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Objective The information of bHLH transcription factor genes from transcriptome dataset of Scutellaria baicalensis was predicted by bioinformatics methods and the gene expression analysis was used to deduce its probable function. Methods The bHLH genes were screened from the transcriptome dataset of S. baicalensis by using BLAST comparison software. Then the open reading frames (ORFs) from the full-length of cDNA of bHLH genes were predicted by ORF Finder online tool, and its protein characteristics were analyzed using bioinformatic method. The expression of bHLH genes was detected by qPCR in different organs and treatments stimulated by Gibberellin A3 (GA3). Results Six genes of bHLH transcription factors were obtained, which belonged to six subfamilies of bHLHs of A. thaliana, two of which had completed ORFs. The results of gene expression showed that: The expression of bHLH2 and bHLH3 increased after 100 μmol/L GA3 treatment, and the expression of bHLH1, bHLH5, bHLH6 and bHLH7 decreased. The bHLH gene had the highest expression level in roots and flowers of S. baicalensis of them, among which bHLH1, bHLH2, bHLH5 and bHLH7 had highest expression in flowers and bHLH3 had highest expression in root. There was a correlation between bHLH gene and expression of biosynthesis and regulation genes of flavonoid. Conclusion These results provided the basis for further improving the molecular regulation network of flavonoids of bHLH genes in S. baicalensis.
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Establishing evaluation system of Dao-di herbs has been a problem to be solved in the field of traditional Chinese medicine (TCM), and is also a difficult problem in restricting the realization of TCM quality. On the basis of national drug standard substance requirements, this paper puts forward to set up reference Dao-di herbs in the first step of the evaluation system of Dao-di herbs, and discusses the properties, evaluation index system and its development requirements of reference Dao-di herbs, aiming at supporting the modern research and characteristics identification of Dao-di herbs in the future.
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Auxin response factor (ARF) is an important transcription factor for auxin signal transduction pathway, which regulates virtually every aspect of plant growth and development from embryogenesis to senescence. Nine full-length genes of ARF transcription factors were obtained from transcriptome dataset of Scutellaria baicalensis Georgi using the bioinformatics methods. The nucleotide and protein characteristics, subcellular localization, senior structural domains and conservative forecasts of those ARF transcription factors were analyzed. The phylogenetic tree showed that the nine ARFs in S. baicalensis were clustered together with ARF transcription factors in Arabidopsis thaliana, Oryza sativa subsp. Japonica and Nicotiana attenuate. The results of gene expression showed that:① The expression levels of ARF1, ARF3, ARF4, ARF8, ARF20 and ARF24 were upregulated after 100 μmol·L-1 GA3 treatment. However, the expression levels of ARF6 and ARF18 were downregulated; ② Those ARF genes were mainly expressed in the flowers of S. baicalensis; ③ There was a significant correlation between ARF genes and the genes involved in flavonoid biosynthesis. Our results provide a basis for further understanding the molecular regulation mechanism of flavonoid biosynthesis in S. baicalensis.
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To establish a new high resolution melting analytical method for identification of Lonicera japonica germplasm, the screening of 7 pairs of SSR (simple sequence repeats) primers, determining the suitable diagnostic primers by the differences of peak pattern and Tm was conducted. Then into the DNA template concentration, annealing temperature and the suitable range of cycle number were investigated. Combined with SIMCA-P software for data processing analysis, the results show that three main germplasm honeysuckle could be divided by four sets of primers. It provides methodology for improving L. japonica germplasm identification.