Tag: horticulture

More comprehensive human intervention studies will be essential in the future to provide insight into the potential influence of dietary polyphenols and their aromatic bacterial metabolites on intestinal microbial communities and their activities.Probiotics are defined as viable microorganisms that, when consumed in sufficient amounts, confer a health benefit on the host.To date, most of the commonly used probiotics are limited to strains of certain Lactobacillus and Bifidobacterium species . Survival during passage through the GI tract is generally considered as the essential feature for probiotics to preserve their active functions in the colon. Indeed, the probiotic strains must overcome biological barriers, including resisting gastric and bile acid secretion and tolerating intestinal lysozyme and toxic metabolites produced during digestion . Various studies found that at least a fraction of probiotic bacteria can be detected in stool for between 1 and 3 weeks after consumption . Probiotic Lactobacillus strains were also found to adapt for survival in the gut and possess gut-inducible genes that are responsive at different sites in the intestine. Interestingly, provision of the probiotic Lactobacillus plantarum to mice fed a Westernstyle diet and to humans resulted in similar gene expression profiles of this strain.As probiotics are delivered via various food vehicles, the complex food matrix should also be viewed as an important factor that may alter the probiotic activity in the gut. To date,pots with drainage holes only a few animal and clinical studies have addressed the functional roles of food on probiotic-conferred health benefits.

The mechanisms of probiotic effects on health are only partially understood but likely function either directly through interactions with host intestinal epithelial and immune cells or indirectly by modulating the indigenous intestinal microbiota. In regard to the latter, several studies have concluded that probiotic consumption does not result in global modifications of the intestinal microbiota in healthy individuals.However, probiotics might confer modest but significant changes to the functional activities of local intestinal bacterial populations. When examined at the meta-transcriptional level, intake of a probiotic fermented milk was associated with the upregulation of microbial genes corresponding to plant polysaccharide metabolism.Similarly, administration of probiotics was shown to induce crosstalk between the probiotics from the diet and the individual bacterial species in the gut and might induce competition for limited substrates that results in fluctuations tof the metabolic profile of the host.The gut microbiome of healthy adults is highly resilient , where the stable native microbiota prohibits the succession of microbes from the diet.In addition, the effect of probiotics on the gut microbiome appears to differ depending on host phenotypes such as age, health status, and chronic conditions. For example, the infant gut microbiome is highly diverse and dynamically changes during development and therefore may be easily influenced by the consumption of probiotics . In individuals with irritable bowel syndrome , probiotic consumption resulted in an increase in the numbers of Bacteroidetes in the intestine.Moreover, intake of two Lactobacillus strains by diet-induced obese mice altered microbial composition and decreased expression of inflammatory genes in the adipose tissue while increasing levels of fatty acid oxidation in the liver.Further studies are needed to investigate the effects of assorted probiotic supplements on the gut microbiome with respect to various host life stages and phenotypes.The premise behind substituting sugar with artificial sweeteners is to maintain the palatability of food at the same time as lowering energy intake.

However, a sufficiently high ingestion of non/low-digestible sugar substitutes stimulates the growth of gut microbiota and can induce transitory diarrhea in humans.In particular, the great proportion of non/low digestible sugar substitutes that reach the distal intestine are subject to fermentation by the colonic microbiota, offering approximately 2 kcal/g of energy.Although discovering and characterizing these compounds within foods is relatively new, it is of interest to note that many of these food ingredients are common in our daily diet. For example, the disaccharide alcohol maltitol is considered a common replacement for sucrose. Urinary and fecal excretions of sorbitol and maltitol after 24 h in conventional rats were shown to be minimal compared with germ-free rats.Likewise, maltitol consumption significantly increased production of SCFAs in addition to nine tested fecal microbes after a 6 week trial, including bifidobacteria, Bacteroides, Clostridium, lactobacilli, eubacteria, Atopobium, Fusobacterium prausnizii, Ruminococcus flavefaciens, and R. bromii. A 12 week administration of Splenda, composed of 1.1% of the artificial sweetener sucralose, increased fecal pH and reduced the amount of fecal bifidobacteria, lactobacilli, Bacteroides, clostridia, and total aerobic bacteria in a rat,whereas isomalt, a widely used low-energy sweetener, was considered to be bifidogenic in a human study.Overall, artificial sweetener fermentation by gut microbiota remains either unexplored or poorly documented, some of which are highlighted in a review by Payne et al.Azo compounds are widely used as coloring agents in foods, beverages, and food packaging.In addition, azo polymer coatings have been specifically designed for colon-selective drug delivery due to the presence of pH-sensitive monomers and azo cross-linking agents in the hydrogel structure. Indeed, azo dyes can be metabolized under anaerobic conditions by intestinal microbial processes and, as a result, produce the reductive cleavage products aromatic amines .

The majority of the toxic effects of azo dyes are exerted through aromatic amines produced by their colonic degradation.Raffi et al. reported that isolated intestinal bacteria in an anaerobic culture system were able to decolorize the dyes in the supernatant, suggesting that some of the azoreductase activities are extracellularly released.Xu et al. demonstrated a variable degree of efficiency in the reduction of Sudan azo dyes and Para Red by 35 prevalent human intestinal microbes in vitro.In contrast, Sudan azo dyes and their metabolites selectively inhibit the growth of some human intestinal microorganisms,which may suggest a potential impact on gut microbiome after long-term exposure. In summary, although there are tantalizing glimpses into the effect of azo dyes on microbes in vitro, more data from animal and human studies are keenly awaited.In the colon, sulfur is present in either inorganic form or organic form .The human GI tract poorly absorbs sulfate, and there is little sulfatase activity in the mucosa of the GI tract; therefore, free sulfate in the colon is likely to be of dietary origin.Dietary sulfate drives the activity of sulfate-reducing bacteria that couple oxidative phosphorylation with reduction of sulfate to produce sulfide.The total inorganic sulfur intake is much higher in the Western diet in comparison to a typical African rural diet.Highly processed foods that are high in sulfate include bread, soy flour, dried fruits, and brassicas, as well as sausage, beers, ciders, and wines.Sulfur-containing amino acids such as cysteine can be found in dietary protein and are a source of sulfur for colonic sulfate-reducing bacteria Desulfovibrio desulfuricans. Native Americans who consume a diet high in resistant starch and low in animal products harbor significantly distinct sulfate-reducing bacterial populations and more diverse and different methanogenic archaea than Americans consuming a typical Western diet.Substrate competition for hydrogen among methanogenic archaea, sulfate-reducing bacteria, acetogenic bacteria, and otherspecies likely occurs in the colon.Because hydrogen is an essential component for the survival of colonic methanogens, removal of the substrate terminates methanogenesis. Given an adequate supply of sulfate, sulfate-reducing bacteria that are more abundant in the right colon out compete methanogenic archaea for H2 due to their higher substrate affinity to produce hydrogen sulfide ,an end-product of dissimilatory sulfate reduction.As a result, the mucosal microbiome may be shaped in part through the availability of toxic sulfide compounds and the differential susceptibility of mucosalistic microbes to the toxins.Furthermore, the activity of methanogenic bacteria can also be disrupted by bile acids.In brief, methane production was thought to occur only when sulfate-reducing bacteria were not active.If sulfate is limited and hydrogen is in relative excess, methanogenic bacteria or perhaps acetogenic bacteria will become essential.Therefore, the levels of sulfate present in the colon are critical for determining which bacterial group gains a better survival advantage.Many people consume alcoholic beverages; however,drainage pot few studies exist on the effect of alcohol consumption on the gut microbiome of healthy individuals. For individuals who consume alcohol to excess, abnormal gut microbiota and bacterial overgrowth can potentially initiate or worsen alcoholinduced impaired gut barrier function and contribute to endotoxemia in patients with alcoholic fatty liver disease.

Yan et al. demonstrated a 3 week acute effect following alcohol administration in mice that resulted in bacterial overgrowth, as well as an expansion of Bacteroidetes and Verrucomicrobia bacteria while decreasing Firmicutes, with no difference observed after only 1 day or 1 weekChronic alcohol consumption induces changes in gut community profiles. For example, daily alcohol consumption for 10 weeks in a rat alters the colonic mucosa-associated bacterial microbiota fingerprint pattern.Similarly, chronic ethanol feeding for 8 weeks increased fecal pH and decreased abundance of both Bacteriodetes and Firmicutes phyla with a remarkable expansion of Proteobacteria and Actinobacteria phyla in mice.In a human trial, chronic alcohol consumption resulted in the alteration of the mucosa-associated colonic bacterial composition in a subset of alcoholics, with lower median abundances of Bacteroides and higher Proteobacteria. Furthermore, measurement of serum endotoxin suggests a change in microbial function, rather than abundance, which may lead to increased levels of gut-derived pro-inflammatory factors in chronic alcohol consumption. It is noted that the inability to detect clear differences between alcoholics with and without liver disease suggests that chronic alcohol consumption, rather than the disease physiology, is the most important event that appears to alter microbiota composition.It is now well established that host diet alters the gut microbiome. Changes in the gut microbiota composition are also considered an important factor in health and disease. Dietary assessment has provided us with a window to discover a way to reconfigure the gut microbiome. In this regard, the nutritional manipulation of the gut microbiome serves as a basis for formulating therapeutic approaches that are feasible and acceptable to the general population as a promising way to promote health in the era of personalized nutrition and medicine. Understanding the impact of foods and nutrients on host− microbe coevolution supports the essential role of a mutualistic relationship for intestinal homeostasis, but there remain challenges for nutritionists and scientific investigators alike to determine the “ideal” diet. This review collectively maintains the emerging view that diet supports a specific bacterial community structure and further suggests that a suboptimal dietary composition/quality may promote the development of diseases through introducing intestinal microbial dysbiosis. Major shifts in intestinal microbial composition are often observed when dietary differences between groups are extreme. Only a few population-wide studies are available to date, but some of them support a role of food diversity as a potential mechanism for altering gut microbial diversity. Although it is difficult to determine the causality of observed fecal microbiota shift with respect to many lifelong changes, generally, an adequate control over influential factors is important for the success of clinical studies to eliminate the drastic effects of unnecessary confounding variables. Many of the studies reviewed here rely on the assumption of equivalence between the term “fecal microbiome” and “intestinal microbiome”. Further studies are necessary to elucidate more clearly the exact impact of the selection of different diets on qualitative changes in the gut microbiota. Some nutrients that have been studied, such as dietary fiber, are a possible option for the maintenance of intestinal homeostasis and improvement of gut health, whereas others may contribute an opposite effect. Therefore, future research must be focused on looking to improve the effectiveness of diets with an underlying long-term “targeted approach” that allows improvement of intestinal microbial composition and functional activities. In other instances, when dietary differences are small and on a short time scale, gut microbiota changes are not as obvious, but that is not to say that changes do not occur. An alteration of the gut microbiota at lower taxonomic levels is still likely to have important functional consequences for the host. Notably, gut microbiota varies dramatically from individual to individual in lower taxonomic levels. Even small dietary changes may have impacts on the gut microbiota and altered metabolic activities in the microbial profile that are not easily detected by the phylogenetic/taxonomic methods. Metabolic alterations induced by diet may result in varying the microbial capability of synthesizing substances in the intestinal tract. It appears that measurement of bacterial enzyme activities may be a more sensitive indicator of diet induced changes in the gut microbiota than taxonomic-based methods.

The total levels of CNGs measured here are much lower than CNG concentrations found in European or American elderberry. In a study of European elderberries evaluated at various growing locations and altitudes found that sambunigrin levels range from 0.08 ± 0.01 to 0.77 ± 0.08 µg g-1 . 6 A nearly 10-fold difference in concentrations between elderberry samples highlights the variation on CNG levels due to differences in growing conditions and environmental factors like sun exposure and temperature fluctuations. Furthermore, evidence of CNGs degrading with thermal processing has been evaluated in European elderberry products: when sambunigrin levels were measured in raw and cooked elderberry juice and other products, heating of elderberry juice reduced the level of sambunigrin, from 18.8 ± 4.3 mg kg-1 to 10.6 ± 0.7 mg kg-1 . 1 Liqueur, tea, and spread also had significantly lower CNG concentrations as compared to the raw and cooked juice. American elderberry was evaluated for concentration of CNGs in the seeds, juice, skin, and stem of two genotypes: Ozone and Ozark.60 Elderberry juice was prepared by thawing previously frozen berries in a plastic bag and gently pressing to release juice. The juice of these elderberries contained amygdalin, dhurrin, prunasin/sambunigrin , and linamarin. Total concentrations of these four CNGs was 4.01 µg g-1 in Ozone and 3.66 µg g-1 in Ozark elderberries. The levels of amygdalin and prunasin/sambunigrin were almost equal in Ozone but in Ozark,vertical gardening in greenhouse prunasin/sambunigrin levels were much higher than amygdalin . These concentrations are much higher than the levels found in the present study, as raw blue elderberry juice had a total CNG concentration of only 0.737 µg g-1 .

