Intergeneric Relationships In Some Bamboo By AFLP

Bamboo is an important cash crop which belongs to the gramineae and is the important forest resources.Because of the different standard adopted and each kind of classification's limitation,both Synonym and homonym are presented very seriously. As a new DNA marker,AFLP(amplified fragment Length Polymorphism) has been widely used in cultivars identification,genetic diversity.For the purpose of providing the essential basis for classified status of the bamboo,the research adopted the technology of AFLP tO analyze the polymorphism of 35 bamboo cultivars and the cooperation of clustering.The results showed as follows.1 Through the comparison and the improvement of plant genome DNA extraction method,a quite suitable bamboo DNA extraction method—improved SDS DNA isolation can be obtained.High-quality DNA which is suitable to AFLP technique can be extracted from bamboo blade.2 Through the system study on AFLP reaction system of bamboo,established optimized system fitted for AFLP analysis.The overall volume of 20U1 digestion system contained purified DNA600ng,the concentration of the restriction endonucleases of EcoRâ… and Mseâ… was 10U/μL,digested at 37℃for 3h,65℃for 3h,add 10μL ligation mixture and then was kept at 16℃for at least 10h;pre-selective amplification was set up with 5μL restriction-ligation sample DNA,0.6μL 20U/μL EcoRâ… and Mseâ… primers,2μL10×buffer and 2μL dNTPs,ddH₂O was added up to 20μL.In 20μL selective amplification,5μL diluted(×20) DNA pre-selective amplification product,0.6μL 20U/μL EcoRâ… and Mseâ… selective primers.Under this optimization system,clear,steady and higher polymorphic of AFLP bands were obtained.3 Ten EcoRâ… /Mseâ… primer pairs showing high level of polymorphism and distinct band were selected out and used for further practical AFLP analysis.872 bands were amplified, each primers can amplify 87 bands and 836 of 872 amplified bands were polymorphic bands,which accounts for 95.87%;All cultivars similarity coefficient varied from 0.528 to 0.936.4 The finger printing data were further analyzed using UPGMA Cluster methods in the software NTSYSpc version 2.10,which clustered the 35 varieties into two main distinct groups,in which the value of the genetic distance was 0.403.