The main effects of lime, aluminium, iron and manganese were studied in field and greenhouse grown tobacco; relations between soil and plant measurements were examined. Ground limestone, ground mixed lime, ground dolomite and slaked lime at rates equivalent to 1,000 and 2,000 lb. CaC0₃/acre increased yield and quality of flue-cured tobacco both on Triassic and granite sands, whether applied early (February/March) or late (September); the highest rate and late application were often best. Yields increased with 4,000 and 6,000 lb. dolomite/acre applied late, but quality decreased when the pH was about 6.0. Lime did not affect leaf maturity as reflected by nitrogen and reducing sugars concentration . Where leaf discolouration (slate) occurred, the best quality and least discoloured leaf had the lowest manganese concentration and was grown on limed soil. On a very acid and probably nitrogen deficient soil, lime, borax and nitrogen (nitrate only tested) reduced the discolouration and improved the quality, but potassium sulphate increased discolouration and decreased quality. Calcium concentration in the leaf was increased by lime, particularly calcitic materials, and magnesium concentration was increased by dolomite. Lime also increased the filling value and petroleum ether extract, but decreased manganese, boron, chloride and sometimes potassium, and had no effect on phosphorus, nitrogen, aluminium, iron, crude fibre, nicotine, reducing sugars and equilibrium moisture. The inorganic composition of greenhouse plants was similar; generally gypsum increased calcium concentration more than calcium carbonate but it did not affect manganese concentration, which was decreased by calcium carbonate. In the stem and roots of field grown plants (dolomite only tested), the concentration of magnesium was increased but the concentrations of calcium, potassium, aluminium and iron were unaffected. Although the concentration of nitrogen was increased and that of phosphorus was decreased in the stem, these were unaffected in the roots. Aluminium and iron behaved differently to other nutrient ions, being more concentrated in the roots than aerial plant parts. Boron and magnesium deficiencies were observed in a dry and wet year, respectively, suggesting that variable mineral deficiencies can affect responses to lime. Initially soil pH was affected more by source of lime, but later mostly by rates. Slaked lime increased the soil pH more than did ground limestone, mixed lime or dolomite. In a glasshouse experiment, pH was more important than calcium supply and in the field, the largest yields were often associated with the highest pH. In pot experiments, aluminium drastically reduced yields in nutrient solution but not in soils, whereas iron was more severe in soils; manganese had little effect on yield. Manganese was readily taken up and translocated to the tops, but aluminium and iron were mainly concentrated in the roots, as was found in field grown plants. Iron decreased manganese concentration in all plant parts and aluminium decreased calcium and manganese in nutrient solution only. Although aluminium and iron generally increased the concentration of phosphorus in the roots, they did not interfere with phosphorus transport in the plant. Manganese caused the leaf to become chlorotic and when no iron was present the upper leaves became yellow, and developed brown and white lesions. However, in soil grown plants, sufficient iron was present in the soil solution to prevent break down of tissue. Yellowing of the upper leaves also occurred when plants were grown in nutrient solution with aluminium and no iron; when both were present, the plants were darker in colour. Although aluminium damaged roots in nutrient solution, high rates of iron severely damaged leaves of plants grown in soil. Since the concentrations of aluminium, iron and manganese were decreased in the soil solution by liming, they were compared with plant growth and composition in 17 different soils, with and without lime. As was expected, lime increased soil pH. It also increased exchangeable calcium, but decreased exchangeable aluminium, iron and manganese; exchangeable magnesium and potassium and resin extractable phosphorus were not affected. As the Ratio Law does not hold for all Rhodesian soils, anion adsorption will be avoided if the soils are equilibrated with O.OOOSM CaC1₂; the concentrations of the cations in solution were affected in the same way as exchangeable cations, but phosphorus was increased. There was no relationship between yield of tobacco and its chemical composition. The correlations between soil solution data and plant composition were poor, except for manganese and phosphorus; the relation between Mn ppm. in plant vsa (superscript)Mn/a (superscript)Ca + Mg (enclosed in square root sign √) in solution, and P% vs pH₂ P0₄ or pH₂ P0₄ +½ pCa, were both curvilinear. On the other hand, all measurements of exchangeable cations were poorly correlated with plant composition. Finally yield was poorly correlated with soil solution data, and pH was as satisfactory as any other measurement tested. Manganese toxicity was observed on three soils, and a probable manganese deficiency on one. It was not possible to define a limit above which manganese toxicity occurred, but manganese deficiency developed at about 63 ppm. manganese. Variations in pH and the availability of aluminium, iron and manganese occurred when soils were incubated at about field capacity, generally the main effects having developed within seven days. In all soils, there was an initial increase in soil pH and a maximum value was reached in one to four days, decreasing by variable amounts with longer periods of incubation. Although the concentration of aluminium was larger than that of iron, the relation between both ions and soil pH was curvilinear, their concentrations increasing with decreasing pH. Increased temperature of incubation increased pH with a resultant decrease in the concentration of aluminium, but in one soil it appreciably increased the availability of iron in the early periods of incubation. Autumn and spring ploughing did not affect subsequent pH or the concentration of aluminium and iron in the soil solution. Manganese concentration varied from soil to soil and was not related to soil pH. In most soils there was a decrease in manganese concentration with length of incubation and it decreased more rapidly the lower the initial concentration. Temperature effects were variable and moisture affected the behaviour of manganese more than temperature. These findings and the distribution of aluminium, iron and manganese in the plant helped to explain the poor correlations.

