sRNA analysis and regulation of photosynthesis and lipid metabolism of Camellia oleifera
[Objective]The types,quantities and characteristics of sRNAs in Camellia oleifera were analyzed and identified,the miRNA's functions and corresponding target genes were revealed,and the theoretical basis for genetic improvement of C.oleifera was provided by miRNAs and their target genes.[Method]The excellent C.oleifera cultivar'Hengchong 89'were used as materials.Its tender leaves and mature seeds were collected to extract total RNA,and separated to enrich mRNA with magnetic beads.The Illumina RNA-seq high-throughput sequencing technology was used to obtain sRNA libraries.The structure characteristics,types,quantities,common and unique sequences of sRNAs were analyzed.Then,sRNAs were located to C.oleifera transcriptome to analyze types and quantities of mapped sRNAs.Mapped analysis and the first base bias analysis of known miRNAs were also carried out.Novel miRNAs were predicted by miREvo and miRDeep2 software.The miRNA family analysis,miRNA expression and differential analysis,clustering analysis were implemented.Finally,prediction of miRNA target genes,GO functional enrichment and KEGG pathway enrichment analysis of target gene candidates were also implemented.The analysis of differentially expressed miRNAs and screening target genes participated in photosynthesis and lipid metabolism were carried out.[Result]The leaf and seed sRNA libraries of C.oleifera were constructed.The clean read counts of 18-40 nt sRNAs were 14 256 189 and 10 822 313,respectively,belonging to types of 6 138 179 and 4 980 518,respectively.Among them,counts of 24-nt sRNA was the largest.The common sequence types of in the two sRNA libraries were 773 647,while the unique sequences were 4 206 871 and 5 364 532,respectively.The total reads aligned to the reference sequences of C.oleifera transcriptome were 5 855 009 and 4 122 070,respectively in the two sRNA libraries.118 miRNA precursors and 78 matures were mapped.It was found that the first nuclotied bias of 18-23 nt known miRNAs was U.54 new miRNAs were identified,22 known miRNAs families,including 105 family members,and 67 differentially expressed miRNAs were also identified.These differentially expressed miRNAs regulated 1346 target genes.In which 26 miRNAs were significantly differentially expressed,regulating 644 target genes.Cluster analysis showed that significant differential expression miRNAs were divided into two groups.One group had lower expression levels,while the other had higher ones in leaves and seeds.The GO analysis revealed that target gene candidates were enriched in 51 terms of biological processes,cellular components and molecular functions.The KEGG analysis indicated that target gene candidates were significantly enriched in metabolic processes of terpenoid backbone biosynthesis,glycerolipid metablism,aminoacyl-tRNA biosynthesis,etc.In addition,it was found that ath-miR5658 and gma-miR169v were differentially expressed in leaves and seeds.Their corresponding target genes NADP-dependent malic enzyme-like,fructose-bisphosphate aldolase 1 in carbon fixation and photosynthesis,monogalactosyl diacylglycerol synthase 2 in glycerolipid metabolism,etc.were also identified.[Conclusion]Our study preliminarily confirmed the phenomenon of"one miRNA regulating multiple target genes,and multiple miRNAs regulating one target gene".It was found that ath-miR5658 could regulate the expression of target genes in pathways of carbon fixation,photosynthesis,glycerophospholipid,glyceride metabolism and fatty acid elongation,and that gma-miR169v could regulate the expression of target genes in glycerophospholipid metabolism.This study has certain theoretical value in revealing the regulatory mechanism of photosynthetic carbon fixation,fatty acid and lipid synthesis of C.oleifera,and cultivating new high-quality and high-yield cultivars.
Camellia oleiferasRNAmiRNAtarget genesregulationphotosynthetic and lipid metabolic metabolism
万方数据
维普
