Transcriptome Analysis Of Anemone Rivularis Var.Flore-Minore Based On High-Throughputsequencing Technology And Flower Development Gene Screening

Anemone rivularis var.flore-minore is basal eudicots,belongs to the genus Anemone rivularis.After several years' survey,a serious of nature floral organs variants of Anemone rivularis var.flore-minore were determined.Based on varation organ and varation level,variants were divided into 5 types: white,green-white,five-flap,green and extreme variation.The transcriptome of Anemone rivularis Var.flore-minore was sequenced by Illumina Hi Seq 2000 platform,and some genes related to flower development were discovered by gene annotation.The m RNA expression levels of these genes were analyzed using real-time PCR.The Analysis of the main indexes of flower morphogenesis based on Principal Component Analysis(PCA).The main results of this study are as follows:The transcriptome of Anemone rivularis Var.flore-minore was sequenced by Illumina Hi Seq 2000 platform.The target sample sequencing a total of 54,513,822 reads fragment contains 7826726115 bp in sequence information were generated.A total of 43767 unigenes were formed by initial sequence splicing,with an average read length of 926 bp.Annotation analysis of unigene indicated that 28130 unigenes had homolog in public protein database.Compared with the NR,Swiss-Prot,KEGG,COG database,the obtained Unigene respectively matches 128051?21101?11574?17440 of the mentioned database,and the result indicated that 64.09%?48.21%?26.44%?39.85% unigenes.CDS prediction is performed to get a total of 30706 CDS,in which we get 27932 CDS by comparing with the Blastx and the public database,while 2774 CDS is obtained by ESTScan software'S prediction.Total 5015 SSRs were found from 43767 unigenes by using the SSR analysis.Frequency of occurrences for SSRs was 11.46%.Tri-nucleotide were major repeated types,accounting for 49.77%?The Di-nucleotide repeats,tetra-nucleotide repeats,penta-nucleotide repeats,hexa-nucleotide repeats account for 27.32%?6.66%?3.05%?13.20% respectively and the highest frequency was AG/CT,followed by AAG/CTT and ACC/GGT.12 MIKC genes closely related to flower development were selected,and the m RNA expression levels of these genes were analyzed using real-time PCR.The expression of AGL6,SEP3,FUL1,PI2 and SEP1 were the main indexes of flower morphogenesis based on Principal Component Analysis(PCA).The transcriptome data of this study enriched the genetic database of Anemone rivularis var.flore-minore,through the transcriptome selected MIKC genes closely related to flower development.Provide the basis for the study of flower organ mutation molecular mechanism of Anemone rivularis var.flore-minore.At the same time,as the basal eudicots,the study on Anemone rivularis var.flore-minore can provide more comprehensive understanding of angiosperm phylogeny and genetic evolution.