Because CNGs are formed from phenylalanine, it is possible that the blue elderberry had limited stock of this key material to create CNGs. An alternative reason may be that blue elderberry may have less expression of the genes needed to form CNGs like sweet almonds compared to bitter almonds.A future study should investigate the impact of freeze thaw cycles on the activity of β-glucosidase in elderberries because elderberries are frequently frozen before processing because they can spoil quickly if only refrigerated. Two cooking temperatures were investigated to understand the impact of temperature on the degradation rates of the phenolic compounds in blue elderberry juice. The pH and soluble solids were evaluated for the five juice replicates to ensure the juices were similar for the cooking process. The average pH value of the juices was 3.76 ± 0.11 and the average Brix reading was 16.2 ± 1.1%. The major phenolic compounds in elderberry juice were measured via HPLC-DAD and include 5-hydroxyprogallol hexoside , which is a novel phenolic compound tentatively identified for the first time by Uhl et al. 202239 chlorogenic acid, rutin, isorhamnetin-3-O-glucoside, cyn 3-sam, and cyn 3-glu. Whereas levels of cyn 3-sam and cyn 3-glu decreased to 82.2 ± 6.9 % and 79.3 ± 6.3 %, respectively , more than 98% of the original concentration of 5-HPG, rutin, isorhamnetin-3-O-glucoside and chlorogenic acid remained after two hours. At the higher cooking temperature , the anthocyanins again experienced significant degradation, retaining only 33.2 ± 4.6 % and 36.8 ± 5.5 % of the original concentration after cooking two hours . In a separate study of the thermal stability of elderberry juice, 15% of cyn 3-sam and cyn 3-glu were retained in juice as compared to control juice.

Szalóki-Dorkó, et al. demonstrated that the more complexly glycosylated anthocyanins cyn 3-sam is more stable during thermal process as compared to cyn 3-glu.The results of our study are similar to Oancea et al. which showed after 90 min at 100 °C, total anthocyanin content degraded 58 %.However, that study also observed an increase in total phenolic and total flavonoid content after 60 min, followed by a gradual decrease, which was not observed herein. If sample vials were sealed well to protect from any loss of moisture, this increase in concentrations may be due to the release of phenolic compounds bound to the cell well or other polysaccharides, which can be released with the assistance of pectinase treatments.The main flavonols in blue elderberry, rutin and isorhamnetin glucoside, were stable during the thermal processing, retaining 100.5% and 99.3%, respectively, of their original concentration even at 95 °C . The high retention rates of rutin and isorhamnetin glucoside match literature reports for the thermal stability of these compounds, which show that rutin has a strong thermal stability at acidic pH. More than 80% of the starting concentration was retained after five hours of cooking at 100 °C at pH 5.148 Our results do not agree with another study in which rutin had an activation energy 107.3 kJ/mol, and the half-life values at 70 and 90 °C were 19.25 and 1.99 h, respectively; however, the rutin was in an aqueous solution at pH 6.6.Other compounds present in blue elderberry juice, in addition to a lower pH, could cause synergistic effects to improve stability of rutin in the present study. Limited information on the thermal stability of isorhamnetin glucoside was found, though a study of black currant juice stability found that during long-term storage at room temperature and at 4 °C, isorhamnetin glucoside concentrations did not change significantly during the 12-month period. In the same study, rutin did not change significantly during storageThe main phenolic acid in blue elderberry juice, chlorogenic acid, was also thermally stable.

This result was unexpected, as another study on the thermal stability of chlorogenic acid in a complex with amylose showed a significant decrease in content after 10-15 minutes, depending on the temperature.Their results also showed that a 10 °C increase in temperature results in a 2.5-fold increase in the rate of degradation of chlorogenic acid. It can be beneficial to maintain levels of chlorogenic acid in anthocyanin-rich matrices, as shown in black carrot extract where chlorogenic acid increased absorbance of cyanidin-based anthocyanins at pH 3.6 and 4.6 due to intermolecular co-pigmentation.Overall, our results show that blue elderberry juice behaves similarly to anthocyanin-rich matrices, in that longer processing at higher temperatures degrades anthocyanins. The two main anthocyanins in blue elderberry, cyn 3-sam and cyn 3-glu, behaves similarly during processing, degrading at about the same rate at 72 °C and 95 °C. Furthermore, the other major phenolic compounds like rutin, isorhamnetin, and chlorogenic acid, were highly stable and can withstand the thermal processing. Our study into the effects of thermal processing on the phenolic composition and cyanogenic glycoside content in blue elderberry juice showed that the main anthocyanins present degrade faster at higher temperatures but other important phenolic compounds like rutin and isorhamnetin 3-glucoside are more thermally stable, retaining over 90% of their original concentrations even after two hours at 95 °C. Furthermore, neoamygdalin and sambunigrin were measured in the blue elderberry juice, which were in lower concentrations compared to European and American elderberry. Perched in the hills above Monterey Bay, the UC Santa Cruz campus looks out on the rich farmland of the Pajaro and Salinas Valleys, home to some of the most successful and productive organic farming operations in the country. UCSC’s own 25-acre farm, managed by the Center for Agroecology and Sustainable Food Systems, has been training organic farmers for nearly four decades. Yet until a year ago, students eating in the campus dining halls seldom had a choice of organically grown food. Today, all five UCSC dining halls and the University Center’s Terra Fresca restaurant serve certified organic produce every day of the week. Growers from the seven local farms that make up the Monterey Bay Organic Farmers Consortium —including UCSC’s farm—pool their produce through the Agriculture and Land-Based Training Association in Salinas to sell to UCSC Dining Services. ALBA’s contract to provide locally grown organic produce is a first among the UC system’s 10 campuses. This transition didn’t come easily. It’s the result of several years of collaboration and hard work by students, staff, and local growers, as well as some fortuitous timing. Thanks to these efforts, UCSC is now positioned to be a national model for a growing movement called “farm-to-college,” which, like the K-12 farm-to-school movement, is bringing fresh,greenhouse vertical farming local produce to student dining halls through direct relationships between farmers and educational institutions. Farm-to-school or farm-to-college arrangements help farmers get more of the food dollar, benefit local economies, and give students access to locally produced, fresh food. Several features distinguish UCSC’s Farm-to-College project: a consortium of organic growers was formed to supply produce to the campus dining halls; sustainable food purchasing guidelines were developed; and education is incorporated from the campus farm fields to the dining halls.

In this article we’ll discuss the history of the farm-to-college work at UCSC, describe some of the strategies and obstacles involved in changing the University’s purchasing practices, and report reactions from participating farmers and campus chefs. We’ll also talk about the key role that students across the UC system are now playing in a campaign to create sustainable food systems at all of UC’s campuses, and offer advice on how other campuses can implement such efforts.In the winter of 2003, UCSC’s Students for Organic Solutions brought together diverse stakeholders of the campus food system at the annual Campus Earth Summit to discuss how to create sustainable change in the system, including the advantages of purchasing organic produce from local farmers. This grassroots effort was largely unsuccessful in garnering support from Sodexho—the largest food and facilities management services company in North America—which was then under contract to provide all the food to UCSC campus dining halls. Sodexho was at the same time dealing with its own challenges. UCSC’s Students for Labor Solidarity—unhappy with the company’s labor practices—had organized to “dump Sodexho” in conjunction with campus labor unions. After a six-month student campaign the UCSC administration ended its 30-year contract with the company in June 2004, enabling Dining Services to contract directly with suppliers for the first time. This transition to an “in-house” service structure opened a crucial avenue to work with the university administration in designing a more sustainable food system. Early in this process, conversations between members of the Center for Agroecology and Sustainable Food Systems and Dining Services director Alma Sifuentes brought staff of Dining Services to the UCSC Farm, an important step in introducing the concept of sustainability to the campus food system. Sifuentes also helped catalyze the effort to bring organic, Fair Trade coffee to campus through the Community Agroecology Network .Several months of meetings followed the 2004 Earth Summit as members of campus and community organizations came together as the Food Systems Working Group to craft the details of a purchasing guidelines proposal. Included were representatives from the Center for Agroecology and Sustainable Food Systems, Community Alliance with Family Farmers, Comercio Justo, the Community Agroecology Network , Students for Organic Solutions, and the Education for Sustainable Living Program —all of whom brought expertise in various aspects of sustainable agriculture and food systems. Under these guidelines, preference is given to price-competitive bids from the primary food vendor that meets the greatest number of criteria. For the local organic contract, all produce must be grown within 250 miles of Santa Cruz and be certified organic. In selecting a local organic produce vendor, preference is given to price competitive bids that are “worker supportive” as defined in the guidelines. In May 2004, the Food Systems Working Group formally presented these guidelines to UCSC’s Dining Services. Students for Organic Solutions also continued to build support for bringing local organic food to the dining halls by putting on educational classroom presentations and organic “taste tests” for students. Timed to coincide with the presentation of the guidelines, a campaign spearheaded by Comercio Justo and CAN generated over 2,000 postcards from students to Dining Services in support of the guidelines’ adoption. Honoring the guidelines, Dining Services selected the local distributing company Ledyard as the prime food vendor in 2004. The sole source organic produce contract with the Monterey Bay Organic Farming Consortium began in late summer 2005 after a year-long struggle to find a way to include “worker supportive” as a criterion in selecting a vendor. During the process, FSWG found that “worker supportive” could not be used as a criterion under UC purchasing regulations because, unlike organic and local, employment practices are not regarded as a characteristic of food. In order to qualify for a sole source contract, local organic farmers formed a consortium and agreed to make their farms available for organic farming and food system research conducted under the auspices of the Center.

European elderberry has been studied for its antioxidant, antimicrobial, anti-inflammatory, anticancer, immunomodulatory, and antidiabetic properties, as well as neuroprotection and cardiovascular protection in vitro and in vivo. These activities have been mainly attributed to the phenolic compounds like cyanidin 3-glucoside and cyanidin 3- sambubioside, but some other compounds have been shown to be bioactive as well, including terpenes, lectins, pectin, peptides, and malic acid. Using data from randomized, controlled clinical trials, a recent review found that elderberry could reduce symptoms from upper respiratory viral infections, providing support for the use of elderberry supplements by consumers to combat colds and flus without the use of antibiotic medicine.In a more unique application, elderberry and elder flower extracts have both shown to be effect in combatting gingival inflammation using a topical herbal patch and elder flower tea, respectively.A study of the mechanism of cyanidin 3-glucoside to treat against the influenza virus showed that elderberry extract had some inhibitory effect during the early stages of virus cycle with stronger impacts during post-infection. The mechanism proposed was that the elderberry extract blocks viral glycoproteins which prevent the virus from attaching or entering cells to replicate, and increases expression of IL-6, IL-8, and TNF. Inflammatory modulating activity of elderberry and elder flower extracts have been investigated.Results showed that quercetin, rutin, and kaempferol are strong inhibitors of nitric oxide production,25 liter pot and metabolites from phenolic degradation including caffeic acid and 3,4-dihydroxyphenylacetic acid were also strong inhibitors without cytotoxicity.

Only a few studies have been done on the bioactivity of S. nigra ssp. canadensis. In one, the fruit was evaluated for anticancer properties, which showed chemo-preventative activity by inducing quinone reductase and inhibiting cyclooxygenase-2, as well as inhibiting ornithine decarboxylase. These activities are attributed to flavonoids and lipophilic compounds. Another study evaluated two Canadian cultivars of this subspecies evaluated the antiproliferative efficacy of the fruit and flowers on glioma and brain endothelial cells and results showed that elderberry and elder flower extracts inhibited the proliferation of cells under normoxic and hypoxic conditions. The elderberry extracts performed the best and the bio-activities were attributed to the synergistic work of cyanidin 3-sambubioside-5-glucoside and rutin content of the berries, though the rutin concentration in the flowers still had beneficial effects. Blue elderberry has only been evaluated as antioxidant activity using the ABTS assay, which indicated that this subspecies has 11.62 ± 0.38 mM Trolox kg-1 FW, roughly one third of S. nigra ssp. nigra evaluated in the same study, where all fruit samples were grown in Slovenia.Further work on elucidating the biological activity of this subspecies through in vivo assays and preferably clinical trials should be explored, especially using blue elderberry plants growing in North America to support its use in supplements. Elder flowers have also been evaluated for their biological activities. A review of antioxidant activity in S. nigra ssp. nigra flowers has recently been published 2 , including ABTS, DPPH, FRAP and CUPRAC assays, therefore it will not be re-summarized here. In general, the data showed that elder flower has higher levels of antioxidant activity compared to the elderberry. Similarly, elder flower extracts had higher nitric oxide inhibition compared to elderberry extracts in vitro.

Elderflower is antidiabetic by increasing insulin-dependent glucose uptake, diuretic, treat respiratory infections, antiviral. While phenolic compounds like flavonols are presumed to be the most active compounds, pectic polysaccharides are also bio-active in elderberry and elder flower, inducing complement fixing and macrophage stimulation.Flowers of the blue elderberry have been evaluated for their antioxidant activity using the ABTS assay, which showed they have 44.87 ± 0.54 mM Trolox kg-1 DW, significantly less than flowers of S. nigra ssp. nigra . Aqueous extracts of wild elder flowers of this subspecies were also found to have neuroprotective effects, especially related to Parkinson’s disease, by increasing the antioxidant response mediated by Nrf2 in cortical astrocytes and improving mitochondrial function in neurons.Unfortunately, that study did not include any growing information about the elder flowers or the concentration of the phenolic compounds in the extract, which would have helped other researchers replicate and expand on the results. While there have been promising studies on the impact of elderberry and elder flower extracts to combat illness and disease, more in vivo studies and clinical trials should be performed to better understand the mechanisms of the bio-activity as well as to determine which compounds are responsible for the bio-activity, particularly in the lesser-known subspecies canadensis and cerulea. This can better inform people involved with the cultivation of elderberry to select for varieties that have the compounds of interest. The market for herbal supplements has been growing in the recent decade and immune system-supporting supplements had a huge spike in sales during the COVID-19 pandemic.

