Expression of Leucaena Leucocephala de Wit Chitinase in Transgenic Koshihikari Rice

The cDNA encoding the 326 amino acids of a Class I basic chitinase gene from Leucaena leucocephala de Wit (KB3, Genbank accession: AAM49597) was cloned under the control of CaMV35S promoter in pCAMBIA 1300 and transferred to Koshihikari. Calli of Koshihikari rice was transformed with agrobacterium with this construct expressing the chitinase and β- glucouronidase (GUS). The frequencies of calli 90 % has been obtained from rice seedlings cultured on NB medium. The high regeneration frequencies, 74% was obtained from calli cultured on regeneration medium containing 4 mg/l BAP, and 7 g/l phytagel at 25°C. Various factors were studied in order to establish a procedure for the transformation of Koshihikari Agrobacterium tumefaciens. Supplementation of 50 mM acetosyringone to the medium during coculivation was important to enhance the frequency to transient transformation. The 4 week-old scutellum-derived calli were excellent starting materials. Selection medium based on NB medium supplement with 40 mg/l hygromycin and 400 mg/l cefotaxime were an optimized medium for selection of transformed rice calli. The percentage of transformation 70 was obtained. Recombinant calli and regenerated rice plants were checked the expression of chitinase and gus by PCR, northern blot gel, southern blot gel, and gus assay. Chitinase and gus were expressed in all parts of recombinant rice. The rice line expressing the KB3 chiitnase was more resistant to the blast fungus Fusarium monoliforme than control line.

Aureobasidium pullulans Used as a Biological Control Agent under Field Conditions Affects the Microbial Quality of Winter Wheat Grain

The biological activity of A. pullulans isolates against species of the genus Fusarium, bacteria of the genus Azotobacter and pseudomonads colonizing wheat kernels was evaluated. A field experiment was carried out in 2009-2011, in north-eastern Poland. Winter wheat (cv. Bogatka) plants were sprayed with a cell suspension of A. pullulans at a density of 106 - 108 per cm3 water at the stem elongation stage and the heading stage. Untreated plants served as control. The abundance of epiphytic yeasts, bacteria of the genus Azotobacter, pseudomonads and Fusarium pathogens on wheat grain was estimated at harvest and after six months’ storage. The average size of yeast communities was significantly greater on wheat kernels treated with a cell suspension of A. pullulans, compared with control samples. In 2010-2011, biological control reduced the abundance of some species of the genus Fusarium.

Microbiological and Physicochemical Studies of Wetland Soils in Eket, Nigeria

The microbiological and physicochemical characteristics of wetland soils in Eket Local Government Area were studied between May 2001 and June 2003. Total heterotrophic bacterial counts (THBC), total fungal counts (TFC), and total actinomycetes counts (TAC) were determined from soil samples taken from four locations at two depths in the wet and dry seasons. Microbial isolates were characterized and identified. Particle size and chemical parameters were also determined using standard methods. THBC ranged from 5.2 (+0.17) x106 to 1.7 (+0.18) x107 cfu/g and from 2.4 (+0.02) x106 to 1.4 (+0.04) x107cfu/g in the wet and dry seasons, respectively. TFC ranged from 1.8 (+0.03) x106 to 6.6 (+ 0.18) x106 cfu/g and from 1.0 (+0.04) x106 to 4.2 (+ 0.01) x106 cfu/g in the wet and dry seasons, respectively .TAC ranged from 1.2 (+0.53) x106 to 6.0 (+0.05) x106 cfu/g and from 0.6 (+0.01) x106 to 3.2 (+ 0.12) x106 cfu/g in the wet and dry season, respectively. Acinetobacter, Alcaligenes, Arthrobacter, Bacillus, Beijerinckja, Enterobacter, Micrococcus, Flavobacterium, Serratia, Enterococcus, and Pseudomonas species were predominant bacteria while Aspergillus, Fusarium, Mucor, Penicillium, and Rhizopus were the dominant fungal genera isolated. Streptomyces and Norcadia were the actinomycetes genera isolated. The particle size analysis showed high sand fraction but low silt and clay. The pH and % organic matter were generally acidic and low, respectively at all locations. Calcium dominated the exchangeable bases with low electrical conductivity and micronutrients. These results provide the baseline data of Eket wetland soils for its management for sustainable agriculture.