Isolation of mutants of the obligate methanotroph Methylomonas albus defective in growth on methane

A strain of Methylomonas albus BG8WM, a type 1 obligate methanotroph, which had been maintained for 2 ycars by serial passage on solid medium containing methanol as a carbon source was found to mutate at a frequency of 10^-5-10^-6 to resistance to dichloromethane (DCM^R), the parental strain BG8 did not give rise to DCM^R colonies. DCM^R strains were no longer capable of growth on methane as a carbon cource and exhibited greatly reduced or undetectable methane mono-oxygenase activity. The mutants fell into three groups on the basis of SDS-PAGE analysis of the polypeptide profiles of the particulate fraction of cell extracts. One or more of four polypeptides of Mr 70,000, 50,000, 25,000 and 23,000 were implicated as being components of the methane mono-oxygenase. Spontaneous reversion to growth on methane and sensitivity to dichloromethane occurred at a frequency of about 10^-8. The loss of methane mono-oxygenase activity was not associated with loss of the resident 55 kb plasmid.Abbreviations DCM^R dichloromethane-resistant - SDS-PAGE sodium dodecyl sulphate polyacrylamide gel electrophoresis - NMS nitrate minimal salts medium     

The alkene monooxygenase from Xanthobacter Py2 is a binuclear non-haem iron protein closely related to toluene 4-monooxygenase

The genes encoding the six polypeptide components of the alkene monooxygenase from Xanthobacter Py2 have been sequenced. The predicted amino acid sequence of the first ORF shows homology with the iron binding subunits of binuclear non-haem iron containing monooxygenases including benzene monooxygenase, toluene 4-monooxygenase (>60% sequence similarity) and methane monooxygenase (>40% sequence similarity) and that the necessary sequence motifs associated with iron co-ordination are also present. Secondary structure prediction based on the amino acid sequence showed that the predominantly α-helical structure that surrounds the binuclear iron binding site was conserved allowing the sequence to be modelled on the co-ordinates of the methane monooxygenase α-subunit. Significant differences in the residues forming the hydrophobic cavity which forms the substrate binding site are discussed with reference to the differences in reaction specificity and stereospecificity of binuclear non-haem iron monooxygenases.     

Evaluating effect of arbuscular mycorrhizal fungal consortia and Azotobacter chroococcum in improving biomass yield of Jatropha curcas

This study was conducted to study the long-term impact of bioinoculants, Azotobacter chroococcum and arbuscular mycorrhizal fungi (AMF) on growth and biomass yield of Jatropha curcas grown in nursery and in field conditions. The experiment was set up in a randomized block design, and the following treatments was designed (T₁ = control, T₂ = Azotobacter, T₃ = inoculation with AMF, and T₄ = inoculation with Azotobacter + AMF). Data on various growth attributes (shoot height and shoot diameter) and biochemical parameters [leaf relative water content (LRWC), sugars, protein, and photosynthetic pigments] were recorded up to 6 months in the nursery and in the field (18 months). Results pertaining to morpho-physiological traits showed Azotobacter and AMF consortia increase shoot height, shoot diameter, LRWC, sugars, proteins, and photosynthetic pigments over control under nursery conditions. Besides enhancing the plant growth, these bioinoculants helped in better establishment of Jatropha plants under field conditions. A significant improvement in the shoot height, shoot diameter, fruit yield/plant, and seed yield (g)/plant was evident in 18-month-old Jatropha plants under field conditions when Azotobacter and AMF were co-inoculated. This work supports the application of bioinoculants for establishment of Jatropha curcas in semi-arid regions.     

Chemical modification of the hydroxylase of soluble methane monooxygenase gives one form of the protein with significantly increased thermostability and another that functions well in organic solvents

Proteolysis of the hydroxylase component of soluble methane monooxygenase (MMO) with trypsin yielded a protein which retained 50% activity in a standard MMO assay. In an H₂O₂-driven assay, in which H₂O₂ replaced two of the protein components, NADH and O₂ used in the standard assay, the proteolysed hydroxylase retained full activity for ethane, propane and propene, but had a 2–3 fold increase with methane as substrate. Several crosslinking reagents have been tested for their ability to stabilise the proteolysed form of the hydroxylase. Using polyoxyethylene bis(imidazolyl carbonyl) (M_r 3350) as the crosslinking agent, increased thermostability of the hydroxylase was observed. Activated methoxypolyethylene glycol (M_r 5000) was used to modify the hydroxylase which was now soluble in organic solvents as well as water and could be activated by H₂O₂. The glycol-modified hydroxylase functioned well in organic solvents in the catalysis of propene oxidation.     

