@article{judd_dong_sun_zhu_li_xie_2023, title={Metabolic engineering of the anthocyanin biosynthetic pathway in Artemisia annua and relation to the expression of the artemisinin biosynthetic pathway}, volume={257}, ISSN={["1432-2048"]}, url={https://doi.org/10.1007/s00425-023-04091-6}, DOI={10.1007/s00425-023-04091-6}, abstractNote={Four types of cells were engineered from Artemisia annua to produce approximately 17 anthocyanins, four of which were elucidated structurally. All of them expressed the artemisinin pathway. Artemisia annua is the only medicinal crop to produce artemisinin for the treatment of malignant malaria. Unfortunately, hundreds of thousands of people still lose their life every year due to the lack of sufficient artemisinin. Artemisinin is considered to result from the spontaneous autoxidation of dihydroartemisinic acid in the presence of reactive oxygen species (ROS) in an oxidative condition of glandular trichomes (GTs); however, whether increasing antioxidative compounds can inhibit artemisinin biosynthesis in plant cells is unknown. Anthocyanins are potent antioxidants that can remove ROS in plant cells. To date, no anthocyanins have been structurally elucidated from A. annua. In this study, we had two goals: (1) to engineer anthocyanins in A. annua cells and (2) to understand the artemisinin biosynthesis in anthocyanin-producing cells. Arabidopsis Production of Anthocyanin Pigment 1 was used to engineer four types of transgenic anthocyanin-producing A. annua (TAPA1-4) cells. Three wild-type cell types were developed as controls. TAPA1 cells produced the highest contents of total anthocyanins. LC-MS analysis detected 17 anthocyanin or anthocyanidin compounds. Crystallization, LC/MS/MS, and NMR analyses identified cyanidin, pelargonidin, one cyanin, and one pelargonin. An integrative analysis characterized that four types of TAPA cells expressed the artemisinin pathway and TAPA1 cells produced the highest artemisinin and artemisinic acid. The contents of arteannuin B were similar in seven cell types. These data showed that the engineering of anthocyanins does not eliminate the biosynthesis of artemisinin in cells. These data allow us to propose a new hypothesis that enzymes catalyze the formation of artemisinin from dihydroartemisinic acid in non-GT cells. These findings show a new platform to increase artemisinin production via non-GT cells of A. annua.}, number={3}, journal={PLANTA}, author={Judd, Rika and Dong, Yilun and Sun, Xiaoyan and Zhu, Yue and Li, Mingzhuo and Xie, De-Yu}, year={2023}, month={Mar} }
 @article{ekelöf_garrard_judd_rosen_xie_kashuba_muddiman_2018, title={Evaluation of Digital Image Recognition Methods for Mass Spectrometry Imaging Data Analysis}, volume={29}, ISSN={1044-0305 1879-1123}, url={http://dx.doi.org/10.1007/S13361-018-2073-0}, DOI={10.1007/s13361-018-2073-0}, abstractNote={Analyzing mass spectrometry imaging data can be laborious and time consuming, and as the size and complexity of datasets grow, so does the need for robust automated processing methods. We here present a method for comprehensive, semi-targeted discovery of molecular distributions of interest from mass spectrometry imaging data, using widely available image similarity scoring algorithms to rank images by spatial correlation. A fast and powerful batch search method using a MATLAB implementation of structural similarity (SSIM) index scoring with a pre-selected reference distribution is demonstrated for two sample imaging datasets, a plant metabolite study using Artemisia annua leaf, and a drug distribution study using maraviroc-dosed macaque tissue. Graphical Abstract ᅟ.}, number={12}, journal={Journal of The American Society for Mass Spectrometry}, publisher={Springer Science and Business Media LLC}, author={Ekelöf, Måns and Garrard, Kenneth P. and Judd, Rika and Rosen, Elias P. and Xie, De-Yu and Kashuba, Angela D. M. and Muddiman, David C.}, year={2018}, month={Oct}, pages={2467–2470} }
 @article{ma_xu_alejos-gonzalez_wang_yang_judd_xie_2018, title={Overexpression of Artemisia annua Cinnamyl Alcohol Dehydrogenase Increases Lignin and Coumarin and Reduces Artemisinin and Other Sesquiterpenes}, volume={9}, ISSN={["1664-462X"]}, DOI={10.3389/fpls.2018.00828}, abstractNote={Artemisia annua is the only medicinal crop that produces artemisinin for malarial treatment. Herein, we describe the cloning of a cinnamyl alcohol dehydrogenase (AaCAD) from an inbred self-pollinating (SP) A. annua cultivar and its effects on lignin and artemisinin production. A recombinant AaCAD was purified via heterogeneous expression. Enzyme assays showed that the recombinant AaCAD converted p-coumaryl, coniferyl, and sinapyl aldehydes to their corresponding alcohols, which are key intermediates involved in the biosynthesis of lignin. Km, Vmax, and Vmax/Km values were calculated for all three substrates. To characterize its function in planta, AaCAD was overexpressed in SP plants. Quantification using acetyl bromide (AcBr) showed significantly higher lignin contents in transgenics compared with wild-type (WT) plants. Moreover, GC-MS-based profiling revealed a significant increase in coumarin contents in transgenic plants. By contrast, HPLC-MS analysis showed significantly reduced artemisinin contents in transgenics compared with WT plants. Furthermore, GC-MS analysis revealed a decrease in the contents of arteannuin B and six other sesquiterpenes in transgenic plants. Confocal microscopy analysis showed the cytosolic localization of AaCAD. These data demonstrate that AaCAD plays a dual pathway function in the cytosol, in which it positively enhances lignin formation but negatively controls artemisinin formation. Based on these data, crosstalk between these two pathways mediated by AaCAD catalysis is discussed to understand the metabolic control of artemisinin biosynthesis in plants for high production.}, journal={FRONTIERS IN PLANT SCIENCE}, author={Ma, Dongming and Xu, Chong and Alejos-Gonzalez, Fatima and Wang, Hong and Yang, Jinfen and Judd, Rika and Xie, De-Yu}, year={2018}, month={Jun} }
 @article{xie_ma_judd_jones_2016, title={Artemisinin biosynthesis in Artemisia annua and metabolic engineering: questions, challenges, and perspectives}, volume={15}, ISSN={["1572-980X"]}, url={https://doi.org/10.1007/s11101-016-9480-2}, DOI={10.1007/s11101-016-9480-2}, number={6}, journal={PHYTOCHEMISTRY REVIEWS}, publisher={Springer Nature}, author={Xie, De-Yu and Ma, Dong-Ming and Judd, Rika and Jones, Ashley Loray}, year={2016}, month={Dec}, pages={1093–1114} }