AFLP Analysis Of Genetic Diversity And Genetic Relationships In 4 Buntings

The principle of Amplified Fragment Length Polymorphism(AFLP) is based on the polymorphism ofDNA fragment which is producted by restricted enzymes and selective amplification. AFLP technologyintegrate the RFLP technology with stabilization and credibility and RAPD with simpleness and highefficiency. The technique involves three steps: (â…°) restriction of the DNA and ligation of oligonucleotideadapters, (â…±) selective amplification of sets of restriction fragments, and (â…²) gel analysis of the amplifiedfragments. Using this method, sets of restriction fragments may be visualized by PCR without knowledgeof nucleotide sequence. The method allows the specific co-amplification of high numbers of restrictionfragments. The number of fragments that can be analyzed simultaneously, however, is dependent on theresolution of the detection system.Typically 50-100 restriction fragments are amplified and detected ondenaturing polyacrylamide gels. So it is suited for the analysis of heredity polymorphism, making hereditymap, identification of genetic relationships, and research of molecular marker.Emberiza belongs to Emberizidae, There are 38 species in the word. Major in Europe, Asia and NorthAfrica, 26 of all in china, 18 of them in Northeast of China.The pre-study of avian taxonomy and phylgenetics was main in exterior and anatomic form. Alongwith the deeply of the research, discover that only use morphology character can not determine itsgenealogy relationship. At this time the method of protein and nucleic detection are used in the study ofavian taxonomy, due to the appear of molecule markers, the research of class has been a high development.Because AFLP technology is more complex than others and with high request, detective method aboutAFLP of the Emberiza wasn't reported at home. Only have some reports about karyotype and NumericalCytotaxonomic. First, the condition on construction of AFLP fingerprinting map is optimized, and thenAFLP analysis system of silver staining with tow cost was set up. Second, 11 primer sets were analyzed byeffect and polymorphism to seek for AFLP primer sets which were suited for bunting.At last, Genetic Diversity and Genetic Relationships of 4 buntings(2 Emberiza cioides;5 Emberizaspodocephala;5 Emberiza leucocephala; 6 Emberiza tristrami)was analysised by AFLP technology.Discussed the feasibility about AFLP applied to Emberiza.The relevant results are as follows:1. Eleven out of thirty-nine AFLP primer paies produced reproducible polymorphic DNA amplificationpatterns. Of the 535 AFLP markers scored, 429 were polymorphic. The proportion of polymorphic loci was80.1ï¼…. 2. An UPGMA tree based on Jaccard similarity index has been made. Four buntings were grouped into onebranch, in which Emberiza cioides and Emberiza leucocephala clustered together, and Emberiza tristramiand Emberiza spodocephala were grouped closely. The results indicated that it was practical to revealgenetic diversity of Emberiza using AFLP markers.