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Microbial environment affects innate immunity in two closely related earthworm species Eisenia andrei and Eisenia fetida.

Dvořák J, Mančíková V, Pižl V, Elhottová D, Silerová M, Roubalová R, Skanta F, Procházková P, Bilej M - PLoS ONE (2013)

Bottom Line: While E. andrei lives in a compost and manure, E. fetida can be found in the litter layer in forests.Genomic DNA analyses revealed significantly higher level of fetidin/lysenins (determined using universal primer pairs) in E. andrei compared to E. fetida.It can be hypothesized that E. andrei colonizing compost as a new habitat acquired an evolutionary selection advantage resulting in a higher expression of antimicrobial proteins.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cellular and Molecular Immunology, Institute of Microbiology of the Academy of Sciences of the Czech Republic, v. v. i., Prague, Czech Republic.

ABSTRACT
Survival of earthworms in the environment depends on their ability to recognize and eliminate potential pathogens. This work is aimed to compare the innate defense mechanisms of two closely related earthworm species, Eisenia andrei and Eisenia fetida, that inhabit substantially different ecological niches. While E. andrei lives in a compost and manure, E. fetida can be found in the litter layer in forests. Therefore, the influence of environment-specific microbiota on the immune response of both species was followed. Firstly, a reliable method to discern between E. andrei and E. fetida based on species-specific primers for cytochrome c oxidase I (COI) and stringent PCR conditions was developed. Secondly, to analyze the immunological profile in both earthworm species, the activity and expression of lysozyme, pattern recognition protein CCF, and antimicrobial proteins with hemolytic function, fetidin and lysenins, have been assessed. Whereas, CCF and lysozyme showed only slight differences in the expression and activity, fetidin/lysenins expression as well as the hemolytic activity was considerably higher in E. andrei as compared to E. fetida. The expression of fetidin/lysenins in E. fetida was not affected upon the challenge with compost microbiota, suggesting more substantial changes in the regulation of the gene expression. Genomic DNA analyses revealed significantly higher level of fetidin/lysenins (determined using universal primer pairs) in E. andrei compared to E. fetida. It can be hypothesized that E. andrei colonizing compost as a new habitat acquired an evolutionary selection advantage resulting in a higher expression of antimicrobial proteins.

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Alignment of E. andrei and E. fetida COI sequences.Alignment of E. andrei and E. fetida COI sequences. Oligonucleotides used as discriminating primers for E. andrei or E. fetida COI are underlined or in bold (refer to the Table 1).
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pone-0079257-g001: Alignment of E. andrei and E. fetida COI sequences.Alignment of E. andrei and E. fetida COI sequences. Oligonucleotides used as discriminating primers for E. andrei or E. fetida COI are underlined or in bold (refer to the Table 1).

Mentions: Molecular differentiation on a basis of polymorphism of mitochondrial gene for COI is widely used for discrimination of closely related animal species. It was previously published that nucleotide sequences of COI differ between E. andrei and E. fetida species [15]. Based on these published sequences, pairs of primers specific for both E. andrei and E. fetida COI were designed. Using these primer pairs with E. andrei or E. fetida cDNA as a template in PCR reaction, fragments of COI of 541 bp were obtained and subsequently sequenced. Alignment of these two sequences confirmed the presence of single- or double-nucleotide mismatches dispersed all along these sequences. In order to obtain the full-length sequences of E. andrei and E. fetida COI genes, RACE amplifications of 5’ as well as 3’ ends were performed. Resulting PCR products were cloned and sequenced. Consequently, the whole coding sequences of E. andrei and E. fetida COI with open reading frames of 1542 bp encoding 514 amino acids were obtained (Figure 1) and submitted to the GenBank databasis (E. andrei COI - NCBI: HQ534065, E. fetida COI - NCBI: HQ534066). The alignment of both sequences revealed a high level of homology (80%).