Elderberry products are a popular option of alternative medicine in hopes of improving and protecting health.Beverages are a popular use of the elderberry, including syrup or other tonics made by soaking the berries in water or alcohol. It can also be found as an ingredient in various kombuchas, juices, energy drinks, wine, and tea. Elderberry is typically mixed with a variety of other ingredients, including but not limited to ginger, honey, echinacea, and other spices.More recent products using elderberry include gummies typically marketed as health supplements, lozenges, tablets, and powdered berries especially as part of a drink mix. Elderberries are also frequently used in jams and jellies. Pomace, the byproduct of juicing, has been studied for its benefits when incorporated into other products just as baked goods.Beyond its potential for bio-active products to benefit consumers, elderberry can also be used as a natural food dye due to the high concentration of anthocyanins, which can be used in place of artificial red or purple food dyes, particularly in acidic foods. Its application in edible films has recently been investigated, explored various bio-polymers that could retain the phenolic compounds of elderberry in the film so that they can remain active to protect foods Active edible films can be an effective solution to reduce plastic packaging and food waste due to spoilage.Cosmetic and skin care applications are also an area of interest, and current products on the market that include elderberry include lip color, toner, face mask, and Epsom salt. Herein, the data available on the other elderberry subspecies of interest are summarized to provide a basis of the expected composition as well as information to compare the subspecies for their composition. Elderberries have a high amount of water, at about 80%. The main sugars in elderberry are glucose and fructose, with some small amounts of sucrose. Sorbitol was also measured, which was very minor compared to the other three sugars and was seen in the highest concentrations in the wild elderberry. Citric acid is the main organic acid in elderberry, with malic acid the next highest acid. Small amounts of shikimic, tartaric, and fumaric acid have been measured in elderberry as well. Only data on European elderberry is available for micro-constituents such as vitamins, minerals, fatty acids, and amino acids. Vitamins found in elderberry include various B vitamins, vitamin C, and vitamin E.The main minerals are magnesium, calcium, and potassium.Because studies of these micro-nutrients have only been performed on the European elderberry, it is important for further work to include other subspecies, including the American and blue elderberry so that better comparisons can be made. An important group of bioactive compounds found in fruit and vegetables is phenolic compounds,raspberry cultivation pot which consist of one or more phenolic groups . Types of phenolic compounds include phenolic acids and flavonoids; flavonoids can be further separated into groups such as anthocyanins, flavonols, flavan-3-ols, and flavones. Phenolic compounds may have some biological activity, although bio-availability can be very low. A common, albeit imperfect, way to measure phenolic content of elderberries is using a colorimetric method like Folin-Ciocalteu which can measure a complex that forms between phenolic compounds and molybdenum-tungsten at 765 nm. Because this method measures all reducing agents in the matrix, reducing sugars and ascorbic acid will also react and increase the absorption thus inflating the total phenolic content . Standard curves are typically constructed using gallic acid, hence the units for TPC are gallic acid equivalents . TPC in European elderberries can vary greatly but reported values include 461 ± 121 49 and 683 ± 49.

In American elderberry, TPC has been reported to be 390 ± 56 50, 593 ± 70. One study has included blue elderberry grown in Slovenia, which had a TPC of 416 ± 31. However, because of the imprecise nature of this assay, it is important to identify and measure the concentration of each phenolic compound present whenever possible, the results of which is explored in the following sections.Anthocyanins are water-soluble pigments in plants, and they give elderberries their bluepurple hue. Total monomeric anthocyanin content is typically measured using the pH differential method, which takes advantage of the change in light absorption of anthocyanins in solutions with different pH and the unit is typically cyanidin glucoside equivalents . European TMA content of European elderberry can range from 170 ± 12 to 343 ± 11 mg CGE 100 g -1 FW. In American elderberry, the range of TMA content is 354 ± 59 to 595 ± 26 mg CGE 100 g -1 FW 52 and 106 ± 2 to 444 ± 14 mg CGE 100 g-1 FW. Analysis of the phenolic compounds via high performance liquid chromatography with UV-Visible light detection or with mass spectrometry have elucidated a variety of molecules present in the European elderberry. Anthocyanins, a type of flavonoid and popular for their red to blue pigments, are of high interest in elderberry. Most studies have found that cyanidin -based anthocyanins are the dominant type in European and American elderberry, including cyn 3-O-sambubioside -β-Dglucopyranoside and cyn 3-O-glucoside. Cyn 3-sambubioside-5-glucoside and cyn 3,5- diglucoside are also commonly seen in the elderberry. The American elderberry has a more unique anthocyanin profile with high presence of acylated anthocyanins compared to the European elderberry, including cyn 3-O-coumaroyl-sambubioside-5-O-glucoside , cyn 3-Ocoumoaryl-sambubioside. These acylated anthocyanins may be more stable during processing, but the authors found that cyn 3-O-coumaroyl-sambubioside was the least stable anthocyanin during storage , whereas cyn 3-O-cou-sam-5- O-glu and cyn-3-O-sam-5-O-glu were more stable. Another major type of phenolic compound in elderberry is flavonol glycosides, which include rutin , isorhamnetin 3-O-glucoside or 3-O-rutinoside, and kaempferol 3-O-rutinoside. Rutin has frequently seen to be the most concentrated flavonol in European elderberry, and often the most concentrated phenolic compound of any present. 8 S. nigra ssp. canadensis also contains higher levels of rutin than other flavonols.Other flavonol glycosides present in elderberry include kaempferol and isorhamnetin derivates, such as kaempferol-rutinoside, isorhamnetin-rutinoside, and isorhamnetin-glucoside. Phenolic acids are also present in high amounts in elderberry, including chlorogenic acid isomers , p-coumaric acid, sinnapic acid, cinnamic acid, and ferulic acid. Flavan-3-ols found in elderberry include -catechin, -epicatechin, and procyanidins than those detected in elderberry juice, found to be 18.8 ± 4.3 mg kg-1 in raw juice and 10.6 ± 0.7 mg kg-1 in cooked elderberry juice, suggesting that thermal processing can reduce CNG levels in elderberry products. American elderberries have been evaluated for their concentrations of CNGs. These include amygdalin, sambunigrin , linamarin, and dhurrin. Specifically, the Ozone and Ozark genotypes were evaluated, giving better insight into how CNG concentrations may be impacted by plant genetics. While the total concentrations of the four CNGs in the two American elderberry genotypes were somewhat similar , the composition of which CNGs made up that total were quite different: Ozone elderberries had similar levels of amygdalin and sambunigrin while Ozark elderberries had much higher levels of amygdalin than sambunigrin . The flavor profile of elderberries is an important factor in the consumer sensory experience with elderberry products. Two of the most common compounds identified as drivers of elderberry aroma identified in multiple studies of the berries or elderberry juice are β-damascenone and dihydroedulan. Nonanol was also identified as a key volatile compound contributing to the characteristic elderberry aroma, while ethyl-9-decenoate was found to be important for the characteristic elderberry aroma by another study. While these volatile compounds can be key to the unique aroma, they are not typically the most concentrated compounds. Studies have found the most concentrated compounds to be linalyl acetate, linalool, phenylacetaldehyde, benzaldehyde, hexanal, 2- and 3-methyl-1-butanol, nonanal and benzaldehyde. However, comparing concentration of compounds across studies can be difficult due to differences in sample preparation, extraction method, and method parameters, to name a few important factors.

Due to the barrier created by type IV enzymes in the free-living condition , horizontal transfer involving Frankia in the soil is likely limited to other actinobacteria whose genomes similarly lack methylation , many of whom are endophytes . Inside the host, however, Frankia is likely more receptive to genes from other endophytic taxa as well. Combined, these factors could result in the preferential acquisition of genes involved in growth and host-microbe interactions in planta. In legume symbioses, exudates from hosts into their rhizospheres have been proposed to promote the conjugative transfer of symbiosis genes from a donor group of rhizobia to others, broadening the range of symbionts available to the host . Down-regulation of type IV restriction genes in actinorhizal nodule symbioses could be another mechanism that enhances horizontal transfer of genes related to interactions with plants by making the recipient Frankia more susceptible in the endophytic environment. M. tuberculosis showed much lower transcription of its annotated type IV methyladenine targeting restriction enzyme than other actinobacteria. M. tuberculosis expresses an adenine methyltransferase in hypoxic conditions that regulates the expression of genes likely involved with survival during macrophage infection . For this reason it is likely that M. tuberculosis responds to methylated DNA differently than other actinobacteria; indeed electrotransformation of M. tuberculosis can be readily achieved with methylated plasmids replicated in E. coli DH5α , suggesting that methylated DNA is not digested in M. tuberculosis. In this study derivatives of broad host-range plasmid pSA3 were capable of replication in F. alni. This shows that the broad host-range origin is capable of replication in Frankia and supports its use as a vector for the manipulation of Frankia spp. The parent plasmid of pSA3, pIP501,grow bucket replicates in a very broad range of bacteria including Streptomyces lividans and E. coli indicating the potential for transformation of additional actinobacteria with these plasmids.

The expression of the egfp gene of plasmid pIGSAFnif was up-regulated in N media compared with expression in N media, at proportional levels to the expression of the nifH nitrogenase gene , demonstrating for the first time that expression of reporter genes can be manipulated in Frankia. This transformation system resulted in the ability to visualize the expression of nitrogen fixation genes in vitro by fluorescence microscopy . Interestingly, fluorescence was detected in both the spherical portion of the vesicle as well as in the stalk that connects to the hyphae, suggesting that nitrogen fixation genes are expressed in both parts of the vesicle. Previous studies have shown that the vesicle envelope is deposited around the stalk as well as the spherical part of the vesicle , supporting the observation that nitrogen fixation can occur in the stalk. Although the fluorescence observed when egfp was expressed under the control of the F. alni nif cluster promoter was predominantly in the vesicles, some fluorescence was occasionally observed in hyphae under nitrogen-fixing conditions whereas in N media there was no observable fluorescence . This suggests that there can be condition-dependent expression of nif genes in the hyphae as well as the vesicles induced by nitrogen limitation. Frankia spp. in symbiosis with members of the Casuarinaceae have been reported to fix nitrogen in hyphae, since no vesicles are differentiated ; this pattern correlated with the formation of a lignified host cell wall in the symbiotic tissue that likely reduces oxygen partial pressure . In liquid culture, there may be zones of low pO2 that develop in portions of a Frankia hyphal colony where nitrogen fixation could be induced. Frankia spp. in symbiosis have been suggested to be more autonomous than rhizobial microsymbionts due to their ability to control the flow of oxygen with the formation of vesicles, and due as well to the expression of more metabolic pathways in the micro-symbiont in symbiosis. These factors potentially allow Frankia to be more metabolically independent from their hosts .

The development of genetic tools for the manipulation of Frankia will allow further exploration into these and other distinctive molecular aspects of actinorhizal symbioses, which will, in turn, further inform analyses of the evolution and diversity of root nodule symbiosis. The transformation methods presented here should be applicable for genetic experiments in other Frankia strains. Because plasmid pIGSAF has a broad host-range origin of replication and expresses egfp under the control of a constitutive promoter, the plasmid is likely to be usable in other strains as well. Even in the absence of selection, plasmid pIGSAF was found to be stable in F. alni cultures for at least 3 weeks and cultures continued to show fluorescence after at least 4 weeks . Three to 4 weeks is a time period that sufficiently spans the stages of nodulation and early nitrogen fixation in a broad-spectrum of hosts. This includes Alnus glutinosa with F. alni ACN14a , and Casuarina cunninghamiana , Discaria trinervis , Shepherdia argentaea , and Datisca glomerata . This suggests that transformants can be used to inoculate plants to study of the role of Frankia and its interactions with hosts during nodule establishment and symbiosis. In future this system could be modified using recombination or site-specific integrases to anchor genes within the genome. Differential regulation of reporter genes such as egfp can be used to localize the expression of genes identified by genomics and transcriptomics in specific Frankia cell types, in different growth conditions, and in symbiosis. Replicating plasmids may also enable the study of gene function by constitutive expression of selected genomic genes, by promoter switching or by knock-down experiments expressing anti-RNAs to genes of interest . Circumventing the natural restriction systems of Frankia will also increase the transformation rate of non-replicating plasmids and enable higher efficiency recombination, which can be combined with CRISPR systems, for gene knock-out experiments as attempted by Kucho et al. . While this manuscript was in review a separate method for the transformation of Frankia spp. utilizing conjugation with a methylation-positive E. coli was reported by Pesce et al. . Conjugative transfer has been shown to evade recipient restriction systems , thus presumably allowing Frankia spp. to circumvent the type IV restriction barrier identified in this study.

