Dollar spot, caused by the pathogen S. homoeocarpa (F.T. Bennett), is a common disease that infects a wide variety of turfgrasses all over the world. Yet it is significant problem on golf course putting greens and fairways consisting of creeping bentgrass (Agrostis stolonifera L.) and annual bluegrass (Poa annua L.). It is active in a wide variety of environmental conditions ranging from 16-30˚C but favors warm, humid days, followed by cool nights. Sclerotinia homoeocarpa overwinters as dormant mycelium in dead plant tissue. In the spring, germinating mycelia begin to infect leaf blades causing foliar lesions, which then spread via mycelium by means of wind, rain, animals and equipment. While there are a number of cultural practices that can reduce disease severity, frequent fungicide applications are required to maintain acceptable playing conditions on a golf course. The repeated use of fungicides with the same mode of action has led to the development of fungicide resistance of S. homoeocarpa to certain fungicide classes. Most notably, demethylase inhibitor (DMI) fungicides have been found to have varying levels of inefficacy against S. homoeocarpa across North America. The cause for reduced efficacy is suspected to the shifted sensitivity levels of many S. homoeocarpa populations, which are resulted from repeated use of the DMI fungicide. Recently, “early-spring fungicide applications” targeting to reduce initial inoculum density of dollar spot have gained popularity in an attempt to reduce dollar spot severity. In addition, preventative fungicide applications (from late October through mid-November) containing DMI fungicides have been traditionally practiced to target snow molds (caused by Microdochium nivale, Typhula spp.) in the northeastern United States. To date, there is not a clear understanding as to what effect, if any, these applications have on S. homoeocarpa DMI sensitivity or residual dollar spot control the following year. Traditional preventative snow mold applications were also investigated on the effect of S. homoeocarpa DMI sensitivity and early-season dollar spot control. The objective of this study was to investigate the effect of early-spring dollar spot application and late-fall snow mold application on S. homoeocarpa population with a bimodal distribution of DMI sensitive and insensitive isolates.

Dollar spot is one of the most common diseases of warm- and cool-season turfgrass stands and is especially devastating on creeping bentgrass (Agrostis stolonifera L.). The fungus Sclerotinia homoeocarpa degrades the foliage by creating silver, dollar-sized depressions of dead and bleached turf. Frequent fungicide applications and cultural management strategies are required throughout the growing season to prevent or reduce severity of this disease. Previous research has demonstrated that ferrous sulfate applied at 48.8 kg ha-1 suppresses dollar spot epidemics without traditional fungicides. In vitro studies showed 100 to 1,000 mg kg-1 of ferrous sulfate directly suppressed S. homoeocarpa growth of an isolate collected from an established, intensively-maintained creeping bentgrass putting green. Genetic diversity of S. homoeocarpa segregates isolates into two groups; strains generally associated with warm-season and cool-season grasses. It is unknown whether isolates of each group react similarly in the presence of ferrous sulfate. Our research explored use rates of ferrous sulfate required to suppress 50% of dollar spot in the field and in vitro. Ferrous sulfate (heptahydrate 20% Fe, Valudor Products Inc) rates in field trials included 0, 4.88, 24.4, 48.8, and 97.6 kg ha-1. Our results indicate a hyperbolic relationship between ferrous sulfate rate and dollar spot reduction. Using this model, 26.4 kg ha-1 reduced dollar spot incidence by 50%. We concluded that ferrous sulfate suppresses 50% of S. homoeocarpa mycelial growth at between 480 and 720 mg L-1 concentration in 0.25 strength potato dextrose agar in vitro, and fungitoxic activity of ferrous sulfate was dependent primarily on historical fungicide inputs at isolate collection sites. The use of ferrous sulfate may supplement traditional fungicide use. Chlorothalonil is the most common fungicide used to suppress dollar spot in