Improved solubility and stability of carp myosin by conjugation with alginate oligosaccharide

ABSTRACT:   Carp myosin was conjugated with alginate oligosaccharide (AO) through the Maillard reaction under low relative humidity, and the functional properties of the myosin-AO conjugate were investigated to clarify the role of myosin in the functional improvement of fish myofibrillar proteins (Mf) by the glycosylation. The findings were as follows. First, myosin became highly solubilized at lower NaCl concentrations by conjugation with AO and NaCl-dependence of the solubility was lost when > 12% of the available lysine residues were reacted with AO and 50 µg/mg of AO was attached to myosin. Second, the thermal stability of myosin was effectively improved by conjugation with AO. Heat-treatment at 50°C for 6 h has no effect on the solubility of the myosin-AO conjugate regardless of the NaCl concentration. Third, the improved functionalities of myosin conjugated with AO remained even at a nearly isoelectric point. The improving effect of AO-conjugation on the characteristics of myosin was almost the same as Mf reacted with AO. Therefore, it is apparent that that improved functionalities of the glycosylated Mf reflect the functional changes of myosin.     

Intragenomic variation in the internal transcribed spacer regions between 16S–23S rRNA genes among the three copies of Sinorhizobium fredii strains

The soybean-nodulating Sinorhizobium fredii strain has been reported to possess three copies of rRNA gene operons. In the present study, we investigated the diversity of the 16S–23S rDNA internal transcribed spacer (ITS) regions of S. fredii strains. Based on the sequences of the ITS regions, we divided the sequences of the S.   fredii strains into two groups, type A and type B. A dot-matrix analysis indicated that the region flanked by tRNA-Ile and tRNA-Ala is longer in type A than in type B, whereas type B sequences possess longer regions upstream of tRNA-Ile and downstream of tRNA-Ala than those of the type A sequence. Restriction fragment length polymorphism of polymerase chain reaction product (PCR-RFLP) of the ITS region in the cloned plasmids as templates could reconstruct the PCR-RFLP pattern from the total DNA as a template. The results of Southern hybridization using the insert sequence between tRNA-Ile and tRNA-Ala in type A as a probe indicated differences in the copy numbers of the type A ITS regions among the strains tested. These results indicated that S. fredii strains possess the type A and type B sequences of the ITS regions at ratios of 3:0, 2:1, 1:2 or 0:3. These S. fredii strains may be useful biological materials for the study of intraspecific variations.     

Colonization and growth promotion characteristics of Enterobacter sp. and Herbaspirillum sp. on Brassica oleracea

We investigated the effects of inoculating a dicot plant, Brassica oleracea , with nitrogen-fixing endophytic bacteria isolated from monocots. The bacteria used were Enterobacter sp. strain 35 isolated from sugarcane and Herbaspirillum sp. strain B501 isolated from wild rice. Under glasshouse conditions, B. oleracea inoculated with strain 35 had a significantly greater fresh weight than uninoculated plants, and the fresh weight of plants inoculated with strain B501 tended to be higher than that of uninoculated plants. We conducted a laboratory-scale experiment to confirm the above results and to investigate the colonization and nitrogen-fixing abilities of these bacteria. Plants inoculated with Enterobacter sp. strain 35-1, a green fluorescent protein (GFP)-labeled strain derived from strain 35, had larger bacterial populations and greater acetylene reduction activity than those inoculated with Herbaspirillum sp. strain B501 gfp 1. Strain 35-1 colonized the intercellular spaces and the junctions of the lateral roots with the parent root. The results indicated that isolates from monocots can successfully colonize B. oleracea (a dicot) and promote plant growth.     

Estimation of nodulation tendency among Rj-genotype soybeans using the bradyrhizobial community isolated from an Andosol

The nodulation tendency of indigenous soybean bradyrhizobia on Rj -genotype soybean cultivars was investigated using approximately 260 bradyrhizobia isolated from an Andosol with 13 soybean cultivars of five Rj -genotypes (non- Rj , Rj ₂ Rj ₃, Rj ₃, Rj ₄ and Rj ₂ Rj ₃ Rj ₄). A dendrogram was constructed based on restriction fragment length polymorphism analysis of the polymerase chain reaction products (PCR-RFLP) of the 16S–23S rDNA internal transcribed spacer (ITS) region. Bradyrhizobium USDA strains were used as a reference. The dendrogram indicated nine clusters based on similarities among the reference strains. The ratio of beta diversity to gamma diversity ( H' _β/ H' _γ), which represents differences in the bradyrhizobial communities by pair-wise comparison between each cultivar, was obtained from Shannon–Wiener diversity indices. The results showed that bradyrhizobial communities among the same Rj -genotype cultivars were similar to each other, whereas bradyrhizobial communities between the Rj ₂-genotype and non- Rj , Rj ₃ or Rj ₄-genotype cultivars were significantly different. These results suggest that the Rj ₂-gene might not only affect the nodulation compatibility between Rj -genotype soybeans and bradyrhizobia, but also the nodulation tendency of the bradyrhizobia.