Nevertheless, the transformation efficiency of Streptomyces spp. by conjugation has been shown to increase over 104 -fold with unmethylated DNA . Thus, in addition to enabling the transformation of Frankia spp. by electroporation, our analysis of restriction systems in Frankia spp. can be further utilized to improve the transformation efficiency of conjugative transfer to Frankia spp. through the use of methylation-deficient E. coli donors. Optimized transformation efficiency will be crucial for future studies involving the generation of recombinants including gene knock-outs. Elderberry is a part of the Viburnaceae family and grows all over the world, including Europe, North America, and Asia.Due to the vast geographic and morphological variety within Sambucus, there have historically been many species within the genus. However, a reorganization by Bolli reclassified some of the most common species under Sambucus into subspecies of S. nigra. More recently, elderberry was moved out of the Adoxoaceae family, which had already been changed before when elderberry was taken out of the Caprifoliaceae family. These changes have impacted the three subspecies most of interest in this work: the European elderberry S. nigra ssp. nigra; the American elderberry S. nigra ssp. canadensis; and the blue elderberry S. nigra ssp. cerulea .However, due to wide acceptance of this naming scheme for the subspecies, it will be used through this work to align with the current naming, but previous works cited may use the former species names. Furthermore, some sources refer to the entire plant as an “elder”,dutch bucket for tomatoes while others refer to the plant as “elderberry”, which is also used to denote the fruit of the plant. In this work, “elderberry” is used to discuss the plant as well as the fruit. “Elderflower” is used to refer to the blossoms of the plant. European elderberry is the most well-studied and widely used subspecies of elderberry in the market. This subspecies grows throughout the European continent, including countries such as Slovenia, Portugal, and Austria. The fruit and flower have been studied for decades for their composition and bio-activity, and while elderberry and elder flower are not new ingredients to the market, they have garnered more attention in the last several years as consumers look for more natural remedies and supplements to support their health. This has been especially true during the COVID-19 pandemic, in which elderberry became a popular ingredient in immunity-supporting supplements. Thus, investigating other elderberry subspecies like the blue elderberry, the focus of future chapters, allows for farmers in the United States to capitalize on this demand, but more information is needed on this particular plant if it is going to be used in consumer products. There is a long, rich history of the use of different parts of the elderberry plant by many cultures. For example, the wood has been used for kindling and musical instruments. Indeed, the name of the plant is derived from various ancient words related to instruments. The flowers and berries have been used in a variety of beverages, foods, and other herbal supplements.\

The plant itself has been revered by many cultures, with a story about the “Elder Mother” living within the plant would protect those near the plant. It was even expected to ask the Elder Mother for the berries or flowers before taking them; without permission, she may seek revenge. The leaves, branches, flowers, and berries were believed to have protective powers for a home and the leaves were also used during burial rituals by some Celtic people. The personification and deep reverence for the elderberry show the importance of the plant through generations. Hippocrates and Pliny the Elder both wrote about elderberry and its medicinal properties. Tribes indigenous to North America used flowers and fruit for medicinal and beverages. Berries were also used as a natural dye for baskets and branches were used to make musical instruments. Elderberry is a perennial, deciduous plant native to many regions of the northern hemisphere. Elderberry plants are neither tree nor bush, as the plant sends new canes up each season, which without pruning, can lead to a large, shrub-like plant that can be several meters tall and wide. They prefer to grow in sunny, riparian climates with moist, well-drained soil, though subspecies in North America can be drought-tolerant. While pruning even down to the ground level of the elderberry can improve yield and accessibility for harvesting, there is a limitation on pruning of the blue elderberry in the Central Valley of California. Due to the threatened status of the Valley Elderberry Longhorn Beetle , which lives only in the elderberry, branches larger than one inch in protected areas should not be pruned or removed from a growing site.Elderberry shrubs typically produce small white flowers with five petals in the spring, though the elder flowers of the blue elderberry are a creamy yellow color. Small, dark blue-purple berries ripen in the summer in large clusters called umbels or cymes, though there are examples of subspecies that have some variation to these morphologies, such as the blue elderberry that has a white bloom on the berries, causing the berries to look blue, and S. racemosa, which are red.Variation can also occur within a subspecies due to growing conditions, such as soil type, precipitation, and temperatures, as well as a key differentiation tool: cultivars or genotypes. There are established cultivars or genotypes of the European subspecies , such as Sambu or Haschberg, as well as of the American subspecies , like Bob Gordon or Wylewood. Cultivars can have more consistent growing patterns, such as blooming or ripening all at once, and desired chemical compositions, such as increased anthocyanins, thus are more desirable to use in large scale growing of elderberry for commercial use. Blue elderberry does not have any established genotypes to date. If commercial interest in this subspecies continues to expand, effort should be made to develop cultivars with consistent quality and improved harvest ability, which is hampered right now due to flowers and berries ripening throughout a season, instead of a smaller window of time like the American and European subspecies. Indeed, starting this work can help increase the commercial interest viability of the blue elderberry. A primary driver in interest in the composition of elderberry and elder flower is for their potential health benefits. Several reviews have recently been published on this topic; thus, it will not be explored in depth here.

The B-number and soot yield are fundamentally robust parameters that may be used in the future as means to classify the flammability of a given warehouse commodity, to strengthen the level of confidence in determining the flammability of a commodity, and to increase the effectiveness of warehouse fire protection and suppression applications. Additionally, the results of this study are useful for the application of sprinkler activation and determining the amount of sprinkler suppression that is necessary as a fire grows larger.Electricity consumption projections place data centers at up to 13% of global electricity demand in 2030 due to the expected growth in the production and use of electronic devices, cloud services, and computer networks. Cooling infrastructure accounts for up to 40% of the total energy delivered to a data center. With business and society relying so much on data centers, there is a greater need for reliable and clean electric power for data centers. Solid Oxide Fuel Cell systems have the potential to provide more reliable and cleaner electrical load-following characteristics compared to other technologies while enabling dynamic operation and control. High temperature exhaust of SOFC can be used to run a bottoming cycle such as cooling system which makes it an attractive integrated system for data center applications. However, cost and durability are major challenges associated with SOFC technology. On the other hand, the biggest challenge for using hydrogen as an energy carrier for SOFC is the very high pressure or very low temperature required for its storage, transmission, and distribution,nft growing system which makes the need for a more dense liquid energy carrier like ammonia inevitable. This dissertation, first, focuses on evaluating the integrated system concept and assesses the achievable air conditioning from SOFC waste heat.

To explore the feasibility of thermally integrating SOFC with liquid desiccant dehumidification , a spatially resolved physical model developed in MATLAB is used to simulate the operating characteristics of this SOFC system. A corresponding physical model is developed to simulate the liquid desiccant air conditioner for dehumidification. This research evaluated SOFC systems for powering demand of a single server rack to powering a row of servers . The LDD operation is based on distributed waste heat from SOFC that powers the servers. This research indicates whether waste-heat based cooling and dehumidification could power the servers and maintain server operating temperatures and humidity in the safe range for different weather conditions. It calculated the yearly storage capacity required for each location to meet the demand of the data center for the entire year. Next, the performance and degradation of a 1.5kW commercial system, that is proposed for source of power and cooling of servers, is evaluated under steady state and dynamic load cycling conditions for over 6000 hours. The degradation rate and performance characteristics of the SOFC system is analyzed to determine the long-term performance and durability of SOFC system under dynamic condition. Finally, to analyze and compare the degradation of single cell SOFC directly fed with ammonia , externally reformed ammonia and pure hydrogen , three durability tests are conducted on anodesupported SOFC. Electrochemical impedance spectroscopy and scanning electron microscopy are conducted to study the performance losses during operation and to observe the microstructure changes of the cell after testing. The rapid growth of internet use, cloud computing and data-driven machines and services is increasing the electric power consumption and carbon footprint of data centers. Data center electricity demand was around 200TWh in 2018.However, due to the expected growth in the production and use of electronic devices, cloud services, and computer networks, electricity consumption projections place data centers at up to 13% of global electricity demand in 2030.

As shown in Figure 1 in the best-case scenario data center electricity consumption doubles in the next 10 years. Associated with this massive electricity consumption are greenhouse gas and criteria pollutant emissions due to the use of conventional fossil energy resources to power data center electric loads. In 2020, the information and communications technology sector represented 2.3% of global GHG emissions, 28.8% of which were associated with data centers. In addition, as a result of the COVID-19 pandemic, dependency upon internet technology, and cloud services, data center demands are only increased further. Data centers will use around 3–13% of global electricity in 2030 compared to 1% in 2010. In order to meet the emissions reduction targets imposed by many IT companies like Microsoft, the power supply for data centers must come from renewable energy sources. The trend of using renewable power is being used widely and likely many data centers can be run with low GHG and pollutant emissions, even if they do not find ways to reduce their absolute electricity usage.In a traditional data center connected to the electric grid, less than 35% of the energy from the fuel source that is supplied to the power plant is delivered to the data center. Power plant generation losses and transmission and distribution losses are the most significant inefficiencies in data center. Figure 2 shows the process of power loss through the transportation and distribution network of the electric grid starting from the fuel source up to the power supplied to the consumer. It is evident that the largest inefficiency is from the power generated at the power plant level with additional losses associated with the transmission and distribution to the data center, where the data center receives roughly 30% of the total energy that could have been supplied ideally from the fuel source. There are further losses associated with the infrastructure required for daily reliable operation systems. The additional power consumed by the cooling, lighting, and energy storage, means approximately less than 17.5% of the energy supplied to the power plant is ultimately delivered to the servers. Information Technology equipment and cooling infrastructure are the two major power consumers. As shown in Figure 3 cooling infrastructure is a large electricity consumer in the data center and can account for up to 40% of the total energy delivered to the data center. Cooling also represents the biggest slice of the total cost in a data center.

With more capacity and higher density there is an increased need for energy-efficient cooling of the IT equipment. While data centers need cooling to protect servers and other equipment, an additional challenge is humidity control. Too much humidity can cause corrosion of metal parts of the servers leading to short circuiting of electronics. On the other hand, low humidity allows the servers to develop electrostatic charges that can cause damage to sensitive equipment. According to Uptime Institute, electric power outages are the main causes of the increasing outage trend in data centers. According to the Eaton Blackout Tracker, the US experienced more than 3,500 utility power outages in 2017, the vast majority of which were attributed to faults in the transmission and distribution system. This number represents a 62% increase in outages from a decade ago. In 2018, a storm interrupted grid power to Microsoft’s San Antonio Azure data center, knocking cooling systems offline, damaging a significant amount of equipment, and bringing down Active Directory and Visual Studio Team Services for almost 24 hour. In some areas, rising temperatures are driving up the cost of data centers. In other areas, extreme rainfall and flooding have damaged equipment, and prevented the fuel deliveries which are so critical to the traditional back-up power solutions. And power costs continuously increase. The global cost of power for data centers is expected to increase 80% in the next five years. At 35% of total costs,vertical hydroponic nft system electric power continues to represent the largest portion of data center operating costs. Every power generation system that supplies a data center must comply with stringent reliability and availability constraints to ensure 99.9999% server uptime. With business and society relying so much on data centers, and data centers growing more and more, there is a greater need for reliable and clean electric power for data centers. Our solution is using small scale Solid Oxide Fuel Cell for rack level power generation, which is capable of delivering uninterrupted, 24×7 power that is resilient and clean and eliminates the need for back up generation and is also able to provide high quality heat to run a liquid desiccant dehumidification system to provide cooled and dehumidified air for servers. Generating power on-site, at the point of consumption, rather than centrally, eliminate the cost, complexity, interdependencies, and inefficiencies associated with electrical transmission and distribution. Providing the required cooling for servers through liquid desiccant dehumidification run by high quality exhaust of SOFC has the potential to decrease the power consumption of data centers by up to 40%.In this research, first, I investigate the integration of rack level fuel cell powered servers with Liquid Desiccant Dehumidifier technology that can be dynamically dispatched to produce electricity and cooling in various amounts to meet power and air conditioning demands of data centers. This thesis focusses first on evaluating the integrated system concept and to assess the achievable air conditioning from SOFC waste heat. To explore the feasibility of thermally integrating SOFC with LDD, a spatially resolved physical model developed in Matlab is used to simulate the operating characteristics of this SOFC system.