turfgrass. Annual site-use limitations of chlorothalonil often prevent turf managers from achieving acceptable dollar spot control throughout the season. It is not known how ferrous sulfate may contribute to a successful chlorothalonil fungicide program. Therefore, we examined whether ferrous sulfate can be used to minimize chlorothalonil requirements through reducing active ingredient concentrations and extending the longevity, while still maintaining acceptable disease control. Chlorothalonil treatments were applied at 0, 2.28, 4.57, 6.86, and 9.16 kg ai ha-1 (Daconil WeatherStik) across plots treated with and without 48.8 kg ha-1 ferrous sulfate applied bi-weekly. Ferrous sulfate reduced the chlorothalonil rates necessary for 80% disease reduction by 36 to 51% across all trials. Additional studies showed that ferrous sulfate applied with chlorothalonil increased duration of disease control by five days and eliminated two seasonal treatments. Our research expands the guidelines for practical ferrous sulfate usage for dollar spot suppression by elucidating the rate-to-disease relationship and providing best management practices involving admixtures with chlorothalonil. / Master of Science / Turfgrass systems offer many environmental and recreational benefits. Managing turfgrass stands that are free of damaging turf pests is essential to providing aesthetically pleasing lawns, golf courses, and sports fields. Creeping bentgrass is one of the most common turfgrass types found on golf course putting greens but is also used on golf course fairways and tee boxes. There are many diseases that can be found on creeping bentgrass when environmental conditions are favorable. Of these diseases, dollar spot is the most common. When dollar spot is present, half-dollar sized spots of bleached turf can be seen. In order to prevent these easily noticeable spots from appearing, fungicide applications are required in a given growing season to prevent the pathogen from infecting. Available fungicides are very effective at providing control but can be very costly. Beyond fungicide use, other research has shown various cultural practices to reduce disease incidence. Previous research has shown that iron sulfate applied to creeping bentgrass can reduce dollar spot epidemics without the use of fungicides. Laboratory studies have shown a similar trend as ferrous sulfate at varying concentrations directly suppressed dollar spot pathogen growth. In both cases, a limited range of ferrous sulfate rates was tested. To obtain more information we explored use rates of ferrous sulfate required to suppress 50% of dollar spot in the field and in vitro. Ferrous sulfate rates in field trials ranged from 0 to 97.6 kg ha⁻¹ . Results from these trials were used to create a hyperbolic regression. Using this model, we were then able to determine that 26.4 kg ha⁻¹ iron sulfate was required to suppress 50% of the dollar spot in the field. For the laboratory studies we concluded that ferrous sulfate suppresses 50% of the dollar spot pathogen mycelial growth between 480 and 720 mg L ⁻¹ iron sulfate concentrated potato dextrose agar. Although there are many different fungicides available for dollar spot control, the active ingredient chlorothalonil is the most common used. Due to the mode of action which chlorothalonil exhibits, it is much less likely that the pathogen causing dollar spot can become resistant. Although resistance is not an issue, governmental annual site-use limitations restrict turf managers from achieving desirable control. The use of iron sulfate in conjunction with chlorothalonil has not been previously studied. Chlorothalonil treatments were applied at a range of labeled use rates across plots treated with and without 48.8 kg ha⁻¹ ferrous sulfate applied bi-weekly. Ferrous sulfate reduced the chlorothalonil rates necessary for disease reduction. If a threshold of 80% is used, up to 50% reduction in chlorothalonil use was observed. Supplemental studies investigating the duration of control achieved by the combination showed an increase of up to 5 days and eliminated the need for two applications across one season. This research fills a huge gap in our knowledge base on the practical use of iron sulfate for dollar spot control.

