De Novo Assembly and Analysis of Polygonatum sibiricum Transcriptome and Identification of Genes Involved in Polysaccharide Biosynthesis

Abstract

<i>Polygonatum sibiricum</i> polysaccharides (PSPs) are used to improve immunity, alleviate dryness, promote the secretion of fluids, and quench thirst. However, the PSP biosynthetic pathway is largely unknown. Understanding the genetic background will help delineate that pathway at the molecular level so that researchers can develop better conservation strategies. After comparing the PSP contents among several different <i>P. sibiricum</i> germplasms, we selected two groups with the largest contrasts in contents and subjected them to HiSeq2500 transcriptome sequencing to identify the candidate genes involved in PSP biosynthesis. In all, 20 kinds of enzyme-encoding genes were related to PSP biosynthesis. The polysaccharide content was positively correlated with the expression patterns of β-fructofuranosidase (<i>sacA</i>), fructokinase (<i>scrK</i>), UDP-glucose 4-epimerase (<i>GALE</i>), Mannose-1-phosphate guanylyltransferase (<i>GMPP</i>), and UDP-glucose 6-dehydrogenase (<i>UGDH</i>), but negatively correlated with the expression of Hexokinase (<i>HK</i>). Through qRT-PCR validation and comprehensive analysis, we determined that <i>sacA</i>, <i>HK</i>, and <i>GMPP</i> are key genes for enzymes within the PSP metabolic pathway in <i>P. sibiricum.</i> Our results provide a public transcriptome dataset for this species and an outline of pathways for the production of polysaccharides in medicinal plants. They also present more information about the PSP biosynthesis pathway at the molecular level in <i>P. sibiricum</i> and lay the foundation for subsequent research of gene functions.