| Apis mellifera and Apis cerana are widespread and raised in China, they are the most important pollinators, providing extremely important economic value. It has been reported that the gut of honeybees harbors a large number of microbiota,, and these microbiota play an important role in host’s development, nutrition, food digestion, immune response and even pathogen protection. The biological genetic variation and adaptation mechanism under the varialble environment can provide a good special gene pool for gut microbiota in A. cerana. We investigated the gut microbial composition of honey bees in China, by surveying 193 individual workers in two species, using high throughput sequencing of 16 S r RNA amplicons. In particular, we addressed the extent of variation in gut communities within and between host species and whether such variation is associated with geographic, castes, or environmental factors. Our results showed:(1)The Gut microbiota type of A. mellifera and A. cerana in China.As reported in A. mellifera from other conuntries, adult workers of A. cerana harbor a characteristic gut microbiota consisting of eight distinct bacterial phylotypes. The four dominant phyla included Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria and Proteobacteria and Firmicutes were the most dominant phyla. The four dominant genera were Lactobacillus, Gilliamella, Snodgrassella and Bifidobacterium.(2)The Comparison of bacterial community between A. cerana and A. mellifera.We compared the variation in gut communities between A. creana and A. mellifera from 5 different provinces(Hainan, Shandong, Gansu, Beijing and Jilin) in the same conditions(area, season and nectar) based on the PCA, PCo A and PLS-DA plots analysis, the results indicated there was a distinct comparison of gut bacterial communities between A. creana and A. mellifera.The abundance of Gilliamella(P=0.024), Bifidobacterium(P=0.003), Flavobacteriaceae*(P<0.001), Acetobacteraceae*(P<0.001) and Bartonella(P<0.001) in A. creana were significant difference than that in A. mellifera. But the richness of the Lactobacillus and Snodgrassella were no significant diference between these two species.(3)The Comparison of bacterial community among queen, worker and drone in A. cerana.We compared the gut microbiota of queen, drone and worker in the same apiary and different apiary based on the PCo A and PLS-DA plots methods, the results showed that the community compositions of the gut microbiota were clearly separated among queen, drone and worker no matter in the same or different apiary. In the genus level, the core bacterial of Gilliamella, Bifidobacterium and Flavobacteriaceae were not significant(P=0.333, 0.057 and 0.404 resepectively) among them. The richness of the Lactobacillus in queen and worker was significant higher than that in male(P=0.003); But the Snodgrassella in drone was significant higher than that in queen(P<0.001)(4)Dynamic of characteristic gut bacteria in A. cerana across different life stageThe bacterial community structure was quite different during the development stages of A. cerana workers. Based on the Mi Seq sequence data we found the bacterial community separated distinctly among the day1, 5 and 10. With the increase of age, the overlapping phenomenon occurred from days 15-30, and the bacterial community of days 30 was easy separated with other days, but not obvious. Using quantitative PCR we characterized four gut bacterial(Bifidobacterium, S. alvi, G. apicola, and Lactobacillus) communities across the life cycle of A. cerana from larvae to workers. Our results indicate that the presence and quantity of these four bacteria were low on day 1, increased rapidly after days 5, and then peaked during days 10-20. They stabilized from days 20-25 or days 25-30, then dropped to a low level at day 30.(5)The Comparison of bacterial community among different season of A. cerana.In order to characterize the influence of the season on the gut microbial diversity in A. creana, we analysed the worker bees from the same colony in spring, summer and autumn during the main nectar flow. We also compared the bacterial community in the different conlonies among different provinces and seasons. For the same colony, the results illustrated a distinct comparison of gut microbiota for all of 18 bee samples, this finding indicated that the season was one of factors influencing these variation of the gut bacterial community in A. cerana. For the core bacteria, the Lactobacillus, Gilliamella, Snodgrassella and Flavobacteriaceae kept stable during different seasons, only the Bifidobacterium was significant(P= 0.030) higher in autumn than in spring. For the different colonies among different locations, our results found that the gut microbial comparison was overlap in the same season but in different location, thus we could conclude that the gut microbial communities were affected by season change of diets, and the geographic area was not mainly influence factor.(6)The Comparison of bacterial community among different hive type of A. cerana.We collected the bee samples from the movable frame hive and unmovable hive, based on the PCA and PLS-DA plots analysis, the results revealed the structure of the communities in these two types were not clearly separated from each other, indicated the hive type may not influence the microbial communities of A. creana individuals.(7)The Heatmap analysis of bacterial community for all groupsWe built the Heatmap based on the genus and OUT level, and found all groups(species, caste, seasons, developmental stage and hive type) were cluster into two distinct community types. The present study is the first report on Gut microbial diversity of honey bees in China based on the High throughput sequencing technologies. And we found that the adult workers of A. cerana harbored a characteristic gut microbiota consisting of eight distinct bacterial phylotypes which were the same as reported in A. mellifera from other conuntries. Microbial composition of the A. cerana gut can be influenced by multi factors, including species, caste, seasons, developmental stage and hive type. We discovered that gut bacterial communities of A. cerana and A. mellifera were cluster into two distinct community types, based on composition at the genus and OUT level, and this type was not affected by species, caste, seasons, developmental stage and hive type. |
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