| Both intraspecific and interspecific relationships of the 25 bamboos of P.leioblastus Nakai in the section were analyzed using ISSR (Inter simple sequence repeats)markers.The species are as follows:P. gramineus, P. anarus, P. gozadakensis,P. chino,P. solidus,P. amarus var. pendulifolius,P. longifimbriatus,P. amarus var. hangzhouensis,P. juxianensis,P. maculates,P. intermedius,P. oleosus,P. gramineus f. monstarispiralis,P. chino f. angustifolius,P. simonii f. albostriatus,P. longosonens f. aureokamuyast,P.rugatus Wen et S.Y.Chen,P. amarus var.subglabratus,P.dokgyoamus,P.leioblastusmaculosoides,P.leioblastusyixingensis,P.yunhoensis, P.qingyuanensis ,P.suichangensis,P.hsienchuensis Wen,18 random primers were selected which produced distinct bands,showing unmistakable polymorphism. By DPS7.05 program, Jaccard's genetic similarity coefficients were generated and dendrogram was constructed using The weighted average method.It is concluded as follows:(1) Established the P.leioblastus Nakai. technique system by ISSR through one-factor experimental design,Amplification reaction system(20uL/tube):10×Buffer2.0μl,Mg2+2.5mM,dNTPs0.2mM,ISSR primer 0.2μM,TaqDNA 1.0U,template DNA50ng,The mixture was joined with super-pure water reach to 20μl.(2)18primers which produced distinct bands, showing unmistakable polymorphism were selected from 100 primers.The polymorphism analysis for 25 bamboo species of P.leioblastus Nakai which pick from bamboo garden of Fujian Agriculture & Forestry University showed a total of 155 bands,among which 137 bands were polymorphic, attaining a rate of 88.3%.(3) With 18 random primer to amplify 25 bamboo species of individual,gains the polymorphism of abundant ISSR product, show the P.leioblastus Nakai taxa between intraspecific and interspecific of complex genetic background, through ISSR markers can be accurate, sensitive and efficient to detect genetic diversity of P.leioblastus Nakai.It showed that ISSR markers can not only detect genetic diversity in interspecific and intraspecific of P.leioblastus Nakai and judgment within its relationships, but also can give the reference of the classification of value.(4) Using the DPS 7.05: software clustering analysis, cluster results are similar with traditional classification, 25 bamboo species for the average genetic distance 0.5006, the variation rate of 0.1486-0.7191, 18 ISSR primer produces 155 calculated the DNA fragments were the genetic similarity coefficient between material constructed by UPGMA, 25 of resources of daming bamboo clustering tree in the genetic distance, for 0.6138 place 25 bamboo species were divided into three groups,The first group(genetic distance 0.1618): P.leioblastus Nakai P. chino,.The second group( genetic distance 0.4702): P.leioblastusmaculosoides, P.leioblastus yixingensis, P. gramineus f. monstarispiralis. The third group(genetic distance 0.5634): P. gramineus, P. anarus, P. gozadakensis ,P. solidus,P. amarus var. pendulifolius,P. longifimbriatus,P. amarus var. hangzhouensis,P. juxianensis,P. maculates,P. intermedius,P. oleosus, P. chino f. angustifolius,P. simonii f. albostriatus,P. longosonens f. aureokamuyast,P.rugatus Wen et S.Y.Chen,P. amarus var.sub glabratus, P.dokgyoamus, P.yunhoensis, P.qingyuanensis ,P.suichangensis,P.hsienchuensis ⑸The digital fingerprint identifier was established about P.leioblastus Nakai. With the primers such as U807,U815,U835,U836,U840,U841 and 844,The digital fingerprint identifiers of 25 provenances were established based ISSR. |
