Functional genomics of O-glucosyltransferases from Concord grape (Vitis labrusca)

Grape (Vitis spp.) is a culturally and economically important crop plant that has been cultivated for thousands of years, primarily for the production of wine. Grape berries accumulate a myriad of phenylpropanoid secondary metabolites, many of which are glucosylated in plantae More than 90 O-glucosyltransferases have been cloned and biochemically characterized from plants, only two of which have been isolated from Vitis spp. The world-wide economic importance of grapes as a crop plant, the human health benefits associated with increased consumption of grape-derived metabolites, the biological relevance of glucosylation, and the lack of information about Vitis glucosyltransferases has inspired the identification, cloning and biochemical characterization of five novel "family 1" O-glucosyltransferases from Concord grape (Vitis labrusca cv. Concord). Protein purification and associated protein sequencIng led to the molecular cloning of UDP-glucose: resveratrollhydroxycinnamic acid O-glucosyltransferase (VLRSGT) from Vitis labrusca berry mesocarp tissue. In addition to being the first glucosyltransferase which accepts trans-resveratrol as a substrate to be characterized in vitro, the recombinant VLRSGT preferentially produces the glucose esters of hydroxycinnamic acids at pH 6.0, and the glucosides of trans-resveratrol and flavonols at 'pH 9.0; the first demonstration of pH-dependent bifunctional glucosylation for this class of enzymes. Gene expression and metabolite profiling support a role for this enzyme in the bifuncitonal glucosylation ofstilbenes and hydroxycinnamic acids in plantae A homology-based approach to cloning was used to identify three enzymes from the Vitis vinifera TIGR grape gene index which had high levels of protein sequence iii identity to previously characterized UDP-glucose: anthocyanin 5-0-glucosyltransferases. Molecular cloning and biochemical characterization demonstrated that these enzymes (rVLOGTl, rVLOGT2, rVLOGT3) glucosylate the 7-0-position of flavonols and the xenobiotic 2,4,5-trichlorophenol (TCP), but not anthocyanins. Variable gene expression throughout grape berry development and enzyme assays with native grape berry protein are consistent with a role for these enzymes in the glucosylation of flavonols; while the broad substrate specificity, the ability of these enzymes to glucosylate TCP and expression of these genes in tissues which are subject to pathogen attack (berry, flower, bud) is consistent with a role for these genes in the plant defense response. Additionally, the Vitis labrusca UDP-glucose: flavonoid 3-0-glucosyltransferase (VL3GT) was identified, cloned and characterized. VL3GT has 96 % protein sequence identity to the previously characterized Vitis vinifera flavonoid 3-0-glucosyltransferase (VV3GT); and glucosylates the 3-0-position of anthocyanidins and flavonols in vitro. Despite high levels of protein sequence identity, VL3GT has distinct biochemical characteristics (as compared to VV3GT), including a preference for B-ring methylated flavonoids and the inability to use UDP-galactose as a donor substrate. RT-PCR analysis of VL3GT gene expression and enzyme assays with native grape protein is consistent with an in planta role for this enzyme in the glucosylation of anthocyanidins,but not flavonols. These studies reveal the power of combining several biochemistry- and molecular biology-based tools to identify, clone, biochemically characterize and elucidate the in planta function of several biologically relevant O-glucosyltransferases from Vitis spp.

The determination of sinigrin in Brassica, and investigations into the use of allyl-isothiocyanate as a nematicide