A corresponding physical model is developed to simulate the liquid desiccant air conditioner for dehumidification. This study considers SOFC systems capable of powering a 1. single server rack and the operation of an LDD for cooling and dehumidification of that same rack, and 2. SOFC and LDD systems designed for a row of 20 servers. The analysis will indicate whether waste-heat based cooling and dehumidification is capable of powering the servers and maintaining server operating temperatures and humidity in the safe range for different weather conditions. Even though SOFC technology offers several advantages such as fuel flexibility, high efficiency, and zero criteria pollutant emissions, cost and durability are major challenges associated with current SOFC technology. Durability of SOFC technology is a key aspect for its commercialization and long-lasting deployment in different applications. Dynamic operating conditions have considerable effects on the long-term performance and durability of SOFC systems. On the other hand, while many studies have focused on green hydrogen produced through electrolysis from sun and wind, as a clean fuel for fuel cells, the biggest challenge for using hydrogen as an energy carrier is the very high pressure or very low temperature required for its storage, transmission and distribution, which makes the need for a more dense liquid energy carrier like ammonia inevitable. I propose that ammonia made in a sustainable way can serve as a sustainable, low-cost, and high-density energy carrier and the fuel for SOFC systems in the Future.In the second part of this research, I first evaluate the performance and degradation of a 1.5kW Alternating Current commercial SOFC system that is proposed for source of power and cooling of servers under steady state and dynamic load cycling conditions for over 6000 hours. I monitored and analyzed the degradation rate and performance characteristics of the SOFC system to determine the long-term performance and durability of SOFC system under dynamic operating conditions. Second, I evaluate the effect of ammonia as fuel on SOFC performance. I study and compare the degradation of an SOFC single cell fed with fed with ammonia , externally reformed ammonia and pure hydrogen . The goal of this thesis in to evaluate the integration of SOFC with Liquid Desiccant Air Conditioning for efficient, reliable, and grid-independent data center power and cooling. The integrated system will supply electricity and cooling to data center applications. This configuration offers the potential for high energy efficiency, environmental and economic benefits. The integrated concept is shown in Figure 4. The SOFC generates electricity from natural gas which is used directly for powering a rack of servers. The SOFC thermal exhaust will be used to produce hot water which is used for regenerating the liquid desiccant. Concentrated liquid desiccant will be stored and is used for dehumidifying the air when there is a cooling demand. The integrated system is expected to have high efficiency, low emission and provide reliable power and cooling for data center applications. This integrated configuration eliminates grid connection and any transmission line for powering servers as well as significantly decreasing the electric power used for providing cooling to servers.Most traditional data centers get their primary electricity from utility electric grid.

Due to concerns of flavanol monomer epimerization during cocoa production which might yield substantially less bioactive -catechin from naturally occurring -epicatechin the specific concentrations of catechin and epicatechin enantiomers of thinned clusters and cocoa powders are also provided in Table 2 using chiral chromatography to facilitate future epidemiological study designs.The same trend was observed for Chardonnay fractions as the less mature seed had the highest flavanols and procyanidins compared to its more mature dark seed fraction . However, for the Chardonnay seedless fractions, the dark fraction had a significantly higher amount than the light fraction . Ivanova et al. have found that the total content of catechin, epicatechin, and procyanidin dimers B1- B4 in Chardonnay and Merlot grown in R. Macedonia were significantly higher in seeds rather than in skins and pulp at veraison. Our data on flavanols and procyanidins DP 1–2 are in accordance with their findings as the highest content was found in Chardonnay seeds , followed by Pinot noir seeds , Chardonnay seedless , and Pinot noir seedless . Overall, 2.2-fold and 1.2-fold variations of DP 1–2 were found in Chardonnay and Pinot noir thinned cluster fractions, respectively. Previous studies also showed that flavanol monomers in white and red grapes slow down or stop accumulating from veraison to ripening while procyanidin dimers B1-B4 increase slightly until the intermediate phase . These all indicated that grape thinned clusters at veraison could be richer sources of monomeric and oligomeric flavanols and procyanidins compared to grapes during berry formation and berry ripening.

With the increasing interest in utilizing specific bioactive catechin and epicatechin enantiomers,hydroponic bucket mainly -epicatechin and -catechin, in epidemiological studies , we separated catechin and epicatechin enantiomers using chiral chromatography and presented the data in Table 2. Like grapes, the predominant forms of flavanol monomers in natural cacao beans are -epicatechin and -catechin. However, Payne et al. have found that various processing techniques used in cocoa production, such as fermentation and roasting at over 70°C, significantly decrease the total catechins amount and generate significant amounts of -catechin from -epicatechin epimerization, respectively. The Dutch processing is widely used in cocoa production where the cocoa is treated with an alkali solution to reduce the acidity and to intensify the chocolaty flavor in the final powder . Compared to natural cocoa powders, Dutch processing also caused a greater loss in both epicatechin and catechin according to Payne et al.As -catechin has been shown to have pharmacological effects including antimicrobial , antioxidative , and had about five times higher bioavailability than -catechin , thinned clusters could complement cocoa ingredients—a small portion of grape thinned cluster as an ingredient would likely make a positive impact on the -catechin and total catechins content of cocoabased products.Drosophila suzukii , also known as the spotted wing drosophila , is a vinegar fly originating from Southeast Asia. SWD was first detected in North America in August 2008 in Santa Cruz County, California, where it was observed infesting strawberries and caneberries.In 2009, SWD was detected in Washington, Oregon, and Florida. By 2010, SWD was detected in Utah, Mississippi, North Carolina, South Carolina, Wisconsin, and Michigan in the United States, and Alberta, Manitoba, Ontario, and Quebec in Canada.

Recent trapping indicates that SWD can be found in virtually any region of North America where host fruit are available. A coincidental invasion of SWD with a genetically distinct population has also been observed in Europe, with initial detections in both Spain and Italy in 2008, followed by its spread throughout the continent. In North America, SWD is primarily a pest of berries and cherries. In Europe, it is reported to also damage a number of stone fruits and grapes. Unlike native vinegar flies in North America and Europe, female SWD possess a serrated ovipositor that can pierce the skin of healthy, soft-skinned fruits to lay eggs. These eggs quickly develop into larvae, which consume the fruit and render it unmarketable. The only other Drosophila species known to oviposit in sound, marketable fruit is Drosophila pulchrella Tan. This species is native to Japan.1 Growers have attempted to mitigate crop damage risk by applying additional insecticide, harvesting more frequently, performing field sanitation, and implementing trapping programs to detect SWD populations. These management practices are costly and many growers still face significant yield losses from SWD infestations. Raspberry producers are perhaps the most affected by SWD’s invasion among California commodities, although producers of blueberries and cherries have experienced substantial losses too. Strawberry producers have experienced lower damage rates and primarily on the lower-value fruit produced for processing. SWD-related losses in these industries vary by year and crop depending on management practices, weather conditions, time of the year, and geographic location. A primary motivation for focusing on the California raspberry industry is that California accounts for the majority of raspberry production in the U.S. and the raspberry industry accounts for the majority of economic losses due to SWD among berry crops.A second motivation is the magnitude of change in pest management practices; few of the SWD control practices used by raspberry producers were needed to prevent injury from other pests prior to its establishment.

Economic losses in the California raspberry industry include the cost of managing SWD and the value of the fruit lost due to SWD infestations despite management efforts. First, we compute the cost of the chemical management programs and the labor-intensive sanitation practices implemented to mitigate SWD-related yield losses. Second, we calculate the industry level yield losses due to infestation. These components form an estimate of the full economic cost of SWD’s invasion into California raspberry production.In 2013, raspberries were estimated to be the twenty-seventh largest crop in California by value of production. The United States is the third largest producer of raspberries in the world, producing 91,300 tonnes, after the Russian Federation and Poland, which produce 143,000 and 121,040 tonnes, respectively. Across all counties, California’s raspberry production was worth an estimated $239 million according to the United States Department of Agriculture’s National Agricultural Statistics Service , and $437 million according to California County Agricultural Commissioners’ Reports.The difference in these estimates reflects that the NASS data report cash receipts to producers while the Agricultural Commissioners’ Reports estimate the total value of production. Figures 1, 2, 3, and 4 plot California raspberry hectares, production, yield per hectare, price per kilogram, and the total cash receipts between 2004 and 2013.Note that raspberry hectares multiplied by yield per hectare is equivalent to production, and production multiplied by price per kilogram is equivalent to total cash receipts. Four counties account for virtually all commercial raspberry production in California: Ventura, Santa Cruz, Santa Barbara, and Monterey.In 2014, Ventura County produced approximately 52% of California’s raspberry crop by value, $241 million, on 1,873 hectares. Raspberries are the third most valuable crop in Ventura County.9 Santa Cruz County produced approximately 28% of California’s raspberry crop by value, $131 million, on 979 hectares. Raspberries are the second most valuable crop in Santa Cruz County.10 Santa Barbara County produced approximately 10% of California’s raspberry crop by value, $45.2 million, on 591 hectares. Raspberries are the ninth most valuable crop in Santa Barbara County.Monterey County produced approximately 10% of California’s raspberry crop by value, $45 million, on 316 hectares. Raspberries are the sixteenth most valuable crop in Monterey County.Table 1 summarizes California raspberry production by county.Counties are listed from north to south along the Pacific Coast. Figure 5 identifies these berry-producing regions with a stylized map of California.Most commercial raspberry plantings in California have had an 18-month lifespan. The crop is planted in the winter and then harvested twice, first in the fall following planting and then in the subsequent summer. Both harvest seasons last approximately three months, with crews harvesting fruit every three days on average. Variations in harvest frequency depend on yields and pest management activities. Yields are low at the beginning and end of a harvest season,stackable planters and peak near the middle of a season. Pesticide applications may require an interval of time, depending on the particular pesticide, before normal harvesting activities can resume. This period is known as the pre-harvest interval , and it is determined by the U.S. Environmental Protection Agency. Occasionally, low yields are realized during the harvest season due to crop damage resulting from weather, pest activity, or other external factors. The summer harvest is typically larger than the fall harvest.Organically produced raspberries represent a significant share of total California raspberry production.

In 2008 and 2011, California’s organic raspberry production was valued at $11.4 million and $8.98 million, respectively, according to the USDA-NASS. In 2012, 408 hectares of California raspberries were organically managed according to the University of California Agricultural Issues Center. Raspberry prices vary throughout the year, but on average organic raspberries are sold at a price premium. In 2015, the national average retail price of organic raspberries over the entire year was $3.52 per six ounce tray according to the USDA Agricultural Marketing Service. The average retail price of conventional raspberries over the same period was $2.55 per tray. The average California terminal market prices for organic and conventional raspberries were $3.29 and $1.97 per tray, respectively. 16 California raspberries are a major export crop. In 2013, the combined category of raspberry, blackberry, mulberry, and loganberry exports was the twentieth largest export crop category by value in California. Raspberries account for the majority of the production volume and the total value of this category. This California export category was valued at $157 million, and accounted for approximately 85% of total US fresh and processed raspberry, blackberry, mulberry, and loganberry exports. 84% of these exports are received by Canada, 6% by Japan, and 5% by the European Union.The presence of SWD has clearly increased production costs and caused yield losses for California raspberry producers through a variety of channels. However, these studies occurred within one or two years of the first SWD infestations in North America when information on the pest was still sparse and management techniques were rapidly evolving. We can improve on these original estimates now that much more is known about SWD biology, risks, and management. We briefly review these original studies before establishing new estimates of the economic cost of SWD in the California raspberry industry. Walsh et al. 1 and Bolda et al. 6 are the first studies to estimate the economic cost of SWD. These studies utilize yield loss estimates and observations for strawberries, blueberries, raspberries, blackberries, and cherries in California, Oregon, and Washington in conjunction with production data to calculate revenue loss estimates for each state and crop pairing. Walsh et al.1 assume a yield loss of 20% for all the listed crops in these states. As a result, the study estimates a total of $511 million in potential damages annually due to SWD. Bolda et al.6 continue the analysis by assuming the maximum reported yield losses of 40% for blueberries, 50% for blackberries and raspberries, 33% for cherries, and 20% for processing strawberries. The study concludes that potential revenue losses across these states and crops could be as large as $421.5 million given current prices.Goodhue et al. refine these estimates of lost revenue for the California raspberry and strawberry industries by including potential price responses into their estimates. This additional assumption reflects that as the production of raspberries and strawberries decreases, the prices of these products may increase in response. The interaction between production and price is quantified with the inverse own-price elasticity of demand for each crop. The elasticity predicts the percentage change in price of a good in response to a 1% increase in quantity demanded. Drawing upon elasticity estimates established in prior studies, the authors conclude that SWD-induced yield losses could decrease California raspberry and processed strawberry revenues by up to 37% and 20%, respectively. The authors also evaluate the cost of different SWD-targeting insecticide applications and the cost of a specific conventional raspberry pest control program in California’s Central Coast region. The insecticide material and application costs are estimated to be $825.33 per hectare. However, these chemical applications may also provide incidental control of other pests. This implies that the estimate represents an upper bound of the potential chemical control costs associated with SWD.The revenue loss and management cost estimates in these prior studies can be substantially improved using current information about SWD-induced yield losses and management practices. Fruit losses due to SWD and SWD management costs have decreased over time as researchers and producers have developed and implemented better techniques for reducing crop losses.

Lewis himself stretches this instant into a paragraph, slowing the action as he describes how “[t]he friar’s eyes followed with dread the course of the dagger,” settling into Ambrosio’s gaze as “his eye dwelt” on Matilda’s nudity, and evoking the enormity of Ambrosio’s fatal abandonment of self-control as “a thousand wild wishes bewildered his imagination.”I will show how it is control that comes to the fore when we compare multiple illustrations of the scene, as different artists explore the indeterminacy of this moment when each character holds power over the other, before Ambrosio submits to Matilda’s will and his own desire. The illustrations of Matilda and Ambrosio’s struggle of wills suggest dramatically different interpretations of the situation through positioning, gesture, facial expression, and visibility of the dagger. While these illustrations of the scene are truer to the novel than the pulp cover’s depiction that I examined earlier, they still sometimes represent it in sentimentalized ways that, at first glance, have little to do with the situation Lewis conveys. As the first editions of The Monk were published without any illustrations, the earliest image I found that represents this scene is a watercolor by British artist Charles Reuben Ryley that the Lewis Walpole Library estimates to be from around the time of the novel’s publication, 1796. From a distance, a viewer may perceive only a man threatening a woman. Ambrosio leans toward Matilda with arms raised menacingly, and she braces herself and holds up what appears to be a defensive arm. On closer examination, a viewer could notice the exposed nipple that is nearly camouflaged against the shadow of Matilda’s robe,hydroponic nft system the fact that one of her hands holds open the robe, and the dagger that partially blends into the background foliage.

