Two-dimensional single strand conformation polymorphism (SSCP) of 16S rRNA gene fragments reveals highly dissimilar bacterial communities in an acidic fen

Genetic fingerprinting methods such as denaturing gradient gel electrophoresis (DGGE) and single strand conformation polymorphism (SSCP) are only able to separate about 20–40 well-distinguishable bands (signals) within each sample. As a result, the diversity of 16S rRNA genes within biological samples may be underestimated, because multiple sequences can migrate at the same rate to form a single band. This study reports a two-dimensional SSCP fingerprinting method that has the capability to resolve hundreds of signals in a single fingerprint by using different gel temperatures in the two dimensions of the separation (20 °C and 30 °C, respectively). Unlike previous two-dimensional approaches, the method presented in this study does not rely on DNA products of variable lengths but is able to separate 16S rRNA gene fragments of the same length. To demonstrate the effectiveness of this new method, DNA samples from oxic and anoxic zones of an acidic fen were examined. Whereas one-dimensional SSCP fingerprints indicated high similarity (>93%) between 16S rRNA gene fragments from oxic and anoxic zones of the fen, the two-dimensional SSCP approach virtually found no similarities.     

Evaluation of the risk of motor neuron disease in horses fed a diet low in vitamin E and high in copper and iron

OBJECTIVE: To determine whether equine motor neuron disease (EMND) could be induced in adult horses fed a diet low in vitamin E and high in copper and iron. ANIMALS: 59 healthy adult horses. PROCEDURE: Horses in the experimental group (n = 8) were confined to a dirt lot and fed a concentrate low in vitamin E and high in iron and copper in addition to free-choice grass hay that had been stored for 1 year. Control horses (n = 51) were fed a concentrate containing National Research Council-recommended amounts of copper, iron, and vitamin E. The hay fed to control horses was the same as that fed to experimental horses, but it had not been subjected to prolonged storage. Control horses had seasonal access to pasture, whereas experimental horses had no access to pasture. Horses that developed clinical signs of EMND were euthanatized along with an age-matched control horse to determine differences in hepatic concentrations of vitamin E, vitamin A, copper, iron, and selenium. RESULTS: 4 experimental horses developed clinical signs of EMND. Plasma concentrations of vitamin E decreased in all 8 experimental horses. There were no significant changes in plasma concentrations of vitamin A, selenium, and copper or serum concentrations of ferritin. There were no significant differences in those analytes between experimental horses with EMND and experimental horses that did not develop EMND. No control horses developed EMND. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that lack of access to pasture, dietary deficiency of vitamin E, or excessive dietary copper are likely risk factors for EMND.     

Rapid identification of betacyanins from Amaranthus tricolor, Gomphrena globosa, and Hylocereus polyrhizus by matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometry (MALDI-QIT-TOF MS)

Natural betacyanins have attracted great attention as food colorants and potential antioxidants. Matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometry (MALDI-QIT-TOF MS) is a new and powerful technique for the identification of low molecular weight compounds. This study is the first to employ MALDI-QIT-TOF MS to rapidly identify, within a few minutes, a great number of betacyanins in crude extracts from Amaranthus tricolor seedlings, Gomphrena globosa flowers, and Hylocereus polyrhizus fruits. The fresh crude extract samples without any purification were directly used for MALDI-QIT-TOF MS analysis with 2,5-dihydroxybenzoic acid as a matrix. The MS2 and MS3 spectrometric data acquired could provide important characteristic information for structural elucidation of the betacyanins. Fourteen free and acylated betacyanins, belonging to amaranthin-type, betanin-type, and gomphrenin-type betacyanins, respectively, were identified. However, the related isomers should be differentiated with the aid of HPLC.     

Phytophthora genome sequences uncover evolutionary origins and mechanisms of pathogenesis

Draft genome sequences have been determined for the soybean pathogen Phytophthora sojae and the sudden oak death pathogen Phytophthora ramorum. O?mycetes such as these Phytophthora species share the kingdom Stramenopila with photosynthetic algae such as diatoms, and the presence of many Phytophthora genes of probable phototroph origin supports a photosynthetic ancestry for the stramenopiles. Comparison of the two species' genomes reveals a rapid expansion and diversification of many protein families associated with plant infection such as hydrolases, ABC transporters, protein toxins, proteinase inhibitors, and, in particular, a superfamily of 700 proteins with similarity to known o?mycete avirulence genes.     