The goal of this thesis was to study factors related to the development of Brassica juncea as a sustainable nematicide. Brassica juncea is characterized by the glycoside (glucosinolate) sinigrin. Various methods were developed for the determination of sinigrin in Brassica juncea tissue extracts. Sinigrin concentrations in plant tissues at various stages of growth were monitored. Sinigrin enzymatically breaks down into allylisothiocyanate (AITC). AITC is unstable in aqueous solution and degradation was studied in water and in soil. Finally, the toxicity of AITC against the root-lesion nematode (Pratylenchus penetrans) was determined. A method was developed to extract sinigrin from whole Brassica j uncea tissues. The optimal time of extraction wi th boiling phosphate buffer (0.7mM, pH=6.38) and methanol/water (70:30 v/v) solutions were both 25 minutes. Methanol/water extracted 13% greater amount of sinigrin than phosphate buffer solution. Degradation of sinigrin in boiling phosphate buffer solution (0.13%/minute) was similar to the loss of sinigrin during the extraction procedure. The loss of sinigrin from boiling methanol/water was estimated to be O.Ol%/minute. Brassica juncea extract clean up was accomplished by an ion-pair solid phase extraction (SPE) method. The recovery of sinigrin was 92.6% and coextractive impurities were not detected in the cleaned up extract. Several high performance liquid chromatography (HPLC) methods were developed for the determination of sinigrin. All the developed methods employed an isocratic mobile phase system wi th a low concentration of phosphate buffer solution, ammonium acetate solution or an ion-pair reagent solution. A step gradient system was also developed. The method involved preconditioning the analytical column with phosphate buffer solution and then switching the mobile phase to 100% water after sample injection.Sinigrin and benzyl-glucosinolate were both studied by HPLC particle beam negative chemical ionization mass spectrometry (HPLCPB- NCI-MS). Comparison of the mass spectra revealed the presence of fragments arising from the ~hioglucose moiety and glucosinolate side-chain. Variation in the slnlgrin concentration within Brassica juncea plants was studied (Domo and Cutlass cuItivars). The sinigrin concentration in the top three leaves was studied during growth of each cultivar. For Cutlass, the minimum (200~100~g/g) and maximum (1300~200~g/g) concentrations were observed at the third and seventh week after planting, respectively. For Domo, the minimum (190~70~g/g) and maximum (1100~400~g/g) concentrations were observed at the fourth and eighth week after planting, respectively. The highest sinigrin concentration was observed in flower tissues 2050±90~g/g and 2300±100~g/g for Cutlass and Domo cultivars, respectively. Physical properties of AITC were studied. The solubility of AITC in water was determined to be approximately 1290~g/ml at 24°C. An HPLC method was developed for the separation of degradation compounds from aqueous AITC sample solutions. Some of the degradation compounds identified have not been reported in the literature: allyl-thiourea, allyl-thiocyanate and diallyl-sulfide. In water, AITC degradation to' diallyl-thiourea was favored at basic pH (9.07) and degradation to diallyl-sulfide was favored at acidic pH (4 . 97). It wap necessary to amend the aqueous AITC sample solution with acetonitrile ?efore injection into the HPLC system. The acetonitrile amendment considerably improved AITC recovery and the reproducibility of the results. The half-life of aqueous AITC degradation at room temperature did not follow first-order kinetics. Beginning with a 1084~g/ml solution, the half-life was 633 hours. Wi th an ini tial AITC concentration of 335~g/ml the half-life was 865 hours. At 35°C the half-life AITC was 76+4 hours essentially independent of the iiisolution pH over the range of pH=4.97 to 9.07 (1000~g/ml). AITC degradation was also studied in soil at 35°C; after 24 hours approximately 75% of the initial AITC addition was unrecoverable by water extraction. The ECso of aqueous AITC against the root-lesion nematode (Pratylenchus penetrans) was determined to be approximately 20~g/ml at one hour exposure of the nematode to the test solution. The toxicological study was also performed with a myrosinase treated Brassica juncea extract. Myrosinase treatment of the Brassica juncea extract gave nearly quantitative conversion of sinigrin into AITC. The myrosinase treated extract was of the same efficacy as an aqueous AITC solution of equivalent concentration. The work of this thesis was focused upon understanding parameters relevant to the development of Brassica juncea as a sustainable nematicide. The broad range of experiments were undertaken in support of a research priority at Agriculture and Agri-Food Canada.

Canada seventy years ago : or, Prince Edward's visit to Niagara /

Verse.

John W. Fisher Fonds, 1937-1961, 1964-1989, 1993, n.d.

