Authors:C. Lizette Del-Toro-Sánchez, S. Villaseñor-Alvarado, Florentina Zurita-Martínez, O. Castellanos-Hernández, Araceli Rodríguez-Sahagún, M. Isabel Torres-Morán, D. Rojas-Bravo, and M. Gutiérrez-Lomelí
Padmalatha, K., Prasad, M. N. V. (2006) Optimization of DNA isolation
Authors:A. Wiater, J. Szczodrak, and M. Pleszczyńska
The strain Streptococcus sobrinus CCUG 21020 was found to produce water-insoluble and adhesive mutan. The factors influencing both stages of the mutan production, i.e. streptococcal cultures and glucan synthesis in post-culture supernatants were standardized. The application of optimized process parameters for mutan production on a larger scale made it possible to obtain approximately 2.2 g of water-insoluble glucan per 1 l of culture supernate - this productivity was higher than the best reported in the literature. It was shown that some of the tested beet sugars might be successfully utilized as substitutes for pure sucrose in the process of mutan synthesis. Nuclear magnetic resonance analyses confirmed that the insoluble biopolymer synthesized by a mixture of crude glucosyltransferases was a mixed-linkage (1?3), (1?6)-a?-D-glucan (the so-called mutan) with a greater proportion of 1,3 to 1,6 linkages.
Authors:A. Behpouri, A. Perochon, F.M. Doohan, and C.K.-Y. Ng
Brachypodium distachyon has emerged as the model species for important temperate grass crops such as wheat and barley and the genome of the B. distachyon community inbred line Bd21 has been sequenced. Methods for tissue culture and Agrobacterium-mediated transformation have been developed for this model grass as a resource for reverse genetics and functional genomic analyses. In order to obtain a high quantity and quality of compact embryogenic callus (CEC) in B. distachyon, it is important to examine and optimize the optimal concentration of the auxin 2,4-D (dichlorophenoxyacetic acid) to use in both callus induction and callus proliferation media. Here, we investigated the effects of different concentrations of 2,4-D on callus induction and callus proliferation of B. distachyon Bd21. Our results showed that 2.5 mg l–1 2,4-D is an optimal concentration to use for both callus induction and proliferation, although 5.0 mg l–1 may also be used for callus proliferation. Additionally, the suitability of hygromycin or bialaphos as selectable markers was examined and results indicated that hygromycin is significantly more efficient than bialaphos when using the Agrobacterium-mediated transformation system.
This study investigated the factors influencing in vitro flowering of gherkin (Cucumis anguria L.). Multiple shoots were efficiently regenerated from cotyledonary node and axillary bud explants of C. anguria within 15 days on MSB5 medium containing 3% sucrose and supplemented with 1.5 mg l−1 6-benzyladinine (BA). The elongated shoots were excised and transferred to MSB5 medium containing 4% sucrose supplemented with 0.5 mg l−1 gibberellic acid (GA3) and 1.0 mg l−1 indole-3-butyric acid (IBA) induced maximum number of flowers (9.5 flowers/plant) and root induction (16.5 roots/plant). Factors that influence the in vitro flowering were optimizing pH, photoperiod and temperature. In vitro flowering was significantly early and higher number of flowers produced at pH (5.8), photoperiod (12/12 h) and room temperature (28 °C). In vitro developed flowers were less viable (80 ± 1.0%) compared to control plants (90 ± 2.0%). Our in vitro flower induction procedures provide an extremely effective method for further research on flowering regulation mechanisms in C. anguria. These plantlets were successfully transferred to the soil where they grew well for 3 to 5 weeks with 90% survivability. Plants grew normally and produced flowers with viable pollen and fertile seeds.
Nineteen fungal strains belonging to different genera were tested for extracellular mutanase production in shaken flasks. The optimal enzymatic activity was achieved by Trichoderma harzianum F-470, a strain for which the mutanase productivity has not yet been published. Some of factors affecting the enzyme production in shaken flasks and aerated fermenter cultures have been standardized. Mandels mineral medium with initial pH 5.3, containing 0.25% mutan and inoculated with 10% of the 48-h mycelium, was the best for enzyme production. A slight mutanolytic activity was also found when sucrose, raffinose, lactose and melibiose were carbon sources. Application of optimized medium and cultural conditions, as well as use of a fermenter with automatic pH control set at pH 6.0 enabled to obtain a high mutanase yield (0.33 U/ml, 2.5 U/mg protein) in a short time (2-3 days). The enzyme in crude state was stable over a pH range of 4.5-6.0, and at temperatures up to 35 °C; its maximum activity was at 40 °C and at pH 5.5.