Genomic-proteomic analysis of a novel Bacillus thuringiensis strain: toxicity against two lepidopteran pests, abundance of Cry1Ac5 toxin, and presence of InhA1 virulence factor.

Bacillus thuringiensis (Bt) is a biological alternative to the indiscriminate use of chemical insecticides in agriculture. Due to resistance development on insect pests to Bt crops, isolating novel Bt strains is a strategy for screening new pesticidal proteins or strains containing toxin profile variety that can delay resistance. Besides, the combined genomic and proteomic approaches allow identifying pesticidal proteins and virulence factors accurately. Here, the genome of a novel Bt strain (Bt TOL651) was sequenced, and the proteins from the spore-crystal mixture were identified by proteomic analysis. Toxicity bioassays with the spore-crystal mixture against larvae of Diatraea saccharalis and Anticarsia gemmatalis, key pests of sugarcane and soybean, respectively, were performed. The toxicity of Bt TOL651 varies with the insect; A. gemmatalis (LC50 = 1.45 ng cm-2) is more susceptible than D. saccharalis (LC50 = 73.77 ng cm-2). Phylogenetic analysis of the gyrB gene indicates that TOL651 is related to Bt kenyae strains. The genomic analysis revealed the presence of cry1Aa18, cry1Ac5, cry1Ia44, and cry2Aa9 pesticidal genes. Virulence factor genes such as phospholipases (plcA, piplc), metalloproteases (inhA), hemolysins (cytK, hlyIII, hblA, hblC, hblD), and enterotoxins (nheA, nheB, nheC) were also identified. The combined use of the genomic and proteomic data indicated the expression of Cry1Aa18, Cry1Ac5, and Cry2Aa9 proteins, with Cry1Ac5 being the most abundant. InhA1 also was expressed and may contribute to Bt TOL651 pathogenicity. These results provide Bt TOL651 as a new tool for the biocontrol of lepidopteran pests.

Morinda citrifolia Essential Oil: A Plant Resistance Biostimulant and a Sustainable Alternative for Controlling Phytopathogens and Insect Pests.

With the growing demand for sustainable and safe agricultural practices, plant compounds emerge as a solution for biological activities. Here, we evaluated the potential of using Morinda citrifolia essential oil to induce plant resistance and to control phytopathogens (Curvularia lunata) and insect pests (Daubulus maidis). We conducted a chromatographic analysis to unveil the essential oil components. We also quantified the activity levels of antioxidant enzymes and chitinase for resistance induction. The antifungal action was evaluated through disease progression and the inhibition of mycelial growth in addition to in silico studies that made it possible to predict the interaction site between the fungal protein and the compounds. We assessed the toxicity and repellent actions towards the D. maidis. Octanoic acid (58.43%) was identified as the essential oil major compound. Preventive treatment with essential oil and octanoic acid (25.0 µL mL-1) increased not only the plant defense activities (i.e., the activity of the enzymes superoxide dismutase, catalase, phenol peroxidase, ascorbate peroxidase, and chitinase) but also controlled Curvularia leaf spot. The stable interactions between octanoic acid and tyrosine-tRNA ligase from C. lunata suggested protein synthesis inactivation. The essential oil inhibited 51.6% of mycelial growth, and this effect was increased to 75.9% with the addition of adjuvants (i.e., angico gum). The essential oil reduced 76% of the population of D. maidis adults and repelled 50% of the number of D. maidis after 48 h under field conditions. The repellency effect in the field reduced the population of D. maidis adults, transmitters of the stunting complex, by 50%. The results highlight the potential of M. citrifolia as a resistance activator, fungicide, insecticide, and an effective biorational alternative.