Matilda appears to be composed, looking directly into Ambrosio’s agitated face. Because Ambrosio is hunching, their heads and toes are on a level with each other, suggesting parity. In this image, Matilda exposes her right breast, not her left, as in the novel. Though she points the dagger near her heart, the fact that she gratuitously reveals her chest on the opposite side implies that she is tempting him with her sexuality, an artistic interpretation founded in other scenes from the novel. Here, it provides more evidence for the perception that though Matilda initially appears helpless, she is in control of their encounter, subtly undermining Ambrosio’s attempts to overpower her. Even illustrations that use similar compositions suggest drastically different readings of the what is at stake in this moment of indeterminacy. In both images below, the figures appear close together, low in the frame, centered or nearly so, of roughly equal height and on a level with each other, surrounded by the garden.The one on the left is an anonymous engraving from the French Maradan edition of 1797, the earliest illustrated version of the novel I found. Though the page features a caption in which Ambrosio blames his impending doom on Matilda, the image suggests a sentimental serenity. The soft folds of the robes and the stylized grace of the figures’ physical attitudes make it appear as if the two are dancing rather than that Ambrosio is attempting to prevent Matilda’s suicide. Matilda gazes at Ambrosio with a look that seems adoring while baring both of her breasts, which the engraver renders in sensual detail. The gently rounded, untrimmed trees and shrubs create an Edenic atmosphere. The lone indicator of peril amid this calm scene is Matilda’s dagger, which is well defined but small, thin, and easily ignored. Ambrosio’s gaze might be resting on her chest , and his heavy lidded eyes negate any sense that he may be alarmed. In contrast, the illustration on the right appears stark and urgent.

Ambrosio, instead of being stunned when Matilda raises the dagger, as he is in the text, is rushing to stop her. With his eyes fastened on Matilda’s large, prominent dagger, he places his hand on the arm with which she holds it. Matilda’s indistinct breast is more legible as the target of the dagger than as the enticement of a lover, and her expression is resolute. Compared to the Maradan illustration’s ripples and curves, the lines of the robes and sculpted trees are stiff and forbidding. The former image emphasizes the sensuality of the scene; the latter, its threat, which, unlike the earlier threatening image, portrays Ambrosio not as an aggressor but as Matilda’s would-be savior. The first engraving seems to suggest that the most significant outgrowth of this moment in the text is Ambrosio’s sexual awakening, an occurrence which the image presents as mutually desired. The second implies that this moment is most important as the seed of the destruction that follows, which it clearly attributes to Matilda. The two final illustrations I’ll consider both highlight the erotic and destructive aspects of the scene, though they imply very different things about Ambrosio’s self-control. One is an illustration from an 1891 American edition, one of many in which the story is reprinted with the more sensational title Rosario; or, The Female Monk. The illustration, by August Leroy, shows Matilda revealing her breast with a guarded expression and Ambrosio grasping her sleeve with what appears to be a smile, as if he doesn’t notice the poniard Matilda holds. Meanwhile, the knife is pointing at his groin. The last is a woodcut from a 1984 Folio edition illustrated by George Tute, who makes Matilda’s voluptuous body the focus of the image.Her white breast against the prevalent darkness draws the eye and makes the point of the dagger pressed to it more conspicuous.

Though Matilda holds the knife and the viewer’s gaze, Tute positions her below Ambrosio and facing away from the viewer, which could imply that she is less powerful and less important than Ambrosio. He appears impassive, with a grave expression and a palm raised to command her to stop. He holds his other hand up defensively, but it blends in with the folds of his robe and does not diminish the impression of his dominance. In these five illustrations, Matilda appears in turn to be victimized, determined, amorous, desperate, and unreadable. Ambrosio seems to be dangerous, pleasant, distressed, admiring, and forbidding. It may seem that some of these visual interpretations must actually be misinterpretations, but each one, whether straightforward or ambiguous, has a basis in the text’s complex negotiation of authority and feeling. Comparing illustrations in this way has several possible benefits for literary scholars writing about novels like The Monk. This practice encourages us to see beyond our own subjective responses to a novel or the limited responses enabled by our current critical norms, because when we are able to view several different interpretations of the same textual moment, we are less likely to conclude that there is only one way to respond to it, as several scholars have done with scenes and characters from The Monk. Though the early marketing of The Monk as a sentimental work failed due to scandal, sentimentality persists in many illustrations of the novels and invites readings that take Lewis’s use of the mode seriously. The visual medium allows us to reconsider the emotional portrayals in the text, because it can vivify passages that we may have dismissed because we find the writing too conventional to be effective, as with scholars’ responses to Lewis’s sentimental and theatrical descriptions. Some scholars have argued that Lewis must have intended these ineffective portrayals to be critiques or parodies, but most illustrations present the text’s moments of terror, indecision, and desperation as sincere, making it clear that recent scholarly ways of interpreting the text’s emotional qualities have been narrowed by current critical lenses. Even the act of noticing what passages have been illustrated can be generative, as it may help us determine where others have located the imaginative energy of the text and which moments distill some of the essential dynamics of a novel, as the encounter between Matilda and Ambrosio does. When the same or similar moments from the text have been illustrated multiple times, they can suggest patterns in the novel that we may not have observed. The illustrations of The Monk that I found often depict moments of danger and desire, which mirrors the critical focus of scholars and online reviewers. However,nft channel seeing multiple visual portrayals of the novel’s violence and sex made it clear that nearly all of them feature two characters, and these two characters are most often positioned in ways that emphasize the balance of power between them. And alongside these sensational illustrations are other representations of power between two characters that are neither aggressive nor sexual. In addition to illustrations of Matilda and Ambrosio’s confrontation, Ambrosio lusting over Antonia, Ambrosio murdering Elvira and Antonia, and Lucifer carrying Ambrosio to his death, there are images of Agnes pleading with Ambrosio to preserve her secret and Lucifer demanding that Ambrosio sign his demonic contract. With these latter scenes, and even in many of the more sensational ones, artists often freeze the scene in a moment of decision.

This tendency of illustrations to emphasize choice and complex relational dynamics in The Monk differs from illustrations of other gothic novels that I could find, which suggests that these motifs are significant in the novel. The illustrations of Ann Radcliffe’s novels that I located are generally of scenes of terror that focus on an individual’s experience—the cloaked monk frightening Vivaldi in The Italian, Emily fleeing from the black curtain or from the castle of Udolpho, and several images of Emily discovering a corpse. The gothic illustrations collected in Maurice Levy’s “Images du Roman Noir” most often portray a clear imbalance of power in which a heroine is imperiled: a woman is carried down a cliffside, threatened atop a gothic castle, menaced by men with weapons, dragged across a room or into a closet.These images of scenes from other gothic novels do not emphasize the struggle in relations of power or the choice in fatal moments the same way images from The Monk do, which makes Lewis’s situations of possibility worthy of further attention. Though scholars are certainly justified in arguing that the accumulation and amplification of scenes of sex and violence are a defining pattern of The Monk, we could also envision the narrative as being built on these moments of choice, in which one character holds tenuous power over the fate of another. Seeking out the moments of decision that illustrators represent allowed me to see many more of them, and to observe that whole scenes contain many moments in which characters vie for control. Thinking in terms of the various illustrated iterations of the same instant prompted me to notice the verbal iterations of control within and among scenes, which made it clear that power in The Monk is more complex than scholars have acknowledged. Considering how similar scenes in the novel contain repetition with difference is essential in emphasizing the novel’s kaleidoscopic power relations, as well as the struggle, precarity, and choice involved in Ambrosio’s downfall, elements that many scholars minimize when they focus on Lewis’s authorial control. Even more importantly, what this reading makes clear is that there are more options for discussing feeling in The Monk than the current scholarly parameters have allowed us to see.Felski’s claim that the prevailing scholarly mood of the last several decades has been a suspicious one is especially helpful in considering the way scholars have focused on issues in The Monk that harmonize with distrust. Multiple studies analyze the role of characters’ various kinds of concealment, which I will address in my discussion of Ann Radcliffe’s Italian in the next chapter. Even more informative for this chapter’s consideration of how scholars have argued that Lewis disallows certain feelings in response to particular scenes is the way scholars have dealt with the issue of control in the novel. Several prominent scholars interpret The Monk’s tightly constructed plot of Satanic manipulation or Lewis’s ostentatious writing style as evidence that Lucifer and Matilda dictate Ambrosio’s actions and Lewis attempts to dictate his readers’ responses to an unusual degree. For instance, Maggie Kilgour contends that Lewis “gives the reader a sense of power and control over the narrative” but in reality “keeps us outside, to make us marvel at his authorial powers.”David Punter argues that he “tries to be more cynical than his audience, and to dominate it by means of this cynicism.”

Claudia Johnson claims that Northanger “does not ridicule gothic novels nearly as much as their readers.”She argues that Austen makes both Catherine and Henry out to be bad readers of gothic novels—one for seeing them as true life and the other for seeing them as mere entertainment, when the reality is that they are somewhere in between. Ultimately, she writes, Austen encourages her own readers to approach both fiction and society “with critical detachment.”Natalie Neill also emphasizes the value of critical distance in Northanger, arguing that this is the skill Catherine needs to learn, and that Austen prompts readers “to assume a detached critical relationship” with her own novel.In different ways, all of these scholars interpret Northanger as a sort of textbook that teaches professional reading. Just as Catherine learns to discriminate, attend to context, and become less emotionally involved, so do attentive readers of Austen, they argue. These techniques allow for a detached, scholarly interest—one that is different from the unconditionally involved interest Parsons models in Wolfenbach, and which I will examine more fully in the final section of this chapter. What these readings of Northanger do, even when they argue for the value of gothic fiction within Austen’s text, is use Austen’s gothic parody to delineate developmental hierarchies of reading and writing and imply that they correspond to mature perception, feeling, and action in everyday life. Within these hierarchies, Parsons’s Wolfenbach and the other Horrid Novels are positioned as immature—aesthetically unrefined, morally indiscriminate, and emotionally inappropriate. Northanger and its criticism thus work together to teach ways of condescending to novels like Wolfenbach, making it likely that readers who come to Parsons’s novel through Austen or Austen scholarship will be aware that in reading one of the Horrid Novels,blueberry box they are reading a work that they ought to think is bad, and that its badness can serve as model for what not to write, read, feel, or do.

For nonprofessional readers, this awareness may result in readings that resist the emotional immersion that Parsons encourages, but it does not necessarily result in cold condemnation. Despite or because of being dismissed and recommended as “horrid” and puerile by Northanger and its criticism, Wolfenbach continues to be read and reviewed by amateur critics. As the first named and first published of the seven Horrid Novels, it is the most reviewed on sites like Goodreads and Amazon, and it is the one most thoroughly covered on blogs. In expressing their responses to the novel, amateur reviewers often adopt an authoritative tone and teach other readers how they ought to feel about the work. This instruction often recommends paying attention to formula more than detail, much like Parsons’s own attention to categories in expressions of emotion. Modern amateur critics, though, like scholars, suggest that every aspect of the novel, emotion included, is equally typical. Parsons’s repetition of sad stories, which seems intended to train her readers to recognize them as emotional conventions and respond appropriately, appears to backfire with modern readers. While a reader who has spent time inhabiting Parsons’s school of affliction and studying the beliefs that support its literary-emotional conventions could have the conditioning necessary to feel with the formula of these repetitious emotional displays, recent reviewers treat them as just another genre cliché. One blogger asks, “How do you know when you’re reading a gothic novel? Characters faint then weep, and then faint some more and then someone comes to their rescue.”Another writes, “The story speeds along, packed with tales of woe, heroes and villains, titles and society, swooning and fainting, weeping and wailing, swooning and fainting.”On Goodreads, a reviewer writes that “Eliza Parsons included damsels in distress, swooning and fainting, weeping and wailing, incest, murder, kidnapping, a haunted castle . . .” In keeping with the codified, disdainful language used for literary clichés, these reviews echo each other when mentioning emotional conventions; this is especially striking in the repetition of “swooning and fainting, weeping and wailing” in the last two examples, as if reviewers are actually supplying each other with the exact language with which to critique the conventions of the novel.

