland tropical rain forest in Sarawak: dynamics of carbon,nutrients, and organic constituents

The effects of tree species on the dynamics of nutrient transformations during leaf-litter decomposition are not well understood in tropical rain forests. To examine differences in the dynamics of C, nutrients, and organic constituents during decomposition among tree species, the leaf-litter decomposition of 15 trees was assessed using a litter-bag method in a lowland tropical rain forest in Sarawak. The dynamics of C was parallel to that of weight loss. The dynamics of nutrients were grouped into three patterns. The dynamics of K was characterized by a high leaching loss in the initial stages, and that of Ca and Mg generally showed a gradual decrease over the course of decomposition. The dynamics of N and P showed highly different patterns with the weight loss, and was characterized by relatively higher remaining mass at the end of the experiment. The variations or exceptions of nutrient dynamics among tree species were considered to be related to the initial concentration of each nutrient. For the dynamics of organic constituents, water-soluble carbohydrates disappeared quickly at the initial stages, and acid-soluble carbohydrates were the second fastest decomposing fraction; the decomposition of acid-insoluble residue (AIS) was the slowest. The release of limiting nutrients (N and P) generally followed the disappearance of AIS, but was independent of the disappearance of AIS when the initial concentrations of these nutrients were very low.     

Powdery mildew resistance gene (<Emphasis Type="Italic">Pm</Emphasis>-<Emphasis Type="Italic">AN</Emphasis>) located in a segregation distortion region of melon LGV

Powdery mildew is one of the most important melon pathogens all over the world. So far, many genes conferring resistance to powdery mildew of melon have been described, but few of these have been finely mapped or cloned. Two F₂ populations derived from Ano2 × Hami413 and Ano2 × Queen were used to map the powdery mildew resistance gene by methods of Bulked Segregation Analysis (BSA), comparative genomics and Resistance Gene Analogues (RGA) mapping. It was found that the resistance to powdery mildew in Ano2 was conferred by a dominant gene, and the gene was named Pm-AN. The genetic analysis revealed that Pm-AN located between two codominant markers RPW and MRGH63B in linkage groupV. The genetic distances between Pm-AN and these two markers were 1.4–1.8 and 1.6–2 cM. No recombination was found between Pm-AN and markers ME/E1, SRAP23. Pm-AN was located in a RGA-rich region and cosegregated with the RGA marker MRGH5 and the resistance gene Vat. Synteny analysis showed that markers in this region were collinear between melon and cucumber. Segregation distortion was found in this region using both Ano2 × Hami413 and Ano2 × Queen F₂ populations, and the distortion was more distinct in Ano2 × Hami413 F₂ population. The center of segregation distortion was located in the RGA rich region harboring Pm-AN.