The curing of flue-cured tobacco (Nicotiana tabacum L.) is an energy intensive process and represents a significant portion of the overall cost of production. Given the goal of the industry to reduce the environmental footprint of tobacco production and the energy demand of curing, attention has been directed to explore options for the use of renewable fuels for heating tobacco barns. A two-year study conducted at the Virginia Tech Southern Piedmont Center evaluated the effectiveness and cost of curing flue-cured tobacco with a wood pellet burner. Additionally, field studies were conducted to evaluate the feasibility of on-farm production of biomass fuel crops as well as on-farm manufacture of biomass fuel pellets. The first time use of a wood pellet burner with an air-to-air heat exchanger in a bulk curing barn proved to be a viable alternative to a conventional propane fueled burner. Curing cost averaged $0.05 with the pellet burner compared to $0.04 per kilogram of tobacco with the propane burner. The increase in cost was offset by a 90 percent reduction of CO2 emissions with the use of wood pellets. The use of low lignin grass varieties did have an impact on biomass pellet properties. Pellet testing revealed high ash and chloride levels which could be problematic using a high efficiency wood pellet burner. Full maturity harvest of annual grasses fertilized with 112 kg per ha N resulted in higher yields. However, fertilizing for maximum yield would increase the CO2 footprint for biomass fuel pellet production. / Master of Science

Flue-cured tobacco (Nicotiana tabacum L.) is an intensively cultivated crop that typically receives four to eight primary tillage passes before being transplanted on a raised row-ridge. Strip-tillage, a conservation tillage system that only requires tilling a small strip before transplanting, has been shown to be effective for tobacco producers in southside Virginia. The cost of fertilizer in recent years and the loss of applied nutrients has brought new attention to the impact of cover crops used in conservation tillage on the nitrogen fertilization of tobacco. A two-year study conducted at the Southern Piedmont Agricultural Research and Extension Center evaluated a strip-tillage production system on agronomic performance of flue-cured tobacco and evaluated the impact of cover crop management on soil nitrogen cycling and nitrogen uptake by plants. Treatments evaluated whether a wheat cover crop was broadcast or strip killed, topdressing a wheat cover crop with 0, 22, or 45 kg ha-1, and tobacco fertilization rates. Additional treatments included a soybean residue treatment, and a conventional tillage control. Topdressing wheat with nitrogen resulted in nitrogen being released late in the growing season. Whether a wheat cover crop was strip or broadcast killed had no effect on yield or cured leaf quality. Soybean residue did not provide adequate soil cover, but was shown to be a suitable ground cover option for tobacco production. Wheat not topdressed with nitrogen and tobacco receiving normal fertilization had adequate soil surface residue cover, good cured leaf quality, and yields that were comparable to those of conventional tillage. / Master of Science

Host resistance has become a cornerstone of sustainable production of flue-cured tobacco in regions where root-knot nematodes present a threat to growers. Resistance to races 1 and 3 of M. incognita, historically the most significant root-knot nematode threat to tobacco production, is now widely available in commercially available flue-cured tobacco varieties, and is imparted by the gene Rk1. The same gene also provides resistance to race 1 of M. arenaria. The widespread deployment of this resistance has fostered a shift in root-knot nematode population dynamics, as a result of which M. arenaria race 2 has become the predominant root-knot nematode threat in Virginia. A second resistance gene known to impart resistance to M. javanica, Rk2, has also been incorporated into numerous released cultivars in combination with Rk1. This combination has been demonstrated to impart increased resistance to M. incognita and M. javanica relative to either gene alone. In the present work, eleven greenhouse trials conducted from 2017-2019, as well as two trials conducted in 2018 and 2019 on a cooperating farm, investigated the efficacy of this stacked resistance against M. arenaria race 2 and compared the effect of stacking both resistance genes to the effect of either gene alone relative to a susceptible