Annotation And Functional Researches Of The Amino Acid Transporter Family Gene Of Bemisia Tabaci And Annotation Of The Carboxylesterases Family Gene Of Bemisia Tabaci

The whitefly(Bemisia tabaci)is one of the most devastating pests for agricultural crops and ornamentals all over the world.The whitefly has the broadest hosts,which is estimated to have 74 families and more than 500 species.B.tabaci can greatly reduce crop yields by feeding on phloem,excreting honey dew,and transmitting plant viruses.Amino acids are essential nutrients and are closely related to the life activities of organisms.The amino acid are essential for protein synthesis,nerve transmission,regulation of cell growth,nitrogen metabolism,hormone metabolism,and production of metabolic energy.Nevertheless,many animals are incapable of synthesizing 10 amino acids that are required for normal growth and development;these amino acids are typically acquired through diet.The amino acid transporter(AAT)genes,which form a large family in organisms,take part in transport of amino acids into and out of cells and play important nutritional roles in insects.Carboxylesterases are important super-family detoxification system,and widely found in microorganisms,insects,plants and animals and other organisms.In the study of insects,the carboxylesterases have been confirmed that take parts in the metabolic detoxification of various insecticides,including carbamates,pyrethroids and organophosphates,and cost insects resistant to a variety of chemical insecticides.Genome-wide and transcriptome-wide investigation of the amino acid/polyamine/organocation(APC)and the eukaryotic-specific amino acid/auxin permease(AAAP)genes were performed in the B.tabaci B.The characteristics of B.tabaci B APC and AAAP family genes were identified,and the function of B.tabaci-specific AAAP family expansion genes BtAAAP15 and BtAAAP21 were researched using RNAi.The results provide a foundation for future functional analysis of APC and AAAP genes in B.tabaci.In addition,the annotation of carboxylesterase were performed in the B.tabaci Q.The results are great significance to study the insecticide resistance and the carboxylesterase function of Bemisia tabaci.The main contents and conclusions are as follows:1.Identification and expression analysis of APC family in B.tabaci BGenome-wide identification and characterization of APC family in B.tabaci B based on the genome sequencing.A total of 14 putative APC superfamily genes were identified,Most B.tabaci B AAT genes have a multi-exon structure with 12-14 exons,the lengths of the encoded proteins of APC transporters range from 486 to 792 amino acids.In our phylogenetic analysis of APC family genes in insects,an 11 paralog aphid-specific expansion of the APC transporter family was detected.The transcriptome data from different development stage of B.tabaci B showed that,the nymph APC clustered together.The nymph APC were more similar to those of males than females.The APC of eggs clustered separately from those of all other samples.Most APC exhibited low levels of expression in eggs than in other stages,and highly expressed in 3rd-instar nymphs.The data from different plant hosts and sexes showed that most APC had similar expression profiles among different hosts but that AAT expression profiles differed between B.tabaci B females and males.2.Identification and expression analysis of APC family in B.tabaci BGenome-wide identification and characterization of AAAP family in B.tabaci B based on the genome sequencing.A total of 25putative APC superfamily genes were identified,Most B.tabaci B AAAP genes have a multi-exon structure with 10-11 exons.The lengths of the encoded proteins of most AAAP transporters range from 442 to 561 amino acids,except BtAAAP5,which encodes a 1081-amino acid protein.In our phylogenetic analysis of AAAP family genes in insects,a 10 paralog whitefly-specific expansion,a 7 paralog A.aegypti-specific expansion,a 16 paralog Lepidoptera-specific expansion.A total of nine members of the B.tabaci B AAAP family expansion shared a similar genomic location and mapped to a 167-kbp region of genomic scaffold 566.The transcriptome data from different development stage of B.tabaci B showed,the nymph AAAP clustered with those of males,3rd-and 4th-instar nymphs were clustered together,while AAAP of 1st-and 2nd-instar nymphs and males clustered together;the AAAP of these stages then clustered with those of females.Finally,the AAAP of eggs clustered separately from those of all other stages.The data from different plant hosts and sexes showed that most AAAP had similar expression profiles among different hosts but that AAAP expression profiles differed between B.tabaci B females and males.3.The molecular evolution analyses of whitefly-specific expansion of AAAP geneTo test whether the whitefly-specific expansion of AAAP gene experienced accelerated evolution rates,branch-based and site-based models was implemented.Branch-based model of molecular evolution was used to detect lineages under accelerated rates of evolution,and site-based model was used to detect specific sites under positive selection.The analysis of evolutionary rates indicated that purifying selection can explain the B.tabaci-specific AAAP expansion.4.The RNAi of BtAAAP15 and BtAAAP21 genesTo investigate the effects of whitefly-specific expansion of AAAP genes on B.tabaci B,BtAAAP15 and BtAAAP21 genes were knockdown by feeding dsRNA of AAAP family genes to adults.The whitefly-specific expansion and highly expressed in nymphs and adults of B.tabaci B genes BtAAAP15 and BtAAAP21 were cloned from adults.Specific dsRNA primers of AAAP 15 and AAAP21 were designed,and dsRNA were synthesized using T7 Ribomax?Express RNAiSystem.The results of RNAi demonstrated that the 2 days mortality and 7 days mortality of B.tabaci B adults fed with dsRNA of AAAP family genes significantly higher than that fed with dsRNA of EGFP.5.Identification and expression analysis of carboxylesterases family in B.tabaci QGenome-wide identification and characterization of carboxylesterases family in B.tabaci Q based on the genome sequencing.A total of 42 putative carboxylesterases family genes were identified.The lengths of the encoded proteins of carboxylesterases range from 302to 932 amino acids.Our investigation of carboxylesterases genes from insects with completely sequenced genomes indicated that B.tabaci B and A.pisum contains more ?-esterase subfamily genes than other species.While B.tabaci B and A.pisum contains fewer a-esterase subfamily genes.The B.tabaci B contains more neuroligin subfamily genes and acetylcholinesterase genes than other insects.The transcriptome data from different development stage of B.tabaci B showed,carboxylesterases of egg and 1st-and 2nd-instar nymphs clustered together,3rd-and 4th-instar nymphs clustered together,the egg and nymphs clustered together,then clustered with males and females.