Diversity of proteobacterial endosymbionts in hemlock woolly adelgid (Adelges tsugae) (Hemiptera: Adelgidae) from its native and introduced range
Version of Record online: 4 MAR 2013
© 2013 John Wiley & Sons Ltd and Society for Applied Microbiology
Thematic Issue: Invertebrate-Microbe Interactions
Volume 15, Issue 7, pages 2043–2062, July 2013
How to Cite
von Dohlen, C. D., Spaulding, U., Shields, K., Havill, N. P., Rosa, C. and Hoover, K. (2013), Diversity of proteobacterial endosymbionts in hemlock woolly adelgid (Adelges tsugae) (Hemiptera: Adelgidae) from its native and introduced range. Environmental Microbiology, 15: 2043–2062. doi: 10.1111/1462-2920.12102
- Issue online: 4 JUL 2013
- Version of Record online: 4 MAR 2013
- Accepted manuscript online: 30 JAN 2013 04:43AM EST
- Manuscript Accepted: 23 JAN 2013
- Manuscript Revised: 21 DEC 2012
- Manuscript Received: 31 AUG 2012
- USDA-FS Cooperative Agreement. Grant Number: #05-CA-11242343-026
- Utah Agricultural Experiment Station. Grant Number: #8463
Table S1. Collection and voucher information for samples used in this study. All host genera are Tsuga.
Table S2. Primers and probes used in this study.
Fig. S1. Maximum-parsimony phylogram of Gammaproteobacteria GS-B (‘Ca. Serratia symbiotica’) sequences amplified from Japanese T. sieboldii populations from the island of Honshu and from eastern North American populations; all other samples yielded no amplifications of that phylotype. Numbers above branches indicate inferred substitutions along the branch. GenBank accessions are indicated in brackets.
Fig. S2. Laser-scanning confocal images from FISH with the general eubacterial probe 1507r (labelled with A-568).
A. Japanese sample from T. sieboldii first-instar stage, showing probed bacteria (in red) in both the body cavity and salivary glands. Inset: high magnification of a probed salivary gland.
B. Same sample as in A, but different insect; no differentiated bacteriome is visible.
C. Japanese sample from T. sieboldii, fourth-instar or adult stage, showing probed bacteria in the haemocoel and the central region of a bacteriome; insets: high magnification of bacteriome-dwelling bacteria and haemocoel bacteria.
D. Western North America sample from T. heterophylla, third-instar stage cross-section, showing probed rod-shaped bacteria (in red) in the haemocoel and completely filling several bacteriocytes.
Green = autofluorescence from insect tissue. b, bacteriome, s, salivary gland, h, haemocoel, n, bacteriocyte nucleus.
Fig. S3. Laser-scanning confocal images of A. tsugae after FISH with the GS-A-specific probe GANA 1118r (labelled with A-568), indicating GS-A (Pseudomonas adelgestsugas; in red) are found only extracellularly in the haemocoel.
A. Japanese sample from T. sieboldii (first instar) showing rod-shaped bacteria in the posterior body cavity.
B. Western North America sample from T. heterophylla (second instar).
C. Japanese sample from T. sieboldii egg showing bacteria free in the posterior cavity and clustered around developing bacteriome (arrows).
D. Japanese sample from T. diversifolia egg, showing bacteria in the posterior region. Green = autofluorescence from insect tissue. b, bacteriome; h, haemocoel.
Fig. S4. Laser-scanning confocal images of A. tsugae after FISH with the GS-C-specific probe GamC440r (labelled with A-568), indicating GS-C (‘Ca. Annandia adelgestsuga’; in red) are harboured in bacteriocytes, are transmitted to eggs, and invade salivary glands in early instars.
A–C, samples from Japan.
A. Sample from T. sieboldii, egg stage, showing ‘Ca. A. adelgestsuga’ clustered in two regions of one pole; not all bacteria appear to be contained within discrete bacteriocytes at this stage (arrow).
B. Sample from T. diversifolia, first-instar stage showing ‘Ca. A. adelgestsuga’ in both bacteriome and salivary gland.
C. Sample from T. diversifolia, first-instar stage, showing ‘Ca. A. adelgestsuga’ in bacteriocytes.
D–F, samples from North America.
D. Sample from eastern North America, second-instar stage, showing ‘Ca. A. adelgestsuga’ in most areas of the bacteriome; some areas are unprobed (arrow).
E. Sample from eastern North America, third-instar stage showing ‘Ca. A. adelgestsuga’ in most areas of the bacteriome; other areas are unprobed (arrows).
F. Sample from western North America, first-instar stage, showing ‘Ca. A. adelgestsuga’ in both bacteriome and salivary gland (arrow).
Green = autofluorescence from insect tissue. b, bacteriocyte/bacteriome, s, salivary gland.
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