cultivar. We also evaluated how these forms of resistance compare with resistance possessed by a breeding line with resistance reportedly derived from N. repanda to determine if additional, novel sources of resistance to root-knot nematodes previously identified from other species in the genus Nicotiana could play a role in expanding the genetic diversity of germplasm available for the refinement of host resistance in flue-cured tobacco. Additionally, in light of potential new rule making from the FDA mandating reduced nicotine content of cured tobacco leaf, we investigated the relationship between alkaloid (nicotine) content of flue-cured tobacco and root-knot nematode parasitism, while also evaluating nematode parasitism effects on carbohydrate content. Despite considerable variability in our results, particularly under field conditions, our results demonstrate that stacking Rk1 and Rk2 imparts greater resistance to M. arenaria race 2 than either gene alone, but that an entry possessing resistance reportedly derived from N. repanda exhibited significantly greater resistance to root-knot nematodes than the combination of Rk1 and Rk2 based on root galling, and egg mass and egg production. The alkaloid content of flue-cured tobacco did not appear to have an effect on root-knot nematode parasitism under greenhouse or field conditions, but the presence of the nematode did lead to increased accumulation of nicotine in the roots of plants, while translocation of nicotine to leaves was reduced. Conversely, root-knot nematode parasitism was reduced accumulation of carbohydrates in roots, while having no significant effects on leaf carbohydrate content. / Ph.D. / Root-knot nematodes (Meloidogyne spp.) are microscopic round worms that can cause considerable damage to flue-cured tobacco (Nicotiana tabacum L.), and while not typically responsible for killing plants outright, can reduce the quality of cured tobacco leaf and may predispose plants to a host of other issues, resulting in challenges and economic burdens on growers. Chemicals that effectively control nematodes, which are animals, pose inherent threats to human applicators and may harm the environment in a number of ways, so the use of tobacco varieties that are resistant to root-knot nematodes is increasingly common and essential to sustainable tobacco production. One form of root-knot nematode resistance, called Rk1, has become common and is found in all commercially grown flue-cured tobacco. This form of resistance is effective against 2 ‘races’ of the root-knot nematode M. incognita, which has historically caused tobacco growers the most issues. However, because this resistance is so widely employed, growers have controlled these nematodes, while another species, M. arenaria, has become more prevalent, particularly ‘race’ 2, which is not controlled by Rk1. We know from previous research that another gene, Rk2, provides resistance to some root-knot nematode that Rk1 does not effect, and that combining both genes seems to provide even greater root-knot nematode control than either gene alone. We investigated whether Rk2 is effective at controlling M. arenaria race 2 when it is combined with Rk1 in greenhouse and field experiments. We also investigated how a different, novel type of resistance, which comes from a species of tobacco related to cultivated tobacco, compares with the Rk1/Rk2 resistance in greenhouse trials. Additionally, the FDA has recently suggested that nicotine levels in tobacco leaf should be dramatically reduced to help mitigate adverse human health consequences associated with tobacco consumption. Nicotine may play some role in resistance to root-knot nematode in tobacco, and conversely, root-knot nematodes may impact levels of nicotine, as well as other important chemical constituents of tobacco. We also investigated these questions in greenhouse and field experiments. Our results ultimately demonstrate that combining both Rk1 and Rk2 gives flue-cured tobacco a higher level of resistance to root-knot nematodes than either gene alone, but also suggests that the form of resistance we evaluated from a related Nicotiana species could be even more effective in controlling these nematodes. We observed that the amount of nicotine present in tobacco did not impact nematode parasitism, but that nematode parasitism could lead to lower levels of nicotine in the leaves of plants because the nematodes, which feed on plants roots, cause damage to the plant that interferes in the movement of nicotine from roots to leaves.

