The Study On Genetic Management Of Reintroducted Hydropotes Inermis In Shanghai

Hydraotes inermis(Chinese water deer)is a primitive small deer animal.Due to the destruction of habitats and the disruption of human activities,quantities of Chinese water deer have dropped sharply,while Chinese water deer in Shanghai have disappeared at the beginning of the 20 th century.In order to protect this species and increase the diversity of Shanghai,the Shanghai department started the project of reintroduction Chinese water deer in 2006.With the expansion of the reintroduced Chinese water deer in Shanghai,the population urgently needs scientific genetic management to ensure the healthy development.Therefore,this study collected blood samples of reintroduced Chinese water deer to tested the blood physiological and biochemical indicators to establish reference intervals for its physical and chemical indicators;useing the mitochondrial DNA molecular markers to evaluate the relationship between the genetic diversity of reintroduced Chinese water deer and wild populations;The microsatellite primers were used to evaluate the genetic diversity of reintroduced Chinese water deer in Shanghai,and to analyze the genetic relationship among individuals and populations at each reintroducting site.The result is as follows: 1.Establish reference intervals for blood physiological and biochemical indicatorsReflecting the health status of Chinese water deerBlood physiological and biochemical indicators are important means to reflect the health of animals.In this study,29 physiological and biochemical indicators of 131 Chinese water deer were tested.According to the nonnormality of blood data,the reference range of each blood index was expressed by the interquartile range(IQR),and reference intervals of the reintroduced Chinese water deer in Shanghai was initially confirmed.The range of health of the indicator and regression analysis of environmental factors(sampling time months and years,captive and artificial breeding)and individual differences(individual gender,age,and gestation period).The result shows that 15 physical and chemical indicators of pregnancy and non-pregnancy was showed significant differences;there were significant differences in the 13 physical and chemical indicators between the populations established in Shanghai and derived Zhoushan supplementary populations.2.Genetic diversity analysis of reintroduced Chinese water deer in Shanghai base on D-loop and Cytb sequences of mitochondrial DNAIn this study,114 Cytb sequences and 102 D-loop sequences of reintroduced Chinese water deer were measured.The 114 Cytb sequences were defined six haplotypes,the Haplotype diversity(Hd)was 0.4673 and the Nucleotide diversity(π)was 0.0091;102 D-loop sequences were defined 9 haplotypes,the haplotype diversity was 0.9794 and the nucleotide diversity was 0.00468.Compared with the study on the genetic diversity of wild Chinese water deer studied in 2006,the level of genetic diversity reintroduced Chinese water deer in Shanghai was slightly reduced,but with reference to other deer’s p and Hd,the reintroduced Chinese water deer genetic diversity in Shanghai is still in the middle-high level.In order to explore the genetic relationship of reintroduction Chinese water deer between the Shanghai and other regions,researchers combined the Cytb and D-loop sequences from Zhoushan,Dafeng,Yancheng,Jishan,and the Korean to construct ML phylogenetic trees.The phylogenetic tree of Cytb sequence and D-loop sequence have the same results.Shanghai and Zhoushan Chinese water deer do not differentiated into two branches;Zhoushan and Yancheng Chinese water deer have close genetic relationship;Jishan has the farthest relationship with Shanghai.The intermediary network map shows that the haplotype of Zhoushan Chinese water deer is the original haplotype.3.Screening of microsatellite primers for analysis genetic relationships of Chinese water deer in ShanghaiIn this study,52 pairs of microsatellite primers were developed from the 55 published literatures about related species of Chinese water deer and Chinese water deer’s genomes.Finally,18 pairs of microsatellite primers and 18 corresponding microsatellite loci were selected for using in this study.To analyse the genetic diversity of the Chinese water deer individuals,and to analyse the genetic relationships among the 4 places where they were introduced.In this study,96 individuals of Chinese water deer were used based on 18 microsatellite primers.The results showed that the average expected heterozygosity of the reintroduced population in Shanghai was 0.626(0.323-0.854),and the average polymorphism information content was 0.576(0.286-0.816),Compared with the reintroducted Chinese water deer whose average heterozygosity is from 0.3736 to 0.5299,and the polymorphic information content is from 0.5654 to 0.7144,the genetic diversity of reintroduced Chinese water deer from Shanghai was higher.The genetic differentiation factor of the reintroduced population in Shanghai was 0.03(Fst<0.05).The STUCTURE software was used to analyze the population structure of four small populations.The reintroduced population in four habitats in Shanghai did not show genetic differentiation.This study analyzed the polymorphism content,linkage disequilibrium,and PI(the probability that there was no genetic relationship or the same genotype between sibling individuals)of the 18 microsatellite locus.The results showed that 11 locus of the polymorphism content of microsatellite locus is high(PIC>0.25),there is no linkage disequilibrium sites,and the cumulative PI is low,that is,11 pairs of microsatellite primers can meet the requirements for recognizing of Chinese water deer.The cumulative exclusion rate(CEP)of 18 microsatellite loci was 99.99%,and 12 groups of mimical child-mother-father relationships were obtained.In summary,this study preliminarily establishes the reference range for the physical and chemical indicators of reintroduced Chinese water deer in Shanghai.Cytb,D-loop,and microsatellite molecular markers indicate that the genetic diversity of Chinese water deer in Shanghai is high;this study screened the 18 pairs of microsatellite primers,they can be used for the individual identification and phylogenetic identification of reintroduced Chinese water deer in Shanghai.In short,managers can deepen the research on the genetic management of reintroduction of Chinese water deer in Shanghai based on the results of this study,and formulate scientific conservation measures and population genetic management plans so that the reintroduced Chinese water deer population in Shanghai can sustainably develop.