Biosynthesis of artemisinic acid in engineered Saccharomyces cerevisiae and its attractiveness to the mirid bug Apolygus lucorum

Artemisia annua is an important preferred host of the mirid bug Apolygus lucorum in autumn.Volatiles emitted from A.annua attract A.Iucorum.Volatile artemisinic acid of A.annua is a precursor of artemisinin that has been widely investigated in the Chinese herbal medicine field.However,little is known at this point about the biological roles of artemisinic acid in regulating the behavioral trends of A.lucorum.In this study,we collected volatiles from A.annua at the seedling stage by using headspace solid phase microextraction(HS-SPME).Gas chromatography-mass spectrometry(GC-MS) analysis showed that approximately 11.03±6.00 and 238.25±121.67 ng hartemisinic acid were detected in volatile samples and milled samples,respectively.Subsequently,a key gene for artemisinic acid synthesis,the cytochrome P450 gene cyp71 av1,was expressed in engineered Saccharomyces cerevisiae to catalyze the production of artemisinic acid.After the addition of exogenous artemisinic alcohol or artemisinic aldehyde,artemisinic acid was identified as the product of the expressed gene.In electroantennogram(EAG) recordings,3-day-old adult A.lucorum showed significant electrophysiological responses to artemisinic alcohol,artemisinic aldehyde and artemisinic acid.Furthermore,3-day-old female bugs were significantly attracted by artemisinic acid and artemisinic alcohol at a concentration of 10 mmol L,whereas 3-day-old male bugs were attracted significantly by 10 mmol Lartemisinic acid and artemisinic aldehyde.We propose that artemisinic acid and its precursors could be used as potential attractant components for the design of novel integrated pest management strategies to control A.lucorum.