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Burkholderia insecticola triggers midgut closure in the bean bug Riptortus pedestris to prevent secondary bacterial infections of midgut crypts


In addition to abiotic triggers, biotic factors such as microbial symbionts can alter development of multicellular organisms. Symbiont-mediated morphogenesis is well-investigated in plants and marine invertebrates but rarely in insects despite the enormous diversity of insect-microbe symbioses. The bean bug Riptortus pedestris is associated with Burkholderia insecticola which are acquired from the environmental soil and housed in midgut crypts. To sort symbionts from soil microbiota, the bean bug develops a specific organ called the “constricted region” (CR), a narrow and symbiont-selective channel, located in the midgut immediately upstream of the crypt-bearing region. In this study, inoculation of fluorescent protein-labeled symbionts followed by spatiotemporal microscopic observations revealed that after the initial passage of symbionts through the CR, it closes within 12–18 h, blocking any potential subsequent infection events. The “midgut closure” developmental response was irreversible, even after symbiont removal from the crypts by antibiotics. It never occurred in aposymbiotic insects, nor in insects infected with nonsymbiotic bacteria or B. insecticola mutants unable to cross the CR. However, species of the genus Burkholderia and its outgroup Pandoraea that can pass the CR and partially colonize the midgut crypts induce the morphological alteration, suggesting that the molecular trigger signaling the midgut closure is conserved in this bacterial lineage. We propose that this drastic and quick alteration of the midgut morphology in response to symbiont infection is a mechanism for stabilizing the insect-microbe gut symbiosis and contributes to host-symbiont specificity in a symbiosis without vertical transmission.

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Fig. 1: Morphological alteration of the constricted region and M4B during symbiont colonization in 2nd instar nymphs of R. pedestris.
Fig. 2: Closing of the constricted region.
Fig. 3: The closure of the constricted region and M4B during symbiont colonization in 3rd instar nymphs of R. pedestris.
Fig. 4: The midgut closure prevents subsequent infections.
Fig. 5: The midgut closure is irreversible.
Fig. 6: Midgut closure requires passage through the constricted region and crypt colonization.


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We thank H. Ooi (AIST) for technical assistance. Part of the fluorescence microscopy was performed at the Imagerie-Gif facilities ( This study was supported by the JSPS-CNRS Bilateral Open Partnership Joint Research Project to YK and PM, the Ministry of Education, Culture, Sports, Science and Technology (MEXT) KAKENHI to YK (18KK0211), and the JSPS Research Fellowship for Young Scientists to TO (20170267 and 19J01106) and SJ (201911493).

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YK, TO, and PM designed the study. YK, TO, and SJ performed inoculation tests, antibiotic treatments, and microscopy observations. TO and PM measured morphological change of gut parts and bottleneck of symbiont infection. YK and PM wrote the paper.

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Correspondence to Yoshitomo Kikuchi.

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Kikuchi, Y., Ohbayashi, T., Jang, S. et al. Burkholderia insecticola triggers midgut closure in the bean bug Riptortus pedestris to prevent secondary bacterial infections of midgut crypts. ISME J 14, 1627–1638 (2020).

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