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Gammaproteobacteria as essential primary symbionts in the striped shield bug, Graphosoma Lineatum (Hemiptera: Pentatomidae)

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ABSTRACT

Many members of suborder Heteroptra harbor heritable symbiotic bacteria. Here we characterize the gut symbiotic bacterium in Graphosoma lineatum (Hemiptera: Pentatomidae) by using molecular phylogeny, real-time PCR analysis as well as light and electron microscopy observations. The microscopy observations revealed the presence of a large number of rod-shaped bacterial cells in the crypts. A very high prevalence (98 to 100%) of the symbiont infection was found in the insect populations that strongly supports an intimate association between these two organisms. Real-time PCR analysis also showed that the Gammaproteobacteria dominated the crypts. The sequences of 16sr RNA and groEL genes of symbiont showed high levels of similarity (93 to 95%) to Pantoea agglomeranse and Erwinia herbicola Gammaproteobacteria. Phylogenetic analyses placed G. lineatum symbiont in a well-defined branch, divergent from other stink bug bacterial symbionts. Co-evolutionary analysis showed lack of host-symbiont phylogenetic congruence. Surface sterilization of eggs resulted in increased pre-adult stage in the offspring (aposymbionts) in comparison to the normal. Also, fecundity, longevity, and adult stage were significantly decreased in the aposymbionts. Therefore, it seems that the symbiont might play a vital function in the host biology, in which host optimal development depends on the symbiont.

No MeSH data available.


Amplification of conserved genes of the symbiont and G. lineatum.(a) Detection of symbiotic bacteria with 16S rRNA gene in adult individuals. (b) COI gene of the G. lineatum. (c) Relative quantification of the bacterial symbionts in different developmental stages of G. lineatum. (d) Composition of bacteria within the V4. (e) Relative quantification of the bacterial symbionts in different developmental stages of G.lineatum. c-: negative control [distilled water], c+: positive control [DNA from midgut crypts of G. lineatum that harbored symbionts].
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f3: Amplification of conserved genes of the symbiont and G. lineatum.(a) Detection of symbiotic bacteria with 16S rRNA gene in adult individuals. (b) COI gene of the G. lineatum. (c) Relative quantification of the bacterial symbionts in different developmental stages of G. lineatum. (d) Composition of bacteria within the V4. (e) Relative quantification of the bacterial symbionts in different developmental stages of G.lineatum. c-: negative control [distilled water], c+: positive control [DNA from midgut crypts of G. lineatum that harbored symbionts].

Mentions: To evaluate the prevalence of the bacterial symbiont in G. lineatum, a total of 100 adult insects and 10 individuals of each nymphal stage were subjected to PCR symbiont detection based on 16S rRNA gene. COI gene of insects was also amplified by using specific primers. Our results showed that nearly all (98–100%) the adult insects harbored the bacterial symbiont (Fig. 3a). The COI genes of adult insects were also amplified and sequenced (Fig. 3b).


Gammaproteobacteria as essential primary symbionts in the striped shield bug, Graphosoma Lineatum (Hemiptera: Pentatomidae)
Amplification of conserved genes of the symbiont and G. lineatum.(a) Detection of symbiotic bacteria with 16S rRNA gene in adult individuals. (b) COI gene of the G. lineatum. (c) Relative quantification of the bacterial symbionts in different developmental stages of G. lineatum. (d) Composition of bacteria within the V4. (e) Relative quantification of the bacterial symbionts in different developmental stages of G.lineatum. c-: negative control [distilled water], c+: positive control [DNA from midgut crypts of G. lineatum that harbored symbionts].
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5016839&req=5

f3: Amplification of conserved genes of the symbiont and G. lineatum.(a) Detection of symbiotic bacteria with 16S rRNA gene in adult individuals. (b) COI gene of the G. lineatum. (c) Relative quantification of the bacterial symbionts in different developmental stages of G. lineatum. (d) Composition of bacteria within the V4. (e) Relative quantification of the bacterial symbionts in different developmental stages of G.lineatum. c-: negative control [distilled water], c+: positive control [DNA from midgut crypts of G. lineatum that harbored symbionts].
Mentions: To evaluate the prevalence of the bacterial symbiont in G. lineatum, a total of 100 adult insects and 10 individuals of each nymphal stage were subjected to PCR symbiont detection based on 16S rRNA gene. COI gene of insects was also amplified by using specific primers. Our results showed that nearly all (98–100%) the adult insects harbored the bacterial symbiont (Fig. 3a). The COI genes of adult insects were also amplified and sequenced (Fig. 3b).

View Article: PubMed Central - PubMed

ABSTRACT

Many members of suborder Heteroptra harbor heritable symbiotic bacteria. Here we characterize the gut symbiotic bacterium in Graphosoma lineatum (Hemiptera: Pentatomidae) by using molecular phylogeny, real-time PCR analysis as well as light and electron microscopy observations. The microscopy observations revealed the presence of a large number of rod-shaped bacterial cells in the crypts. A very high prevalence (98 to 100%) of the symbiont infection was found in the insect populations that strongly supports an intimate association between these two organisms. Real-time PCR analysis also showed that the Gammaproteobacteria dominated the crypts. The sequences of 16sr RNA and groEL genes of symbiont showed high levels of similarity (93 to 95%) to Pantoea agglomeranse and Erwinia herbicola Gammaproteobacteria. Phylogenetic analyses placed G. lineatum symbiont in a well-defined branch, divergent from other stink bug bacterial symbionts. Co-evolutionary analysis showed lack of host-symbiont phylogenetic congruence. Surface sterilization of eggs resulted in increased pre-adult stage in the offspring (aposymbionts) in comparison to the normal. Also, fecundity, longevity, and adult stage were significantly decreased in the aposymbionts. Therefore, it seems that the symbiont might play a vital function in the host biology, in which host optimal development depends on the symbiont.

No MeSH data available.