发芽提高黑小麦可溶性和结合性提取物的抗氧化活性和多酚含量

发芽是提高种子生物活性成分和功能的一种极具前景的生物加工技术。本课题主要研究了发芽对黑小麦提取物体外抗氧化活性及多酚含量和组成的影响。总体上,发芽和未发芽黑小麦的结合性提取物比可溶性提取物具有更高的抗氧化活性和总酚含量。另外,发芽3-8天显著提高黑小麦可溶性和结核性提取物的抗氧化活性和总酚含量。在发芽黑小麦中,可溶性提取物可能富含多种黄酮化合物,而结合性提取物主要含有对香豆酸和阿魏酸两种酚酸。并且,这两种结合酚酸在发芽3-8天的过程中明显增加。上述结果表明,发芽能够提高黑小麦种子的抗氧化多酚水平;而且,具有较高抗氧化多酚含量的发芽黑小麦是抗氧化剂的一个天然来源,具有潜在的健康效益。     

Properties of Starch Noodles as Affected by Sweetpotato Genotype

Starch was extracted from 14 sweetpotato genotypes from the Philippines. The Rapid Visco-Analyzer (RVA) viscoamylographs of the starches showed Type A pasting curves, characterized by a high pasting peak followed by a high degree of shear-thinning. The major difference among genotypes was in the sharpness of the peak, with some showing a very sharp peak while others showed a broad peak. This difference was related to time from onset of pasting to peak viscosity, and to stability ratio (holding viscosity/peak viscosity), which were also highly correlated (r = 0.84, P < 0.01) to each other. Stability ratio was also correlated to noodle firmness (r = 0.95, P < 0.01), rehydration (cooked weight) (r = -0.89, P < 0.01), and swelling volume of the starch (r = -0.62, P < 0.05). The amylose content was correlated significantly only to peak viscosity (r = -0.84, P < 0.01). Significant differences in texture and cooking quality of the starch noodles produced from the different genotypes was found. It was shown that the RVA viscoamylographs could be used to detect differences in pasting characteristics of sweetpotato starch which are related to quality of noodle produced.     

Potential Use of Raman Spectroscopy for Determination of Amylose Content in Maize Starch

An analytical method using Raman spectroscopy was developed for the determination of amylose concentration in maize starches. FT-Raman spectra of four maize starches with amylose content varying from 3.3 to 66% were obtained. A Raman band at ≈1657 cm^-1 correlated linearly with amylose concentration in the four maize starches, and a calibration curve for Raman band intensity versus amylose content was developed. The linear correlation of the I₁₆₅₇/I₉₀₀ integrated areas with amylose content was r = 0.997. The Raman-based calibration curve allows fast and nondestructive determination of the amylose content in maize starches with minimal sample preparation.     

Genetic Diversity in Physical Properties of Starch from a World Collection of Amaranthus

Physical and functional properties of starches isolated from 93 noncultivated genotypes of nine Amaranthus species from a world germ plasm collection and an additional 31 cultivated Amaranthus genotypes obtained from China were tested. A wide variation was found in the properties tested among the Amaranthus species and among genotypes within the same species. When comparing starches from cultivated and noncultivated genotypes, it was generally found that amylose was lower; starch pasting profiles were more consistent with higher peak viscosity, lower breakdown, and lower setback; the gelatinization temperature was lower; and energy of enthalpy was higher. Under cool storage, the hardness of cultivated starch pastes was lower and the adhesiveness was higher. As expected, amylose content was a primary factor affecting the physical and functional properties of Amaranthus starch. Compared with reference maize, rice, and wheat starches, Amaranthus starch tended to have lower hot paste viscosity and lower cool paste viscosity; and higher gelatinization temperatures and higher energy of enthalpy. Furthermore, Amaranthus starch pastes showed less change of gel hardness and adhesiveness after cold storage. The environmental effect on the different properties of starch varied among Amaranthus species. It is suggested that Amaranthus starches can be developed for a wide range of food uses.