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Study On The Diversity Of Symbionts In Different Wing Morphs Of Brown Planthopper,Nilaparvata Lugens(St(?)l)

Posted on:2022-10-14Degree:MasterType:Thesis
Country:ChinaCandidate:J X FanFull Text:PDF
GTID:2543306347499674Subject:Biochemistry and Molecular Biology
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Brown planthopper(BPH),Nilaparvata lugens(St(?)l)is a severely destructive migratory monophagous rice pest.Its control is a huge difficult challenge in pest management.The structure of microbial community has a highly important impact on the internal environment of insects and is an indispensable content that reflects the internal environment of insects.Wing dimorphism is a strategy for insects to balance flight ability and reproductive ability.In order to clarify the diversity of symbionts in the different wing morphs of BPH,In this research,female BPHs with different wing morphs were used to study the changes of symbionts in diversity with different wing morphs at different development times.A metagenomic library was constructed based on 16 S r RNA and ITS sequences,and the species diversity and community structure of symbionts in the gut and fat body of different wing morphs of BPHs were compared and analyzed.The specific research results are as follows:1 、 The total number of symbiont of different wing morphs at different developmental stages was detected by fluorescence quantitative PCR technology,and it was found that the number of unit gut symbiotic bacteria of different wing morphs from the 1st day to the 4th day of emerged female adults decreased first and then increased.The number of symbiotic bacteria in the unit gut of the macropterous BPH was significantly higher than that of the brachypterous BPH on the 1st and 2nd days of emergence.There was no significant difference in the number of symbiotic bacteria unit gut on the 4d-emerged female adults.The number of symbiotic bacteria unit fat body of different wing morphs gradually increases from day 1 to day 4 of emergence.The number of symbiotic bacteria unit fat body of brachypterous BPH was significantly higher than that of macropterous BPH on the 1st and 2nd days of emergence.There was no significant difference in the number of symbiotic bacteria unit fat body on the4 th day of emergence.On the 1st,2nd,and 4th days of emergence,the number of symbiotic fungi in the unit gut of different wing morphs BPH was gradually increased and the unit fat body increased first and then decreased.The number of symbiotic fungi in the gut and fat body of the brachypterous BPH on the 2nd and 4th days of emergence was significantly higher than that of macropterous BPH.2、A metagenomic library was constructed based on the 16 S r RNA sequence,and the gut and fat body symbiotic bacteria of different wing morphs of BPHs were sequenced by high-throughput sequencing.At a similarity level of 97%,OTU clustering was performed on the optimized sequence of each sample.The results showed that he gut bacteria of macropterous BPH(brachypterous BPH)obtained a total of 345(293)OTUs and 15(16)phyla,30(32)classes,78(75)orders,137(115)families and 245(184)genera of bacteria were found.The fat body bacteria of macropterous BPH(brachypterous BPH)obtained a total of 150(183)OTUs and 13(11)phyla,20(18)classes,43(48)orders,71(77)families and 111(121)genera of bacteria were found.The dominant bacteria phyla of gut and fat body of the two wing morph BPH were Proteobacteria.The dominant bacteria at the genus level in the gut and fat body of the macropterous BPH and the brachypterous BPH were Serratia and Acinetobacter,respectively.According to the Ace index,Chao index,Shannon index and Simpson index of Alpha diversity,the species richness of the gut bacteria of the brachypterous BPH was higher than that of the macropterous BPH,the species diversity of the gut bacteria of the macropterous BPH was significantly higher than that of the brachypterous BPH,and the species richness and diversity of the fat body bacteria of the brachypterous BPH were both higher than that of macropterous BPH.According to the COG function prediction of the gut and fat body bacteria of the BPH,the main pathways involved in the gut and fat body bacteria of the macropterous BPH and the brachypterous BPH were amino acid transport and metabolism.In addition,the abundance of the bacteria of the brachypterous BPH was mostly higher than that of the macropterous BPH.In addition,the bacterial abundance of the macropterous BPH in the extracellular structure was higher than that of the brachypterous BPH.The results of the study preliminarily indicated that there were differences in the symbiotic bacterial diversity and colony composition of the gut and fat body of the different wing morphs of BPH.3、A metagenomic library was constructed based on the ITS sequence,and the gut and fat body symbiotic fungi of different wing morphs of BPHs were sequenced by high-throughput sequencing.At a similarity level of 97%,OTU clustering was performed on the optimized sequence of each sample.The results showed that the gut fungi of macropterous BPH(brachypterous BPH)obtained a total of 63(165)OTUs and 5(9)phyla,13(26)classes,17(42)orders,25(62)families and 30(73)genera of fungi were found;The fat body fungi of macropterous BPH(brachypterous BPH)obtained a total of 60(57)OTUs and 4(5)phyla,13(15)classes,16(17)orders,23(24)families and 28(29)genera of fungi were found.The dominant fungi phyla of gut and fat body of the two wing morph BPH were Ascomycota.The dominant fungi at the genus level of gut and fat body of the two wing morph BPH were unclassified_Hypocreales.According to the Ace index,Chao index,Shannon index and Simpson index of Alpha diversity,the species richness as well as diversity of the gut fungi of the brachypterous BPH were both higher than that of the macropterous BPH,and the species richness and diversity of the fat body fungi of the macropterous BPH were both higher than that of brachypterous BPH.According to FUNGuild function analysis,the main nutrient types in the gut and fat body fungi of different winged BPHs were Saprotroph,the main functional groups were Undefined Saprotroph,and the relative abundance of macropterous BPH was higher than the brachypterous BPH.The results of the study preliminarily indicate that the gut and fat body symbiotic fungi of different wing morphs of BPH may have an essential influence on the wing differentiation of BPH.This study provides a theoretical basis for further research on the regulation of the BPH symbiotic bacteria on the host.The results of the study preliminarily indicated that there were differences in the symbiotic fungi diversity and colony composition of the gut and fat body of the different wing morphs of BPH.4、Combining the results of fluorescence quantitative PCR and high-throughput sequencing methods,the differences on the number of dominant symbionts of different wing morphs of BPH were analyzed.The absolute number of Serratia,Brucella and Stenotrophomonas in the gut and fat body of the macropterous BPH were higher than those of the brachypterous BPH.The absolute number of Microbacterium,Staphylococcus and Acinetobacter in the gut and fat body of the brachypterous BPH were higher than that of the macropterous BPH.The absolute number of Sphingommonas,Sphingobacterium as well as Bosea in the gut of the macropterous BPH were higher than that of the brachypterous BPH,but the results of fat bodies showed the opposite trend.The absolute number of Talaromyces and Candida in the gut and fat body of the macropterous BPH were greater than that of the short-winged.The absolute number of unclassified_Hypocreales,Cutaneotrichosporon and unclassified_Fungi in the gut and fat body of the brachypterous BPH were greater than that of the macropterous BPH.The absolute number of Fusarium in the gut of the macropterous BPH was greater than that of the brachypterous BPH.This study enriched the connotation of the symbionts of the BPH and laid a theoretical foundation for the further study of the interaction between BPH and symbionts as well as the function of symbionts in the host.
Keywords/Search Tags:brown planthopper Nilaparvata lugens(St(?)l), wing dimorphism, symbionts, diversity, gut, fat body, high-throughput sequencing
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