@article{ma_li_alejos-gonzalez_zhu_xue_wang_zhang_li_ye_wang_et al._2017, title={Overexpression of a type-I isopentenyl pyrophosphate isomerase of Artemisia annua in the cytosol leads to high arteannuinB production and artemisinin increase}, volume={91}, DOI={10.1111/tpj.13583}, abstractNote={SummaryWe recently characterized a gene–terpene network that is associated with artemisinin biosynthesis in self‐pollinated (SP) Artemisia annua, an effective antimalarial plant. We hypothesize that an alteration of gene expression in the network may improve the production of artemisinin and its precursors. In this study, we cloned an isopentenyl pyrophosphate isomerase (IPPI) cDNA, AaIPPI1, from Artemisia annua (Aa). The full‐length cDNA encodes a type‐I IPPI containing a plastid transit peptide (PTP) at its amino terminus. After the removal of the PTP, the recombinant truncated AaIPPI1 isomerized isopentenyl pyrophosphate (IPP) to dimethyl allyl pyrophosphate (DMAPP) and vice versa. The steady‐state equilibrium ratio of IPP/DMAPP in the enzymatic reactions was approximately 1:7. The truncated AaIPPI1 was overexpressed in the cytosol of the SP A. annua variety. The leaves of transgenic plants produced approximately 4% arteannuin B (g g−1, dry weight, dw) and 0.17–0.25% artemisinin (g g−1, dw), the levels of which were significantly higher than those in the leaves of wild‐type plants. In addition, transgenic plants showed an increase in artemisinic acid production of more than 1% (g g−1, dw). In contrast, isoprene formation was significantly reduced in transgenic plants. These results provide evidence that overexpression of AaIPPI1 in the cytosol can lead to metabolic alterations of terpenoid biosynthesis, and show that these transgenic plants have the potential to yield high production levels of arteannuin B as a new precursor source for artemisinin.}, number={3}, journal={Plant Journal}, author={Ma, D. M. and Li, G. and Alejos-Gonzalez, F. and Zhu, Y. and Xue, Z. and Wang, A. M. and Zhang, H. and Li, X. and Ye, H. C. and Wang, H. and et al.}, year={2017}, pages={466–479} }
 @article{ma_wang_wang_alejos-gonzales_sun_xie_2015, title={A Genome-Wide Scenario of Terpene Pathways in Self-pollinated Artemisia annua}, volume={8}, ISSN={["1752-9867"]}, DOI={10.1016/j.molp.2015.07.004}, abstractNote={Scenarios of genes to metabolites in Artemisia annua remain uninvestigated. Here, we report the use of an integrated approach combining metabolomics, transcriptomics, and gene function analyses to characterize gene-to-terpene and terpene pathway scenarios in a self-pollinating variety of this species. Eighty-eight metabolites including 22 sesquiterpenes (e.g., artemisinin), 26 monoterpenes, two triterpenes, one diterpene and 38 other non-polar metabolites were identified from 14 tissues. These metabolites were differentially produced by leaves and flowers at lower to higher positions. Sequences from cDNA libraries of six tissues were assembled into 18 871 contigs and genome-wide gene expression profiles in tissues were strongly associated with developmental stages and spatial specificities. Sequence mining identified 47 genes that mapped to the artemisinin, non-amorphadiene sesquiterpene, monoterpene, triterpene, 2-C-methyl-D-erythritol 4-phosphate and mevalonate pathways. Pearson correlation analysis resulted in network integration that characterized significant correlations of gene-to-gene expression patterns and gene expression-to-metabolite levels in six tissues simultaneously. More importantly, manipulations of amorpha-4,11-diene synthase gene expression not only affected the activity of this pathway toward artemisinin, artemisinic acid, and arteannuin b but also altered non-amorphadiene sesquiterpene and genome-wide volatile profiles. Such gene-to-terpene landscapes associated with different tissues are fundamental to the metabolic engineering of artemisinin.}, number={11}, journal={MOLECULAR PLANT}, author={Ma, Dong-Ming and Wang, Zhilong and Wang, Liangjiang and Alejos-Gonzales, Fatima and Sun, Ming-An and Xie, De-Yu}, year={2015}, month={Nov}, pages={1580–1598} }
 @article{alejos-gonzalez_qu_zhou_saravitz_shurtleff_xie_2011, title={Characterization of development and artemisinin biosynthesis in self-pollinated Artemisia annua plants}, volume={234}, ISSN={["1432-2048"]}, DOI={10.1007/s00425-011-1430-z}, abstractNote={Artemisia annua L. is the only natural resource that produces artemisinin (Qinghaosu), an endoperoxide sesquiterpene lactone used in the artemisinin-combination therapy of malaria. The cross-hybridization properties of A. annua do not favor studying artemisinin biosynthesis. To overcome this problem, in this study, we report on selection of self-pollinated A. annua plants and characterize their development and artemisinin biosynthesis. Self-pollinated F2 plants selected were grown under optimized growth conditions, consisting of long day (16 h of light) and short day (9 h of light) exposures in a phytotron. The life cycles of these plants were approximately 3 months long, and final heights of 30-35 cm were achieved. The leaves on the main stems exhibited obvious morphological changes, from indented single leaves to odd, pinnately compound leaves. Leaves and flowers formed glandular and T-shaped trichomes on their surfaces. The glandular trichome densities increased from the bottom to the top leaves. High performance liquid chromatography-mass spectrometry-based metabolic profiling analyses showed that leaves, flowers, and young seedlings of F2 plants produced artemisinin. In leaves, the levels of artemisinin increased from the bottom to the top of the plants, showing a positive correlation to the density increase of glandular trichomes. RT-PCR analysis showed that progeny of self-pollinated plants expressed the amorpha-4, 11-diene synthase (ADS) and cytochrome P450 monooxygenase 71 AV1 (CYP71AV1) genes, which are involved in artemisinin biosynthesis in leaves and flowers. The use of self-pollinated A. annua plants will be a valuable approach to the study of artemisinin biosynthesis.}, number={4}, journal={PLANTA}, author={Alejos-Gonzalez, Fatima and Qu, Guosheng and Zhou, Li-Li and Saravitz, Carole H. and Shurtleff, Janet L. and Xie, De-Yu}, year={2011}, month={Oct}, pages={685–697} }
 @article{mitchell_alejos-gonzalez_gracz_danehower_daub_chilton_2003, title={Xanosporic acid, an intermediate in bacterial degradation of the fungal phototoxin cercosporin}, volume={62}, ISSN={["0031-9422"]}, DOI={10.1016/S0031-9422(02)00517-4}, abstractNote={The red fungal perylenequinone phototoxin cercosporin is oxidized by Xanthomonas campestris pv zinniae to a non-toxic, unstable green metabolite xanosporic acid, identified via its lactone as 1,12-bis(2′R-hydroxypropyl)-4,9-dihydroxy-6,7-methylenedioxy-11-methoxy-3-oxaperylen-10H-10-one-2-carboxylic acid. Xanosporolactone was isolated in approximately 2:1 ratio of M:P atropisomers.}, number={5}, journal={PHYTOCHEMISTRY}, author={Mitchell, TK and Alejos-Gonzalez, F and Gracz, HS and Danehower, DA and Daub, ME and Chilton, WS}, year={2003}, month={Mar}, pages={723–732} }