This echoing recalls the way Matilda learns to speak in the aphorisms of her mentors by the end of the story, but if these reviews provide education in the recognition of and response to emotional categories, it is far from the kind that Matilda learns. In each of these examples, the reviewer includes the emotional conventions alongside genre conventions of plot, character, or setting . It seems as if, due to different literary-emotional training, today’s readers see the cues for feeling as simply conventions. The first review of Wolfenbach, from 1794, provides a good comparison to this, as the writer inserts an emotional response in the midst of a listof plot events: “he [Matilda’s uncle] also discloses the measure of his crimes; restores her [Matilda] to her mother, from whom he had likewise taken her when an infant and burying himself in a convent . . .” Though this writer also places emotion alongside other literary features, he phrases the plot in specifics while making the emotion general and even minimizing it in parenthesis. This eighteenth-century professional critic suggests through his sentence structure that his own emotional response has a subordinate place in a review, as a sort of embarrassed aside that interrupts the real work of relating the situations from the novel in detail. He carefully elucidates the circumstances in the novel while minimizing a personal emotional response to those circumstances, implying, like Parsons, that situations are more relevant than personal feelings to well-trained audiences of stories of suffering. In contrast, the modern amateur critics generalize about every aspect of the novel, making both situations and emotions unimportant. The earlier review and the later ones judge Wolfenbach as inferior to good novels, but they model different forms of judgment. Parsons’s contemporary makes distinctions, detailing the plot, separating plot summary from emotional response, and characterizing the work as not “one of the first order, or even of the second” but possessing “sufficient interest” to command some degree of engagement.The online reviewers quoted here have inherited the critical tendency to flatten distinctions among formulaic gothic works that began in the decade of Wolfenbach’s publication and was likely enhanced by the pedagogical dimensions of Northanger and its scholarship.

Going beyond the modeling of conventional disdain that the amateur critics above demonstrate, some of the online commentary about Wolfenbach is explicitly emotionally didactic, but rather than employing categorical thinking to achieve sympathy, as Parsons does, this emotional education uses categorical thinking to achieve amusement. This trend is especially obvious in blog entries and comments from the Gothic Lit Classics Circuit Tour of October 2011 . The tutor of this particular read, in addition to educating her “student” on eighteenth-century culture and gothic conventions, also educates the reader on how to enjoy the novel, by advising her student to count the number of times a female character bursts into tears. They both continue to mockingly quote these dramatic passages.It is worth noting that ironically tracking weeping in novels is not a twentieth or twenty-first-century phenomenon. Henry Morley famously edited an edition of Mackenzie’s The Man of Feeling in 1886 that included an “Index to Tears” for the amusement of a Victorian audience distant from the conventions of eighteenth-century sensibility.Many of these amateur critics, however,blueberry package go further than offering humorous commentary on the novel by instructing their community of readers on how to feel. The kind of emotional curriculum shown in the “so bad it’s good” blog post and comments or the LibraryThing exchange, while teaching to feel against the formula of Wolfenbach, is still feeling—but substituting pleasure for pity. Scholars generally do not express pleasure in what they present as failed writing—pleasure for scholars is most expressible in terms of interest, as I will discuss in the following section. As scholars, we value critical emotion—emotion that requires labor, or specialized taste, or that resists what appear to be the intended emotions of the work—more than feeling with the formula, which, when it lines up with the intended emotions, appears passive, naïve, and boring. Nonprofessional reviewers are not sanctioned by their community for enjoying formulaic fiction, but when a novel like Wolfenbach comes prejudged as “horrid” and unsophisticated, they may be less inclined to express simple enjoyment, even as they are also disinclined to aloofly dismiss it. By modeling for other readers how to enjoy what they call bad writing, they teach a way of valuing formula that incorporates critical judgment, even if it favors categorical responses over careful discernment—which, as I will demonstrate in the next section, is not entirely without its own categorical tendencies. Even if scholars believe that The Castle of Wolfenbach possesses no engaging qualities in and of itself, they may express their engagement with the project of literary recovery that it allows. Deidre Lynch, writing about scholarly affects, describes the “pleasures of laboriously recovered, recondite information” that literary historicism allows.

Devendra Varma mentions this feeling in his introduction to The Northanger Set of Jane Austen’s Horrid Novels in 1968, when Wolfenbach had been out of print for more than a century. He writes, “[T]his piece of research has brought to me many moments of delight and given me the thrill and pleasure of literary archaeology.”At that time, Varma was one of only a handful of modern scholars to assess Wolfenbach and the other Horrid Novels, and the opportunity he had to help bring the books back into print and inform modern readers about the authors and works that played a large role in Jane Austen’s novel is one that few scholars today ever get to experience the like of. Diane Long Hoeveler was in a comparable position in 2006 when she wrote the introduction to the first major edition of Wolfenbach since Varma’s, and she makes a similar statement, though a less fervent one. She writes, “In a decade that saw the fall of a King and the rise of Emperor Napoleon, this novel spoke about the attraction and allure of France and the French people to the British bourgeois imaginary. Unpacking that allure even today is an engaging exercise in literary archaeology.”The pleasures of literary archaeology are difficult to share with others— Varma’s is for him alone, and Hoeveler’s is for readers with a strong background in history. Thus, scholars expressing the joys of recovery work with Wolfenbach are not likely to transmit their enjoyment to others, but they may transmit their less positive attitudes. The attitudes scholars express are important because the authoritative nature of scholarly discourse could influence other readers, instructing those readers on how a text ought to be approached. Rita Felski writes that “the authority of critique is often conveyed implicitly . . . via inflexions of manner and mood, timbre and tone.”A critical mood spreads because, she explains, “the student learns by imitating the teacher, adopting similar techniques of reading and reasoning, learning to emulate a style of thought,” which includes attending to “displayed dispositions” and “emulation of both tone and technique.”This emulation becomes apparent in the way several amateur reviewers paraphrase Hoeveler’s introduction to the modern edition, repeating her assertion that the novel is important as a window into history. While they endorse her interest, they also copy her condescension. Hoeveler shows disdain for Parsons’s tendency to seem “blatantlynationalistic in her celebration of British superiority” and writes that in her “bourgeois agenda” Parsons “goes so far as to say in this novel that middle-class people are more attractive than aristocrats because they do not stay out late at night partying, drinking, and losing their looks.”Similarly, a reviewer on Goodreads expresses patronizing amusement when noting, “The book also has a very strong patriotic bent and wouldn’t you know it, England is the best country in the world” and “If [ . . . ] the rich gave up late nights and gambling, there would be nothing to improve.”Of course, readers possess agency in how they respond to any novel, but we cannot discount the fact that scholarly experts act as teachers, especially in their introductions to novels. When these teachers introduce the uninitiated to a novel like Wolfenbach, they draw readers’ attention to certain features, and they encourage certain kinds of responses.

However, as Felski observes, “Much work in the New Historicist vein . . . leans toward diagnosis rather than dialogue.”Many of these studies imply that to read gothic novels without ascribing their features to specific historical forces is to read them badly. Felski explains that New Historicism was an important corrective to “grand narratives”— the oversimplification of history in service of a particular perspective—but she argues that it has over corrected by keeping texts bounded in their historical periods.She writes that “standard ways of thinking about historical context are unable to explain how works of art move across time.In this dissertation I am interested in the manner in which the work of novelists and critics reaches us across time—for example, in chapter 1, how present-day fans of Jane Austen learn about The Castle of Wolfenbach from her list of “horrid novels,” or in chapter 2, how mid-twentieth-century paperback covers of The Monk present Lewis’s novel to readers as a sexy, modern thriller, and how these mediations facilitate different responses to the texts. In my inclusion of these sorts of mediations, I am drawing on Bruno Latour’s theories to some extent. Latour emphasizes the importance of the human and nonhuman mediators that enable certain ways of knowing and interacting.In order to account for particular features in the current reception of the novels I write about in my dissertation, I will attempt to trace different lines of influence. These lines are not unbroken, as my reconstruction of them relies on the work that historicist scholars have done and the texts that digitizers have preserved. I would argue, though, that these selections themselves are significant,pe grow bag as they help to demonstrate the priorities that have shaped our critical inheritance.

In gauging the importance of different critical voices, I have paid special attention to which early responses appear again and again in bibliographies and introductions and which quotations from those critical sources have served as a reference point for recent scholars. Often, scholars differ in the ways they contextualize the same response, as I examine in chapter 4 when discussing the selection and interpretation of Thomas Gray’s letter to Walpole about The Castle of Otranto, in which he writes that it “makes some of us cry a little.”I note that critics use Gray’s words variously as evidence that Walpole fooled his contemporaries into reading sentimentally, that earlier readers found the novel effective in a way that is no longer possible, that Walpole’s inner circle understood that he meant the work to be satirical, or that Gray was simply being polite. I, in turn, read the frequent use of this quotation as evidence that critics have attempted for years to resolve the emotional indeterminacy of Otranto by using Gray’s letter as a window into Walpole’s intentions. A trans-historical approach is especially relevant when considering how minute details of older works affect readers today. Wai Chee Dimock, in “A Theory of Resonance,” calls for “diachronic” rather than synchronic historicism. She argues that “the text, as a diachronic object, yields its words differently across time, authorizing contrary readings across the ages and encouraging a kind of semantic democracy.”Attending to “the changes in the webs of meaning surrounding individual words,” she writes that “[t]hese semantic webs, broadening, contracting, acquiring new overtones and inflections, bear witness to the advent and retreat of social norms.”For instance, in chapter 3, I examine changing historical associations of the word sweet that could inflect different readings of Radcliffe’s portrayals of femininity in Udolpho. I also use this technique when thinking about changes in the grammar of emotional expression in chapter 1 and the punctuation of dialogue in chapter 4.

Even the smallest details of fictional texts can accrue affective associations that transform over time and alter readers’ engagements with novels. Historicizing these associations allows us to see the specific ways these texts are in flux and helps some of the historical variability of responses. That is to say, a professional critic immersed in the age of sensibility reads a gothic novel differently from a mid-twentieth-century magazine critic with a taste for minimalism, a twenty-first-century scholar comparing electronic reproductions different editions of a gothic novel in an office reads differently than a college student on break curled up with a Kindle, etc. Though I do not conduct experiments or surveys to determine how people read, I approach reception with an awareness of how different circumstances structure reading and how public literary judgments can implicitly or explicitly recommend ways of feeling about novels. A crucial goal of my approach to reception is to make legible the emotional conventions of recent scholars and amateur critics. As Lynch notes, the popular stereotype of scholars as emotionless is false, and this stereotype often opposes scholars to amateur readers “not yet subjected to the affective deformation that supposedly comes with formal education.”In Lynch’s view, we are all formed by historical constructions of literature as an object of love, but as I discussed above, the modern history of reading literature has also constructed nonprofessional readers as a threat to themselves or to the broader culture. Willis describes the historical construction of reading as involving “the division between ‘good’ and ‘bad’ forms of reading, and the way these map on to gender, class and race”—ideas that she argues “continue to structure both critical and popular notions of audience and reception down to the present day.”The discourses of good and bad, tasteful and tasteless do not explicitly invoke gender, class, and race today, as they did in the eighteenth, nineteenth, and early twentieth centuries, but the way they deal with emotion can be a coded way of reinforcing the construction of certain groups of readers as bad. Many of the twentieth-century writers who helped establish scholarly approaches to criticism employ language or methods that place emotion within a hierarchy.

Lynne Pearce writes that the wording of W. K. Wimsatt and C. M. Beardsley’s mid-twentieth-century concerns that the “affective fallacy” “ends up in impressionism and relativism” allows us to “speculate that their real anxiety is with the fact that such indeterminacy signals a reader’s lack of control over both the text and the reading process; and such lack of control is, in modern Western culture, a mark of both the feminine and the uneducated, working class.”Though the formative work of reception theorists like Wolfgang Iser, Stanley Fish, and Roland Barthes allows more importance to the role of the reader than previous critics did, Pearce writes that their emphasis on interpretation of the text rather than engagement with it still draws on “the gendering of knowledge” and “the class politics which has made ‘the ability to interpret’ the sign of bourgeois status.”By focusing on the emotional dimensions of responses from different kinds of critics, including scholarly ones, I am contributing to efforts to explore and validate previously devalued modes of reading and responding to literature. Scholars have become more interested in the differences and similarities of lay and professional readers, as John Guillory terms them. Writing in 2000,growing bags he considers their forms of reading to be two entirely different practices. His description of professional reading is similar to Felski’s, as one that is “vigilant,” “stands back from,” and is “wary” of pleasure.For Guillory, though, this is preferable to lay reading, which he describes as solitary consumption for enjoyment or diversion alone.Felski, however, argues that “in spite of their differences,” academic methods of reading and common methods of reading “share certain affective and cognitive parameters.”In response to fears about the current status of reading outside academia and concerns about paranoid reading inside academia, Magnus Persson argues that “we should become more curious about our own and others’ impassioned reading.”To act on this curiosity, Persson recommends studying the way more people read with more entirety. He writes, “Exploring the diversity of reading practices entails several perspectives: investigating reading practices available outside the educational system; viewing individual reading practices as a multitude of complex entities of which text interpretation is only one; and paying closer attention to the social dimensions of reading.”These are three of the objectives I attempt to achieve in my dissertation by focusing on the responses of amateur critics, the emotional qualities of lay and professional responses, and the ways critics influence each other, including in online forums. The responses I use in my dissertation include periodical reviews contemporary to the novels I analyze, criticism from nineteenth-century essays or studies of literature, private correspondence, twentieth-century magazine articles, conference papers, dissertations, and literature blogs, but I devote the most attention to scholarly monographs and articles of the last four decades and online review criticism.

