Biochemical and Multiplex PCR Analysis of Toxic Crystal Proteins to Determine Genes in Bacillus thuringiensis Mutants

The Egyptian Bacillus thuringiensis isolate (M5) produce crystal proteins that is toxic against insects was irradiated with UV light to induce mutants. Upon testing 10 of the resulting mutants for their toxicity against cotton leafworm larvae, the three mutants 62, 64 and 85 proved to be the most toxic ones. Upon testing these mutants along with their parental isolate by SDS-PAGE analysis of spores-crystals proteins as well as vegetative cells proteins, new induced bands appeared in the three mutants by UV radiation and also they showed disappearance of some other bands as compared with the wild type isolate. Multiplex PCR technique, with five sets of specific primers, was used to detect the three types of cryI genes cryIAa, cryIAb and cryIAc. Results showed that these three genes exist, as distinctive bands, in the wild type isolate (M5) as well as in mutants 62 and 85, while the mutant 64 had two distinctive bands of cryIAb and cryIAc genes, and a faint band of cryI Aa gene. Finally, these results revealed that mutant 62 is considered as the promising mutant since it is UV resistant, highly toxic against Spodoptera littoralis and active against a wide range of Lepidopteran insects.

Effects of Entomopathogenic Nematodes on Suppressing Hairy Rose Beetle, Tropinota squalida Scop. (Coleoptera: Scarabaeidae) Population in Cauliflower Field in Egypt

The potential of entomopathogenic nematodes in suppressing T. squalida population on cauliflower from transplanting to harvest was evaluated. Significant reductions in plant infestation percentage and population density (/m2) were recorded throughout the plantation seasons, 2011 and 2012 before and after spraying the plants. The percent reduction in numbers/m2 was the highest in March for the treatments with Heterorhabditis indica Behera and Heterorhabditis bacteriophora Giza during the plantation season 2011, while at the plantation season 2012, the reduction in population density was the highest in January for Heterorhabditis Indica Behera and in February for H . bacteriophora Giza treatments. In a comparison test with conventional insecticides Hostathion and Lannate, there were no significant differences in control measures resulting from treatments with H. indica Behera, H. bacteriophora Giza and Lannate. At the plantation season is 2012. Also, the treatments reduced the economic threshold of T. squalida on cauliflower in this experiment as compared with before and after spraying with both the two entomopathogenic nematodes at both seasons 2011 and 2012. This means an increase in the marketability of heads harvested as a consequence of monthly treatments.