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Winter injury of bermudagrass (Cynodon spp.) continues to be a problem across the transition zone. In an attempt to delay or induce winter dormancy while maintaining cold hardiness, applications of seaweed extract (SWE) (0.54 kg ha-1), ethephon (16 L ha-1), Fe (1 kg ha-1), and N (49 kg ha-1) took place every three weeks during the fall of 2001 and 2002. Cultivars examined included 'Riviera', 'Midiron', 'Princess', and 'Tifway'. Tifway exhibited greatest fall color retention in both years of the study. Ethephon promoted early senescence and turfgrass quality during fall ratings in both years of the study while N, Fe, and SWE increased quality over the control in 2001 and only N showed better quality and color retention over the control in 2002. Samples removed from cold acclimated plots were artificially frozen as a measure of cold hardiness. Treatments did not have an effect on post freeze regrowth, however, cultivar was significant in both years. Midiron showed best regrowth followed by Riviera, Tifway, and Princess. In both years Riviera and Midiron displayed the quickest and greatest amount of spring greenup followed by Tifway and then Princess. Ethephon reduced greenup in both years and SWE, Fe, and N showed no differences from the control in 2001 and Fe showed significantly better greenup in 2002. Proline and Linolenic acid levels were highest in Midiron, followed by Riviera, Tifway, and Princess. Nitrogen, SWE, and Fe generally did not have an effect on linolenic acid and no consistent effects were noted on proline concentration. Ethephon treatments did not have an effect on linolenic acid levels, however, there was a negative effect on proline concentrations. The results of this study indicate that judicial N applications during the fall can promote color retention and do not have a negative effect on bermudagrass cold-tolerance. Linolenic acid and proline findings also help to explain differences in cold-tolerance between different bermudagrass cultivars. / Ph. D.

One of the principle factors that limits the areas to which seeded bermudagrass can be adapted is low temperature. Therefore, increasing the winter tolerance of seeded bermudagrass cultivars has been a goal of turfgrass breeders for many years. Design of an efficient breeding method for developing cultivars with increased tolerance to cool and freezing temperatures could be enhanced by having heritability estimates for cold tolerance traits. Additionally, the identification of correlated traits can be useful in improvement of cultivar development. Heritability estimates for winter tolerance can be obtained from cold treatments imposed artificially or from observations made in the field. Parental clones and their respective polycross half-sib families were established in a randomized complete block design with four replications in 2004 in Lexington, KY. Differences in spring green up and fall dormancy measured in 2006 were detected between genotypes within the progeny and parental groups. These same lines were subjected to 15 replications of a freezing treatment in a freeze chamber. Differences in spring green up and fall dormancy were detected between genotypes within the progeny and parental groups. Differences in freeze response were also detected using the artificial freeze treatment. Broad-sense heritabilities were estimated to be 0.895 and 0.573 for spring green-up and fall dormancy respectively. Narrow-sense heritability estimates were found to be 0.885 and 0.265 for these same traits. These results indicate that this population could be improved for cold tolerance using phenotypic recurrent selection. Freeze response was found to be positively correlated to winter hardiness and seed yield in the parent group.

Diss. (sammanfattning). Umeå : Sveriges lantbruksuniv., 2003. / Härtill 4 uppsatser.

Master of Science / Department of Agronomy / S.V. Krishna Jagadish / Low temperature is one of the most limiting stresses to crops that are adapted to tropical and subtropical regions, such as maize (Zea mays L.) and sorghum [Sorghum bicolor (L.) Moench], when introduced into temperate regions. However, no studies have compared the chilling tolerance of maize and sorghum grown together. Therefore, the objective of this research was to screen maize hybrids and sorghum genotypes for chilling tolerance at the germination and seedling stages. With the hypothesis that grain composition of maize and sorghum could lead to varying chilling tolerance, the seeds were analyzed for concentrations of protein, starch, and amylose. Five commercial hybrids of maize and 18 genotypes of sorghum were maintained in growth chambers for 31 days at two temperatures: a control temperature (25/20 °C, day/night) and at chilling temperatures (11/8 °C for 14 days; 12.5/9.5 °C for 14 days, and 14/11 °C for 3 days). Emergence and seedling height were measured during the experiment. At the end of the experiment, shoot dry weight, root dry weight, and leaf area were determined. Emergence of sorghum under the chilling temperature regime was low (18%). Average height of the emerged sorghum seedlings in the cold temperatures at the end of the experiment was 1.4 cm compared to 55.5 cm in the control treatment. All maize hybrids emerged, but emergence and growth were slowed by the cold temperatures, and average height at the end of the experiment was 4.6 cm compared to 96.1 cm in the control treatment. Shoot dry weight, root dry weight, and leaf area of the sorghum under the chilling temperatures were too small to measure, and, for maize, they were greatly reduced. The results showed that, for sorghum, temperatures should be above 14 °C for emergence, while maize could emerge at lower temperatures. The analyses of the sorghum seeds showed that Redbine 60 and RTx430 had the highest protein concentrations (15.71% and 15.35%, respectively), and Segaolane had the lowest protein concentration (9.83%). Segaolane had the highest starch concentration (72.71%), and RTx430 had the lowest starch concentration (65.31%). There was an inverse relationship between protein and starch concentrations in the sorghum seeds (R2 = 0.69). Amylose concentrations did not vary significantly among the sorghum seeds. The analyses of the maize seeds showed that Dekalb 51-20 and Pioneer 1151 had the highest protein concentrations (10.98% and 10.95%, respectively), and Pioneer 1105 had the lowest protein concentration (9.26%). Starch and amylose concentrations did not vary significantly among the maize seeds.

