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 b...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.展开更多
Plants of Artemisia annua produce artemisinin, a sesquiterpene lactone widely used in malaria treatment. Amorpha-4,11-diene synthase （ADS）, a sesquiterpene synthase, and CYP71AV1, a P450 monooxygenase, are two key e...Plants of Artemisia annua produce artemisinin, a sesquiterpene lactone widely used in malaria treatment. Amorpha-4,11-diene synthase （ADS）, a sesquiterpene synthase, and CYP71AV1, a P450 monooxygenase, are two key enzymes of the artemisinin biosynthesis pathway. Accumulation of artemisinin can be induced by the phytohormone jasmonate （JA）. Here, we report the characterization of two JA-responsive AP2 family transcription factors-AaERF1 and AaERF2-from A. annua L. Both genes were highly expressed in inflorescences and strongly induced by JA. Yeast one- hybrid and electrophoretic mobility shift assay （EMSA） showed that they were able to bind to the CRTDREHVCBF2 （CBF2） and RAVlAAT （RAA） motifs present in both ADS and CYP71AV1 promoters. Transient expression of either AaERF1 or AaERF2 in tobacco induced the promoter activities of ADS or CYP71AV1, and the transgenic A. annua plants overexpressing either transcription factor showed elevated transcript levels of both ADS and CYP71AV1, resulting in increased accumulation of artemisinin and artemisinic acid. By contrast, the contents of these two metabolites were reduced in the RNAi transgenic lines in which expression of AaERF1 or AaERF2 was suppressed. These results demonstrate that AaERF1 and AaERF2 are two positive regulators of artemisinin biosynthesis and are of great value in genetic engineering of arte- misinin production.展开更多
基金supported by the National Natural Science Foundation of China (31772176 and 31972338)the National Key Research and Development Program of China (2019YFD0300100)
文摘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.
基金This research was supported by State Key Basic Research Program of China （2007CB108800）, the National Natural Science Foundation of China （30630008）, and the National HighTech Program of China （2007AA021501 ）.ACKNO WLEDGMENTS We thank CYP71AV1. discussions Ke-Xuan Tang for supplying the promoter sequence of We thank Ji-Rong Huang and Gao-Jie Hong for he pfu No conflict of interest declared
文摘Plants of Artemisia annua produce artemisinin, a sesquiterpene lactone widely used in malaria treatment. Amorpha-4,11-diene synthase （ADS）, a sesquiterpene synthase, and CYP71AV1, a P450 monooxygenase, are two key enzymes of the artemisinin biosynthesis pathway. Accumulation of artemisinin can be induced by the phytohormone jasmonate （JA）. Here, we report the characterization of two JA-responsive AP2 family transcription factors-AaERF1 and AaERF2-from A. annua L. Both genes were highly expressed in inflorescences and strongly induced by JA. Yeast one- hybrid and electrophoretic mobility shift assay （EMSA） showed that they were able to bind to the CRTDREHVCBF2 （CBF2） and RAVlAAT （RAA） motifs present in both ADS and CYP71AV1 promoters. Transient expression of either AaERF1 or AaERF2 in tobacco induced the promoter activities of ADS or CYP71AV1, and the transgenic A. annua plants overexpressing either transcription factor showed elevated transcript levels of both ADS and CYP71AV1, resulting in increased accumulation of artemisinin and artemisinic acid. By contrast, the contents of these two metabolites were reduced in the RNAi transgenic lines in which expression of AaERF1 or AaERF2 was suppressed. These results demonstrate that AaERF1 and AaERF2 are two positive regulators of artemisinin biosynthesis and are of great value in genetic engineering of arte- misinin production.
