title: [Study on Genetic Stablity of Cryopreservation Regeneration Plant of Gentiana straminea with ISSR-PCR].
abstract: OBJECTIVE
  To establish ISSR-PCR system of cryopreservation regeneration plant of Gentiana straminea, and to select appropriate primers and analyze the genetic stability.
  
  
  METHODS
  DNA was extracted by CTAB, the optimal ISSR-PCR system was established by orthogonal experiment,and genetic stability was analyzed.
  
  
  RESULTS
  The optimal ISSR-PCR system (25 μL) was established: dNTPs 0.50 μL, Mg2+ 1.00 μL, 10 x PCR Buffer 2.00 μL, primer 0.60 μL, Taq DNA polymerase 1.25 μL, template DNA 1.30 μL, and ddH2O 18.35 μL. The amplification program was devised: 94 degrees C for 5 min, denaturing at 94 degrees C for 30 s, annealing of 1 min due to denaturing temperature of different primer,extension at 72 degrees C for 1.5 min, 35 cycles, last extension at 72 degrees C for 7 min, conservation at 4 degrees C . The DNA mutation rate of cryopreservation regeneration plant of Gentiana straminea was 1.05%.
  
  
  CONCLUSIONS
  The cryopreservation regeneration plant of Gentiana straminea retains very good genetic stability, there is little variation between each plant, so the cryopreservation can be used as a feasible method for resource protection of Gentiana straminea.
authors.forename: Su-fang
authors.initials: SF
authors.lastname: Gao
authors.forename: Hong-gang
authors.initials: HG
authors.lastname: Chen
authors.forename: Yan-hong
authors.initials: YH
authors.lastname: Zhang
authors.forename: Hui-lin
authors.initials: HL
authors.lastname: Li
authors.forename: Ling
authors.initials: L
authors.lastname: Jin
mesh_terms: D015925:Cryopreservation; D017931:DNA Primers; D018744:DNA, Plant; D042822:Genomic Instability; D030012:Gentiana / Q000235:genetics* / Q000254:growth & development; D016133:Polymerase Chain Reaction; D012038:Regeneration
chemical_list: D017931:DNA Primers; D018744:DNA, Plant