R-banding And Restriction Endonucleases Banding Of Cervus Nippon Hortulorum Chromosomes

Objectives:Sika deer is subordinated to Artiodactyla,Cervidae,Muntiacus. Northeasten sika deer(Cervus nippon hortulorum),one of the three survival subspecies in China,has important value of medicine and sanitarian value.It has generally focused on physiochemiacal identities,energetic metabolization,feeding management,diseases prevention and cure in Cervus nippon hortulorum researches,but the researches about the nuclear genes and chromosomes are much less.The analysis of high resolutionR-banding,Restriction Endonucleases banding with EcoRâ…¢and Hindâ…¢and the kindred of genomic DNA using with the technique AFLP will provide the basic information for cytogenetic researches of C.n.hortulorum,also provide some evidences about the relationship of high resolution R and G banding and make the basis for the following locating QTLs about producton of velvet antler using with AFLP and FISH.Methods:(1)The peripheral blood was obtained while sawing the velvet antler. The thymidine(TdR) was taken off after the lymphocytes were synchronized by it.Add 5-bromodexyuridine(BrdU) to start up the cell cycle.Collect lymphocytes after 72-hours culture.The chromosomes were dealed with R-banding technique,and the result of R-bands of C.n.hortulorum chromosomes was analysed.(2) The peripheral blood was obtained while sawing the velvet antler.The lymphocytes was collected after 72-hours culture and the chromosomes were dealed with Restriction endonucleases EcoRâ…¢and Hindâ…¢,then the banding was analysed.(3) The peripheral blood was obtained through ear vein.Then genomic DNA from Northeastern Sika Deer was extracted from whole blood and purified using with AxyPreP ^TM Blood Genomic DNA Midiprep Kit and the kindred of the sample Northeastern Sika Deer was analysed.Results and conclusions:(1)The number of the bands for haploid chromosomes was 400 according to Reading Conference,International System for Cytogenetic Nomenclature of Domestic Animals(ISCNDA),and chromosome bands of C.n. hortulorum and the high resolution R-bands idiogram were achieved.(2)The R-banding of metacentric chromosomes No1,No2,The longest telocentric chromosomes No3,No4,sex chromosome X,the shortest and the only submetacentric chromosome Y were just opposite to the G-banding of them besides the width of the bands is a little different.The terminals of the chromosomes were dyed dark except those of chromosomes No21,No24,No28.The terminal of chromosome Ywas dyed dark the same as those of Cervus unicolor and Vietnamese sika deer,but The terminals of chromosome Y were dyed light in Ovis ariesL and Capra hircus.All these manifested the stability in the same specie and the differences between the different species.(3) The metaphase chromosomes of Northeastern Sika Deer displayed G-like bands and C bands by treatment with restriction endonucleases Haeâ…¢and Hindâ…¢followed by Giemsa.However not all constitutive heterochromatin was dyed dark. eleven pairs of autosome(No1,No2,No4,No6,No 11,No 15,No27,No28,No 29,No 31,No 32 and two sex chromosome X and Y of Northeastern Sika Deer manifested the heterogeneity of constitutive heterochromatin when they were treated with Haeâ…¢and Hindâ…¢.(4)The analogical coefficient of Cervus nippon hortulorum genomic DNA fragment after treaten with AFLP was from 0.6809 to 0.8648,The average analogical value is 0.7841.The most analogical was between C₂₂ and C₂₀,from Yongchuan.The analogical value was high to 0.8648,The relation of C₂₂ and C₂₀ was paternity,Furthermore,the production of velvet antler in C₂₂ was much higher than the other male parents,That of it in C₂₀ was much higher than the generation F₁ of the other cross-bred compounding.