Diversity Analysis Of Bacterial Community Structure In Different Developmental Stages Of Andraca Bipunctata And Preliminary Analysis On The Metabolism Of Caffeine

Andraca bipunctata Walker,who is a lepidopteran pest in tea plantation.In recent years,its damage has become more and more extensively and seriously,and it has gradually become a potential major pest.Microorganisms often play an important role in the growth and development of many insect species.A.bipunctata harbor many bacterial species that may have important physiological roles in host nutrition,detoxification,immunity and protection.So far,research on A.bipunctata has mainly focused on pesticide control,and the structure and dynamics of gut-associated bacterial communities have not been understood,nor has the ability to metabolize caffeine.In this study,we performed high-throughput sequencing of the V3-V4 hypervariable region of the 16 S r RNA gene of the tea silkworm at different developmental stages,and analyzed the diversity of the gut microbiota in the whole A.bipunctata developmental stage under laboratory feeding conditions.and abundance.And by using high performance liquid chromatography and bacterial colony transplantation,the ability of tea silkworm to tolerate caffeine was understood.The main research results of this paper are as follows: Taking A.bipunctata eggs,larvae(newly hatched larvae to fifth instar larvae),pupa and adult worms and four different stages and nine different instars as samples,a total of48 phyla,87 classes,191 Order,383 families,880 genera were identified.Proteobacteria and Firmicutes dominated the microbial community structure in the whole life cycle of A.bipunctata.The relative abundance of the two in all stages of A.bipunctata larvae was above 64.72%,and the highest could reach98.78%.showing a negative correlation.The colony structure and diversity of intestinal flora will change with different developmental stages.At the genus level,the relative abundance of the genus Acinetobacter(12.98%)at the egg stage is the highest;The highest relative abundance in the larval stage was Sphingomonas(6.20%);the highest relative abundance in the first instar larvae was Pseudomonas(26.78%);the second instar larvae(89.54%)Enterococcus has the highest relative abundance in the third instar larvae(94.71%),fifth instar larvae(41.60%)and adult(38.03%);the fourth instar larvae has the highest relative abundance in Staphylococcus(24.73%);the highest relative abundance in pupal stage was Methylobacterium(22.70%).Among them,Enterococcus bacteria exist in each instar,and show dynamic changes with the different developmental stages of A.bipunctata,which are the core dominant bacteria for the growth and development of A.bipunctata.The body fluid of A.bipunctata has the ability to metabolize caffeine,and the degradation rate in the body fluid of A.bipunctata is 87.5% within one hour.This result was verified by in vitro experiments.The degradation rate within one hour was 59.79%,and the intestinal flora also Can metabolize caffeine without significant effect.The results of this study will help to better understand the correlation between gut bacteria and A.bipunctata biology,and provide reference and research for the development of new insect control strategies in the future.Understanding the correlation of A.bipunctata metabolism to caffeine will provide an important theoretical basis for future research on the mechanism of A.bipunctata metabolism of caffeine.