Utilizing resistant cultivars is a main control strategy for root-knot nematodes in flue-cured tobacco (Nicotiana tabacum L.). Most commercial cultivars possess the Rk1 gene, providing resistance to races 1 and 3 of Meloidogyne incognita and race 1 of M. arenaria. This initiated a shift in root-knot populations to other species and races, creating a need for resistance to those populations. Numerous cultivars possess a second resistance gene, Rk2. Greenhouse experiments investigated whether possessing both Rk1 and Rk2 increases resistance to a variant of M. incognita race 3 compared to either gene alone, and if high soil temperatures impact their efficacy. Root galling, numbers of egg masses and eggs, and the reproductive index were compared from roots of Coker 371-Gold (susceptible), NC 95 and SC 72 (Rk1Rk1), T-15-1-1 (Rk2Rk2), and STNCB-2-28 and NOD 8 (Rk1Rk1 and Rk2Rk2). The same data were analyzed from plants in open-top root zone cabinet growth chambers set to 25ºC, 30ºC, and 35ºC to examine if resistance is temperature sensitive. Despite variability, Rk1Rk2 entries conferred greater resistance than entries with Rk1 or Rk2 alone. Entries with Rk1 alone reduced galling and reproduction compared to the susceptible control, whereas T-15-1-1 (Rk2) did not, but often suppressed reproduction. An apparent reduction in nematode reproduction was observed at 25ºC and 30ºC on entries possessing Rk1 and Rk1Rk2 compared to the control and Rk2. However, no apparent differences in reproduction occurred on Rk1 and/or Rk2 entries at 35ºC compared to the control, indicating parasitism increased on resistant entries at higher temperatures. / Master of Science in Life Sciences

Flue-cured tobacco (Nicotiana tabacum L.) is a high value-per-acre crop that is intensively managed to optimize the yield of high quality cured leaf. Aerial remote sensing, specifically unmanned aerial vehicles (UAVs), present flue-cured tobacco producers and researchers with a potential tool for scouting and crop management. A two-year study, conducted in Southside Virginia at the Southern Piedmont Agricultural Research and Extension Center and on commercial farms, assessed the potential of aerial remote sensing in flue-cured tobacco. The effort encompassed two key objectives. First, examine the use of the enhanced normalized difference vegetation index (ENDVI) for separating flue-cured tobacco varieties and nitrogen rates. Secondly, develop hyperspectral indices and/or machine learning classification models capable of detecting Phytophthora nicotianae (black shank) incidence in flue-cured tobacco. In 2017, UAV-acquired ENDVI surveys demonstrated the ability to consistently separate between flue-cured tobacco varieties and nitrogen rates from topping to harvest. In 2018, ENDVI revealed significant differences among N-rates as early as 34 days after transplanting. Two hyperspectral indices were developed to detect black shank incidence based on differences in the spectral profiles of asymptomatic flue-cured tobacco plants compared to those with black shank symptoms. Testing of the indices showed significant differences between the index values of healthy and symptomatic plants (alpha = 0.05). In addition, the indices were able to detect black shank symptoms pre-symptomatically (alpha = 0.09). Subspace linear discriminant analysis, a machine learning classification, was also used for prediction of black shank incidence with up to 85.7% classification accuracy. / Master of Science / Unmanned Aerial Vehicle’s (UAVs) or drones, as they are commonly referred to, may have potential as a tool in flue-cured tobacco research and production. UAVs combined with sensors and cameras provide the opportunity to gather a large amount of data on a particular crop, which may be useful in crop management. Given the intensive management of flue-cured tobacco, producers may benefit from extra insight on how to better assess threats to yield such as under-fertilization and disease pressure. A two-year study was conducted in Southside Virginia at the Southern Piedmont Agricultural Research and Extension Center and on commercial farms. There were two objectives to this effort. First, assess the ability of UAV-acquired multispectral near-infrared imagery to separate flue-cured tobacco varieties and nitrogen rates. Secondly, develop hyperspectral indices and machine learning models that can accurately predict the incidence of black shank in flue-cured tobacco. Flue-cured tobacco nitrogen rates were significantly different in 2017 from 59 days after transplanting to harvest using UAV-acquired near-infrared imagery. In 2018, heavy rainfall may have led to nitrogen leaching from the soil resulting in nitrogen rates being significantly different as early as 34 days after transplanting. The imagery also showed a significant relationship with variety maturation type in the late stages of crop development during ripening. Two hyperspectral indices were developed and one machine learning model was trained. Each had the ability to detect black shank incidence in fluecured tobacco pre-symptomatically, as well as separated black shank infested plants from healthy plants.