ABSTRACT Dollar spot is a fungal disease that affects grass, and is caused by the fungus Sclerotinia homoeocarpa. It was discovered for the first time in Scandinavia in 2013. Dollar spot is a very serious disease of grass in the United States and a lot of money is spent to control the fungus. In the present study, dollar spot development in potted turfgrass, caused by four geographically distinct, Scandinavian isolates of S. homoeocarpa, was investigated at three different temperatures; 6 °C, 15 °C and 21 °C. The objective was to identify the lowest temperature of development of disease, and to establish possible differences among the isolates with regard to disease development. The isolates used originated from Denmark, Norway and Sweden. The grass samples that were inoculated with S. homoeocarpa came from two golf courses in Sweden, located in Uppsala and in Gävle, respectively. Turf grass was dug out and placed in pots, and subsequently inoculated with fungal hyphae from cultures of S. homoeocarpa. These pots were individually covered with plastic bags and placed in different climate chambers at set temperatures. At three time points (day 7, 14 and 21), the ratio of area infected grass to total grass-covered area was estimated. After all the results had been analysed, it was concluded that dollar spot had the most severe disease development at 6 °C. Differences in disease development of dollar spot were also observed among the four isolates at different temperatures. There was a link between geographical origin of an isolate and its optimum temperature for disease development, which may suggest that the isolates can have adapted to the environment in which they have established themselves. S. homoeocarpa is apparently able to proliferate at low temperatures. The present study demonstrates it should be able to infect grass and establish in turf grass even in a colder climate, and thus it is very likely that dollar spot has gained a foothold in Scandinavia, and that it is here to stay. With climate change taking place, i.e. increasing temperatures and precipitation, which in turn results in elevated humidity, the disease will be able to establish and spread yet more, because the fungus thrives in humid conditions. If the climate zones will continue to move northward in the Nordic countries, it will increase the potential for continued establishment and spread of dollar spot. / SAMMANFATTNING Dollar spot är en svampsjukdom som drabbar gräs och som orsakas av svampen Sclerotinia homoeocarpa. Under 2013 upptäcktes dollar spot för första gången i Norden. Dollar spot är en mycket allvarlig grässjukdom i USA och man spenderar massor med pengar för att bekämpa svampen. I denna uppsats har sjukdomsutvecklingen av dollar spot, från fyra geografiskt skilda isolat, undersökts vid olika temperaturer; 6 °C, 15 °C och 21°C. Detta för att hitta den lägsta tillväxttemperaturen hos svampen som orsakar sjukdomen och även för att upptäcka eventuell skillnad mellan de olika isolatens sjukdomsutveckling i gräs. Isolaten som använts i experimenten kom ifrån Danmark, Norge och Sverige. Gräsproverna som svampen S. homoeocarpa odlades på kom ifrån två golfbanor, från Uppsala respektive Gävle. Krukor med gräs inokulerades med S. homoeocarpa och dessa krukor placerades i klimatskåp som höll olika temperatur. Vid tre tidpunkter (dag 7, 14 och 21) lästes den infekterade gräsarean av. Efter att alla resultat hade sammanställts visade det sig att dollar spot hade den snabbaste och mest aggressiva sjukdomsutvecklingen vid 6 °C. Det fanns även skillnader i sjukdomsutveckling av dollar spot mellan de fyra isolaten vid olika temperaturer. Det fanns en viss koppling mellan geografisk härkomst och optimumtemperatur för sjukdomsutveckling hos isolaten, vilket kan tyda på att de kan ha anpassat sig till den omgivning som de etablerat sig i. Med de låga tillväxttemperaturer som svampen S. homoeocarpa klarar för infektion och etablering i grönytor är det mycket troligt att sjukdomen dollar spot fått fäste i Norden, och kommit för att stanna. Med de klimatförändringar som sker, dvs. ökade temperaturer och ökad nederbörd, som i sin tur medför förhöjd luftfuktighet, kommer sjukdomen att kunna breda ut sig ännu mer, eftersom svampen trivs vid hög luftfuktighet. Flyttas klimatzonerna högre upp i Norden kommer det att bidra till att utbredningen av dollar spot ökar.

The effect of potassium silicate fortified with cow manure compost tea and seaweed extract on dollar spot (Sclerotinia homoeocarpa) was studied in creeping bentgrass (Agrostis stolonifera L.) and perennial ryegrass (Lolium perene L.). Application of potassium silicate fortified compost tea increased the silicon content in bentgrass and ryegrass at 5 days after treatment, but there was only a minimal effect on silicon content in the grass at 15 days after treatment. However, the fortified treatments did not affect the susceptibility of grass to dollar spot disease in the greenhouse experiments. Fortified treatments did not have a significant effect on the chlorophyll content, total phenol content and phenylalanine ammonia lyase activity in creeping bentgrass. Similarly, the treatment did not affect area of the lesion and mycelial growth of the fungus. Although the synergistic effect of silica fortification was not investigated in detail, preliminary results of the study indicate potassium silicate fortified with compost tea or seaweed extract was not different from that of non-fortified treatments in reducing dollar spot disease in the greenhouse trial.

