Analysis Of Characteristics Of Quality And The Transcriptome Of Three Tuckahoe Varieties(Poria Cocos)

Tuckahoe?Poria cocos? is a traditional Chinese medicine. Polysacchrides of tuckahoe have many biological functions such as anti-aging, anti-neoplastic, anti-inflammatory, and so on. The thesis mainly focuses on the analysis of characteristics of quality and transcriptome of three tuckahoe cultivars?"5.78" is a traditional wild type; "XIANGJING 28" comes from "5.78"; "JINGHANGYIHAO" is originated from the "XIANGJING 28" after being carried to the "Number ten Shenzhou spaceship"?. Our work is to screen out some key genes that are releted with the tuckahoe quality, and establish the foundation for the further study, popularization and application of the tuckahoe.The major results related with the characteristics of quality analysis are as follow:?1? We compared the processing performance of the three tuckahoe cultivars. The results showed that the drying rate of "JINGHANGYIHAO" is the highest, and the second is "5.78", and that of "XIANGJING 28" is the lowest. The production rate of "JINGHANGYIHAO" is the highest, and the second is "XIANGJING 28", and that of "5.78"is the lowest. The ratio of skin and flesh of "JINGHANGYIHAO" is the highest, and the second is"XIANGJING 28", and that of "5.78"is the lowest. Thus it can be concluded that "JINGHANGYIHAO" has the most significant advantages in production and economic benefits.?2? We analyzed the common active ingredients of tuckahoe. The total protein content of "5.78" is the most and the second is "XIANGJING 28". The content of polysaccharide of "XIANGJING 28" is the highest, and the second is "JINGHANGYIHAO", and "5.78" is the lowest. The content of polysaccharide of "JINGHANGYIHAO" and "XIANGJING 28" have few differences, but the content of polysaccharide of "JINGHANGYIHAO" is ten percent higher than that of "5.78". Because the dry weight ratio of fresh weight of the "JINGHANGYIHAO" is higher than that of the "XIANGJING 28", the content of polysaccharide in the fresh "JINGHANGYIHAO" is much higher than that of the "XIANGJING 28".The analyses of transcriptome of three varieties of tuckahoe are as follow:?1? After the extraction of RNA, preparation of the cDNA library, sequencing and control of the quality of the sequencing, eventually we got 13.04 Gb Clean Data in total.?2? Based on the query with the COG, GO, KEGG, Swiss Prot, nr databases, the numbers of genes annotated are 4261,5418,2712,7711,2712, respectively. According to the COG database, the genes were divided into 25 different gene functional annotations, among which the gene involving general function prediction makes up most. The most genes are enriched in the categories involved in amino acid transport and metabolism, replication recombination and repair, secondary metabolites biosynthesis transport and catabolism, carbohydrate transport and metabolism. According to the GO database,15191 genes are related with the biological processes, accounting for 52% of all, and 8331 genes with the molecular function, accounting for 28%, and 6163 genes with the cell components, accounting for 20%. Pathway enrichment analysis showed that the genes were mainly mapped onto 92 KEGG standard metabolic pathways.?3? The encoded peptide chains that are too short ?less than 50 amino acid residues? or only contain a single sequence of exon are filtered out. Compared with the original genome annotation information, this research discovered 673 new genes of three varieties of tuckahoe. Based on the query with the COG, GO, KEGG, Swiss Prot, nr databases, the numbers of new genes annotated are36,59,8,51,247, respectively.?4? Based on the RPKM method, there are 947 differentially expressed genes?DEG? between "5.78" and "XIANGJING28", among which there are 549 up-regulated genes and 398 down-regulated genes. There are 1194 DEGs between "5.78" and "JINGHANGYIHAO"; among which there are 540 up-regulated genes and 654 down-regulated genes.There are 1097 DEGs between "XIANGJING28" and "JINGHANGYIHAO", among which there are 548 up-regulated genes and 549 down-regulated genes.?5? Based on the query with the COG, GO, KEGG, Swiss Prot, nr databases, the numbers of DGEs between "XIANGJING 28"and "5.78" annotated are 372,539, 289,705,880, respectively. Based on the query with the COG, GO, KEGG, Swiss Prot, nr databases, the numbers of DGEs between "XIANGJING 28"and "5.78" annotated are 476,601,343,820,1085, respectively. Based on the query with the COG, GO, KEGG, Swiss Prot, nr databases, the numbers of DGEs between "JINGHANGYIHAO"and "XIANGJING 28" annotated are 458,531,244,705,995 respectively.?6?The further analysis on the differentially expressed genes in starch and sucrose metabolic pathway and galactose metabolism pathway was carried on. The results showed that in the starch and sucrose metabolism pathway, compared with "5.78" and "XIANGJING 28", gene₁0432 controlled the synthesis of polygalacturonase expressed significantly up-regulated in "JINGHANGYIHAO" and compared with "XIANGJING 28" gene₂561, gene₃259 controlled the synthesis of a-glucosidase expressed up-regulated in "JINGHANGYIHAO". In the Galactose metabolism pathway, compared with "5.78" and "XIANGJING 28", gene₅42 controlled the synthesis of a-galactosidase expressed significantly up-regulated in "JINGHANGYIHAO". We selected eight DEGs that are related to the synthesis of polysaccharides for qRT-PCR validation. The results showed consistent expression patterns are consistent with the observed findings in transcriptome analysis. That indicate the dependability of the transcriptome sequencing data. It's speculated that the content of polysaccharide in fresh "JINGHANGYIHAO" is more than that of fresh "5.78" and "XINANGJING 28", which may be due to these four differentially expressed genes in two pathways.