Study Of Genetic Relationships Among The Part Of Bamboos In Bambusa Using Random Amplified Polymorphic DNA(RAPD)

Both intraspecific and interspecific relationships of the 28 ornamental bamboos of Bambusa in the section were analyzed using RAPD(Randomly Amplified Polymorphic DNA)markers.The species are as follows: Subgen.Bambusa:Bambusa sinospinosa,B,rutila,B.subaequalis,B.gibba,B.ventricosa;Subgen.Lingnania:B. remotiflora,B.cerosisssima,B.textilis cv.Maculata,B.textilis var.gracilis;Suben.Leleba(Nakai)Keng f.:B.tulda, B.eutuldoides McClure var.viridi-vittata,B,pervariabilis,B.longispiculata,B.tuloides,B.subtruncata,B.boniopsis, B.gibboides,B,albo-lineata,B.lenta,B.contracta,B.vulgaris and two cultivars B.vulgaris cv.Vittata andB. vulgaris cv.Wamin,B.multiplex and four cultivars B.multiplex cv.Alphonse-Karr,B,multiplex cv.Silverstripe, B.multiplex cv.Fernleaf.,B.multiplex var.riviereorum,16 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 minimum distance method.It is concluded as follows:(1)Amplification reaction system(20uL/tube):10×Buffer2.0μl,Mg²⁺3.5mM,dNTPs0.4mM,RAPD primer 0.6μM,TaqDNA 3.0U,template DNA40ng,The mixture was joined with super-pure water reach to 20μl.Amplification reaction program:After initial denaturation for 2min at 94℃.Then the reaction was continued for another 38 cycles through the following procedure:94℃for 45s,36℃for 30s and70℃for90s; Finally it ended at 72℃for7min for extension and 4℃for conservation.(2)16primers which produced distinct bands,showing unmistakable polymorphism were selected from 96 primers.The polymorphism analysis for 28 bamboo species of Bambusa showed a total of 218bands,among which 211 bands were polymorphic,attaining a polymorphic rate of 96.8%.The number of base pairs rage of 190 to 3000bp.Per primer produced 13.6 bands,including 13.2 polymorphic bands.(3)The average genetic distance of 28 bamboo species is 0.5597,the range of variation is from 0.2139 to 0.7647,The average genetic distance and the range of variation are both large,there are 211 polymorphic bands in 218 bands,attaining a polymorphic rate of 96.8%.all these show the 28 bamboo species have high genetic diversity.(4)Combined with morphological classification,the results are:the genetic distance of the cultivated type bamboos belonging to the same bamboo is small,while the genetic distances are big between different bamboos.such as 4 cultivated type bamboos of B.multiplex for 0.2139 to 0.4013;the 2 cultivated type bamboos B.vulgaris cv.Vittata and B.vulgaris cv.Wamin belonging to B.vulgaris as 0.3986;the genetic distance of the species between different subgenus is 0.7521 from B.subaequalis to B.subtruncata.(5)Analysing the PCR products using RAPD,integrating the bands,the molecular checking index of 28 bamboo species is establishmented,this can provide a shortcut to identificate bamboos from the term of gene in theory,and reveal the inner relationship of the 28 bamboo species.It has guiding meaning in theory to identificate the relationship in term of gene.(6)The PCR products using sixteen random primers to amplify 28 accessions DNA of Bambusa showed the complex genetic base in intraspecific and interspecific and RAPD markers can sensitively detect the genetic diversity.The clustering results according to genetic similarity coefficients and genetic distance were in consistence with that of the analysis of morphology.It showed that RAPD can not only detect genetic diversity in interspeeific and intraspecifie of Bambusa and show genetic relationships,but also apply in the systematic study of Bambusa.