Expression Analysis Of Growing Related Genes In Rehmannia Root

Rehmannia glutinosa Libosch to scrophulariaceae rehmannia is a perennial herbaceous plants,is one of the famous "four huaiqing Chinese medicines",one has Ziyinqingre,blood and stop bleeding and other effects.Rehmannia widely used for a variety of prescription drugs and proprietary compatibility to the main raw material.Rehmannia root mainly in medicine,the resources depend on cultivation,but the molecular mechanism is unknown to root development,medicinal material production are susceptible to natural condition such as disease,the effect of continuous cropping obstacle,etc,which reduces the yield and quality of medicinal materials.Therefore it is necessary to research on molecular biology,explore rehmannia root growth mechanism,guide rehmannia production practice.To this end,we first carried out rehmannia internal research,fluorescence quantitative PCR screening in stable internal genes expressed in different tissues and organs.The experimental application BestKeeper,GeNorm and NormFinder 3 kinds of software on rehmannia reference gene stability were evaluated.Based on the comprehensive analysis of the three types of software as a result,we found that the Tip41,UCH2,rGAPDH,GAPDH four genes are expressed in different tissues and organs of rehmannia stability is good,of which the most Tip41 stability,can choose as rehmannia quantitative internal gene expression research.Rehmannia internal genetic screening for relevant genes of rehmannia root development in the next step to lay the foundation,also for the future of rehmannia other molecular biology research provides the convenience.Then,in order to explore the related mechanism of rehmannia root development,screening and rehmannia root development related genes.This study by adopting Illumina Hiseq2000 high-throughput sequencing technology platform for radix rehmanniae seven organs,namely:the adventitious roots(AR),the starting period root(TAR),expands period root(DTR),mature root(MR)and stem(ST)and mature leaves(L),flower(F)conducted the transcriptome sequencing and functional analysis.Won the 220000 rehmannia unigene and use Nr,Nt,Interpro,SwissProt,KEGG,COG,and GO to annotation unigene database.On this basis,we according to each gene RPKM worth to different tissues and organs and radix rehmanniae radix rehmanniae root growth amount of gene expression in different periods,the method of using Z-score in aerial parts of radix rehmanniae,for reference,the root development of differentially expressed genes screening(parameter Z-score>3.75 and thewire>2.0),and screened 8393 unigenes enrichment in rehmannia root.Then according to the result of radix rehmanniae unigene comments,and GO,COG functional classification and metabolism of KEGG pathway analysis,screening transcription factors associated with rehmannia root development in 61,involved 111 related uni genes.The main response factors including plant hormones,cell division-related protein,cellulose synthase,ethylene response transcription factor nuclear factor and mitogen-activated protein kinase.According Z value<-1.5 and ratio<0.5,we screened 1048 Unigenes down at four different times in Rehmannia root.Then according to the gene functional annotation,May 26 transcription factors associated with rehmannia root development,involving the Unigenes 29.Including AUX/IAA plant hormones inducing factor,C3HL family of transcription factors,MADS family of transcription factors and LOB family of transcription factors.Through the screening of differentially expressed genes related to rehmannia root development,explore the molecular mechanisms of root development,molecular mechanisms in the development of rehmannia root to clarify the theory basis and variety improvement.