Transcriptome Analysis Of Mikania Micrantha Infected By Mikania Micrantha Wilt Virus At Different Time

Mikania micrantha is one of the major exotic plants in southern China.At present,the method of controlling the invasion of Mikania micrantha is mainly about artificial removal,chemical control and biological defense.However,these methods are expensive and just available within a small range.It still needs to do a lot of in-depth research before being applied to practice.In this paper,the invasive weed Mikania micrantha is chosen as the object of our study.By analyzing the RNA-Seq results of Mikania micrantavir wilt virus(MMWV)infected Mikania micrantha at different time,This paper mainly discusses the response mechanism of Mikania micrantha to MMWV infection and uncovers related functional genes.Meanwhile,this study provides an important theoretical basis for developing new methods to prevent the invasion of Mikania micrantha by using MMWV.The results are as follows: 1.Construction sequence library and high-throughput sequencing and assembly of MMWV of infecting Mikania micrantha at different timeMMWV can induce the wilting and shrinkage of Mikania micrantha as well as inhibit the growth of Mikania micrantha.But at present research for the virus' genetic diversity and pathogenesis is still blank.In this study,MMWV is infected by Mikania micrantha for 30 days and 15 days,and normal control(non-infective)treatment.RNA-Seq results are obtained by Illumina HiSeq 4000.The final C lean Reads is about 1.39 × 108 bp,and then by rearranging the C lean Reads of three sequence samples through the Trinity software,a total of the total group transferred out of the potential transcripts is 169497 and the total potential gene is 121379.It is found that there are 10960 genes that can be read at the same time as Uniprot,NR and N T databases.In the frame of Unigene,in the classification of biological processes,the GO classification of Mikania micrantha unigene is mainly about the cellular process and the metabolic process.In the classification of cell components,the cell part is involved in the cell part with the highest proportion of unigene.In the molecular functional classification,genes with binding and catalytic activity function account for the highest proportion.All the Unigene are compared with COG databases,and the result shows that the largest number of genes is overall functional class with 2425 genes,followed by replication,recombination,repair,translation,ribosome structure and biotransformation,post-translational modification,protein Metabolic and other categories.2.Analyzing the differentially expressed genes of MMWV infecting Mikania micrantha at different timeComparing gene expression in infection under different time period,Under Infected for 15 days and 30 days the condition of differentially expressed genes number 356 and 18272 respectively with compared normal control group There are 87 distribution on cross section of the two contrast,namely the differentially expressed in every conditions,there are 56 raised expression,13 down expression.O n this basis,the focus on the response MMWV infection related genes involved in adaptive mechanism,After infection15 days,unsaturated fatty acid biosynthesis raised five gene expression,the four genes appear lower expression of cell cycle;Infection.After infection 30 days,164 genes appear to raise expression and1 gene down expression of transporters,;Plant-pathogen interaction of 67 genes raised expression,17 genes down expression.These activities may be related to plant growth and development,regulation and control of resistance,defense reaction.3.Real-time quantitative PCR validation analysisFour genes related with the secondary metabolism of plant are chosen and they are polyphenol oxidase(PPO),chalcone synthase(CHS),ascorbate peroxidase(APX)and dihydro Flavonol-4-reductase(DFR).Their gene expressions changes infected of MMWV on days of 5 days,10 days,15 days,20 days,25 days and 30 treatments are investigated by real-time fluorescence quantitative PC R.It can be seen from the quantitative results of fluorescence,although there are some differences in the degree of expression changes,from the expression of four genes,MMWV infection significantly increases the resistance of Mikania micrantha and resistance-related gene expression.The results of real-time fluorescence quantitative analysis are consistent with those of the RNA-Seq results,The results Confirmed the inhibition by MMWV infect f mikania micrantha.In conclusion,Through the variance analysis of the transcriptome sequencing results,this study obtained 87 genesof differences significant expression of Mikania micrantha infected by MMWVat different time,56 genes are up expression,13 genes are down expression;These genes enrich in 48 metabolic pathways,the main metabolic pathways are related with plant defense responses,secondary metabolism;The results of real-time fluorescence quantitative analysis are consistent with those of the RNA-Seq results,indicating the reliability of the RN A-Seq results data obtained by Illumina sequencing.