| The paper has evaluated analysised123SSR primer5and7sugar beet material, in order to establish a stable, accurate, economical and efficient beet Germplasm by SSR identification system.The paper has identificated57sugar beet material, They can Provide effective use of scientific methods and theoretical basis for beet germplasm.The results were as follows:(1) Based on PCR and polyacrylamide gel electrophoresis technique,This paper has established accurate, efficient and economic beet germplasm resources evaluation of SSR identification system. the template and primer concentration,DNA,Mg2+,dNTPs And Taq enzyme have been Compared with different dosage on PCR amplification effect and established57sugar beet varieties Germplasm Resources Identification of PCR reaction, PCR amplification reaction system:2.0μL Mg2+(25.0mM),2.5μL10×buffer,2.0μL dNTPs (2.5mM),0.25U Taq enzyme (2.5U/μL), SSR Reverse primer1.0μL (100μM),2.0μLTemplate DNA,9.40μL sterile water。different annealing temperature have been Compared on PCR amplification effect, they determinated the sugar beet varieties Germplasm Resources Identification of PCR reaction amplification procedures, PCR response procedures:94DEG C predegenerated5min, PCR amplification includes35loops,Each cycle at94degrees C degeneration0.5min, Appropriate temperature (to be) annealing1min,72degrees of extension0.5min, After closing the loop72deg.c5min, Stored at4℃.3kinds of silver staining technique have been compared, PCR amplification product from the6%PAGE glue separation, fast silver staining techniques for staining has been applied, Time is short, low cost,With a PCR band and background contrast, Can get clear strip chart.10pairs of sugar beet germplasm identification primer were identified from123SSR primers, The author have defined2pair of proprietary primer in10pairs of sugar beet germplasm identification primer, the2pair of sugar beet proprietary primers tested randomly selected seven sugar beet materials, Fingerprinting and statistical results showed that the strains between the banding patterns, They confirmed that the experimentally determined beet SSR identification system accuracy and repeatability of the advantages.(2) Two pairs of SSR proprietary primer PCR products on6%polyacrylamide gel electrophoresis showed extensive polymorphism. I applied the marker map of the two pairs of sugar beet SSR-specific primers To confirmate of57sugar beet varieties with abundant genetic diversity.2pairs of SSR primers in proprietary57sugar beet varieties were detected30allelic variants, Each primer detection can detect18allele and12, an average of15. SSR molecular tag proprietary primers can been used stinguish the57sugar beet material. SSR loci polymorphism information have been applied, PIC value of0.86and0.89, an average of0.875. The author Analyzed by POPGENE the Version1.31software to calculate the effective alleles (NE), Proportion of polymorphic loci (PP), Shannon index (SI). The results show that the germplasm resources of57sugar beet material variance, the polymorphic information content, the Shannon index was relatively large, confirmed the57sugar beet tested materials germplasm resources with a wealth of genetic diversity.(3) The author has detected genetic diversity map57beet material by two pairs of sugar beet proprietary primers. molecular biology to professional statistical software NTsys system was applied, The authors tested materials in accordance with the UPGMA method cluster, Using30polymorphic SSR markers were calculated for57sugar beet varieties between genetic similarity coefficient (J),The author cluster analysis cluster indicate that I class sugar beet material49,47,43,40and32,other sugar beet material is divided into Part II class. Type I is disease resistance to the closely linked to the genetic material, Part II class is closely linked to the genetic material with sugar and production, from agronomic characteristics of the known material. |
PDF Full Text Request