Norah and Fred Fisher welcomed John Fisher into the world on November 29, 1912, not knowing what an influential role he would play in shaping Canada's history. John Fisher grew up as the middle child of five brothers and sisters in Frosty Hollow, New Brunswick, close to today’s town of Sackville. Sackville’s main industry was the Enterprise Foundry which the Fisher family owned and operated; however, Fisher had no plans of going into the family business. He was more inspired by his maternal grandfather, Dr. Cecil Wiggins, who lived with the family after retiring from the Anglican ministry. Wiggins encouraged all his grandchildren to be well read and to take part in discussions on current events. There were often visitors in the Fisher household taking part in discussions about politics, religion, and daily life. Fisher forced himself to take part in these conversations to help overcome his shyness in social settings. These conversations did help with his shyness and also in forming many opinions and observations about Canada. It put Fisher on the road to becoming Mr. Canada and delivering the many eloquent speeches for which he was known. Fisher did not venture far from home to complete his first degree. In 1934 he graduated from Mount Allison University in Sackville, NB with an Arts degree. The same year Fisher enrolled in Dalhousie’s law school. During his time at Dalhousie, Fisher discovered radio through Hugh Mills. Mills or “Uncle Mel” was on CHNS, Halifax’s only radio station at the time. Fisher began by making appearences on the radio drama show. By 1941 he had begun writing and broadcasting his own works and joined the staff as an announcer and continuity writer. In 1936 the Canadian Broadcasting Corporation was formed, the first National radio station. Fisher joined the CBC shortly after it’s beginning and remained with them, as well as the Halifax Herald newspaper, even after his law school graduation in 1937. By 1943 Fisher’s talks became a part of the CBC’s programming for a group of maritime radio stations. Fisher once described his talks as follows “my talks weren’t meant to be objective. . . they were meant to be favourable. They were ‘pride builders’” He began his famed John Fisher Reports at CBC Toronto when he transfered there shortly after the war. This program brought emmence pride to the fellow Canadians he spoke about leading to approximately 3500 requests per year to speak at banquets and meeting throughout Canada and the United States. Fisher was a well travelled indivdual who would draw on personal experiences to connect with his audience. His stories were told in simple, straight forward language for anyone to enjoy. He became a smooth, dynamic and passionate speaker who sold Canada to Canadians. He became a renowned journalist, folk historian, writer and broadcaster. Fisher was able to reach a vast array of people through his radio work and build Canadian pride, but he did not stop there. Other ways Fisher has contributed to Canada and the Canadian people include: Honoured by five Canadian Universities. 1956, became the Director of the Canadian Tourist Association. 1961, was appointed Special Assistant to the Prime Minister of Canada. 1963, Commissioner of the Centennial Commission (the Federal Agency Responsible for Canada’s 100th birthday) 1968, received the Service Medal , a coveted Order of Canada. President of John Fisher Enterprises Ltd., private consultant work, specializing in Centennial planning, broadcasts, lectures and promotion. John Fisher continued recording radio broadcasts even after his diagnosis with cancer. He would record 3 or 4 at a time so he was free to travel across Canada, the U.S., Europe and Mexico in search of treatments. Fisher passed away from the disease on February 15, 1981 and he is buried at Mount Pleasant Cemetery in Toronto.

The New-York Weekly Museum Newspaper, Vol. II, No.24- October 16, 1813

A weekly paper that was published from 1805 to 1814. War related news includes: Page 95 - Weekly Retrospect: relief effort in London, England for those affected by the war in British North America; naval battles on Lake Ontario by Commodore Chauncey in late September; movements of Major Grant, De Rottenburgh's regiment, General Wilkinson, Commodore Yoe's fleet, General Hampton; the capture of Fort Malden by William Harrison. The rest of the newspaper contains literary works (poems and translations), marriages, deaths and other anecdotes. The motto on the front page states: "Visiting Every Flower with Labour Meet, and Gathering all its Treasures, Sweet by Sweet."