Microbial environment affects innate immunity in two closely related earthworm species Eisenia andrei and Eisenia fetida.

Dvořák J, Mančíková V, Pižl V, Elhottová D, Silerová M, Roubalová R, Skanta F, Procházková P, Bilej M - PLoS ONE (2013)

Alignment of E. andrei and E. fetida COI sequences.Alignment of E. andrei and E. fetida COI sequences. Oligonucleotides used as discriminating primers for E. andrei or E. fetida COI are underlined or in bold (refer to the Table 1).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3815151&req=5

pone-0079257-g001: Alignment of E. andrei and E. fetida COI sequences.Alignment of E. andrei and E. fetida COI sequences. Oligonucleotides used as discriminating primers for E. andrei or E. fetida COI are underlined or in bold (refer to the Table 1).
Mentions: Molecular differentiation on a basis of polymorphism of mitochondrial gene for COI is widely used for discrimination of closely related animal species. It was previously published that nucleotide sequences of COI differ between E. andrei and E. fetida species [15]. Based on these published sequences, pairs of primers specific for both E. andrei and E. fetida COI were designed. Using these primer pairs with E. andrei or E. fetida cDNA as a template in PCR reaction, fragments of COI of 541 bp were obtained and subsequently sequenced. Alignment of these two sequences confirmed the presence of single- or double-nucleotide mismatches dispersed all along these sequences. In order to obtain the full-length sequences of E. andrei and E. fetida COI genes, RACE amplifications of 5’ as well as 3’ ends were performed. Resulting PCR products were cloned and sequenced. Consequently, the whole coding sequences of E. andrei and E. fetida COI with open reading frames of 1542 bp encoding 514 amino acids were obtained (Figure 1) and submitted to the GenBank databasis (E. andrei COI - NCBI: HQ534065, E. fetida COI - NCBI: HQ534066). The alignment of both sequences revealed a high level of homology (80%).

Bottom Line: While E. andrei lives in a compost and manure, E. fetida can be found in the litter layer in forests.Genomic DNA analyses revealed significantly higher level of fetidin/lysenins (determined using universal primer pairs) in E. andrei compared to E. fetida.It can be hypothesized that E. andrei colonizing compost as a new habitat acquired an evolutionary selection advantage resulting in a higher expression of antimicrobial proteins.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cellular and Molecular Immunology, Institute of Microbiology of the Academy of Sciences of the Czech Republic, v. v. i., Prague, Czech Republic.

ABSTRACT
Survival of earthworms in the environment depends on their ability to recognize and eliminate potential pathogens. This work is aimed to compare the innate defense mechanisms of two closely related earthworm species, Eisenia andrei and Eisenia fetida, that inhabit substantially different ecological niches. While E. andrei lives in a compost and manure, E. fetida can be found in the litter layer in forests. Therefore, the influence of environment-specific microbiota on the immune response of both species was followed. Firstly, a reliable method to discern between E. andrei and E. fetida based on species-specific primers for cytochrome c oxidase I (COI) and stringent PCR conditions was developed. Secondly, to analyze the immunological profile in both earthworm species, the activity and expression of lysozyme, pattern recognition protein CCF, and antimicrobial proteins with hemolytic function, fetidin and lysenins, have been assessed. Whereas, CCF and lysozyme showed only slight differences in the expression and activity, fetidin/lysenins expression as well as the hemolytic activity was considerably higher in E. andrei as compared to E. fetida. The expression of fetidin/lysenins in E. fetida was not affected upon the challenge with compost microbiota, suggesting more substantial changes in the regulation of the gene expression. Genomic DNA analyses revealed significantly higher level of fetidin/lysenins (determined using universal primer pairs) in E. andrei compared to E. fetida. It can be hypothesized that E. andrei colonizing compost as a new habitat acquired an evolutionary selection advantage resulting in a higher expression of antimicrobial proteins.

Show MeSH
Related in: MedlinePlus