Aims: Cloning and expression of a new cry1Ia-type gene of Bacillus thuringiensis. Methods and Results: PCR amplification, using gene cry1I-specific primers revealed the presence of such a gene in the strain BNS3 of Bacillus thuringiensis subsp. kurstaki. The cloning and sequencing from BNS3 of the cry1Ia-type gene, called crybns3-3, showed an open reading frame of 2160-bp, encoding a protein of 719 amino acid residues. Both nucleotide and amino acid sequences similarity analysis revealed that the crybns3-3 is a new cry1Ia-type gene, presenting several differences from the cry1Ia-type genes. The study of the expression of crybns3-3 by Northern blot and RT-PCR showed that it was transcribed. The expression of crybns3-3 under the control of BtI and BtII promoters revealed that Crybns3-3 would co-crystallize with the endogenous delta-endotoxins. Conclusions: crybns3-3 is a novel cry1Ia gene isolated from B. thuringiensis subsp. kurstaki strain BNS3. Significance and Impact of the Study: The characteristics of crybns3-3 indicate that it is a new cry1Ia-type gene. Amino acid residue substitutions presented in Crybns3-3 could be exploited for both toxicity and specificity studies. Crybns3-3 would interact and co-crystallize at least partially with the endogenous delta-endotoxins of BNS3, and then participate in the formation of the parasporal crystal inclusions.

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Bacillus species are attractive due to their potential use in the biological control of fungal diseases. Bacillus amyloliquefaciens strain BLB369, Bacillus subtilis strain BLB277, and Paenibacillus polymyxa strain BLB267 were isolated and identified using biochemical and molecular (16S rDNA, gyrA, and rpoB) approaches. They could produce, respectively, (iturin and surfactin), (surfactin and fengycin), and (fusaricidin and polymyxin) exhibiting broad spectrum against several phytopathogenic fungi. In vivo examination of wheat seed germination, plant height, phenolic compounds, chlorophyll, and carotenoid contents proved the efficiency of the bacterial cells and the secreted antagonist activities to protect Tunisian durum wheat (Triticum turgidum L. subsp. durum) cultivar Om Rabiia against F. graminearum fungus. Application of single bacterial culture medium, particularly that of B. amyloliquefaciens, showed better protection than combinations of various culture media. The tertiary combination of B. amyloliquefaciens, B. subtilis, and P. polymyxa bacterial cells led to the highest protection rate which could be due to strains synergistic or complementary effects. Hence, combination of compatible biocontrol agents could be a strategic approach to control plant diseases.

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B. methylotrophicus strain 39b exhibited antibacterial activity against phytopathogenic A. tumefaciens C58 and B6 both in vitro and in vivo. Lipopeptides are the main compounds that confer the biocontrol ability. This strain has the potential to be developed as a biological control agent for crown gall disease. © 2016 Society of Chemical Industry.

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