Diagnostic PCR detection of the symbiotic bacteria from dissected tissues of acanthosomatid stinkbugs. The results of Elasmostethus humeralis (A and B), Elasmostethus nubilus (C and D), and Elasmucha putoni (E and F) are shown. (A, C, and E) Detection of mitochondrial COI gene of the host insects. (B, D, and F) Detection of 16S rRNA gene of the symbiotic bacteria. Lane 1, head; lane 2, flight muscle; lane 3, foregut; lane 4, midgut first section; lane 5, midgut second section; lane 6, midgut third section; lane 7, midgut fourth section with crypts; lane 8, hindgut; lane 9, abdominal tip containing the lubricating organ; lane 10, ovary; lane 11, egg before hatching; lane 12, no template control; lane 13, positive control (template DNA from midgut crypts of E. humeralis TK used for phylogenetic analyses); lane M, DNA size markers (1500, 1000, 900, 800, 700, 600, 500, 400, 300, and 200 bp from top to bottom).