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Artemisia annua  produces artemisinin, an effective antimalarial drug. In recent decades, the later steps of artemisinin biosynthesis have been thoroughly investigated; however, little is known about the early steps of artemisinin biosynthesis. Comparative transcriptomics of glandular and filamentous trichomes and  13 CO 2  radioisotope study have shown that the 2-C-methyl-  D  -erythritol-4-phosphate (MEP) pathway, rather than the mevalonate pathway, plays an important role in artemisinin biosynthesis. In this study, we have cloned three  1-deoxy- D -xylulose 5-phosphate synthase  ( DXS ) genes from  A .  annua  ( AaDXS1 ,  AaDXS2 , and  AaDXS3 ); the DXS enzyme catalyzes the first and rate-limiting enzyme of the MEP pathway. We analyzed the expression of these three genes in different tissues in response to multiple treatments. Phylogenetic analysis revealed that each of the three  DXS  genes belonged to a distinct clade. Subcellular localization analysis indicated that all three AaDXS proteins are targeted to chloroplasts, which is consistent with the presence of plastid transit peptides in their N-terminal regions. Expression analyses revealed that the expression pattern of  AaDXS2  in specific tissues and in response to different treatments, including methyl jasmonate, light, and low temperature, was similar to that of artemisinin biosynthesis genes. To further investigate the tissue-specific expression pattern of  AaDXS2 , the promoter of  AaDXS2  was cloned upstream of the  β-glucuronidase  gene and was introduced in arabidopsis. Histochemical staining assays demonstrated that  AaDXS2  was mainly expressed in the trichomes of Arabidopsis leaves. Together, these results suggest that  AaDXS2  might be the only member of the DXS family in  A. annua  that is involved in artemisinin biosynthesis.

Molecular Characterization of the 1-Deoxy-D-Xylulose 5-Phosphate Synthase Gene Family in Artemisia annua