LiGAPS-Beef,a mechanistic model to explore potential and feed-limited beef production 1: model description and illustration

The expected increase in the global demand for livestock products calls for insight in the scope to increase actual production levels across the world. This insight can be obtained by using theoretical concepts of production ecology. These concepts distinguish three production levels for livestock: potential (i.e. theoretical maximum) production, which is defined by genotype and climate only; feed-limited production, which is limited by feed quantity and quality; and actual production. The difference between the potential or limited production and the actual production is the yield gap. The objective of this paper, the first in a series of three, is to present a mechanistic, dynamic model simulating potential and feed-limited production for beef cattle, which can be used to assess yield gaps. A novelty of this model, named LiGAPS-Beef (Livestock simulator for Generic analysis of Animal Production Systems – Beef cattle), is the identification of the defining factors (genotype and climate) and limiting factors (feed quality and available feed quantity) for cattle growth by integrating sub-models on thermoregulation, feed intake and digestion, and energy and protein utilisation. Growth of beef cattle is simulated at the animal and herd level. The model is designed to be applicable to different beef production systems across the world. Main model inputs are breed-specific parameters, daily weather data, information about housing, and data on feed quality and quantity. Main model outputs are live weight gain, feed intake and feed efficiency (FE) at the animal and herd level. Here, the model is presented, and its use is illustrated for Charolais and Brahman × Shorthorn cattle in France and Australia. Potential and feed-limited production were assessed successfully, and we show that FE of herds is highest for breeds most adapted to the local climate conditions. LiGAPS-Beef also identified the factors that define and limit growth and production of cattle. Hence, we argue the model has scope to be used as a tool for the assessment and analysis of yield gaps in beef production systems.     

The anaerobic decomposition of benzoic acid during methane fermentation

Anaerobic rupture of the benzoic acid ring was investigated. Carbon 4 was converted primarily to carbon dioxide. Following ring rupture during methane fermentation, propanoic acid is an intermediate, and carbon 4 of benzoate becomes its carboxyl.Contribution No. 1285-j, Division of Biology, Kansas State University, Manhattan, KS 66506. This work was supported in part by funds from the Kansas Agricultural Experiment Station, Kansas State University, Manhattan, KS 66506. Paper II of this series is Fina and Fiskin (1960)     

Variation of Pleurotus ostreatus (Jacq. Ex Fr.) P. Kumm. (1871) performance subjected to differentdoses and timings of nano-urea

Supplementation of the growing substrate by nitrogenous additives has been known to improve the production of oyster mushroom (Pleurotus ostreatus (Jacq. ex Fr.) P. Kumm. (1871)). However, the application of nano-additives has not been reported in such cultivation yet. The study investigated the effect of nano-urea added in two different doses (3 g and 5 g per kg substrate), once (at spawning or after first flush) or twice (at spawning and after first flush) to the growing substrate consisting of wheat straw and spent oyster substrate (1:1, w/w). Results showed that the application of nano-urea once has induced the highest number of mushroom flushes (four flushes) despite the dose applied. Contrarily to early findings, where high doses of nitrogen have caused inhibition of mushroom growth and production, nano-urea application has had better effects when applied twice. With 5 g/kg, it induced the shortest period between the first and the third flush (15 days). With 3 g/kg, it resulted in the highest biological and economic yields at the third flush (332.7 g/bag and 283.1 g/bag respectively), in total (973.4 g/bag and 854.0 g/bag respectively), the highest biological efficiency (109.6%), and pileus diameter/stipe length ratio (2.8). Experimental findings of the current study may be potentially applied at commercial scale.     

EFFECTS OF ARSENATE ON GROWTH OF NITROGEN-AND PHOSPHORUS-LIMITED CHLORELLA VULGARIS (CHLOROPHYCEAE) ISOLATES1

The effects of arsenate on the growth characteristics of five isolates of the freshwater alga, Chlorella vulgaris Beij., were examined. Two field isolates originated from arsenic-contaminated sites in Yukon, Canada and Kyushi, Japan; two reference isolates were obtained from the University of Texas Culture Collection. One isolate was selected for arsenic-tolerance in the laboratory. All five strains survived in culture solutions containing high arsenate concentrations. Arsenate (1–25 mM As) reduced photosynthesis and cell growth, as reflected by induced lag periods, slower growth rates, and lower stationary cell yields. Field isolates had shorter lag periods, higher growth rates, and enhanced cell yields compared to lab isolates when exposed to the same arsenic concentrations. Growth of the phosphorus-limited field strains was stimulated by the addition of arsenic. The cell yield of phosphorus-limited C. vulgaris Yukon, when treated with arsenic, was two times that of the phosphorus-limited control. This pattern was not evident when photosynthesis was used as a measure of cell response.     

Instrumental test to estimate a model of forecast yield: a non-parametric application to the pollen index in South Italy

A statistical test is described to verify the characteristics of the biological information contained in the dynamics of the flowering process. The test focuses on interactions between the pollen index and climatic variables to investigate if the biological indicator can synthesise the information of the pre-flowering phases. The multiple-regression model is built upon two pre-flowering climate macro-indicators extracted by Principal Component Analysis (PCA) and the optimised pollen index is obtained by non-parametric estimation. The empirical analysis is applied to 15 stations located in southern Italy in regions that have a longstanding tradition of olive production. Using the variance explained, we find that an optimised pollen index is fairly well predicted by the pre-flowering climatic data. We conclude that the optimised pollen index makes more parsimonious the modelling for predicting olive production.