Biological invasions have been observed throughout the world for centuries, often with major consequences to biodiversity and food security. Tying invasion to species identity and associated traits has led to numerous hypotheses on why, and where, some species are invasive. In recent decades, attention to intraspecific variation among invaders has produced questions about their adaptation to climate, land use, and environmental change. I examined the intraspecific variation of invasive Johnsongrass's (Sorghum halepense (L.) Pers.) seedling stress response, propagule cold tolerance, and large-scale niche dynamics for correlation with populations' climatic and ecotypic (i.e., agricultural vs. non-agricultural) origin. Overall, I found a greater number of home climate effects than ecotypic effects on various traits. Non-agricultural seed from cold climates and agricultural seed from warm climates germinated more and faster, while non-agricultural seedlings showed uniform chlorophyll production regardless of home soil carbon origin, unlike their agricultural counterparts. Neither seedling stress response nor propagule cold tolerance interacted with ecotype identity; however, drought stress varied with population origins' aridity and soil fertility, and seed from warm/humid and cold/dry climates was most germinable. Comparison of seed and rhizome cold tolerance also suggested that the latter is a conserved trait that may be limiting S. halepense poleward range expansion. This physiological limit, an unchanged cold temperature niche boundary between continents and ecotypes, and a narrowed niche following transition to non-agricultural lands all imply low likelihood of spread based on climatic niche shift. Instead, evidence points to range expansion driven primarily by climate change and highlights agriculture's role in facilitating invasibility. This tandem approach to climate and land use as drivers of intraspecific variation is transferable to other taxa and can help refine our conception of and response to invasion in the Anthropocene. / Doctor of Philosophy / Exotic invasive species are a global problem, threatening biodiversity and biosecurity now and in the future. In the last several decades, ecologists have studied many individual invaders and their traits to understand what drives their spread. More recently, abundant differences in traits between populations within an invasive species have raised questions about humans' role in facilitating invasion through climate change, land use, and other disturbances. I studied the invasive Johnsongrass's (Sorghum halepense (L.) Pers.) response to drought, nutrient limitation, and freezing to detect differences between populations based on their climate and ecotype (agricultural vs. non-agricultural) origin. I also tracked differences in the climates the species occupied across the globe and North America and projected its future distribution under climate change. Overall, I found a greater number of home climate effects than ecotypic effects on various traits. Non-agricultural seed from cold climates and agricultural seed from warm climates germinated the most, while non-agricultural seedlings performed consistently regardless of soil carbon origin, unlike their agricultural counterparts. In addition, drought stress varied with population origins' rainfall and soil fertility, and seed germination favored warm/humid and cold/dry origin. Rhizome (underground stem) cold tolerance appears to be a trait that limits S. halepense poleward range expansion. Along with no change in the coldest climates occupied worldwide and no spread to new climates with transition to non-agricultural lands, this implies that Johnsongrass is unlikely to expand its range without external forces. Instead future range expansion will likely be driven by climate change. This coupled approach to climate and land use affecting invasion is transferable to other species and can help refine both our concepts and response strategies.

