Construction Of The Molecular Genetic Linkage Map For Betula Luminifera

Betula luminifera H.Wink.is one endemic tree of China in Betula.Betulace .It is naturally distributed in south area of the Qinling Mountains and the Huai River, witch has strong adaptability and widespread application, such as in aerospace, papermaking and so on. Based on the techniques of Amplified Fragment Length Polymorphism (AFLP) and Simple Sequence Repeat (SSR) markers, a molecular genetic linkage map of F1group by LA05-1×JX0401 was constructed in this research. The major results are described as follows:1. The method for extracting DNA of Betula luminifera is selected through comparing the method of CTAB﹢SiO2 , CTAB and SDS. Using EcoRI and MseI enzyme-cut combination, and the best DNA amount, enzyme-cut times and dilution multiple of pre-expending product were determined by comparison of experiments. The AFLP molecular labeling technique system for Betula luminifera is optimized in the first time. Forty-one pairs of primers were selected using parents of map group for constructing genetic map.2. In this study, the factors of infecting PCR reaction were optimized by changing one factor respectively. The SSR molecular labeling technique system for Betula luminifera is established in the first time using primers from betula. The Amplified products were detected by PAGE, then three primers'segments were selected from thirty-six pairs of primers at random and were sequenced for further checking the universality of the primer. The blast results show that the value of Max ident are all above 95%. Seven pairs of good primers (3 from Betula pendula and 4 from Betula platyphylla var. japonica) were selected using parents of map group for constructing genetic map.3. Phenotypic traits in F1group have big separation, most of that show intermediate type and show more mother trait than father. The phenotypic of leaf, growth traits and stem form traits are all normal distribution, which indicate that they are controlled by polygons. The correlation among the phenotypic described by SPSS 17.0. It indicates that the leaf width, the petiole length ,the distance between neighboring leaves and the leaf length have highly significant correlation; the leaf width and the petiole length have highly significant correlation; the leaf width and the distance between neighboring have significant correlation; the leaf width and length-width ratio have highly significant negative correlation; the distance between neighboring ,the seeding height, the ground diameter and the leaf width have significant correlation; and the seeding height and the ground diameter have highly significant correlation, which the correlation is 0.849.4. After the marker was analyzed by FsLinkageMap 2.0, the result shows that Linan 05-1 was constructed based on 63 AFLP, 16 groups including 9 triplets were identified in this map. This map covers 651.08 centiMorgan (cM) of genome length, and the average distance between two markers is 10.33cM, where the most length distance between two markers is 37.7cM and the most shortage distance between two markers is 1.5cM.(except 0 cM ). JX0401 was constructed based on 37 AFLP, 8 groups including 3 triplets were identified in this map. This map covers 547.48cM of genome length, and the average distance between two markers is 14.80cM, where the most length distance between two markers is 36.8cM and the most shortage distance between two markers is 2.9cM.(except 0 cM ); two SSR markers which is from mother's seven SSR markers were identified in the 20th doublet of Linan 05-1's map ,and the other one composes a doublet with an AFLP marker ;but the father's six SSR markers can't identified in the map.