De novotranscriptomic analysis of Chlorella sorokiniana:Pathway description and gene discovery for lipid production

Abstract:

Abstract:［Objective］ The paucity of genomic information limits the metabolic engineering of non-model microalgae Chlorella sorokiniana．Our study aimed to elucidate the fatty acid，triacylglycerol and starch biosynthetic pathways in the microalgae C．sorokiniana based on de novo transcriptomic analysis．［Methods］ We cultured C．sorokiniana with different nitrogen concentrations (KNO3:8g/L and 2g/L)，then sequenced the transcriptomeusing Illumina Hiseq2000 platform．We used Trinity to de novo assemble the reads so as to obtain transcripts，aligned all the transcripts with Nr database，UniProtKB/Swiss-Prot database and COG database to annotate the function and classify using BLASTx algorithm，and assigned the transcript with metabolic pathway by aligning with KEGG database． Then we used ＲSEM to calculate FPKM value，and used it for preliminary analysis of different gene expression in the related pathways．［Results］Over 49M high quality raw reads were produced with the length of 100bp，We used Trinity to assembled these reads into 49885 transcripts with an N50 of 1941bp，ranging from 300bp to 14100bp． 26479 transcripts were annotated through BLASTx similarity search，2357 transcripts were assigned with EC number，and 207 metabolic pathways were assigned in total．Based on these analyses，we reconstructed the fatty acids，triacylglycerol and starch biosynthetic pathways in C.sorokiniana．We also identified preliminarily different geneexpression in the pathways．［Conclusion］ Using RNA-seq technology，we reconstructed the metabolic pathways involving in the fatty acid，triacylglycerol and starch biosynthesis in non-model microalgae C.sorokiniana without genomic data，which is consistent with those in model microalgae Chlamydomonas reinhardtii，and compared the gene expression level under different conditions．These information is very useful for the metabolic engineering of C．sorokiniana and other microalgae to enhance the production of lipids．