Transcriptome analysis of Saposhnikovia divaricata and mining of bolting and fowering genes

Objective: Early bolting of Saposhnikovia divaricata has seriously hindered its medicinal value and sustainable development of resources. The molecular mechanism of bolting and fowering of S. divaricata is still unclear and worth of research. In our study, we explored the transcriptome of the genes related to the bolting and fowering of S. divaricata.Methods: The transcriptome library was constructed, sequenced, assembled and annotated from the bolting and unbolting leaves of S. divaricata by high-throughput sequencing at the bud and fowering stage.Focus on the pathways related to bolting and fowering in plants, and exploring genes. The expression of seven candidate genes was verified by real-time fuorescence quantitative PCR(qRT-PCR).Results: Transcriptome results showed that 249 889 422 high-quality clean reads were obtained. A total of 67 866 unigenes were assembled with an average length of 948.1 bp. Trinity de Novo assembly produced 67 866 unigenes with an average length of 948.1 bp. Among 993 differentially expressed genes,484 genes were significantly up-regulated and 509 genes were down-regulated in the SdM group. A total of 79 GO terms were significantly enriched for differentially expressed genes. KEGG results showed that 11 154 unigenes were enriched in 89 pathways. And 21 candidate genes related to bolting and fowering of S. divaricata were excavated. The qRT-PCR results showed that expression trends of HDA9, PHYB, AP2,TIR1, Hsp90, CaM, and IAA7 were consistent with transcriptomic sequencing results. In addition, RNA-seq had identified 10 740 transcription factors and classified them into 58 families by their conserved domains. Further studies showed that the transcription factors regulating the fowering of S. divaricata were mainly distributed in the NAC, MYB_related, HB-other, ARF, and AP2 families.Conclusion: Based on the results of this study, it was found that the plant hormone signal transduction pathway was one of the decisive factors to control bolting and fowering. Among them, auxin related genes IAA and TIR1 are the key genes in the bolting and fowering process of S. divaricata.

Specification and grade of Saposhnikoviae Radix(Saposhnikovia divaricata)

Objective:Saposhnikoviae Radix(Fangfeng in Chinese),the roots of Saposhnikovia divaricata,lacks commodity specification and grade standardization in the current market.This study investigated the existing specifications and grades of Saposhnikoviae Radix to provide a standardized scientific reference for its market use.Methods:Based on a textual research of Chinese herbal medicine from the Han Dynasty to the present,medicinal materials of different specifications and grades obtained from Saposhnikoviae Radix in the main producing areas of China were collected and the markets for these materials were investigated.Field investigations were performed in the major producing areas such as Northeast China,Hebei Province,and Inner Mongolia.Four major Chinese herbal medicine markets in China were investigated.Sensory indices were used to categorize the two specifications(wild and cultivated)according to the shape,color,texture,and cross-section.High-performance liquid chromatography was performed to determine the active components.Vernier calipers and measuring tape were used to measure the diameter and length,respectively,of 41 samples.Using Excel and the R Language software,cluster analysis and descriptive statistical analysis were performed to assist in the application of new specifications and grades based on physical characteristics,pharmacological activity,and chemical composition.Results:The two specifications(wild and cultivated)of Saposhnikoviae Radix were divided into three grades each based on the length and diameter.Prim-O-glucosylcimifugin,5-O-methylvisamminoside,and the length of Saposhnikoviae Radix can be used as a basis for classifying the commodity specifications and grades.The specifications and grade standards of Saposhnikoviae Radix were established based on the following eight aspects:shape,surface characteristics,texture,cross section,taste,prim-Oglucosylcimifugin content,5-O-methylvisamminoside content and length.Conclusion:The formulation of this standard stipulates the commodity specification level of Saposhnikoviae Radix.It is also suitable for the evaluation of commodity specifications in the process of production,circulation and use of Saposhnikoviae Radix.