Juvenile Hormone III R Stereoisomer Is Specifically Synthesized by Honeybees (Apis mellifera ligustica) and Shows a Higher Biological Activity in Regulating Their Social Behavior

Abstract

The juvenile hormone (JH) plays a key role in the development of honeybee larvae and the alteration of adult behavior. Verification methods of types and stereoisomers of JHs in honeybees were established in this study. The regulatory modes of different stereoisomers of JH III on the social behaviors of honeybees were revealed by the disparity of interaction and RNA-seq. This result represented the first assessment of the effects of R-JH III and S-JH III in honeybee interactions; the former (367 times in total) was significantly higher than the latter (235 times in total); honeybees with high JH titers are always welcome in the colony because the effect of JH III on bees involves the sensing and signaling of hormones, and R-JH III is much more active than S-JH III in this regulation. Efficient R-JH III may be the insurance for bees to establish their social system advantages.

Simultaneous Detection and Distribution of Five Juvenile Hormones in 58 Insect Species and the Absolute Configuration in 32 Insect Species

“…The supplier had already performed nuclear magnetic resonance (NMR) validation of the JH I, JH II, JHSB 3 , and JHB 3 standards, so the absolute configurations of the main peaks of these four juvenile hormones had already been determined, as shown in Figure . Separation and identification of JH III were performed by our group . JH III standard was separated and identified by HPLC and chiral column (IG00CE-XL022) and is shown in Figure S1.…”

“…The predicted ECD spectrum was calculated by time-dependent density functional theory (TDDFT). By comparing the similarity between the experimental ECD spectra and the predicted ECD spectra in Figure S2, the absolute configuration of this pair of stereoisomers was determined …”

“…By comparing the similarity between the experimental ECD spectra and the predicted ECD spectra in Figure S2, the absolute configuration of this pair of stereoisomers was determined. 22 Five JH standards (0.25 mg JH I, 0.5 mg JH II, 100 mg JH III, 0.5 mg JHB 3 , and 0.5 mg JHSB 3 ) were accurately weighed, and the standards were dissolved in 10 mL of acetonitrile to obtain stock solutions. Thus, standard solutions of five JHs were prepared at a concentration of 2 μg/mL by acetonitrile dilution, and then a 0.2 μg/ mL mixed standard solution with acetonitrile was made.…”

“…Separation and identification of JH III were performed by our group. 22 JH III standard was separated and identified by HPLC and chiral column (IG00CE-XL022) and is shown in Figure S1. A J815 circular dichroism spectrometer was used to determine the experimental electronic circular dichroism (ECD) of the JH III stereoisomers.…”

“…We experimentally measured electron circular dichroism (ECD) spectra and computationally calculated the time-dependent density functional theory in our previous study. 22 ECD spectra of stereoisomers of 10R-JH III and 10S-JH III indicated that in Figure 1D the first peak (retention time 12.1 min) was 10S-JH III, and the other peak (retention time 13.1 min) was 10R-JH III. In Figure 1E, the fourth peak (retention time 14.1 min) was (10R, 11S)-JH II, and the remaining peaks were other isomers of JH II.…”