| Ants are the social insects which have the largest number and widely distributed on the earth. They have established a close relationship with numerous microorganisms, and these microorganisms played an important role in the trophic evolution and population multiplication of ants. Camponotus is one of the most abundant and successful evolutionary ant genus, and Blochmannia is the widespread obligate intracellular endosymbiotic bacteria in all Camponotus species which can provide nitrogenous substances for host ant(mainly amino acid, vitamin and sterols substances) and enhance the host immune response. Recently studies have revealed that the digestive tract of C. japonicas harbor not only the obligate intracellular endosymbiotic but also many other gut microbes. In order to further reveal the impact of the food and living environment on the gut microbes in Camponotus ants, in this study we use T-RFLP and PCR-DGGE method to compare the gut bacterial composition and diverisity in different development phase from the lab-raised and the field colonies.The results are as follows:1.The T-RFLP analysis results showed that there was no significant differences(p>0.05) in bacterial diversity among larvae, pupae and workers. In comparison, Shannon index of larval stage were highest but that of pupal stage were the lowest. Similarity analysis showed that the similarity coefficients of gut microbes between larvae and workers was 0.53; the similarity coefficients of gut microbes between larvae and pupae was 0.51 and that between larva and pupa was 0.36; while PCA and cluster analysis results showed that there were certain differences in the bacterial community structure of larvae, pupae and workers, but the differences was not significant, and they didn’t form an independent bacterial colonies each other.2. Digestive tract bacteria in different development phase(egg, larva, pupa, worker and queen) from lab-raised colonies were detected by PCR-DGGE and 33 DGGE bands were found. Sequence alignment analysis showed that these bacteria mainly belonged to Proteobacteria, Firmicutes and Crenarchaeota, including eleven genera and two unknown species. Sequence analysis of DGGE bands showed that the dut bacterial diversity indexes were the highest in egg and queen and both contained seven genus of bacteria, five genuses of bacteria were found in larval and pupal stage respectively, and four genuses of bacteria was found in workers. The similarity coefficient was 0.89 between worker and larva, and it is 0.89 beteween worker and pupa, but the similarity coefficient between queen and larva was the least and was only 0.57.3. The gut bacteria of different development stages from the field colonies was detected by PCR-DGGE analysis and 52 DGGE bands were found.The sequecnce blasting results in the GenBank database showed that the detected bacteria belonged to Proteobacteria and Firmicutes and Actinobacteriaone, including 11 genus and 1 species in Acetobacteraceae and 5 other unknown species.There was obvious difference in the detected bacterial species of different development stages, and 5, 8, 4, 2, 3, 6 and 3 bacterial genus were detected from the egg, larva, pupa, big workers, small workers, males and the queen, respectively.4. Comparative analysis of gut bacterial composition among different stages between the lab-raised and the field colonies showed that the gut bacterial diversity in egg, pupa, worker and queen of the field colonies was lower than those of lab-raised colonies, however, the gut bacterial diversity in larvae was higher than that of lab-raised colonies; there was a remarkable difference between lab-raised and the field colonies, but both all contained Blochmannia and Wolbachia, other bacterial species were quite different and this may be caused by the food and the change of environment conditions. |
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