| The genus Acer L.belongs to the Aceraceae Family,China is the main distributing centres in the world.There are abundant precious genentic resource in the China.but, study on the genetic relationships of the genus Acer L .are less for a long time.This study developed the SRAP molecular marker for the genus Acer L.To elucidate the genentic relationshipsof species of the genus Acer L.at the DNA molecular level and provide the scientific and efficient of the persimmon germplasm.Using the leaves of The genus Acer L.as materials, different ways of isolating genomic DNA were tested to find the best one. At the same time, the optimal Sequence-Related Amplin Polymorphism(SRAP)techniques for The genus Acer L.were established and applied to 22 leaves. The results were as follows:1. In this study, Comparison conventional CTAB with improved CTAB methods in the results of DNA extraction the genus Acer L.Using conventional CTAB method could isolate only a little amount of DNA,the band was Weak, protein was exist in the point-like hole, the towing more serious situation, the DNA integrity was bad, When the SRAP amplified at late ,the effects of secondary substances lead to the amplification was instable or failed.The results of using improved CTAB method was neat band, the DNA quality was higher, lower protein, non-towing situation, the genome integrity was better.Using improved CTAB method was an economic, easy and fast way to isolate genomic DNA.It could obtain qualified DNA from plants containing high secondary substances,such as polyphenols,pigment and protein.The DNA was qualified for SRAP analysis.2. The optimized SRAP marker suitable for The genus Acer L. DNA analysis was established.Taking genus Acer L.genome DNA as template, the major components of SRAP, such as concentrations of Mg , dNTPs, Primer, DNA, Taq polymerase were optimized by orthogonal design. The results showed that the optimum SRAP reaction system includes: Mg2+ 2.5 mmol·L,dNTPs 0.25mmol·L-,prmier 0.8 ng,Taq Polymerase1.5 U,DNA 40 ng in the 25μl volume reaction. The most suitable protocol was as follows: pre-denaturation at 94℃for 5 min followed by 5 cycles of denaturation at 94℃for 1 min, anneal at 35℃for 1 min and extension at 72℃for 1 min; then 35 cycles of 94℃for 1 min, 50℃for 1 min and 72℃for 1 min; and a final extension at 72℃for 5 min; kept at 4℃. The program and system could meet the demands for genome SRAP marker in The genus Acer L.3. Polymorphic primer pairs selected.Seletion was carried among the 64 primer pairs in the SRAP-PCR reaction program and system optimizationed. The results showed that there were 24 primer pairs displaying polymorphism. Each combination has 4～13 clear amplified fragments and the total was169, average was 7.04; Each primer pairs has3～10 polymorphic fragments and the total was 107, average was 4.46. The percentage of polymorphic fragments was 54%～83%, average was 63%.4. Using selected parts primer pairs amplified for 22 plants of The genus Acer L.it gets a stable band.These primers amplified gets the characteristic band reflects the specificity of the total genomic DNA sequence,It can be used for cluster analysis.5. The result of cluster analysis by using UPGMA method showed that 22 genotypes could be classified into five types in genetic distance 3.32. which reflected the genetic relationships among species and were almost agreement with the results of traditional morphologic taxa,but hypotaxis statusof very few species was slightly changed.
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