| Paeonia suffruticosa was sheepberry and be named as the king. It is one of ten famous Precious flowers, and has longer than2000years cultivation history in China. In this study,we aimed to breed new varieties of Paeonia that according to some special needs by cross breeding. Sequence related amplified polymorphism (SRAP) was conducted to assess the genetic diversity among the13Fi progenies and to analyze the phylogenetic relationship between F1progenies and parental generation of Paeonia suffruticosa and took16Hybrid parents as materials, which provide scientific foundation for seed selection and breeding of Paeonia suffruticosa in future.The main results were as follows:1. This experimentation adopted hybridization by hand pollination and established hybridized combination by the special breeding destination. The setting percentage of101hybridized combinations was more than80%, and total received2380seeds. The seedling percentage of2380seeds was36.9%, and total received879seedlings.2. Morphological identification of13hybrid generation showed that, F1can be divided into3types:Partial maternal inheritance type, such as Zl, Z3, Z7, Z8, Z11, Z12; Partial paternal inheritance type, such as Z2, Z5, Z6; Intermediate type, such as Z4, Z9, Z10, Z13;3. This test reaction system selected for25μl PCR reaction system. The total25μl reaction volume including2.5μl of10×PCR buffer,2.0mmol/L of Mg2+,0.25mmol/L of dNTPs,1.5U of Taq enzyme,0.25μmol/L of each primer,2.0ng/μL of DNA template. Amplification program was denatured for5min at94℃; the primer5reaction cycles were1min at94℃,1min at35℃,1min at72℃; the following35cycles were1min at94℃,1min at50℃,1min at72℃;then10min at72℃and stored at4℃.4. SRAP analysis of phylogenetic relationship between parental generations and assess the genetic diversity among the F1progenies.9primer pairs were selected from64pairs, which were applied to study the genetic diversity and relationships of the materials. By using the GENESCAN software9primer pairs produced1344loci of which1330were polymorphic, the percentage of polymorphic loci was98.95%, each primer combination produced149.3loci.16parental Nei’s genetic similarity coefficient (GS) ranged from0.2925-0.4638, the average of the Nei’s genetic diversity (H) was0.1655, and an average of the Shannon’s information index (I) was0.3047.13F1progenies Nei’s genetic similarity coefficient (GS) ranged from0.1651-0.4292, the average of the Nei’s genetic diversity (H) was0.1695, and an average of the Shannon’s information index (Ⅰ) was0.3077.5. According to the results of SRAP amplification, UPGMA method is used to build a tree clustering figure,16cultivars were divided into three categories, and variety’Qing XiangBai’ has the farthest genetic relationship from other varieties. Variety’Qing XiangBai"s flowers are white crown-shaped, high plant, leaf shape are the medium round leaf on morphology.13a hybrid offspring can be divided into partial paternal inheritance type, partial maternal inheritance type and Intermediate type.6. According to the analysis of the similarity coefficient of16hybridization between parents and the setting rate of hybrid combinations. The results showed that:parents with a similarity coefficient of0.31-0.35tended to have a high setting percentage. Considering the characters collocation and the fertility of their parents, in order to improving the affinity of the hybrid. Hybridization should based on single flower varieties as the maternal parent, choose far relatives of varieties. Varieties meet the requirements of traits, genetic relationship were not too close can also chosen. Ultimately filtered out’Qing XiangBai’,’HeiHaiSaJin’,’Zhu ShaLei’,’Cai Hui’and other four outstanding maternal varieties.7.13Fl progenies of the genetic similarity distance was0.2641, the average of the Nei’s genetic diversity (H) and the Shannon’s information index (Ⅰ) was0.1695and;0.3077higher than the16parental generations’genetic similarity distance of0.1713, and the average of the Nei’s genetic diversity (H)=0.1655and the Shannon’s information index (Ⅰ)=0.3047. Hybrids between genetic variation showed increases compared to the parental rich genetic diversity.8. Analysis between F1progenies and their parental generations’amplified polymorphic bands with cluster analysis results obtained Paeonia suffruticosa was partial maternal inheritance and showed cytoplasmic genes played an important role in the genetic traits expression of Paeonia suffruticosa. And combined morphological identification results obtained partial paternal hybrid offspring2,4,6,13, predicted their adult traits may tend to paternal traits; obtained partial maternal hybrid offspring1,3,5,7, predicted their adult traits may tend to maternal traits; obtained intermediate hybrid offspring8,9,10,11,12, predicted their adult traits may have common parental traits or may have specific traits which were different from parental traits, those need to pay attention to nurture;9. Research showed that SRAP markers identification combined with morphological identification can conveniently identified hybrid offspring, conduciving predict adult offspring phenotypic traits and discover the advantages of both parental traits in new varieties. |
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