Indiana University-Purdue University Indianapolis (IUPUI) / Polyacetylenic natural products, compounds containing multiple carbon-carbon triple bonds, have been found in a large collection of organisms. Radiochemical tracer studies have indicated that these bioactive metabolites are synthesized from fatty acid precursors through a series of uncharacterized desaturation and acetylenation steps. To date, there are three main pathways believed to be involved in acetylenic natural product biosynthesis. However, it is apparent that the crepenynic acid pathway is the origin of a vast majority of the known plant and fungal acetylenic products. This investigation provides concrete evidence that the polyacetylenic natural products found in the fungus Fistulina hepatica and the medicinal plant species Echinacea purpurea are biosynthesized from crepenynic acid. Through heterologous expression in Yarrowia lipolytica, two acetylenases capable of producing crepenynic acid were identified from E. purpurea. Furthermore, heterologous expression of two diverged desaturases isolated from F. hepatica, uncovered a ∆12-acetylenase and the first multifunctional enzyme capable of ∆14-/∆16- desaturation and ∆14-acetylenation.

The goals of this research project were to identify the various insects observed to visit inflorescences of Echinacea angustifolia DC, and to rank these visitors according to their importance as pollinators of E. angustifolia in Saskatchewan. Studying nectar and the nectary is essential to understanding the interaction of disc florets with pollinators. Nectar-sugar production by disc florets of E. angustifolia and E. purpurea (L. Moench) was quantified from anthesis to cessation with production per disc floret peaking in the afternoon of the staminate phase (191.7 µg) and at midday of the first day of the pistillate phase (156.6 µg), respectively. Morphology of the disc-like floral nectaries of both Echinacea species was studied, as well as the ultrastructure of the nectary of E. purpurea. Modified stomata on the nectary rim are the most likely exits for nectar, but creases in the epidermis may also participate. The nectary of E. purpurea is vascularized by phloem alone, which occurred adjacent to the epidermis. Companion cells possessed wall ingrowths, and these cells may unload arriving sugar destined for either an apoplastic or symplastic pathway. Lobed nuclei were a key feature of secretory parenchyma cells, as was a predominance of mitochondria, suggesting that energy-requiring eccrine secretion predominates in E. purpurea. E. angustifolia exhibited a generalist pollination system, with pollinating insects belonging to the orders Coleoptera, Diptera, Hymenoptera, and Lepidoptera. The pollination efficiency of visitors was determined by single insect visits to bagged, virgin inflorescences followed by quantifying pollen tubes at the bases of receptive styles and/or calculating the percentage of shrivelled styles. It was determined that bumble bees (Bombus spp.) were efficient pollinators, indicating that they would likely contribute much to the pollination of E. angustifolia. Grasshopper bee flies (Systoechus vulgaris Loew) were plentiful but individually were not efficient pollinators, but taken together, they provided much pollination. Golden blister beetles (Epicauta ferruginea Say) were efficient pollinators but where yellow-petalled flowers occurred, their numbers on E. angustifolia decreased. Honey bees (Apis mellifera L.) were efficient pollinators and were present in low numbers without managed introduction. Pierid (2003) butterflies were regular visitors and efficient pollinators, and likely contributed significantly to E. angustifolia pollination. When introduced, the alfalfa leafcutter bee (Megachile rotundata Fabr.) preferred not to forage on E. angustifolia and as such, these solitary bees were not suitable as managed pollinators. In large agricultural plantings of E. angustifolia, however, native insects may not be capable of providing sufficient pollination for seed production when floral competition occurs.

The goals of this research project were to identify the various insects observed to visit inflorescences of Echinacea angustifolia DC, and to rank these visitors according to their importance as pollinators of E. angustifolia in Saskatchewan. Studying nectar and the nectary is essential to understanding the interaction of disc florets with pollinators. Nectar-sugar production by disc florets of E. angustifolia and E. purpurea (L. Moench) was quantified from anthesis to cessation with production per disc floret peaking in the afternoon of the staminate phase (191.7 µg) and at midday of the first day of the pistillate phase (156.6 µg), respectively. Morphology of the disc-like floral nectaries of both Echinacea species was studied, as well as the ultrastructure of the nectary of E. purpurea. Modified stomata on the nectary rim are the most likely exits for nectar, but creases in the epidermis may also participate. The nectary of E. purpurea is vascularized by phloem alone, which occurred adjacent to the epidermis. Companion cells possessed wall ingrowths, and these cells may unload arriving sugar destined for either an apoplastic or symplastic pathway. Lobed nuclei were a key feature of secretory parenchyma cells, as was a predominance of mitochondria, suggesting that energy-requiring eccrine secretion predominates in E. purpurea. E. angustifolia exhibited a generalist pollination system, with pollinating insects belonging to the orders Coleoptera, Diptera, Hymenoptera, and Lepidoptera. The pollination efficiency of visitors was determined by single insect visits to bagged, virgin inflorescences followed by quantifying pollen tubes at the bases of receptive styles and/or calculating the percentage of shrivelled styles. It was determined that bumble bees (Bombus spp.) were efficient pollinators, indicating that they would likely contribute much to the pollination of E. angustifolia. Grasshopper bee flies (Systoechus vulgaris Loew) were plentiful but individually were not efficient pollinators, but taken together, they provided much pollination. Golden blister beetles (Epicauta ferruginea Say) were efficient pollinators but where yellow-petalled flowers occurred, their numbers on E. angustifolia decreased. Honey bees (Apis mellifera L.) were efficient pollinators and were present in low numbers without managed introduction. Pierid (2003) butterflies were regular visitors and efficient pollinators, and likely contributed significantly to E. angustifolia pollination. When introduced, the alfalfa leafcutter bee (Megachile rotundata Fabr.) preferred not to forage on E. angustifolia and as such, these solitary bees were not suitable as managed pollinators. In large agricultural plantings of E. angustifolia, however, native insects may not be capable of providing sufficient pollination for seed production when floral competition occurs.