Transcriptomics Study Of Sugar Beet M14 Line Under Salt Stress Based On RNA-seq Technology

Northeast region of our country is one of the major distribution areas of salinization of soil,It has become one of the highlights in agro-scientific research of the northeast area crops that researching the salt tolerance ability of plants.Plants can maintain normal growth,increase crop yield by improving the ability of salt tolerance.Sugar beet M14 is special beet germplasm resources,that because addite a wild beet chromate on the basic of the diploid genome of cultivational sugar beet,with the more salt tolerance.So it was regarded as a good material to study the salt tolerance mechanism of plants.RNA-seq technology is regarded as the representative of the second generation sequencing technology,which can complete the transcriptome sequencing fast and accurately,but we do not refer to the data of transcriptome.In the present study,based on RNA-seq technology for the first time using the sequencing platform of Illumina to finish the transcriptome sequencing of sugar beet M14 lines,combined with the upgraded expression profile sequencing technology.In order to analysis and research the salt tolerance mechanism of sugar beet M14 lines under different treatments,equal amounts of leaves and roots expect with 0,200,400 mM NaCl treatment respectively of the function of differential expression gene.The main results were as follows:1.Based on RNA sequencing of the hybrid strains of sugar beet M14 samples,and we build the conferance transcriptome bases of sugar beet M14 lines.After removal of low quality and sub-sequences were 54089906 clean reads,in which the quality of more than 20(Q20)occupy more than 97% of the total base.Assembled by joining together,we get the average of 54843 Unigenes length is 663 nt,and the value of N50 is 1141 nt.2.All the Unigenes sequencing for Nr,GO(Gene Ontology)and COG(Clusters of Orthologous Groups)and KEGG(Kyoto Encyclopedia of Genesand Genomes)comments,function classification and pathway.A total of 33542 Unigenes get comments,which was classified that there is 25 COG function classification and 121 in the KEGG pathway.3.By using Digtial Gene expression profile sequencing,we get the Unigenes annotations of 10591,10613 and 10636 under 0 mM,200 mM and 400 mM NaCl treatment respectively.Also has the Unigenes have comments of 10708,10620 and 10698 under 0 mM,200 mM and 400 mM NaCl treatment at the root respectively.Gene expression is obtained by using the RPKM method.4.Determine the FDR less than 0.001(FDR?0.001)and multiple differences in 2 times or more(|log2(B-RPKM/A-RPKM)|?1).When the data of RPKM more than 10 of Unigenes,which represent of the differentially expression genes under salt stress of sugar beet M14 lines,also analyzes its function and metabolic pathways.Annotation analysis involves some important metabolic pathways which is related to the resistance of plant salt.These comments above those results provide some valuable resources,which is benefit to study some specific process,function and path of crucial substance metabolism in sugar beet M14 lines.According to the information of present study and the literature about the salt tolerance mechanism,we can determine the main 10 research object which is related to the gene of adjustment under salt stress,also draw the metabolic pathways graph of sugar beet M14 lines under salt stress.5.According to the information of transcriptome database of sugar beet M14 lines,we research 20 differential expression genes and related to salt stress gene by using Real Time PCR to verify its data reliability.And select 12 differential expression genes of the related to ROS way by using Real-Time PCR to test related gene expression regulation in the way.