The Transcriptome Analyses Of Rudbeckia Hirta L. Under Lead Stress And Establish The Callus Receptor System Of Arabidopsis

Lead is a toxic heavy metal,which is not only harmful to the ecological environment,will also enter our body through the food chain,serious harm to human health.Restoration heavy metals polluted soil by using hyperaccumulator with it's traits of excess absorbing heavy metals from the soil and transporting to the aerial parts.Rudbeckia hirta L is a plant with a higher coefficient of lead transport,through seedlings were treated by lead stress under the added 1500mg/kg lead acetate solution,after 6 months,we collected the whole Rudbeckia hirta L as the sample and sequenced it by the high-throughput sequencing technology––Illumina Hi Seq,to sequence the transcriptome of Rudbeckia hirta L,a great deal of sequencing information was obtained.The transgenic experiment was carried out by Agrobacterium tumefaciens,the selected target gene Rh NRAMP1 is transferred into the recipient plant Arabidopsis,through the identification of PCR transgenic Arabidopsis,demonstrated that target gene has been integrated into the genome of Arabidopsis,establishment of Arabidopsis callus receptor system.The results were as follows:(1)After the extraction of the sample RNA about Rudbeckia hirta L,preparation of c DNA libraries,Illumina Hi Seq sequencing,filter unqualified sequences and assembly steps,finally we got 147131 of Rudbeckia hirta L Uni Gene,the total length is 86268038 bp,the average length is 586 bp and the length more than 1000 bp has 44653.(2)Compared with the Nr,Nt,Pfam,KOG,Swiss-prot,KEGG,GO database,the obtained Unigene respectively matches 51915?18447?36337?16381?36016?15455?36957 of the mentioned database.(3)Go and Pathway enrichment analysis of the Uni Gene was made to obtian 46 different functional categories and 32 metabolic pathways.The major of Go enrichment analysis is cellular process,metabolic process.The major of Pathway enrichment analysis is the transduction,carbohydrate metabolism,translation,etc.(4)We make a CDS forecast to get a total of 111320 CDS,and get 51853 CDS by comparing with the Blastx and the public database,while 59467 CDS is obtained by ESTScan software's prediction.(5)The optimum medium for callus induction in Arabidopsis was MS + 6-BA 1.0mg/L+NAA 0.1mg/L when leaves were used as explant.Transgenic plant of Arabidopsis which modification with Rh NRAMP1 gene were carried out by agrobacterium tumefaciens mediated method.The of higher efficiency for bud and root differentiation medium were MS+6-BA 2.0mg/L+NAA 0.1mg/L and MS + 6-BA 1.0mg/L+NAA 0.1mg/L respectively.