The New York Weekly Museum Newspaper, Vol. II, No.25- October 23, 1813

A weekly paper that was published from 1805 to 1814. War related news includes: Page 99 - Weekly Retrospect: American report on British naval movements near Louisiana; General William Harrison reports on: a victory at Moravian Town (Moraviantown) on October 5; Ottawas, Chippewas, Wyandots Miamies and Delaware First Nations (Indians) leaving the British side; Tecumseh remaining with the British; American army and navy attempt for a reduction of Mackinac and St. Joseph's; Colonel Chapin attacking a British picket guard near Fort George on October 6; Report that the British have abandoned the Fort George area for Kingston and is being pursued by General McClure; Forces in Sackett's Harbour ready for an attach on Canada; General Provost believed to have left Kingston for Montreal with 1000 men; Report from Burlington (Lake Champlain) of an attack on St. Armands against Colonel Clark on October 15; Report of the American's taking of the Packet Morgiana by the Schooner Saratoga under Captain Adderton by Newport; Announcement by the New York City Common Council for a celebration of Perry and Harrison's victories on the evening of October 23. The rest of the newspaper contains literary works (poems and translations), marriages, deaths and other anecdotes. The motto on the front page states: "Visiting Every Flower with Labour Meet, and Gathering all its Treasures, Sweet by Sweet."

The New York Weekly Museum Newspaper, Vol. I, No.25- October 24, 1812

A weekly paper that was published from 1805 to 1814. The motto on the front page states: "Visiting Every Flower with Labour Meet, and Gathering all its Treasures, Sweet by Sweet."

The New York Weekly Museum Newspaper, Vol. II, No.26- October 30, 1813

A weekly paper that was published from 1805 to 1814. War related news includes: Page 103 - Weekly Retrospect: Reports on the Napoleanic Wars in Europe; Possible French and English troop movements from Europe to America and Canada; Mention of British vessels captured by American privateers Scourge, Rattlesnake and Lion; Report on New York celebration of Commodore Perry and General Harrison's victories on Lake Erie and in Canada respectively; Creek Indians attack on a supply convoy near Savannah, the provisions were recovered by General Floyd; General Harrison's account on Moravian Town (Moraviantown) and Munsey; Movements of General Wilkinson's army. Page 104 - an editorial about the madness of the times and its reflection on humanity. The rest of the newspaper contains literary works (poems and translations), marriages, deaths and other anecdotes. The motto on the front page states: "Visiting Every Flower with Labour Meet, and Gathering all its Treasures, Sweet by Sweet."

The New York Weekly Museum Newspaper, Vol. I, No.26- October 31, 1812

A weekly paper that was published from 1805 to 1814. War related news includes: Page 103 - Weekly Retrospect: Troop movements of Captain Morgan and a company of light dragoons from Niagara to Greenbush; report on the Battle of Queenston Heights including a casualty listing and a mention of the death of Major General Isaac Brock and Colonel McDonald and the American's respect for Brock; British attach of Fort Erie and the Village of Black Rock; the privateer Globe under Captain Murphy returning to Baltimore after capturing a British letter of marque; First Nation / Indian attacks near St. Louis; movements of General W. Harrison and army from Fort Defiance to the rapids of Miama (Miami). The rest of the newspaper contains literary works (poems and translations), essays (including on celibacy), marriages, deaths and other anecdotes. The motto on the front page states: "Visiting Every Flower with Labour Meet, and Gathering all its Treasures, Sweet by Sweet."

Collective violence : a study of the gendered and socio-economic factors behind early modern Italian and English witch hunts