Soil application of a known activator of Systemic Acquired Resistance (SAR), benzo(1,2,3)thiadiazole-7-carbothioic acid-S-methyl ester (BTH), and Harmonizer, a polychlorinated copper (II) phthalocyanine pigment, reduced severity of Colletotrichum orbiculare in Nicotiana benthamiana by 99% and 38%, respectively. BTH induced expression of nine SAR/progammed cell death-related genes and primed expression of two Induced Systemic Resistance (ISR)-related genes, while Harmonizer induced expression of only one SAR-related gene. Soil application of Harmonizer also reduced severity of Sclerotinia homoeocarpa in Agrostis stolonifera up to 39%, whereas BTH was ineffective. Next generation sequencing identified over 1000 genes in A. stolonifera with two-fold or higher increased expression following Harmonizer treatment relative to a water control, and induced expression of three defense-related genes was confirmed by relative RT-PCR. These results demonstrate that Harmonizer can activate systemic resistance in a dicot and a monocot, but changes in expression of genes indicated that it differed from BTH-activated SAR. / Petro-Canada, Natural Sciences and Engineering Research Council of Canada, Ontario Turfgrass Research Foundation

Creeping bentgrass (Agrostis stolonifera L.) is commonly used on golf course greens and fairways in cool-humid regions but is plagued by numerous fungal diseases, one of which is dollar spot disease (Sclerotinia homoeocarpa F. T. Bennett). Dollar spot occurs frequently throughout the growing season requiring biweekly fungicide applications for complete control. The objective of this study was to investigate methods of reducing the number of fungicide applications needed to maintain dollar spot at acceptable levels through dew removal and potential mechanisms of resistance in bentgrass. In the first study, a combination of mowing three times a week and dragging by hose the remaining four days to remove dew was used in an attempt to reduce disease severity. The main effect of this combination treatment was not significant (p>0.05) and did not reduce the number of fungicide applications compared to normal mowing three times a week. However, dollar spot was managed curatively with 20-80% fewer applications compared to a normal preventative fungicide program. In the second experiment, two experimental germplasms with varying disease resistance were tested for the possible production of antifungal compounds known as phytoanticipins. Preliminary results indicate the resistant line may contain compounds not present in the susceptible line.

Annual bluegrass (Poa annua L.) is the most problematic weed to control in creeping bentgrass (Agrostis stolonifera L.) putting greens.  The objective of this study was to transition a mixed putting green stand of annual bluegrass and creeping bentgrass to a monoculture by using fertilizers and plant growth regulators that selectively inhibit annual bluegrass.  A 25 year old loamy sand rootzone research green, planted with \'Penn-Eagle\' creeping bentgrass, with roughly 45% initial annual bluegrass coverage was utilized.  The biweekly application of ammonium sulfate (4.8 kg ha-1) with treatments of ferrous sulfate at rates of 0, 12.2, 24.4, and 48.8 kg ha-1 and in combination with seaweed extract (12.8 L ha-1) or paclobutrazol (0.37 L ai ha-1 spring and fall; 0.18 L ai ha-1 summer) were applied March to October, 2011 and 2012.  Plots receiving the highest rate of ferrous sulfate resulted in annual bluegrass infestation declines from an early trial amount of 45% to a final average of 20% but also resulted in unacceptable late-summer events of annual bluegrass collapse.  The ferrous sulfate medium rate resulted in a smooth transition from early-trial annual bluegrass infestation of 45% to an end of trial infestation of 20% and had the highest putting green quality.  Previous research has reported that consistent use of paclobutrazol can effectively and safely reduce annual bluegrass infestations.  In this trial annual bluegrass was reduced to 9% infestation after three months of application.  Two unexpected observations from this trial were that ferrous sulfate, applied at medium to high rates, significantly reduced silvery thread moss (Bryum argentum Hedw.) populations and occurrences of dollar spot (Sclerotinia homoeocarpa F. T. Bennett) disease.  