Transcriptome Analysis Of Pinellia Ternata Mutant Based On High Throughput Sequencing

Pinellia ternate(Thunb.)Breit is a perennial herb of Pinellia ternate genus of Araceae.In China,as one of the important Chinese traditional medicine,the dry tuber of Pinellia ternate is often used medicinally.As the lower product,It is first recorde din Shen Nong,s Herbal Sutra.Compared to other Chinese traditional medicines,it is more toxic and cannot be used for long time.According to the Pharmacopoeia,it has a significant treatment for cough,phlegm,vomiting,cough,headache,vertigo,relieving bodily pain and swelling for external use and antitumor potential.As one kind of weedy plants,the bulbil breeding is one of the important breeding methods of Pinellia ternate.It has the advantages of high efficiency and high yield by the bulbil multiplication,which greatly increases the survival rate of Pinellia ternate in nature and agricultural production.Therefore,the quality of Pinellia ternate bulbilis an important factor that determines the quality and yield of Pinellia ternate,and the important factor of Pinellia ternate planting.In order to cultivate high quality and high yield varieties of Pinellia ternate,genetic engineering has become one of the solutions to alleviate the shortage of Pinellia ternate resources.Because the wild Pinellia ternate yield is decreasing and the demand of market is increasing.At present,the research of Pinellia ternate is mainly focused on the screening of high quality varieties,high yield,pharmacological effects and clinical studies,but the research on the function genes of Pinellia ternate bulbil is less and progress lower.In the study,Illumina Hi SeqTM 4000 high throughput sequencing technology was used to the two samples of leaf-base-bulbil Pinellia ternate(BY)and stem-base-bulbil Pinellia ternate(ZC),repeated three biological repetitions.The transcriptional group was sequenced,and obtained 52.44 G Raw Data in total,in which Valid Data 48.47 G and Valid% accounted for 92.43%.After data was assembled and spliced by Trinity,28395 unigenes were obtained.The total sequence length was 20314911 bp,the average length was 715 bp,and the length of N50 sequence was 1063 bp.The obtained unigenes was analyzed by GO,KEGG and COG.The results showed that 11351 unigenes obtained functional annotations in GO classification,including three aspects of molecular biological function,biological pathway and cell component.a total of 7474 unigenes were assigned to 242 KEGG metabolic pathways.17130 unigenes was annotated to COG classification.The annotated unigenes covers most of the life activities.The predicted CDS of unigenes is obtained 18316 CDSs and CDS prediction ratio is 64.50%.SNP analysis showed that 36146 SNP polymorphic locis were detected in BY,and 40133 SNP polymorphic locis were detected in ZC.SSR analysis showed a total of 4807 SSR markers,including single nucleotide repeat,di-nucleotide repeat,tri-nucleotide repeat,tetra-nucleotide repeat,five nucleotide repeat and six nucleotide repeat.Screening differentially expressed genes can be used to analyze functional genes.These genes were used to improve the genetic breeding of the species,and to improve the ability of stress resistance and molecular mechanism.Differential expression of genes refers that the differential expression of functional proteins was caused by physiological stress reaction in a specific environment,which leads to the difference in the growth phenotype.By using transcriptome sequencing analysis,we can select and unearth genes of non-reference function genome species in bulk,and exploring differential gene expression of different samples.In this study,the 13656 DEGs were screened efficiently from 28395 unigenes of the Pinellia ternate transcriptional group.The DEGs was included in 50 functional subclasses by GO classification,which were concentrated in the characters expression of the growth and development of the Pinellia ternate as well as the secondary metabolism.According to the analysis of KEGG pathway,3551 differentially expression genes enrich in the process of physiological metabolism and secondary metabolites in the bulbil of Pinellia ternate,which accounted for 26% of the total amount of DEGs.Comparative analysis of the DEGs of leaf-base-bulbil Pinellia ternate(BY)and stem-base-bulbil Pinellia ternate(ZC)and screening the DEGs which affected the biosynthesis and signal transduction of Pinellia ternate bulbil.It provided a basis for the in-depth study of the cloning and functional analysis of the related functional genes of Pinellia ternate.It is significant to study the gene regulation mechanism of Pinellia ternate bulbil character,and provide scientific basis for the breeding and improvement of high quality and high yield.