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.

Annual Variation in Bee Community Structure in the Context of Disturbance (Niagara Region, South-Western Ontario)

This study examined annual variation in phenology, abundance and diversity of a bee community during 2003, 2004, 2006, and 2008 in recovered landscapes at the southern end of St. Catharines, Ontario, Canada. Overall, 8139 individuals were collected from 26 genera and sub-genera and at least 57 species. These individuals belonged to the 5 families found in eastern North America (Andrenidae, Apidae, Colletidae, Halictidae and Megachilidae). The bee community was characterized by three distinct periods of flight activity over the four years studied (early spring, late spring/early summer, and late summer). The number of bees collected in spring was significantly higher than those collected in summer. In 2003 and 2006 abundance was higher, seasons started earlier and lasted longer than in 2004 and 2008, as a result of annual rainfall fluctuations. Differences in abundance for low and high disturbance sites decreased with years. Annual trends of generic richness resembled those detected for species. Likewise, similarity in genus and species composition decreased with time. Abundant and common taxa (13 genera and 18 species) were more persistent than rarer taxa being largely responsible for the annual fluctuations of the overall community. Numerous species were sporadic or newly introduced. The invasive species Anthidium oblongatum was first recorded in Niagara in 2006 and 2008. Previously detected seasonal variation patterns were confirmed. Furthermore, this study contributed to improve our knowledge of temporal dynamics of bee communities. Understanding temporal variation in bee communities is relevant to assessing impacts caused on their habitats by diverse disturbances.