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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.


Phylogenetic placement of the symbiotic bacterium based on the 16S rRNA gene.ML: maximum likelihood method with 100 replications based on GTR + G + I model. NJ: neighbor-joining method with 1000 replications. On the nodes, bootstrap probabilities neighbor-joining/maximum likelihood analysis are shown. Accession numbers of sequences are shown in brackets.
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f4: Phylogenetic placement of the symbiotic bacterium based on the 16S rRNA gene.ML: maximum likelihood method with 100 replications based on GTR + G + I model. NJ: neighbor-joining method with 1000 replications. On the nodes, bootstrap probabilities neighbor-joining/maximum likelihood analysis are shown. Accession numbers of sequences are shown in brackets.

Mentions: BLAST (Basic Local Alignment Search Tool) analyses of 16S rRNA and groEL gene sequences exhibited the most similarity with Erwinia herbicola [AB008150] (with 95% and 92% identity for 16S rRNA and groEL gene, respectively) and Pantoea agglomeranse [LC007456] (with 95% and 93% identity for 16S rRNA and groEL gene, respectively). AT contents were 47.45% for 16S rRNA on 1452 bp and groEL gene AT contents were 47.03% on 1516 bp. The gene sequences were subjected to molecular phylogenetic analyses together with 16S rRNA and groEL gene sequences of other insect symbionts. In phylogenetic trees based on 16S rRNA genes, the G. lineatum symbiont was placed with the bacterial symbionts of Pentatomidae and Scutelleridae bugs in a clade with high bootstrapping values (Fig. 4). In groEL phylogenetic trees of different bacterial symbionts, the G. lineatum symbiont grouped with bacterial symbionts of Pentatomidae, as well (Fig. 5). The phylogenies of the insect host deduced based on COI sequence data placed G. lineatum within Pentatomidae clade with high bootstrap support value (Fig. 6).


Gammaproteobacteria as essential primary symbionts in the striped shield bug, Graphosoma Lineatum (Hemiptera: Pentatomidae)
Phylogenetic placement of the symbiotic bacterium based on the 16S rRNA gene.ML: maximum likelihood method with 100 replications based on GTR + G + I model. NJ: neighbor-joining method with 1000 replications. On the nodes, bootstrap probabilities neighbor-joining/maximum likelihood analysis are shown. Accession numbers of sequences are shown in brackets.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f4: Phylogenetic placement of the symbiotic bacterium based on the 16S rRNA gene.ML: maximum likelihood method with 100 replications based on GTR + G + I model. NJ: neighbor-joining method with 1000 replications. On the nodes, bootstrap probabilities neighbor-joining/maximum likelihood analysis are shown. Accession numbers of sequences are shown in brackets.
Mentions: BLAST (Basic Local Alignment Search Tool) analyses of 16S rRNA and groEL gene sequences exhibited the most similarity with Erwinia herbicola [AB008150] (with 95% and 92% identity for 16S rRNA and groEL gene, respectively) and Pantoea agglomeranse [LC007456] (with 95% and 93% identity for 16S rRNA and groEL gene, respectively). AT contents were 47.45% for 16S rRNA on 1452 bp and groEL gene AT contents were 47.03% on 1516 bp. The gene sequences were subjected to molecular phylogenetic analyses together with 16S rRNA and groEL gene sequences of other insect symbionts. In phylogenetic trees based on 16S rRNA genes, the G. lineatum symbiont was placed with the bacterial symbionts of Pentatomidae and Scutelleridae bugs in a clade with high bootstrapping values (Fig. 4). In groEL phylogenetic trees of different bacterial symbionts, the G. lineatum symbiont grouped with bacterial symbionts of Pentatomidae, as well (Fig. 5). The phylogenies of the insect host deduced based on COI sequence data placed G. lineatum within Pentatomidae clade with high bootstrap support value (Fig. 6).

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.