Abstract
The plant-phloem-feeding Blissus insularis possesses specialized midgut crypts, which harbor a dense population of the exocellular bacterial symbiont Burkholderia. Most individual B. insularis harbor a single Burkholderia ribotype in their midgut crypts; however, a diverse Burkholderia community exists within a host population. To understand the mechanism underlying the consistent occurrence of various Burkholderia in B. insularis and their specific association, we investigated potential gut symbiont transmission routes. PCR amplification detected a low titer of Burkholderia in adult reproductive tracts; however, fluorescence in situ hybridization assays failed to produce detectable signals in these tracts. Furthermore, no Burkholderia-specific PCR signals were detected in eggs and neonates, suggesting that it is unlikely that B. insularis prenatally transmits gut symbionts via ovarioles. In rearing experiments, most nymphs reared on St. Augustinegrass treated with cultured Burkholderia harbored the cultured Burkholderia strains. Burkholderia was detected in the untreated host grass of B. insularis, and most nymphs reared on untreated grass harbored a Burkholderia ribotype that was closely related to a plant-associated Burkholderia strain. These findings revealed that B. insularis neonates acquired Burkholderia primarily from the environment (i.e., plants and soils), even though the possibility of acquisition via egg surface cannot be excluded. In addition, our study explains how the diverse Burkholderia symbiont community in B. insularis populations can be maintained.
Highlights
In heteropterans, exocellular gut symbiont bacteria can be acquired by aposymbiotic neonates from symbiont-contaminated egg chorion [1,2,3,4], specialized egg capsules [5,6], and symbiontcontaining feces [7,8,9]
Fluorescence in situ hybridization (FISH) analyses using both epifluorescence and confocal microscopy revealed that Burkholderia bacteria colonized the midgut crypts rather than other regions of the digestive tract of B. insularis adults (Fig 1)
Dense populations of Burkholderia were confined to the lumen of crypts, supporting the previous findings that the midgut crypts are the specialized symbiotic organ in B. insularis [15,16]
Summary
Exocellular gut symbiont bacteria can be acquired by aposymbiotic neonates from symbiont-contaminated egg chorion [1,2,3,4], specialized egg capsules [5,6], and symbiontcontaining feces [7,8,9]. These transmission routes rely primarily on the symbiont-infected mother, whose gut symbionts are transmitted postnatally to their offspring. In addition to vertical transmission, transmission via the ambient environment is PLOS ONE | DOI:10.1371/journal.pone.0161699 August 22, 2016
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