In this study, I build upon my previous research in which I focus on religious doctrine as a gendered disciplinary apparatus, and examine the witch trials in early modem England and Italy in light of socio-economic issues relating to gender and class. This project examines the witch hunts/trials and early modem visual representations of witches, and what I suggest is an attempt to create docile bodies out of members of society who are deemed unruly, problematic and otherwise 'undesirable'; it is the witch's body that is deemed counternormative. This study demonstrates that it is neighbours and other acquaintances of accused witches that take on the role of the invisible guard of Bantham's Panoptic on. As someone who is trained in the study of English literature and literary theory, my approach is one that is informed by this methodology. It is my specialization in early modem British literature that first exposed me to witch-hunting manuals and tales of the supernatural, and it is for this reason that my research commences with a study of representations of witches and witchcraft in early modem England. From my initial exposure to such materials I proceed to examine the similarities and the differences of the cultural significance of the supernatural vis-a.-vis women's activities in early modem Italy. The subsequent discussion of visual representations of witches involves a predominance of Germanic artists, as the seminal work on the discernment of witches and the application of punishment known as the Malleus Meleficarum, was written in Germany circa 1486. Textual accounts of witch trials such as: "A Pitiless Mother (1616)," "The Wonderful Discovery of the Witchcrafts of Margaret and Philippa Flower (1619)," "Magic and Poison: The Trial ofChiaretta and Fedele (circa 1550)", and the "The Case of Benvegnuda Pincinella: Medicine Woman or Witch (1518),"and witchhunting manuals such as the Malleus Melejicarum and Strix will be put in direct dialogue with visual representations of witches in light of historical discourses pertaining to gender performance and gendered expectations. Issues relating to class will be examined as they pertain to the material conditions of presumed witches. The dominant group in any temporal or geographic location possesses the tools of representation. Therefore, it is not surprising that the physical characteristics, sexual habits and social material conditions that are attributed to suspected witches are attributes that can be deemed deviant by the ruling class. The research will juxtapose the social material conditions of suspected witches with the guilt, anxiety, and projection of fear that the dominant groups experienced in light of the changing economic landscape of the Renaissance. The shift from feudalism to primitive accumulation, and capitalism saw a rise in people living in poverty and therefore an increased dependence upon the good will of others. I will discuss the social material conditions of accused witches as informed by what Robyn Wiegman terms a "minoritizing discourse" (210). People of higher economic standing often blamed their social, medical, and/or economic difficulties on the less fortunate, resulting in accusations of witchcraft.

Minute Books and Tyler’s Register of Plantagenet Preceptory No.8 Knights Templar

The Sovereign Great Priority of Canada is a national Masonic organization which consists of seventy-six preceptories that are organized within fifteen districts. The no.8 Plantagenet, St. Catharines is listed under the Hamilton District .The warrant (document issued to authorize formation of a lodge) was issued to this preceptory on November 14th, 1866. This preceptory is still active and meets on the first Monday of every month excepting June, July, August and September. with information from the website Sovereign Great Priory of Canada Meaning of the Tyler/Tiler’s Register: Historically speaking, medieval craft guilds guarded their trade secrets. They placed a guard outside their doors. This person would generally be a junior apprentice who was not entitled to attend trade discussions. The Masons continued this use of doorkeepers. In 1723 in The First Book of Constitutions Dr. James Anderson mentioned “another brother to look after the door, but shall not be a member of it” and in regulation XXVI charged the use of “doorkeepers”. In the minutes of June 8, 1732 this person’s specific title was referred to as “the Tyler”. The word “tyler” appeared in print in new regulation XXVI of the 1738 Second Book of Constitutions. The Masonic ritual of today calls him “a brother without the door”. The Tyler is usually a Past Master who is very knowledgeable in Masonic law and customs. He does not need to be a member of the lodge. He greets brethren and assures that they are “duly qualified”. He gives the first impression of the lodge and insures that visitors and members sign the Tyler’s Register. with information from www.masonicsites.org

