The Development Of Forsythia SSR Molecular Markers And Transcriptome Analysis Based On High-throughput Sequencing Technology

Forsythia is an important medicinal plant with a variety of active ingredients. It is distributed widely and is belonging to new resources deserving to develop and exploit. With the development of science and technology, the study of forsythia is deepened gradually from physical level to the genetic level. However, molecular markers of other species are often utilized and few of forsythia markers were adopted in the research, which caused the basic level of the study about forsythia transcriptome. In the present study, the advanced high-throughput sequencing technology-Illumina HiSeqTM2500was adopted for transcriptome sequencing of forsythia cultivated in Shanxi Agricultural University campus, and the result showed23.16M’s reads, after assembling,45,112Unigenes were got. The sequencing results was noted for Unigenes bioinformatics, including the comparison of NR, SwissProt, KEGG, COG, GO database, and there were28,699annotation results of Unigenes totally, which constructed the Unigenes forsythia genome database with good sequencing saturation and randomness. The Unigenes structural analysis of sequencing results suggested getting44,970points of Unigene ORF, of which the total length was24,056,832bp.The active ingredients of forsythia are regulated by various genes, and the technology of molecular markers is an effective way to analyze the function of these genes. Development of molecular marker loci is the basis of the gene function analysis. The study combined the technology of EST-SSR molecular markers, and SSR3,199points of marker loci were obtained totally,1,388,1266,605,21and9of which were mononucleotide repeat, di-nucleotide repeat, tri-nucleotide repeat, tetra-nucleotide repeat and penta-nucleotide repeat respectively. In order to verify the feasibility of these SSR loci, the present study was designed76pairs of primers and selected forsythia varieties from16regions, including Shanxi, Henan, Shaanxi and others. Then, PCR amplification of DNA extracted from leaves was conducted. Four primer pairs with obvious polymorphism stripes were screened, which can be used for analyzing the genetic evolution of forsythia species in different regions.The obtaining of biological information of forsythia transcriptome provided the basis of the molecular level research on forsythia. The designed molecule marker site would build a foundation for the classification of forsythia grown in different regions.