Studying of the promotion mechanism of Bacillus subtilis QM3 on wheat seed germination based on β-amylase

The response of β-amylase in early stage germination of wheat seeds to Bacillus subtilis QM3 was mainly focused on to elucidate the promotion mechanism of B. subtilis QM3. The results showed that the changes in apparent activity of amylase and endosperm liquefaction after the strain QM3 treatment were much more obvious than that of the control group; the activity of β-amylase treated with the different concentrations of the strain QM3 increased significantly (P < 0.05) by 4% (107 CFU/mL) and 18.5% (106 CFU/mL) at the germination 6 h. Moreover, after presoaking with α-cyclodextrin, the activity of β-amylase increased significantly (P < 0.05) by 18.5% (107 CFU/mL) and 23.4% (106 CFU/mL) at the same stage of germination; the electrophoretogram of β-amylase isoenzymes showed that there is a growing trend in brightness and width of the band during the early germination from 3 to 6 h of wheat seed treated by the strain QM3 (106 CFU/mL). The increase in activity and isoenzyme expression of β-amylase may be one of the important reasons to promote the germination of wheat seeds after being treated by B. subtilis QM3.

Izozyme polymorphism of alpha-amylase in soft wheat

Aim. Study of alpha amylase polymorphism in winter wheat varieties. Methods. Isoenzymes of alpha-amylase were detected by electrophoretic protein separation in a polyacrylamide gel. The fall number on the Falling Number 1700 was determined. Results. Strains from various collections were explored: Institute of Plant Physiology and Genetics of the National Academy of Sciences of Ukraine, Kiev, The Plant Breeding and Genetics Institute – National Center of Seed and Cultivar Investigation of the National Academy of Agricultural Sciences of Ukraine, Odesa and german strains. The polymorphism of wheat grain alpha-amylase was detected by the method of electrophoretic protein separation. We searched associations between variants of alpha-amylase and resistance to pre-harvest germination, the falling number in the examined samples was determined. Conclusions. The frequency of the prevalence of variants for isoenzymes of alpha-amylase in soft wheat varieties was estimated. It was shown that genotypes containing the variant of the isoenzymes АbC are more resistant to pre-harvest germination. Among the studied varieties of PBGI NAAS and IPPG NAS of Ukraine, the variant of alpha-amylase AbC. Keywords: α-amylase, isoenzymes, soft wheat, electrophoresis, falling number.

Glycogen catabolism enzymes and protein fractions in the third and fourth larval stages ofAnisakis simplex

Extracts ofAnisakis simplexthird (L3) and fourth (L4) larval stages were assayed for protein content and activity and properties of α-amylase, glucoamylase and glycogen phosphorylase. Protein content in L4 was twice that in L3. SDS–PAGE applied to both larval stages revealed 22 protein fractions in each, including five stage-specific fractions in each larval stage. The L3 extracts contained three amylase isoenzymes: α1, α2 and α3; their molecular weights were 64, 29 and 21 kDa, respectively. Only one amylase isoenzyme (64 kDa) was found in the L4 extracts. Glycogen in L3 was found to be broken down mostly by hydrolysis because of low glycogen phosphorylase activity. The α-amylase activity in L4 was higher than that in L3 by half and the glycogen phosphorylase activity was ten times higher. In addition, the same enzymes isolated from L3 and L4 were found to differ in their properties. These differences could be manifestations of metabolic adaptations ofA. simplexlarvae to host switch from fish (L3) to mammals (L4), i.e. adaptations to a new habitat.