Figure 1.
Adult Females of the Plataspid Stinkbugs and Their Posterior Midgut
(A and B) M. cribraria; (C and D) B. subaeneus; (E and F) C. parvipictum. Bars, 1 mm. AEM, anterior enlarged midgut section; BEM, brownish enlarged midgut end section; REC, rectum; SCM, swollen crypt-bearing midgut section [28].
Figure 2.
Phylogenetic Placement of the Symbiotic Bacteria from the Plataspid Stinkbugs in the γ-Subclass of the Proteobacteria on the Basis of 16S rRNA Gene Sequences
A total of 1,213 aligned nucleotide sites were subjected to the analysis. A Bayesian tree is shown, while MP tree and ML tree exhibited substantially the same topologies. On each of the nodes, posterior probabilities in the Bayesian analysis are shown above, and bootstrap probabilities (MP analysis/ML analysis) are shown below. Asterisks indicate bootstrap values lower than 50%. In brackets are sequence accession numbers. In parentheses are collection localities for the stinkbug symbionts and host insect names for the other symbionts, respectively.
Figure 3.
Effects of Symbiont Elimination on the Adult Emergence Rate of the Plataspid Stinkbugs
(A) M. punctatissima; (B) M. cribraria; (C) B. subaeneus; (D) C. parvipictum. Means and standard deviations are shown. Open columns, with capsules; filled columns, without capsules. Sample sizes and p values of Wilcoxon test are indicated.
Table 1.
Developmental Time of Stinkbug Nymphs from Experimental Egg Masses with and without Capsules
Figure 4.
Effects of Symbiont Elimination on Adult Body Size and Phenotype of the Plataspid Stinkbugs
(A) Adult females of M. punctatissima (left) and M. cribraria (right) emerged from the control egg masses with capsules (top) and those from the treated egg masses without capsules (bottom). (B) Thorax width of M. punctatissima and (C) thorax width of M. cribraria. Means and standard deviations are shown. Open columns, with capsules; filled columns, without capsules. Sample sizes are labeled on the columns. Asterisks indicate statistically significant differences (median test; *** p < 0.001 after Bonferroni correction).
Figure 5.
Phylogenetic Congruence between the Plataspid Stinkbugs and Their Symbiotic Bacteria
(A) A Bayesian phylogeny of the host insects on the basis of mitochondrial 16S rRNA gene sequences (847 aligned nucleotide sites). In addition to the plataspid stinkbugs, four pentatomomorphan species were examined as out-group taxa. In parentheses are taxonomic affiliations and in brackets are sequence accession numbers.
(B) A Bayesian phylogeny of the plataspid symbionts on the basis of the same data of Figure 2 (16S rRNA gene, 1,213 aligned nucleotide sites). Dots indicate codivergence events inferred by the jungles algorithm [34]. Posterior probabilities in the Bayesian analysis and bootstrap values in the MP/ML analyses are presented as shown in Figure 2.
Table 2.
Relative Rate Tests for Comparing the Molecular Evolutionary Rate of 16S rRNA Gene between the Lineages of Gut Symbionts of Plataspid Stinkbugs, Endocellular Symbionts of Aphids (B. aphidicola), and Their Free-Living Relatives
Figure 6.
PFGE of the Symbiont Genomic DNA Prepared from the Posterior Midgut of a Female Adult of the Plataspid Stinkbugs
Lane 1, M. punctatissima; lane 2, M. cribraria; lane 3, C. parvipictum; lane 4, yeast PFGE marker; lane 5, lambda PFGE marker. Marker sizes are shown on the right side.