Niagara Falls Park: Notice to Treat to Sutherland Macklem, Esquire, 1885

The Niagara Parks Commission administrative headquarters are located in Oak Hall which is on the cliff above Dufferin Islands. In 1798 this land was granted by the crown to United Empire Loyalist James Skinner until 1898. A century later it was the home of the Clarks, Streets and Macklems. These families controlled the mills of Bridgewater which was a pioneer industrial village located at Dufferin Islands. Then, it was known as Clark Hill. Colonel Thomas Clark, commander of the Second Lincoln Militia in the War of 1812 is the earliest known occupant of the house. When Clark died in 1837, the house went to Thomas Clark Street who was the son of the Colonel’s partner. Mr. Street was a bachelor and his sister, widow of Dr. T.C. Macklem, managed his household. Mrs. Macklem had 2 sons. The eldest son drowned in the Niagara River at the age of 8 and the younger son, Sutherland became heir to the estate. Mr. Macklem opened Cynthia Islands and Cedar Island to the public and had roads built to reach them. Two suspension bridges connected them to the mainland and tolls were charged on the bridges. The improvements to the land cost Macklem $18,962. He called the bridges “Bridge Castor” and “Bridge Pollux”. There was also an office built at the end of Bridge Castor. Macklem also spent $454 fixing up the Burning Spring Building (the burning spring is enclosed in a barrel which collects gas and lets it out through a tube at the top). Macklem received a yearly income of $56,378.79 from tourists and visitors. In 1887 Cynthia Islands and Cedar Island were deeded to the crown and became part of Queen Victoria Park. The name Cynthia was changed to Dufferin in honour of Lord Dufferin. Sources: www.niagarafrontier.com/parks.html www.niagarafrontier.com/burningsprings.html

Bee Communities in Restored Landfill Sites of Niagara Region

This study examined the impact of habitat restoration on bee communities (Hymenoptera: Apidae) of the Niagara Region, Ontario, Canada. Bee abundance and diversity was studied in three restored landfill sites: the Glenridge Quarry Naturalization Site (GQNS) in St. Catharines, Elm Street Naturalization Site in Port Colborne, and Station Road Naturalization Site in Wainfleet during 2011 and 2012. GQNS represented older sites restored from 2001-2003. Elm and Station sites represented newly restored landfills as of 2011. These sites were compared to control sites at Brock University where bee communities are well established and again to other landfills where no stable habitat was available before restoration. The objective of this study is to investigate the impact of restoration level on bee abundance and diversity in restored landfill sites of the Niagara Region. Based on the increased disturbance hypothesis (InDH) and the intermediate disturbance hypothesis (IDH), I hypothesized that bee abundance and diversity will follow two patterns. First pattern according to InDH suggest that as the disturbance decrease the bee abundance and diversity will increased. Second pattern according to the IDH bee abundance and diversity will be the highest at the intermediate level of disturbance. A total of 7 173 bees were collected using pan traps and flower collections, from May to October 2011 and 2012. Bees were classified to five families, 21 genera and sub-genera, containing at least 78 species. In 2011 bee abundance was not significantly different among restoration levels while in 2012 bee abundance was significant difference among restoration level. According to family there were no significant difference in Halictidae and Apidae abundance among restoration level while Colletidae and Megachilidae abundance were varied among restoration levels. The bee species richness was highest in the newly restored sites followed by restored control sites, and then the control site. The current study demonstrates that habitat restoration results in rapid increases in bee abundance and diversity for newly restored sites, and, further, that it takes only 2-3 years for bee assemblages in newly restored sites to arrive at the same levels of abundance and diversity as in nearby control sites where bee communities are well established.

Advertisement - Thomas Cowan

An advertisement for Thomas Cowan, dealers in boots, shoes, trunks & valises. The ad is bright in colour and shows a shoe sitting on a shell with a fan and dragonfly and flower. It reads "compliments of the season" in the top corner.

Niagara Falls Seminary [school for young Ladies] advertisement, ca. 1840

One broadside advertising the Niagara Falls Seminary [school for young Ladies], run by Mrs. Shotter of Drummondville, ca. 1840. The advertisement contains information about fees and courses, including a complete English education, music, French, drawing and flower painting, and dancing. The bottom of the page contains a list of references for Mrs. Shotter, including Ex-Consul Buchanan, Colonel Delatre, Rev. W. Leeming, Rector of Chippawa (all of Drummondville); Dr. Mewburn (Stamford); Rev. T.B. Fuller, Rector of Thorold; John Stayner, Esq., P.M. Queenston; William Gamble, Esq., Milton, near Toronto; and Hon. John Hamilton, Kingston.