Dollar spot control with ferrous sulfate has not previously been reported in the literature, so additional studies were designed to investigate this phenomenon further.  A creeping bentgrass putting green study was conducted to determine if sulfur, iron, or the two combined as ferrous sulfate decreases dollar spot activity.  Ferrous sulfate resulted in the highest turf quality and suppressed S. homoeocarpa infection, even during high disease pressure.  Fe-EDTA suppressed dollar spot infection as well as ferrous sulfate but quality declined to unacceptable levels during the summer, due to Fe-EDTA only.  Sulfur did not affect or increased S. homoeocarpa infection, indicating that a high and frequent foliar rate of iron is responsible for dollar spot control.  An in-vitro study was conducted to determine if agar pH in combination with iron concentrations affects mycelial growth of S. homoeocarpa.  Results from this trial indicated that 5.4 agar pH is an optimal pH for mycelial growth.  The 10 to 100 mg iron kg-1 concentration had little effect on mycelial growth at 5.0 and 5.5 pH, but increased growth at 4.5 and 6.5 pH.  As the iron concentration was increased from 10 to 100 to 1000 mg kg-1, mycelial growth decreased or stopped.  Our final conclusions are that seasonal biweekly foliar applications of the medium rate of ferrous sulfate (24.4 kg ha-1) safely and effectively reduced annual bluegrass infestation out of a creeping bentgrass putting green, while also effectively suppressing silvery thread moss and dollar spot incidence. / Master of Science

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Master of Science / Department of Horticulture, Forestry, and Recreation Resources / Jack D. Fry / Dollar spot, caused by Sclerotinia homoeocarpa F.T. Bennett, and silvery-thread moss (Bryum argenteum Hedw.) are pests affecting creeping bentgrass (Agrostis stolonifera L.) that typically require pesticide inputs. New strategies for pest management may reduce chemical inputs. The objectives of these 2009-2010 field studies were to evaluate: 1) creeping bentgrass cultivars for dollar spot susceptibility; 2) alternative chemical controls for silvery-thread moss; and 3) the response of silvery-thread moss to nitrogen (N) sources. During peak dollar spot development, ‘Declaration’, ‘A-4’, and ‘Crenshaw’ had 7.5, 139.4, and 288.9 infection centers m[superscript]-2 under fairway and 2.1, 27.2, and 106.9 infection centers m[superscript]-2 under putting green conditions, respectively. Two spring and two fall spot applications of sodium or potassium bicarbonate (45 g a.i. L[superscript]-1), premixed essential oil, and broadcast applications of carfentrazone-ethyl at 0.09 kg a.i. ha[superscript]-1 suppressed moss 39% to 55% compared to untreated in 2009. Spot sprays of sodium or potassium bicarbonate, and essential oil, were phytotoxic to creeping bentgrass and required up to 8 or 18 days, respectively, to return to acceptable quality. Fertilization with liquid urea (N at 16.3 kg ha[superscript]-1 biweekly, 210 kg ha[superscript]-1 annually) resulted in 147%, 150%, and 155% more moss than fertilization with IBDU, organic N, and granular urea, respectively, and 156% more moss compared to untreated. Fertilization with urea (liquid or granular) resulted in the best creeping bentgrass color. Averaged across the entire season, plots treated with organic N had unacceptable color in 2009. Nitrogen concentrations in moss tissue ranged from 0.4% to 1.0% and were always significantly lower than N concentrations observed in creeping bentgrass (1.1% to 2.1%), regardless of treatment. In 2010, moss treated with liquid urea had higher tissue N concentrations (1.0%) than untreated moss (0.5%) or that fertilized with IBDU (0.4%). In summary, use of dollar spot-resistant creeping bentgrass cultivars could reduce fungicide requirements. Bicarbonate and essential oil products can reduce moss severity at a similar level to carfentrazone-ethyl, but rates and/or application methods need to be optimized to avoid injury to creeping bentgrass. Applications of liquid urea enhanced moss coverage in creeping bentgrass compared to other N sources.