Relationship Between Behavior And Gene Polymorphism Of BDNF And 5-HTR In Male Père David's Deer(Elaphurus Davidianus)

Animal behavior is the most important way for animals to adapt to the ambient environment.The diversity of environment animal living in determines the diversity of animal behavior.Behavioral ecologists have long found stable individual differences in behavior(or a set of behavioral traits),and this difference has an important genetic basis.However,animal behavior is regulated by multiple loci of multiple genes,so the research on behavior genetics has always been a difficulty in animal behavior.In recent years,with the rapid development of genomics and post genomics,it is possible to study behavioral genetics.At present,the research on the behavior of Père David's deer refers to the behavior mode,behavior seasonal difference,and the relationship between Père David's deer and human disturbance.At the same time,researchers have also studied the physical stress on Père David's deer in captivity,as well as the behavioral countermeasures and endocrinological mechanism of Père David's deer adapting to the artificial environment.However,the physiological basis of Père David's deer behavior and its relationship with gene polymorphism have not been studied.To avoid affecting the daily behavior of Père David's deer,the differences in social behavior patterns of different individuals in male Père David's deer population were studied under the condition of no physical contact with randomly selected male Père David's deer;the gene polymorphisms related to animal pain,neurotransmission,tension,and antagonism were detected.The purpose of this study is to study the relationship between the behavior and the gene polymorphism of different individuals,and to reveal the behavior adaptation and evolution of Père David's deer under the background of low genetic diversity.In view of the above,we used non-invasive sampling method to collect the feces of Père David's deer,observed the individual behavior characteristics,and summarized the behavior according to the PAE coding classification system,that is,the animal behavior classification coding system based on the ecological function with the posture action environment as the axis.Through the literature,we screened out the genes related to animal pain,neurotransmission,tension,and antagonism,and analyzed them by the methods of molecular biology and statistics.We used 5-HT receptor(5-HTR)gene and brain-derived nerve growth factor(BDNF)gene as the target gene.Based on the RefSeq data of cattle 5-HTR and BDNF in the ensemble database,the effects of gene polymorphism on behavior differences among different individuals were studied by means of statistics,bioinformatics and PCR techniques and methods.The relationship between social behavior and gene polymorphism was analyzed,and basic information for BDNF and 5-HTR genes and proteins of male Père David's deer was supplemented.The main results are as follows:1.The quality of fecal DNA samples extracted from non-invasive samples was qualified.The target genes BDNF and 5-HTR were successfully amplified by the combination of landing PCR and nested PCR.2.Three genotypes of BDNF gene were obtained by PCR-RFLP,and 8 SNPs were found by sequence alignment.However,no polymorphism was found in 5-HTR by sequence alignment.3.By using bioinformatics software to splice the amplified sequence successfully,and by referring to the gene annotation of cattle,the complete splicing sequence of Père David's deer BDNF exon 2 and 5-HTR5 a exon 1 were obtained by sequence information prediction,and the protein prediction structures of BDNF exon 2 and 5-HTR5 A exon 1 were obtained by prediction software.The amino acid sequences of BDNF exon 2 and 5-HTR5 A exon 1 of Père David's deer were obtained by sequence information prediction.The same protein sequence of BDNF exon 2 was found in NCBI database by sequence comparison with blast tool of NCBI.The similarity between HTR5 A protein sequence and HTR5 A amino acid sequence of Bos taurus was 95%.The similarity between HTR5 A protein sequence and HTR5 A amino acid sequence of White-tailed deer(Odocoileus virginianus texanus)was 100%.On this basis,we obtained Père David's deer HTR5 A exon 1 protein prediction structure by prediction software.4.8 SNPs loci of BDNF exon 2 were analyzed.Chi square test showed that they were in accordance with Hardy Weinberg equilibrium.Eight SNP loci were completely linked to each other without recombination.5.Through statistical software,there was no significant difference in behavior among Père David's deer individuals,and it was found that there was no significant difference in behavior among BDNF three genotypes.