AFLP Analysis Of Part Of Tree Peony (Paeonia Suffruticosa Andr.) Cultivars In Pengzhou

Paeonia suffruticosa,as cross-pollinated plant,has low rate of self-fertility,and the feature provides abundant Source of variation for artificialselection and natural selection. For long-term domestication and cultivation, form various type of varieties, genetic relationship among varieties are complex.Therefore,discussing classification and determination and genetic relationship about heredity,not only be of great significance to reveal species evolution,but also to the benefit of guiding tree peony breeding of new variety and protection conventional varieties,and so on.The experiment study genetic relationship and genetic diversity of 28 tree peony varieties in molecular level,adopting AFLP(Amplified Fragment Length PolymorpHism) technique.First, extract genome DNA of high quality from tree peony spire,adopting improved CTAB,to be used for AFLP double digestion,and Restriction enzyme digestion completely.Then, optimize AFLP Selective amplification system, establish the best reaction system,screen out primer combination for amplification that adaptive tree peony AFLP analyse. Final,statistical analysis AFLP map and data of sample, the consequence as follhgows:1. The experiment extract genome DNA from tree peony spire adopting improved CTAB, and high yield, OD260/OD280 is among 1.8～2.0. After wipeing off RNA, agarose gel electropHoresis detection, master tape is clear, DNA with high purity,no degradation, no enzyme reaction inhibitors, can be digested completely by restriction enzymes Msel and EcoRI.2. Optimize tree peony double digestion and AFLP selective amplification system, get the best reaction system, optimum temperature is 37℃, the dosage of restriction enzyme is 3U, and the time is 3h;the best selective amplification system is:pre-amplification product 2μl,E-primer 0.5μl, M-primer 0.5μl, Taq enzyme 0.25μl, MgCl2 1.2μl, dNTPs1.6μl, 10×Buffer 2μl, ddH2O 11.95μl 3 Adopt optimization system,screen 64 primer combinations of AFLP, after three replications, screen out 6 primer combinations with many amplification bands and high polymorpHism.4.Make use of 6 choosed selective amplification primer combinations,study 28 tree peony with AFLP.6 primer combinations amplified total 2562 belts that could be counted(among 100～750bp),and 2142 belts are of polymorpHism,take up 83.61%.E-AAC/M-CTA(88.19%)is of the most polymorpHism belts, and E-AAC/M-CAg(73.42%) is of the lowest.The average of every primer combination amplified 427 belts,and 357 belts are of polymorpHism.5.Adopt UPGMA cluster analysis,28 tree peony are clustered into 2 groups.From the result we can find,most varieties from the same producing area showed more closely pHylogenetic relationship;and in the circumstances of same producing area,most varieties are of the same or close planttypes and flower colour cluster together firstly,but there is cross phenomenon; the influence of phenotypic character such as florescence and flowertypes to cluster analysis result are not obvious.