The Issr Analysis Of Genetic Diversity And Research On Garden Application Of Named Species Of Bambusoideae

This study's research object is30named species of Bambusoideae from the Fujian Agriculture AndForestry University in bamboo garden as the,to bagin with the DNA polymorphism analysis of ISSRmolecular marker method,using DNA molecular marker to study bamboo's germplasm resources whosegenetic background is very complicated,to study the genetic diversity,phylogenetic relationship,to carryout bamboo's germplasm resources molecular identification,and establish effective DNA fingerprinting,at the same time Research on the application of bamboo in landscape.This study uses ISSR molecular marker technique to analyze China's30named species ofBambusoideae,The species are as follows：Dendrocalamopsis oldhami,Brachystachyum densiflorum,Semiarundinaria fastuosa,Indocalamus tessellatus,C.marmorea,Neosinocalamus Keng f.,Dendrocalamusstrictu,Icrassiflora McClure,Guadua amplexifolia J.S.Presl,Sinobambusa tootsi,Acidosasa chinensis,Gigantochloa atter,Pseudosasa japonica,Oligostachyum sulcatum,Bambusa arundinacea,Phyllostachysviridis,Pseudosasa amabilis,Melocanna baccifera,Schizostachyum pseudolima,Thyrsostachys siamensis,Pseudostachyum Ploymorphum,Monocladus saxtilis,Cephalostachyum fuchsianum,Bashania fargesii,Drepanostachyum falcatum,Shibataea kumasasa,Pleioblastus. gramineus,Fargesia spathacea Franch.,Yushania niitakayamensis,Sasa longiligulata McClure.Analyzing their genetic diversity, Thisexperiment study the DNA extraction method of30named bamboo species,studying bamboo plantsgenerally applicable ISSR-PCR optimization reaction system,screening out of18bands of clear and goodprimers, using Popgene32software to analyze genetic diversity,Getting genetic similarity and geneticdistance informations. Using SPSS13.0software,Generating polygenetic tree of genetic relationship of30bamboo species. The experimental results are as follows：(1) With the one-single and multi-level gradient testing method,To optimize system in five factors ofMg2+,template DNA,TaqDNA polymerase,dNTPs,and primer. in a total volum of20μL system whichcontained40ng template DNA,0.6μmol/L primer1.0U TaqDNA polymerase,2.5mmol/L Mg2+,0.25mmol/L dNTPs,1×buffer. Through experiments,screening the corresponding primer's annealingtemperature.(2) screened18primers from the100ISSR random primers which can amplify clear and specific ands,With the analyzation of30named species' genetic diversity,there are253polymorphic bands in253bands,a polymorphic rate of100%. Each primer amplified14polymorphic bands on everage,the polymorphic rate are all of100%.(3)30bamboo species polymorphism percentage change in the range of23.72%～44.27%. PPBlargest is Fargesia spathacea Franch, PPB minimum is Dendrocalamopsis oldhami, the two groups 'sadaptability is different. Other bamboo species polymorphism are much higher,30bamboo species havewide genetic basis, and rich genetic diversity.(4)30bamboo species observed number of alleles (Na) is2, the effective number of alleles (Ne) is1.5756, Nei ' s gene diversity index (H) is0.3437, Shannon diversity index (I) is0.5178, the percentageof polymorphic loci (PPB,%) is100%polymorphic loci is253, and30bamboo species' genetic diversityHt=0.3463.This description30species bamboo's genetic diversity is quite rich, the bamboo diversityprovides a molecular basis.(5) With the software of Popgene32,analyze genetic distance of30named bamboo species between0.104~0.6207,the average is0.4414,the largest genetic distance is0.6207,that is between No.9Guaduaamplexifolia J.S.Presl and No.27P. gramineus,the smallest genetic distance is0.104,that is between28Fargesia spathacea Franch and29Yushania niitakayamensis.(6) the DNA cluster analysis results and the traditional classification in many places were compared,in this study of bamboo species, such as the traditional classification of Phyllostachys subfamily ofIcrassiflora McClure, Sinobambusa tootsik, Brachystachyum densiflorum, Phyllostachys viridis, havinga close phylogenetic relationship, gathered in the sixth group; and Dendrocalamus strictus subfamily'sShibataea kumasasa, Semiarundinaria fastuosa, Chimonolamluca marmoreal, gathered in the sixthgroup; and the smallest genetic distance is Fargesia spathacea Franch. And Yushania niitakayamensisalso appeared in the fifth group; the traditional classification Shibataea kumasasa subfamily: theexperimental results appear Semiarundinaria fastuosa and Chimonolamluca marmoreal be all inShibataea kumasasa subfamily. Melocanna baccifera subfamily are concentrated in the second and fifthgroups.The experimental results and the traditional classification results are basically consistent, therefore,DNA molecular markers for the analysis of bamboo species phylogenetic relationships and geneticdiversity, it has higher accuracy and reliability, and can be used as the reference basis of bambooclassification.(7) The experiment uses18primers,as follow,UBC810,UBC811,UBC825,UBC827,UBC835,UBC836,UBC841and UBC857primers all can independent constructe a digital fingerprint identificationcode of30named bamboo species based on ISSR-PCR.(8) Using the traditional method to classify, mainly according to the bamboo outside significantmorphological features, using two ambiguous classification method to classify and naming, easy, intuitive,and has general applicability, appear earlier. ISSR molecular marker is a method completely different fromthe traditional classification, revealing30bamboo's immanent connection, fully reflects the bamboo genetic diversity. And the academic strong, pertinence strong, applicable groups is single, the classificationhas higher reliability and accuracy. ISSR molecular marker classification method is a new identificationtechnique means, opened up a new path to bamboo research.(9)The experiment based on bamboo in landscape studies, from genetic diversity, aestheticcharacteristics,space relations,application forms,cosplay,city green space and so on to analyse, thetraditional gardening thought about"suit the local conditions,obey the nature rule"and modern designconcepts about"ecological,low carbon"combined the two aspects together,pay attention to because ofHabitat home bamboo and biodiversity protection,broaden the horizons of bamboo landscape design,inorder to open a new way of bamboo garden using. But nowadays it need to explore the new design thoughtand design technique actively,to further development,it still has a long way to go.