In chapters, I often separate the responses of scholars and amateur critics not only because I expect my scholarly audience will view these groups differently but also because of the notable distinctions between their respective review conventions, subject matter, or tone that allow for insights into their respective affective approaches to the texts. Within those groupings, when scholars or amateurs write different kinds of emotional responses, I further subdivide them, and when the responses of scholars and amateurs are similar in some ways, I set them alongside each other. The responses I have chosen are ones that represent trends I have noticed in reception, ones that have been especially influential, and ones that offer particularly rich phrasing for analysis. Though I have been very selective in the amateur responses I quote, I attempted to read as many online reviews as possible for each novel. Most of the online reviews I use come from Amazon, Goodreads, and LibraryThing. Megan Milota, who has contributed to establishing methodology for the quantitative study of reader response in online book reviews, has judged these three sites to be similar enough to use without differentiating among them because, in addition to their common use of a five-star rating system, they all allow “the chance to participate in the literary field, a sense of community, an opportunity to present oneself, and finally, the chance to gain or display status” .All three sites aggregate ratings and reviews for particular books across most editions but have separate ratings and reviews for a handful of editions, such as ones where a novel is included in a collection. There are differences among the platforms, though: Amazon is an online megastore, whereas Goodreads and LibraryThing are social networking sites that let users record the books they’ve read or want to read and connect with other readers. Amazon allows ratings only with reviews, Goodreads allows ratings without reviews, and Library Thing allows ratings with reviews but also tracks “mentions” and “conversations” in its forum postings. Goodreads is the most active site for amateur criticism, with its main listing for Udolpho featuring 11,300 ratings and 839 reviews as of March 7, 2018, whereas Amazon has 139 aggregated ratings and reviews and LibraryThing has only 45 ratings and reviews with 378 mentions and 2 conversations. Milota observes that amateur reviews tend to use review conventions, though more loosely than professional reviews, writing that “while there may have been a degree of flexibility in the format, register, and formality of reviews, they still followed an established template of what was deemed appropriate by the broader community of readers.”These standards are maintained through features that allow users to rate or comment on each other’s reviews. Milota is one of a growing number of scholars who use empirical methods to study reception, part of a broader turn toward the sciences in studying literature. Though I initially tried to analyze amateur reviews quantitatively, the correlations I found between numbers of stars and positive or negative assessments were minimal, and more often, amateur critics used mixed assessments and emotional language that demanded interpretation. Like the affectively inflected assessments of recent scholars and critics from older historical periods, these reviews present complex engagements with the text, the broader critical culture, and narratives of history that need to be more closely analyzed and considered in conjunction with the emotional features of the text and its historical and present-day positioning. In her article about the British reception of a novel about Caribbean immigrants, Anouk Lang analyzes the responses of nonprofessional readers to an online survey and in transcriptions of recorded book group discussions, selecting and interpreting these responses in a qualitative way that is more similar to my approach.

Two-thirds of the octoploid strawberry germplasm accessions screened for resistance to Fusarium wilt race 1 in the present study had disease symptom ratings in the resistant range , where ̄y is the estimated marginal mean among replicates and years, y = 1 plants were symptomless, and y = 2 plants were nearly symptomless . The severity of symptoms increased as ̄y increased on our ordinal scale . The race 1 resistance phenotypes observed among resistant and susceptible checks in the present study were consistent with those previously reported. The repeatability of race 1 resistance phenotypes among clonal replicates of resistant and susceptible checks was 0.81. One-fourth of the individuals screened in the present study were symptomless, classified as highly resistant , and appeared to be immune to AMP132 infections . This confirmed our suspicion that resistance to race 1 was widespread in natural and domesticated populations of octoploid strawberry. We suspected this because the only ‘non-California’ individuals screened in our previous study were highly resistant to AMP132 infection, had non-FW1 SNP marker haplotypes, and were presumed to carry novel R-genes . Moreover, the individuals screened in the present study were more diverse than those previously screened from the California population . Slightly more than half of the F. chiloensis and F. virginiana ecotypes screened for resistance to race 1 in the present study were classified as resistant . We did not observe geographic or phylogenetic trends—highly resistant ecotypes were found throughout the natural geographic ranges of both species . Eleven out of 62 F. chiloensis and 12 out of 40 F. virginiana ecotypes were classified as highly resistant . Moreover,plastic square flower bucket highly resistant ecotypes were identified for each of the seven subspecies of F. chiloensis and F. virginiana apart from one ecotype of F. virginiana subsp. platypetala, which was nevertheless classified as resistant .

We did not screen ecotypes of F. chiloensis subsp. sandwichensis, the subspecies found in Hawaii , because none were available when our study was undertaken. Approximately two-thirds of the F. × ananassa individuals screened in the present study were resistant to AMP132 infection . Of these, 20 originated in the California population and were either known or predicted to carry FW1 from SNP marker haplotypes . Other than Wiltguard , the other 141 are heirloom and historically important F. × ananassa individuals originating in North America, Europe, and Japan between 1880 and 1987 . We extracted the pedigree records for Wiltguard and the 141 non-CA individuals from the database described by Pincot et al. to show that the genetic relationships among most of these individuals and their ascendants were complex and intertwined . Of 603 individuals in the pedigree network, 101 were identified to be founders and 180 were phenotyped for resistance to Fusarium wilt race 1 . Twentyone individuals either lacked pedigree records or only had a single-generation of pedigree records with one or both parents known . The orphans are identified in the pedigree database but not shown in Fig. 2, which displays interconnections among the other 120 non-UC individuals and Wiltguard . The pedigree network diagram and database show that the alleles found in these resistant individuals have flowed through several common ancestors . Although the presence of multiple R-genes cannot be ruled out—partly because some of the individuals are orphans or extinct and phenotypes are only known for a subset of the ascendants of resistant individuals—there is a high probability that the number of unique alleles is small and that many of the R-alleles found in cultivars worldwide are identical-by-descent .

Our studies targeted two heirloom cultivars that share three resistant common ancestors , had unique haplotypes for SNPs in LD with FW1, were genetically distant from one another and individuals in the California population, and were predicted to have a high probability of carrying novel R-genes . The pedigrees for these cultivars are shown in Online Resource 7 .Selection of individuals for constructing a host differential panel was informed by insights gained from screening public germplasm collections for resistance to race 1 and insights gained from population structure analyses in strawberry . The host differential panel was assembled to maximize the probability of differentiating races and identifying sources of resistance to different Fof isolates . The phenotypes for resistance to AMP132, MAFF727510, and four other Fof isolates were previously reported for 25 octoploid individuals on the host differential panel: one F. chiloensis ecotype, one F. viriginiana ecotype, and 23 F. × ananassa individuals . MAFF727510 is an Fof race 2 isolate found in Japan . To broaden insights into the frequency and distribution of race 2 resistance sources, the phenotypes for resistance to MAFF727510 are reported here for an additional 116 individuals: 10 F. chiloensis ecotypes, 16 F. viriginiana ecotypes, and 93 F. × ananassa individuals . The latter included cultivars and other individuals selected to broadly sample allelic diversity in California and non-California populations worldwide. Similarly, the ecotypes were selected to sample allelic diversity across the natural ranges of F. chiloensis and F. viriginiana. The race 1 and 2 resistance phenotypes observed in these studies were highly repeatable: estimates of broad-sense heritability were ̂H2 = 0.98 for resistance to AMP132 and ̂H2 = 0.91 for resistance to MAFF727510 . Forty-one individuals on the host differential panel were classified as resistant to race 2 . Thirty-four of these individuals were symptomless and classified as highly resistant , and 34 of the race 2 resistant accessions were resistant to race 1 . Of the 78 F. × ananassa individuals from the California population, only four were resistant to MAFF727510. Conversely, of the 38 F. × ananassa individuals from the non-California population, 21 were resistant to MAFF727510. Slightly more than half of the F. chiloensis and F. virginiana ecotypes were resistant to race 2 , which was comparable to the frequency observed for race 1 resistance .

Three individuals on the host differential panel were resistant to every Fof isolate tested from California, Japan, Australia, and Spain . According to historical breeding records , Royce S. Bringhurst developed 61S016P006 , an S1 descendant of 43C001P036, by selecting for resistance to Verticillium wilt in Davis, California, nearly a half century before Fusarium wilt was discovered in California . Using the host differential panel as the study population and 50K Axiom SNP array genotypes, we searched the genome for associations between SNP marker loci and race 2 resistance phenotypes. Statistically significant GWAS signals for loci affecting resistance to race 2 were not observed . We repeated this analysis with the host differential panel using race 1 resistance phenotypes and reproduced the strong GWAS signal associated with the segregation of FW1 in the California population . The absence of a significant GWAS signal for race 2 resistance has several possible explanations. First, resistance to race 2 might not be governed by gene-for-gene resistance. Second, resistance to race 2 could be governed by gene-for-gene resistance but undetectable in a highly diverse population where multiple alleles and loci are segregating and the resistant alleles are uncommon. Those alleles, however,plastic plant pots could almost certainly be uncovered and identified by forward genetic analyses of segregating populations developed from crosses between resistant and susceptible parents, as described below for the race 1 resistance loci identified in the present study. Third, the sample size may have been insufficient to detect the presence of gene-for-gene resistance to race 2. This seems unlikely because R-genes have large effects, and we have consistently observed strong GWAS signals for FW1 in small samples of California population individuals, including the host differential panel .The FW1 locus was previously identified and physically mapped using diploid genome-informed GWAS in a closed UCD or ’California’ population of 565 individuals genotyped with a SNP array populated with diploid genome-anchored SNPs . To revisit our original analyses using octoploid genome-informed GWAS, 356 of these individuals were genotyped with either the 50K or 850K Axiom SNP arrays . This substantially increased the density and uniformity of SNPs in the FW1 haploblock and facilitated a search for genes with plant defense annotations because the SNP markers on both arrays were physically anchored in silico to octoploid reference genomes developed since the original study was reported . The physical positions of FW1-associated SNPs in the ’Camarosa’ and ‘Royal Royce’ genomes are shown in Online Resource 8. GWAS and de novo genetic mapping pinpointed the location of the FW1 locus to a near-telomeric haploblock on the upper arm of chromosome 2B spanning approximately 3.3 Mb . The 3.3 Mb haploblock was populated with 1,725 SNP markers from the 50K and 850K Axiom SNP arrays, of which 460 were signifcantly associated with race 1 resistance phenotypes . SNP markers with the strongest GWAS signals on chromosome 2B were AX-184226354 and AX-184176344 . KASP markers were developed for SNPs predicted by GWAS to be in LD with FW1 .

These were genotyped in the Fronteras and Portola S1 populations, shown to be tightly linked to the FW1 locus on chromosome 2B, and estimated to predict race 1 resistance phenotypes with 98.7 to 98.8% accuracy in the Fronteras and Portola S1 populations and 95.2 to 97.6% accuracy in the California population . Even though GWAS signals were observed between resistance phenotypes and SNP markers predicted to reside on homoeologous chromosomes, 84% of the 50K and 80% of the 850K Axiom array SNP markers with statistically significant GWAS signals were predicted in silico to reside on chromosome 2B proximal to the previously genetically mapped FW1 locus . Nevertheless, the strongest GWAS signals were observed for SNP markers that had previously been physically assigned in silico to chromosome 2D: AX-89872358 , AX-184098127 , and AX-184513679 on the 50K SNP array and AX-184055143 on the 850K SNP array . The GWAS signals observed on chromosomes other than 2B were almost certainly caused by incorrect in silico physical assignments of Axiom SNP array probe DNA sequences to positions in octoploid reference genomes . This conclusion was supported by several observations and analyses. First, QTL mapping in the Fronteras and Portola S1 populations only uncovered statistically significant signals for race 1 resistance on chromosome 2B . Second, GWAS was repeated for race 1 resistance phenotypes observed in the California population by fitting AX-184226354 as a fixed effect, then searching the genome for significant GWAS signals—AX-184226354 was the SNP marker on chromosome 2B that was most strongly associated with the FW1 locus . GWAS with AX-184226354 incorporated as a fixed effect eliminated signals on other chromosomes, in addition to eliminating signals for other tightly linked SNPs on chromosome 2B . Hence, fitting and testing multilocus genetic models with SNPs selected from single locus genome-wide searches is a powerful approach for resolving physical address assignment errors, a particular problem in polyploids, and generating more accurate estimates of genetic parameters . A certain percentage of erroneous GWAS signals are expected in octoploid strawberry because a small percentage of the short 71-nt DNA probe sequences for Axiom array SNP markers are incorrectly assigned in silico to physical positions in the reference genome, e.g., Hardigan et al. estimated that approximately 74% of QC-passing 850K Axiom SNP array probes could be assigned to the correct homoeolog in the ‘Camarosa’ genome. That percentage was virtually identical to the percentage of Axiom array SNPs with significant GWAS signals on chromosomes other than 2B in our analyses . Genetic mapping of Axiom array SNPs in the present and previous studies have shown that the percentage of Axiom array SNPs with physical positions that were incorrectly assigned in silico are found on homoeologous chromosomes . This GWAS complication is bound to arise in strawberry and other polyploid and paleopolypoid species with complex repetitive DNA landscapes, especially outbred species with whole genome duplications where homoeologous DNA variation complicates the physical assignment of short DNA sequences to subgenomes . The assignment of highly accurate long-read DNA sequences, by contrast, is straightforward . The octoploid genome-informed GWAS analyses described here were initiated in 2017 immediately after we assembled the ‘Camarosa’ reference genome . We have since expanded our understanding of the complexity of the octoploid genome, built superior haplotype-phased genome assemblies , and unequivocally shown that homologous and homoeologous DNA variation can be differentiated nearly genome-wide in strawberry .