The La Crosse virus (LACV) is an emerging pathogen in the Appalachian region of the United States. The virus maintains a complex natural cycle through horizontal transmission with sciurid rodents and Aedes mosquitoes in Virginia. Transovarial transmission also occurs in this host-parasite system and has evolved for the virus to persist through winter. The virus perpetuates in diapause induced embryos and infects naive rodents the following spring. As global temperatures rise, it is imperative we evaluate how the virus and its vectors overwinter. We conducted experiments to evaluate and determine the diapause induction, or prediapause stage, in Aedes japonicus japonicus, and the ecophysiology and low temperature biology of how LACV affects Aedes triseriatus and Aedes albopictus embryos at low temperatures. We found that the prediapause stage of Ae. j. japonicus is the maternal stage, the developing adult female that lays diapause eggs. This more closely resembles Ae. albopictus and not Ae. triseriatus. As measured in the field and laboratory, LACV has a clear negative effect on the ability of Ae. triseriatus and Ae. albopictus to survive the winter. There was no major effect of LACV infection on the ability of these two species to enter diapause or their critical photoperiods, but there was a significant negative effect of LACV infection on survivorship of embryos that were placed in their natural habitat and in their susceptibility to low temperatures. LACV infection had a more significant negative effect on Ae. albopictus than on Ae. triseriatus and suggests that Ae. albopictus is a subordinate vector relative to Ae. triseriatus in the maintenance of the virus over the winter season. Our findings highlight the need for the increased surveillance of LACV as temperatures continue to rise due to climate change. / PHD / In Virginia, mosquitoes may transmit the La Crosse virus (LACV) that can cause swelling of the brain in humans. This virus infects mosquitoes which is then transmitted to chipmunks. When a mosquito bites an infected chipmunk and then bites a human, the virus be transmitted to the human. Adult female mosquitoes can also pass the virus on to their developing offspring, which is the way it is maintained throughout the winter. Certain infected mosquitoes will spend the winter as eggs and the mosquitoes that hatch carry the virus and can infect chipmunks the following spring. The experiments conducted here looked at how one mosquito that can spread the LACV begins the process to survive the winter. We also did experiments to look at how the virus affects two other mosquito’s ability to begin the same process and then if they can survive the winter and cold temperatures while infected. We found that the Asian Bush Mosquito enters diapause in a different way than we suspected by the female mosquito sensing shorter days at the end of the summer; at this point she will then lay eggs that survive the winter. We discovered that viral infection will hurt the chances of the Asian Tiger Mosquito and Eastern Tree-hole Mosquito to survive the winter. We also found that virus-infected mosquitoes cannot survive temperatures as lower than those that are not infected. This means that the risk of the virus to humans can be greater as temperatures rise due to global warming.

A protocol for cauliflower micro-propagule production was developed and optimised for both micropropagation and artificial seed production techniques using meristematic tissues from cauliflower curd. All steps in the protocol were empirically optimised including: blending, sieving, culture methods, liquid culture media composition and plant growth regulator combinations and concentrations. The cost of the micro-propagules could be reduced by as much as 50% on the initial costings reported previously since treatments doubled the number of microshoots produced per culture unit. The research confirmed the suitability of cauliflower microshoots to be encapsulated as artificial seeds and an effective protocol for microshoot encapsulation was designed through the optimization of 1) the production of cauliflower microshoots suitable for encapsulation, 2) encapsulation procedures, 3) artificial seed artificial endosperm structure, 4) conversion materials. The possibility of culturing cauliflower artificial seeds in commercial substrates such as perlite, sand, vermiculite and compost was confirmed. The use of plant preservative mixture (PPM) for the control of contamination in cauliflower culture media and artificial seeds was optimised and the effect of this material on the development of plant material was assessed. It was confirmed that cauliflower artificial seed could be stored in a domestic refrigerator for up to 6 months which could have a great impact in cauliflower breeding programmes. The huge number of cauliflower microshoots that could be produced using this protocol and the homogeneity of the culture system, provided a tool for the molecular analysis of cauliflower microshoots (and artificial seed) abiotic stress tolerance analysis. Various treatments were conducted to improve microshoot cold tolerance and the up-regulation of the CBF/DREB1 transcription factor including low temperature acclimation, mannitol, ABA (abscisic acid) and Mo (molybdenum). Microshoots were confirmed to acclimate successfully using low temperature. Mo was shown to improve the cold tolerance of cauliflower microshoots and to up-regulate CBF/DREB1 in the absence of low temperature acclimation. Acclimation did not increase the accumulation of dehydrin proteins and it is concluded that dehydrins do not play a significant role in the cold tolerance of cauliflower microshoots. Since cauliflower breeding and seed multiplication protocols make extensive use of micropropagation, the studies reported in this research could make a significant impact by decreasing the cost of micropropagation and increasing its reliability. It also opens new perspectives for further research for cauliflower artificial seed production and the possibility of sowing these seeds directly in the field. Furthermore, this research helps to facilitate cauliflower breeding programmes by improving the understanding of abiotic stress tolerance mechanisms and the relationship between different types of abiotic stresses such as cold and drought.

