IiAGL6 participates in the regulation of stamen development and pollen formation in Isatis indigotica.

The AGL6 (AGMOUSE LIKE 6) gene is a member of the SEP subfamily and functions as an E-class floral homeotic gene in the development of floral organs. In this study, we cloned IiAGL6, the orthologous gene of AGL6 in Isatis indigotica. The constitutive expression of IiAGL6 in Arabidopsis thaliana resulted in a late-flowering phenotype and the development of curly leaves during the vegetative growth period. Abnormal changes in floral organ development were observed during the reproductive stage. In woad plants, suppression of IiAGL6 using TRV-VIGS (tobacco rattle virus-mediated virus-induced gene silencing) decreased the number of stamens and led to the formation of aberrant anthers. Similar changes in stamen development were also observed in miRNA-AGL6 transgenic Arabidopsis plants. Yeast two-hybrid and BiFC tests showed that IiAGL6 can interact with other MADS-box proteins in woad; thus, playing a key role in defining the identities of floral organs, particularly during stamen formation. These findings might provide novel insights and help investigate the biological roles of MADS transcription factors in I. indigotica.

Extraction optimization and identification of anthocyanins from Nitraria tangutorun Bobr. seed meal and establishment of a green analytical method of anthocyanins.

This study aimed to extract and identify anthocyanins from Nitraria tangutorun Bobr. seed meal and establish a green analytical method of anthocyanins. Ultrasound-assisted extraction of anthocyanins from N. tangutorun seed meal was optimized using response surface methodology. Extraction at 70°C for 32.73 min using 51.15% ethanol rendered an extract with 65.04mg/100g of anthocyanins and 947.39mg/100g of polyphenols. An in vitro antioxidant assay showed that the extract exhibited a potent DPPH radical-scavenging capacity. Eight anthocyanins in N. tangutorun seed meal were identified by HPLC-MS, and the main anthocyanin was cyanidin-3-O-(trans-p-coumaroyl)-diglucoside (18.17mg/100g). A green HPLC-DAD method was developed to analyse anthocyanins. A mixtures of ethanol and a 5% (v/v) formic acid aqueous solution at a 20:80 (v/v) ratio was used as the optimized mobile phase. The method was accurate, stable and reliable and could be used to investigate anthocyanins from N. tangutorun seed meal.

Applications of HPLC/MS in the analysis of traditional Chinese medicines.

In China, traditional Chinese medicines (TCMs) have been used in clinical applications for thousands of years. The successful hyphenation of high-Performance liquid chromatography (HPLC) and mass spectrometry (MS) has been applied widely in TCMs and biological samples analysis. Undoubtedly, HPLC/MS technique has facilitated the understanding of the treatment mechanism of TCMs. We reviewed more than 350 published papers within the last 5 years on HPLC/MS in the analysis of TCMs. The present review focused on the applications of HPLC/MS in the component analysis, metabolites analysis, and pharmacokinetics of TCMs etc. 50% of the literature is related to the component analysis of TCMs, which show that this field is the most populär type of research. In the metabolites analysis, HPLC coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry has been demonstrated to be the powerful tool for the characterization of structural features and fragmentation behavior patterns. This paper presented a brief overview of the applications of HPLC/MS in the analysis of TCMs. HPLC/MS in the fingerprint analysis is reviewed elsewhere.