Damage ratings of corn lines infested with ACB at V6 whorl stage.

<p>Damage ratings of corn lines infested with ACB at V6 whorl stage.</p

Damages of corn lines infested with ACB at R1 silking stage.

<p>Damages of corn lines infested with ACB at R1 silking stage.</p

Synergistic effects between Cry1Ac and Cry1Ie toxins on ACB.

<p>Synergistic effects between Cry1Ac and Cry1Ie toxins on ACB.</p

Mortality of ACB larvae of four strains fed on plant tissues of Bt corn and non-Bt corn hybrids.

<p>Mortality of ACB larvae of four strains fed on plant tissues of Bt corn and non-Bt corn hybrids.</p

Differentially-expressed genes identified between male and female <i>Ostrinia furnacalis</i> antennae.

<p><b>A</b>. Scatter plot of pairwise normalized abundance of each transcript among reads obtained from male- and female-specific antennal cDNA libraries. <b>B.</b> MA-plot-based method estimates of differential gene expression using a random sampling model, where the red data points represented as red dots indicate genes that show a comparatively significant level of up-regulation in females (above horizontal) and male antennae (below horizontal) (<i>P</i> ≤ 0.001). “M” is the binary logarithm of the intensity ratio and “A” is the average log intensity for a dot in the plot. <b>C.</b> Genes up-regulated in male antennae; <b>D.</b> Genes up-regulated in female antennae. All comparisons of differential gene expression based on log2(fold_change) normalized > 2 or log2(fold_change) normalized > 5.</p

MOESM4 of Binding affinity of five PBPs to Ostrinia sex pheromones

Additional file 4: S4. The sequences of the wild type OfurPBP3 and the mutants. The nucleotide sequences of wild type OfurPBP3 and the mutants (OfurPBP3-m1, OfurPBP3-m2, OfurPBP3-m3 and OfurPBP3-m4)

Table1.pdf

<p>Pheromone binding proteins (PBPs) play an important role in olfaction of insects by transporting sex pheromones across the sensillum lymph to odorant receptors. To obtain a better understanding of the molecular basis between PBPs and semiochemicals, we have cloned, expressed, and purified two PBPs (CpunPBP2 and CpunPBP5) from the antennae of Conogethes punctiferalis. Fluorescence competitive binding assays were used to investigate binding affinities of CpunPBP2 and CpunPBP5 to sex pheromone and volatiles. Results indicate both CpunPBP2 and CpunPBP5 bind sex pheromones E10-16:Ald, Z10-16:Ald and hexadecanal with higher affinities. In addition, CpunPBP2 and CpunPBP5 also could bind some odorants, such as 1-tetradecanol, trans-caryopyllene, farnesene, and β-farnesene. Homology modeling to predict 3D structure and molecular docking to predict key binding sites were used, to better understand interactions of CpunPBP2 and CpunPBP5 with sex pheromones E10-16:Ald and Z10-16:Ald. According to the results, Phe9, Phe33, Ser53, and Phe115 were key binding sites predicted for CpunPBP2, as were Ser9, Phe12, Val115, and Arg120 for CpunPBP5. Binding affinities of four mutants of CpunPBP2 and four mutants of CpunPBP5 with the two sex pheromones were investigated by fluorescence competitive binding assays. Results indicate that single nucleotides mutation may affect interactions between PBPs and sex pheromones. Expression levels of CpunPBP2 and CpunPBP5 in different tissues were evaluated using qPCR. Results show that CpunPBP2 and CpunPBP5 were largely amplified in the antennae, with low expression levels in other tissues. CpunPBP2 was expressed mainly in male antennae, whereas CpunPBP5 was expressed mainly in female antennae. These results provide new insights into understanding the recognition between PBPs and ligands.</p

Table2.pdf

<p>Pheromone binding proteins (PBPs) play an important role in olfaction of insects by transporting sex pheromones across the sensillum lymph to odorant receptors. To obtain a better understanding of the molecular basis between PBPs and semiochemicals, we have cloned, expressed, and purified two PBPs (CpunPBP2 and CpunPBP5) from the antennae of Conogethes punctiferalis. Fluorescence competitive binding assays were used to investigate binding affinities of CpunPBP2 and CpunPBP5 to sex pheromone and volatiles. Results indicate both CpunPBP2 and CpunPBP5 bind sex pheromones E10-16:Ald, Z10-16:Ald and hexadecanal with higher affinities. In addition, CpunPBP2 and CpunPBP5 also could bind some odorants, such as 1-tetradecanol, trans-caryopyllene, farnesene, and β-farnesene. Homology modeling to predict 3D structure and molecular docking to predict key binding sites were used, to better understand interactions of CpunPBP2 and CpunPBP5 with sex pheromones E10-16:Ald and Z10-16:Ald. According to the results, Phe9, Phe33, Ser53, and Phe115 were key binding sites predicted for CpunPBP2, as were Ser9, Phe12, Val115, and Arg120 for CpunPBP5. Binding affinities of four mutants of CpunPBP2 and four mutants of CpunPBP5 with the two sex pheromones were investigated by fluorescence competitive binding assays. Results indicate that single nucleotides mutation may affect interactions between PBPs and sex pheromones. Expression levels of CpunPBP2 and CpunPBP5 in different tissues were evaluated using qPCR. Results show that CpunPBP2 and CpunPBP5 were largely amplified in the antennae, with low expression levels in other tissues. CpunPBP2 was expressed mainly in male antennae, whereas CpunPBP5 was expressed mainly in female antennae. These results provide new insights into understanding the recognition between PBPs and ligands.</p

MOESM3 of Binding affinity of five PBPs to Ostrinia sex pheromones

Additional file 3: S3. The primers used for site-directed mutants. The lists of primers used for site-directed mutants, and the digestion sites were underlined

MOESM1 of Binding affinity of five PBPs to Ostrinia sex pheromones

Additional file 1: S1. The primers used for the PBPs expression. The lists of primers used for PBPs expression, and the digestion sites were underlined