Winter wheat has several advantages over spring varieties, higher (25 % more) yield, efficient use of spring moisture, reduction of soil erosion by providing ground cover during the fall and early spring, rapid initial spring growth to out - compete weeds and circumvent the peak of Fusarium head blight infections by flowering early. Winter wheat is planted in early autumn when it germinates and developing seedlings acclimate to cold. The crown survives under snow cover and in spring rapidly grows into a vigorously growing plant for grain to be harvested in summer. However, the harsh Canadian prairie winters require that winter wheat has increased cold hardiness and improved winter survival to reduce losses from sudden cold snaps during winter and spring. Low temperature (LT) tolerance is one of the major components of cold hardiness. Genetic mapping studies have revealed a major quantitative trait locus (Fr-A2) at wheat chromosome 5A which can explain at least 50 % of LT tolerance in wheat. Physical mapping of 5A LT QTL in a hardy winter wheat cv Norstar revealed a cluster of at least 23 C - repeat binding factors (CBF) coinciding with peak of Fr-A2 QTL. The objective of this study is biochemical, and molecular characterization of CBF co - located at Fr-A2 to identify key CBF participating in conferring LT tolerance in winter wheat. A comparative analysis of CBF gene cluster at the Fr-A2 collinear region among Poaceae members showed an expansion in the number of CBF genes with increased LT tolerance. Rice, a cold sensitive member, had only three CBF genes, whereas cold hardy winter wheat cv Norstar has 23 CBF genes. Amino acid sequence - based cluster analysis of complete CBF genes, or their major functional components such as the AP2 - DNA binding domain and C - terminal trans - activation domain, divide Norstar CBF into Pooideae specific clades. However, analyses of Norstar CBF amino acid sequences of different functional groups revealed a shift in clade members. These results suggest divergence of CBF functions which could lead to possible differences / similarity in the regulon activated by a CBF in a specific group. The 15 CBF genes from winter wheat cv Norstar were expressed in E. coli to produce recombinant TrxHisS - CBF fusion proteins in adequate quantities for structural and functional assays. All CBF fusion proteins could be recovered in the E. coli soluble phase of cell extract, except that the CBF17.0 fusion protein could only be recovered with 6 M urea extraction. Eleven of the 15 CBF fusion proteins were very stable in heat (98 oC), 10 % SDS and 6 M urea treatment. The five other CBF members were very labile under native conditions, but were stable in E. coli cell extracts or when extracted under denaturing conditions. Most of the CBF recombinant proteins in denaturing gel electrophoresis migrated slower than expected from their predicted molecular mass, based on amino acid sequence. The slow migration could be associated to their elongated protein structure as determined by dynamic light scattering (DLS). CBF 12.2 and CBF 17.0 were highly resistant to denaturation and retained their secondary structure in these conditions as determined by circular dichroism (CD) spectra. The high stability of these two CBF proteins may be important for cold acclimation or maintenance of cold hardiness in wheat. CBF proteins are transcription factors that bind to the dehydration-responsive element / C-repeat element (DRE / CRT) motif (CCGAC). Ten of the 15 Norstar recombinant CBFs whether purified under native or denaturing conditions showed in vitro binding to the CRT motif. Within hours of cold exposure (4 oC) the native CBF increased their affinity to CRT interaction which could be due to changes in the CBF secondary structures. Some of the CBF for binding preferred the core GGCCGAC motif while others preferred TGCCGAC. Similarly binding assays with truncated CBF revealed that for some CBF proteins, the second signature motif (DSAWR) and remaining C - terminal were not needed, while for others a considerable portion of the C -terminal region was needed for binding. Norstar CBF 12.1 has a memory of cold experience, and upon exposure to cold, has a high and immediate affinity to CRT elements. A homolog CBF12.2 in less cold - hardy winter wheat cv Cappelle - Desprez had a non - functional protein due to a R → Q substitution in a highly conserved residue within the AP2 domain. Several of the cv Norstar CBFs showed increased activity under LT and denaturing conditions, which may be the reason for the greater cold hardiness in Norstar. In conclusion, detailed and extensive analyses of CBF in this study characterized their structure and function relationships, which are important for understanding and improving LT tolerance in plants. The identification of specific CRT binding motifs and two CBFs which were very stable under adverse conditions may be prime candidates for further study to improve LT tolerance in plants.