Specific and sensitive detection of Alcaligenes species from an agricultural environment

A quantitative real‐time PCR assay to specifically detect and quantify the genus Alcaligenes in samples from the agricultural environment, such as vegetables and farming soils, was developed. The minimum detection sensitivity was 106 fg of pure culture DNA, corresponding to DNA extracted from two cells of Alcaligenes faecalis. To evaluate the detection limit of A. faecalis, serially diluted genomic DNA from this organism was mixed with DNA extracted from soil and vegetables and then a standard curve was constructed. It was found that Alcaligenes species are present in the plant phytosphere at levels 10²–10⁴ times lower than those in soil. The approach presented here will be useful for tracking or quantifying species of the genus Alcaligenes in the agricultural environment.     

Insights into the structural stability of nuclear matrix ribonucleoprotein,LMG160: thermodynamic and spectroscopic analysis

Low-mobility group nonhistone chromatin protein, LMG_160, is a nuclear matrix ribonucleoprotein particle (RNP) which has a RNA molecule with approximately 300 bases. In this study, structural stability of the intact LMG₁₆₀ (I-LMG₁₆₀) was investigated at different ionic strength and in the absence of its RNA moiety (T-LMG₁₆₀) employing spectroscopic and thermodynamic techniques. The UV absorption spectra showed hypochromicity and red shift under increasing ionic strength for both forms of LMG₁₆₀ but in different extents. The fluorescence emission intensity was decreased as ionic strength was increased and the Stern–Volmer quenching constant (K_sv) for T-LMG₁₆₀ was 3.7 times less than for I-LMG₁₆₀. In the absence of sodium chloride, I-LMG₁₆₀ exhibited a very stable structure against the temperature change compared to T-LMG₁₆₀. The thermodynamic parameters showed that the positive values of ΔH_m and ΔS_m increased by increasing ionic strength in both forms of LMG₁₆₀. Removal of the RNA moiety altered secondary structure: as T-LMG₁₆₀ showed more helical content than I-LMG₁₆₀. From the results, it is concluded that I-LMG₁₆₀ is more sensitive to alteration of environment and the RNA has an important role in this RNP conformation. Also, interaction of both I- and T-LMG₁₆₀ with sodium chloride is entropy driven and is usually accompanied by surface hydrophobicity.     

The Baseline Cosinus Function: A Periodic Regression Model for Biological Rhythms

Abstract

Both tryptophan oxygenase and tyrosine aminotransferase of rat liver show diurnal variations in the inducibility by quinolinic acid. The maxima of effectiveness of quinolinic acid precede the maxima of normal enzyme activity. In the case of tyrosine aminotransferase, the induction kinetics and the dose response curve were also greatly depending on the time of day. No rhythmicity could be detected in the activities of 3‐hydroxyanthranilate oxygenase and ornithine aminotransferase.     

Fusarium langsethiae – a HT‐2 and T‐2 Toxins Producer that Needs More Attention

Fusarium langsethiae is a toxigenic fungus that was formally described as a new species in 2004. This fungus was first detailed in the 1990s but was initially referred to as ‘powdery Fusarium poae’ having a spore morphology similar to F. poae but a mycotoxin profile like that of Fusarium sporotrichioides. The species has been isolated from infected oat, wheat and barley grains but has been reported as more problematic in the former crop rather than the latter two. Whilst the epidemiology of F. langsethiae remains unclear, the fungus has been shown to produce high levels of type‐A trichothecenes HT‐2 and T‐2 toxins in small‐grain cereals. HT‐2 and T‐2 toxins are two of the most potent trichothecenes capable of inhibiting protein synthesis in eukaryotes. In this regard, mycotoxin contamination caused by F. langsethiae is clearly a food and feed safety hazard. With the European Commission considering legislation of HT‐2 and T‐2 toxins, more information is required not only on the producer and conditions favouring mycotoxin production, but also on reliable methods of pathogen detection and reduction of cereal contamination. This review describes recent research concerning the known epidemiology of F. langsethiae and suggestions of what needs to be known about the fungus in order to be able to understand and employ measures for preventing its infection and contamination of cereals with HT‐2 and T‐2 toxins.     

Current Status of Cereal Root Diseases in Western Australia Under Intensive Cereal Production and Their Comparison with the Historical Survey Conducted During 1976–1982

Two separate surveys of root diseases of cereals in the Western Australian (WA) cereal belt were conducted: the first conducted annually for wheat and barley during 1976–1982 and the second for wheat during 2005–2007. For the 1976–1982 survey, the cereal belt was divided into 15 zones based on the location and rainfall. Sampling was representative of the actual cropping area, with both wheat and barley sampling sites selected by zone as a percentage of total sites. Over 31 000 plants were assessed from a total of 996 fields. Average take‐all incidence ranged from 3% in the northern low rainfall zone to 57% in the southern high rainfall zone. Other root diseases assessed included rhizoctonia root rot, fusarium crown rot and subcrown internode discolouration. During the 2005–2007 survey, around 20 000 plants from a total of 210 fields being intensively cropped with cereals were surveyed for take‐all, rhizoctonia root rot, fusarium crown rot, common root rot, root lesion nematode and cereal cyst nematode. The 2005–2007 survey results indicated that root and crown diseases prevailed in paddocks frequently cropped with cereals and occurred at damaging levels across all WA cropping districts surveyed. The more recent root disease survey identified that the fungal diseases rhizoctonia root rot and fusarium crown rot and the root lesion nematode were the most serious impediments to intensive cereal production, particularly in the southern region of WA. Comparing the 2005–2007 results with the previous survey of 1976–1982, the relative importance of take‐all appears to have declined over the past 30 years.     

Comparative characterization of novel ene‐reductases from cyanobacteria

The growing importance of biocatalysis in the syntheses of enantiopure molecules results from the benefits of enzymes regarding selectivity and specificity of the reaction and ecological issues of the process. Ene‐reductases (ERs) from the old yellow enzyme family have received much attention in the last years. These flavo‐enzymes catalyze the trans‐specific reduction of activated C?C bonds, which is an important reaction in asymmetric synthesis, because up to two stereogenic centers can be created in one reaction. However, limitations of ERs described in the literature such as their moderate catalytic activity and their strong preference for NADPH promote the search for novel ERs with improved properties. In this study, we characterized nine novel ERs from cyanobacterial strains belonging to different taxonomic orders and habitats. ERs were identified with activities towards a broad spectrum of alkenes. The reduction of maleimide was catalyzed with activities of up to 35.5 U mg^?1 using NADPH. Ketoisophorone and (R)‐carvone, which were converted to the highly valuable compounds (R)‐levodione and (2R,5R)‐dihydrocarvone, were reduced with reaction rates of up to 2.2 U mg^?1 with NADPH. In contrast to other homologous ERs from the literature, NADH was accepted at moderate to high rates as well: Enzyme activities of up to 16.7 U mg^?1 were obtained for maleimide and up to 1.3 U mg^?1 for ketoisophorone and (R)‐carvone. Additionally, excellent stereoselectivities were achieved in the reduction of (R)‐carvone (97–99% de). In particular, AnabaenaER3 from Anabaena variabilis ATCC 29413 and AcaryoER1 from Acaryochloris marina MBIC 11017 were identified as useful biocatalysts. Therefore, novel ERs from cyanobacteria with high catalytic efficiency were added to the toolbox for the asymmetric reduction of alkenes. Biotechnol. Bioeng. 2013; 110: 1293–1301. © 2012 Wiley Periodicals, Inc.