Analysis Of Transcriptome In Pomegranate Peel And Aril And Cloning Of MYB Gene

Pomegranate(Punica granatum L.)is an ancient tree of the family Lythraceae due to the long history of cultivation and known as a functional fruit with genetic diversity in our country.It is rich in flavonoids,punicalagincas and other antioxidants,which has a high economic value.The value of pomegranate merchandise mainly depends on the appearance of the fruit quality and intrinsic quality.The color of peel not only affects the appearance of the fruit traits and market value,but also is related to the fruit of the intrinsic quality and nutritional value to a large extent.The molecular mechanism of anthocyanin synthesis in pomegranate peel can not only lay an important theoretical foundation for the study of the coloring of pomegranate peel,but also improve the appearance quality of pomegranate by using the important gene of cloning to improve the molecular quality of pomegranate.At present,research with molecular mechanism of anthocyanin synthesis of fruit trees such as apple and grape and so on is very deep.In recent years,the molecular mechanism of anthocyanin synthesis of pomegranate has become a hotspot in pomegranate research.Currently,there is no any research in the transcriptome sequence in pomegranate in our country.In this study,the peel and aril of pomegranate were sequenced by Illumina Hi Seq 2500,and analysis of transcriptome of peel and aril comparitively,then research the metobolic pathways such as anthocyanin synthetical metabolism,clone MYB which is regulatory factors and predict its function.The main results are as follows:1?The assembly of the transcriptome sequence in peel and aril of pomegranate.The trancriptome of pomegranate was assembled using Velvet,SOAP Denovo and Trinity.Comparing the result,we find the length of N50 is 1483 bp when using Trinity,which effects was better than Velvet and SOAP Denovo.After assembling the aril,105444 pieces of contigs and 34227 Unigenes were obtained.After assembling the peel,76993 contigs and 24992 Unigenes were obtained.g The transposon data of pomegranate grains were spliced by using Soap Denovo,Velvet and Trinity software,and the results showed that Trinity software had the best effect on the data.2?Functional annotation of Unigenes obtained from pomegranate peel and aril assembled.A total of 16212 Unigenes sequences were homologous to the genes in the COG database.These Unigenes were classified into 25 categories according to the predicted function.The GO classification classifies 14427 Unigenes sequences in the 56 group classification.In the KEGG metabolic pathway analysis,a total of 6108 Unigenes sequences were found in 259 metabolic pathways.A total of 23089 Unigenes sequences were homologous to the genes in the COG database.These Unigenes were classified into 27 classes according to the predicted function.GO classification The 19768 Unigenes sequences were grouped into 54 groups.In the KEGG metabolic pathway analysis,a total of 8278 Unigenes sequences were found to be involved in265 metabolic pathways.3?Cloning and Bioinformatics Analysis of MYB Gene of Pomegranate.We cloned the pomegranate MYB gene by RACE technique and RT-PCR technique to obtain a 1015 bp sequence,which contain a 735 bp opening-reading frame(ORF).The ORF encodes 244 amino acids.We can identify the gene we cloned is MYB gene,because the MYB gene of Kiwi fruit,peach,Purple Ye Li,Litchi has high homology with we cloned.The entire peptide chain of the pomegranate MYB gene is located outside the cell membrane,and the encoded protein is hydrophilic and does not have a signal peptide and has two conservative domains.While the structural elements of the secondary structure are mostly in the alpha helix and random coil proteins,and the extended strand and beta turn are scattered throughout the protein.Tertiary structure of the MYB protein consists of six helves that pass through the connecting ring.