地黄dna条形码序列的筛选与鉴定研究生物学遗传学专业论文.docxVIP

ABSTRACTDNA ABSTRACT DNA barCode is classified tool which discern species fast and accurately,and has a significant impact the analysis of biodiversity．The COI gene is the ideal DNA bareode to identify animals，such fish，insects，and birds，while DNA barcode fit in plant still in exploring stage．In this study,ITS，ITS2， matK,rbcL and psbA．trnH sequences and their combination arc compared and evaluatal in the discriminability of Rehmannia to provide theoretical foundation for screening of plant DNA barcode as well species identifcation．The main results as follows： 1．ITS，ITS2 and psbA-trnH fragments have better primer universality in PCR amplification．Due to amplification length，poor primer universality,and low efficiency of amplification，etc．，the amplification primers of rbcL and matK screened，eventually primer 1f0724r,636f-1368r is used to amplify rbcL sequence from Rehmannia glutinosa，and matk I F·tmk2R is used to amplify mat／(sequences from R· glutinosa．After screening primers and PCR amplification，primers of five sequence in this study have good commonality,amplification and sequencing，with 1 00％effective rate．Among them，ITS sequence needs to be combined with sequencing results to proofread due to poly structure． 2．ITS2 and ITS in Rehmannia glutinosa have high mutation rates and more information sites，such the base．pair substitution between purine and purine well pyrimidine and and pyrimidine，while rbcL and matK in R．glutinosa have strong conservatism．Furthor more ITS2 in R．glutinosa shows the higllest species diversity and low intraspecific variations，whether compared within the species genetic distance， ITS2 sequence has more effect than other sequences，but rbcL and mark have such small differences between species within species and could not be used to discern Rehmannia species．Inaddition， psbA．trnH sequence in R．glutinosa has gap of 0．4％·0．6％，though ITS，ITS2 and matK in R．glutinosa have obvious gap，the distance between the species within the species focuses bot