Table 1.
ABC subfamilies in L. hesperus and ten other species.
Table 2.
Bioinformatics analysis of putative LhABC transporters.
Figure 1.
Phylogenetic analysis of ABCA transporters from L. hesperus and five metazoan species.
Putative L. hesperus ABCA sequences and full-length ABCA proteins from five additional species were aligned using MUSCLE [159] and analyzed using the FastTree2 approximate likelihood method [160]. Numbers at the branch point of each node represent support values. Species abbreviations and color coding are: Bm, Bombyx mori (teal); Dm, Drosophila melanogaster (pink); Hs, Homo sapiens (blue); Lh, Lygus hesperus (green); Tc, Tribolium castaneum (red); and Tu, Tetranychus urticae (orange). Accession numbers are indicated in parentheses. The scale bar represents 0.7 amino acid substitutions per site. A full listing of the accession numbers for the five metazoan sequences is available in Table S11. LhABC transporter sequences are available in Table S12.
Figure 2.
Phylogenetic analysis of ABCB transporters from L. hesperus and five metazoan species.
Clades corresponding to full-transporters and half-transporters are shaded light yellow and light red, respectively. The scale bar represents 0.4 amino acid substitutions per site. Analyses, abbreviations, and color-coding are as in Figure 1.
Figure 3.
Phylogenetic analysis of ABCC transporters from L. hesperus and five metazoan species.
The scale bar represents 0.6 amino acid substitutions per site. Clades corresponding to SUR-like sequences and CFTR are shaded light yellow and light red, respectively. Analyses, abbreviations, and color-coding are as in Figure 1.
Figure 4.
Phylogenetic analysis of ABCD, ABCE, and ABCF transporters from L. hesperus and five metazoan species.
Clades corresponding to the three subfamilies are shaded light red (ABCE), light yellow (ABCD), and green (ABCF). The scale bar represents 0.7 amino acid substitutions per site. Analyses, abbreviations, and color-coding are as in Figure 1.
Figure 5.
Phylogenetic analysis of ABCG transporters from L. hesperus and five metazoan species.
Clades corresponding to transporters involved in eye pigmentation have been shaded light red (scarlet), light yellow (brown), and green (white). The scale bar represents 0.9 amino acid substitutions per site. Analyses, abbreviations, and color-coding are as in Figure 1.
Figure 6.
Phylogenetic analysis of ABCH transporters from L. hesperus and four metazoan species.
The scale bar represents 0.5 amino acid substitutions per site. Analyses, abbreviations, and color-coding are as in Figure 1. Because the ABCH subfamily is largely restricted to the arthropod lineage, no representative sequences for H. sapiens are available and thus the analyses were performed using only arthropod sequences. L. hesperus ABCH sequences that have undergone gene expansion are shaded light red.
Figure 7.
Transcriptional expression profile of 13 LhABC transcripts.
A) Developmental profile. Expression profile of 13 LhABC transporters in eggs through 12-day-old adults was examined by end-point PCR using primers designed to amplify a ∼500 bp fragment of each transcript. Abbreviations: E, eggs; 1st, first instars; 2nd, second instars; 3rd, third instars; 4th, fourth instars; 5th-E, early stadium fifth instars; 5th-L, late stadium fifth instars; I, reproductively immature 1-day-old mixed sex adults; M, mature 12-day-old mixed sex adults. Products were analyzed on 1.5% agarose gels and stained with SYBR Safe. Actin was used as a positive control. Leftmost image - Representative gel image. For clarity, the negative image is shown. Rightmost image – Visual aid depicting semi-quantitative analysis of amplimers of interest compared to actin in the representative gel. Relative to the actin amplimer of each developmental stage, cells with amplimer intensity ≥50% are indicated in green, while cells <50% are indicated in yellow. Red cells indicate no detectable amplimer. Numbers inside individual cells denote the percentage of amplimer intensity compared to actin. The primer set used for LhABCC1 profiling amplifies a shared region of LhABCC1A and LhABCC1B. B) Adult tissue profile. The expression profile of the 13 LhABC transcripts was examined as above in adult body segments and various abdominal tissues prepared from 7-day-old adults. Abbreviations: H, head; T, thorax; A, abdomen; E, epidermis, MG, midgut; HG, hindgut; MT, Malpighian tubules; O, ovary; SD, seminal depository; AG, accessory glands (lateral and medial); T, testis. Top image - Representative gel image. For clarity, the negative image is shown. Lower image – Visual aid depicting semi-quantitative analysis of amplimers of interest compared to actin in the representative gel. Color shading is as above but relative to the intensity of actin amplimers within each respective tissue. C) Three-way Venn diagram comparing the transcriptional expression profile of LhABC transcripts ubiquitously expressed throughout L. hesperus development (blue) with those expressed in female (red) and male (green) tissues.
Table 3.
Oligonucleotide primers used in expression profiling and cloning.