Metabolic, biochemical and molecular profiling of Catharanthus roseus flower cultivars and transformed hairy roots for monoterpenoid indole alkaloid accumulation /

Catharanthlls rosellS (L.) G Don is a commercially significant flower species and in addition is the only source of the monoterpenoid indole alkaloids (MIA) vinblastine and vincristine, which are key pharmaceutical compounds that are used to combat a number of different cancers. Therefore, procurement of the antineoplastic agents is difficult but essential procedure. Alternatively, CatharanthllS tissue cultures have been investigated as a source of these agents; however they do not produce vindoline, which is an obligate precursor to vinblastine and vincristine. The interest in developing high MIA cultivars of Catharantlws rosellS has prompted metabolic profiling studies to determine the variability of MIA accumulation of existing flowering cultivars, with particular focus on the vindoline component ofthe pathway. Metabolic profiling studies that used high performance liquid chromatography of MIAs from seedlings and young leaf extracts from 50 different flowering cultivars showed that, except for a single low vindoline cultivar (Vinca Mediterranean DP Orchid), they all accumulate similar levels of MIAs. Further enzymatic studies with extracts from young leaves and from developing seedlings showed that the low vindoline cultivar has a IO-fold lower tabersonine-16-hydroxylase activity than those of CatharanthllS rosellS cv Little Delicata. Additionally, studies aimed at metabolic engineering ofvindoline bios}l1thesis in Catharanthus rosellS hairy root cultures have been performed by expressing the last step in vindoline biosynthesis [Dcacetylvindoline-4-0- acetyltransferase (DAT)]. Enzymatic profiling studies with transformed hairy roots have confirmed that over-expressing DAT leads to lines with high levels of O-acetyltransferase activity when compared to non-expressing hairy roots. One particular DA T over111 expressing hairy root culture (line 7) contained 200 times the OAT activity than leaves of control lines. Additional MIA analyses revealed that DAT over-expressing hairy roots have an altered alkaloid profile with significant variation in the accumulation of h6rhammericine. Further analysis of transformed hairy root line 7 suggests a correlation between the expression of OAT activity and h6rhammericine accumulation with root maturation. These studies show that metabolic and selective enzymatic profiling can enhance our ability to search for relevant MIA pathway mutants and that genetic engineering with appropriate pathway genes shows promise as a tool to modify the MIA profile of Catharanthus roseus.

Educational Technology Decision-Making: A Case Study on Technology Acquisition for 746,000 Ontario Students

Very little research has examined K–12 educational technology decision-making in Canada. This collective case study explores the technology procurement process in Ontario’s publicly funded school districts to determine if it is informed by the relevant research, grounded in best practices, and enhances student learning. Using a qualitative approach, 10 senior leaders (i.e., chief information officers, superintendents, etc.) were interviewed. A combination of open-ended and closed-ended questions were used to reveal the most important factors driving technology acquisition, research support, governance procedures, data use, and assessment and return on investment (ROI) measures utilized by school districts in their implementation of educational technology. After participants were interviewed, the data were transcribed, member checked, and then submitted to “Computer-assisted NCT analysis” (Friese, 2014) using ATLAS.ti. The findings show that senior leaders are making acquisitions that are not aligned with current scholarship and not with student learning as the focus. It was also determined that districts struggle to use data-driven decision-making to support the governance of educational technology spending. Finally, the results showed that districts do not have effective assessment measures in place to determine the efficacy or ROI of a purchased technology. Although data are limited to the responses of 10 senior leaders, findings represent the technology leadership for approximately 746,000 Ontario students. The study is meant to serve as an informative resource for senior leaders and presents strategic and research-validated approaches to technology procurement. Further, the study has the potential to refine technology decision-making, policies, and practices in K–12 education.