Enhanced production of calycosin-7-O-β-D-glucoside and astragaloside Ⅳ from adventitious root cultures of Astragalus membranaceus var. mongholicus by green leaf volatiles

Elicitation of (E)-2-Hexenal and 2,3-Butanediol on the Bioactive Compounds in Adventitious Roots of Astragalus membranaceus var. mongholicus

“…15 By performing the 2and 6-week root cultures, Sun et al have revealed that the presence of hexanol, hexanal, and (E)-2-hexenal, especially the first chemical, increases the production of CG and astragaloside IV in A. mongholicus adventitious roots. 16 Their work also shows a short-term elicitation of (E)-2-hexenal on the production of astragaloside IV, which deserves further investigation. Therefore, 2,3-butanediol is one of the promising VOCs produced by plant microbes including Bacillus subtilis GB03, Bacillus amyloliquefaciens 1N937a, and Enterobacter aerogenes.…”

“…A. mongholicum adventitious roots were used as plant material, and its induction was performed by proliferating apical buds of A. mongholicus shoots preserved in our laboratory on the solid Murashige and Skoog (MS) medium containing 0.7% agar, 30 g/L sucrose, and 1 mg/L naphthylacetic acid (Bio Basic Inc., Toronto, Canada) by the method described previously. 16 Subsequently, the adventitious roots were cultured in the liquid half-strength MS medium containing 1 mg/L indole-butyric acid (IBA) (Jintan Maosheng, Changzhou, China) and 30 g/L sucrose at 25 °C on a shaker (100 rpm) under dark conditions. The subculture of the root tissue was performed twice before the application of the volatile chemicals.…”

“…Extraction of total RNA and synthesis of first-strand cDNA were performed by the methods, the reagents, and the kits described. 16 The synthesized cDNA was diluted 10 times with sterile water as the template for qPCR. Quantification of the gene expression was performed in a CFX96TM Real-Time System and C1000 Touch Thermal Cycler (Bio-Rad) using the TB Green Premix Ex TaqTM II kit (Tli RNaseH Plus) (Takara, Dalian, China).…”

“…Normalization was carried out by using cold-regulated 413 plasma membrane protein 1 (COR413-PM1) as the internal reference. 16 Details of the genes and primers involved in the biosynthesis of astragaloside IV are presented in Table 2, while the ones associated with the CG biosynthesis were the same as described. 16 Corresponding melt curves from qPCR of the genes are presented in Figure S1, suggesting that the amplification products were single and the primer pairs were specific.…”

“…16 Details of the genes and primers involved in the biosynthesis of astragaloside IV are presented in Table 2, while the ones associated with the CG biosynthesis were the same as described. 16 Corresponding melt curves from qPCR of the genes are presented in Figure S1, suggesting that the amplification products were single and the primer pairs were specific. The quantification of the gene expression was based on a comparative cycle threshold (CT) value.…”