Table 1.
Biological response and enzymatic activities in acephate-treated tarnished plant bug.
Table 2.
Identification of 107 significantly up-regulated (≥2-fold) genes in LLR using microarrays and analyzed with ArrayStar and Blat2go protocol (www.blast2go.org).
Table 3.
Identification of 118 significantly down-regulated gene (≥2-fold) in LLR using microarrays and analyzed with ArrayStar and Blat2go protocol (www.blast2go.org).
Figure 1.
Analysis of microarray data and comparison of 6,688 gene expression levels between acephate-susceptible (LLS) and –resistant (LLR) tarnished plant bugs using ArrayStar software.
A: Hierarchical clustering analysis of 329 up-regulated and 333 down-regulated genes (≥2-fold, P<0.05) in the LLR. Each column represented a sample (LLS or LLR) and each row represented a gene. The gray scale depicted the relative levels of gene expressions from low to high with corresponding grayscale from light (for lower expression) to dark (for higher levels). B: Scatter-plot comparison of 6,688 gene expression levels between LLS and LLR. The dots of the scatter plot in the upper left corner represented up-regulated genes, and the dots in low right corner represented down-regulated genes. Dots above line 2a and below line 2b represent up- and down-regulated genes by 2-fold; Dots above line 4a and below line 4b represent up- and down-regulated genes by 4-fold; Dots above line 8a and below line 8b represent up- and down-regulated genes by 8-fold.
Figure 2.
Annotation and functional analysis of 329 up-regulated and 333 down-regulated genes (≥2-fold) in the LLR.
A: Proportion of up-regulated genes (left side) and down-regulated genes (right side) categorized based on their involvement in biological process at GO level 2 (Blast2go). B: Proportion of up-regulated genes (left side) and down-regulated genes (right side) categorized based on different molecular function at GO level 2 (Blast2go).
Figure 3.
Predicted amino acid sequence of two new esterases from LLS (LLSE1 and LLSE4) and LLR (LLRE1 and LLRE4) aligned with a previously reported esterase (AAT09370) from L. lineolaris using Clustal W method (gap penalty: 3.0, gap length penalty: 0.2) of DNAStar MegAlign (Ver. 8).
GenBank accession: LLSE1: JQ964230; LLRE1: JQ964231; LLSE4: JQ964232; LLRE4: JQ964233. Three catalytic center residues (S213, E342, and H468) were boxed. Amino acid substitutions between LLS and LLR are marked with ♦; Amino acid substitutions between AAT09370 and LLE1 are marked with 0. Hyphens represent sequence alignment gaps. Identical residues among all esterases are shaded with black background.
Figure 4.
Comparison and verification of transcriptional levels of two up-regulated (detected by microarray) esterase genes (LLE1 and LLE4) using absolute estimating method in real-time PCR.
LLS: laboratory susceptible strain; Lula CK: field population collected from Lula MS as an aging control; Lula1000: field population collected from Lula MS and selected with 1,000 mg/L acephate (90WP).
Figure 5.
Synergistic effect of two esterase inhibitors, S,S,S-tributylphosphorotrithioate (DEF) and triphenyl phosphate (TPP) on acephate toxicity against a field population and an acephate-selected colony (LLX) of L. lineolaris.
DEF and TPP were used at 1%; Field population from Leland was treated with acephate at 72 mg/L and acephate-selected colony (LLX) was treated with acephate at 144 mg/L.
Figure 6.
Correlation of survival rates between acephate-treated (OrRate) and imidacloprid-treated (ImRate) L. lineolaris collected in September, 2011 in northwest Mississippi.