Global climate models predict that the current trend of warming in the Arctic will continue over the next century. The productivity of arctic plants is often limited by short growing seasons with relatively low temperatures such that a warmer climate could have large impacts on plants and plant communities. This study characterised alpine plant responses to changes in temperature at an alpine tundra site near Whitehorse, Yukon, Canada. I examined relationships between plant productivity and natural temperature variations and assessed responses of plants exposed to an experimental warming treatment. Non-destructive measurements of reproductive and growth characteristics of four target species (Dryas octopetala, Lupinus arcticus, Polygonum viviparum, and Salix arctica) were taken annually from 1999 to 2008. There was no significant effect of the warming treatment (OTCs) on average daily mean temperatures as midday warming of up to 1.4 °C was largely offset by night time cooling in the OTCs. Vegetative measurements of target species showed no significant responses to OTC treatments. However, peduncles of D. octopetala and sections of P. viviparum inflorescences that produced bulbils were an average of 34.6 % and 64.7 % longer in OTCs than in controls, respectively. These treatment responses were likely due to plants responding to a factor other than temperature that was modified by the chamber. One vegetative and five reproductive characteristics were significantly related to annual variation in temperature. The summer of 2004 was exceptionally hot, and some species that did not respond to smaller fluctuations in temperature showed large changes in growth or reproduction in this year, perhaps indicating a non-linear response to temperature. Among the larger responses to the warm summer of 2004 was a shift in P. viviparum allocation from predominantly asexual to sexual means of reproduction. Measurements of plant community composition assessed at five-year intervals showed no differences in community composition between experimental plots and controls, and changes in composition over the study period were not uni-directional. In general, both individual plants and community composition were highly resilient to observed variation in summer temperatures. Other factors, such as nutrient availability, may be more important in determining plant responses to environmental change at this site than the direct effects of summer temperature variation.

Plants often live in association with growth-promoting bacteria, which provide them with several benefits. One such benefit is the lowering of plant ethylene levels through the action of the bacterial enzyme 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase that cleaves the immediate biosynthetic precursor of ethylene, ACC. The plant hormone ethylene is responsible for many aspects of plant growth and development but under stressful conditions ethylene exacerbates stress symptoms. The ACC deaminase-containing bacterium Pseudomonas putida UW4, isolated from the rhizosphere of reeds, is a potent plant growth-promoting strain and as such was used, along with an ACC deaminase minus mutant of this strain, to study the role of ACC deaminase in plant growth-promotion. Also, transgenic plants expressing a bacterial ACC deaminase gene were used to study the role of this enzyme in plant growth and stress tolerance in the presence and absence of nickel. Transcriptional changes occurring within plant tissues were investigated with the use of an Arabidopsis oligonucleotide microarray. The results showed that transcription of genes involved in hormone regulation, secondary metabolism and the stress response changed in all treatments. In particular, the presence of ACC deaminase caused genes for auxin response factors to be up-regulated in plant tissues suggesting a de-repression of auxin signaling in the absence of high levels of ethylene. Also, transgenic plants had longer roots and grew faster than the non-transformed plants and genes involved in the stress response and secondary metabolism were up-regulated. Plants inoculated with bacteria had lower levels of secondary metabolism gene expression and slightly higher stress response gene expression than uninoculated plants. Yet, inoculation with the ACC deaminase-expressing bacterium caused less up-regulation of plant genes involved in stress and defense responses and the down-regulation of genes involved in nitrogen metabolism in comparison to plants inoculated with the ACC deaminase minus mutant. Nickel stress caused the down-regulation of genes involved in photosynthesis and carbon fixation and the up-regulation of genes involved in stress responses, and amino acid and lipid breakdown suggesting energy starvation. When transgenic plants expressing ACC deaminase in the roots were exposed to nickel stress, plant stress symptoms were significantly lower and biomass was significantly higher suggesting that lowering the level of ethylene relieved many of the stress symptoms. In fact, genes involved in photosynthesis, secondary metabolism and nitrate assimilation were up-regulated in transgenic plants compared with non-transformed plants in the presence of nickel, suggesting that ACC deaminase is effective at reducing the severe effects of this metal stress.

Plants often live in association with growth-promoting bacteria, which provide them with several benefits. One such benefit is the lowering of plant ethylene levels through the action of the bacterial enzyme 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase that cleaves the immediate biosynthetic precursor of ethylene, ACC. The plant hormone ethylene is responsible for many aspects of plant growth and development but under stressful conditions ethylene exacerbates stress symptoms. The ACC deaminase-containing bacterium Pseudomonas putida UW4, isolated from the rhizosphere of reeds, is a potent plant growth-promoting strain and as such was used, along with an ACC deaminase minus mutant of this strain, to study the role of ACC deaminase in plant growth-promotion. Also, transgenic plants expressing a bacterial ACC deaminase gene were used to study the role of this enzyme in plant growth and stress tolerance in the presence and absence of nickel. Transcriptional changes occurring within plant tissues were investigated with the use of an Arabidopsis oligonucleotide microarray. The results showed that transcription of genes involved in hormone regulation, secondary metabolism and the stress response changed in all treatments. In particular, the presence of ACC deaminase caused genes for auxin response factors to be up-regulated in plant tissues suggesting a de-repression of auxin signaling in the absence of high levels of ethylene. Also, transgenic plants had longer roots and grew faster than the non-transformed plants and genes involved in the stress response and secondary metabolism were up-regulated. Plants inoculated with bacteria had lower levels of secondary metabolism gene expression and slightly higher stress response gene expression than uninoculated plants. Yet, inoculation with the ACC deaminase-expressing bacterium caused less up-regulation of plant genes involved in stress and defense responses and the down-regulation of genes involved in nitrogen metabolism in comparison to plants inoculated with the ACC deaminase minus mutant. Nickel stress caused the down-regulation of genes involved in photosynthesis and carbon fixation and the up-regulation of genes involved in stress responses, and amino acid and lipid breakdown suggesting energy starvation. When transgenic plants expressing ACC deaminase in the roots were exposed to nickel stress, plant stress symptoms were significantly lower and biomass was significantly higher suggesting that lowering the level of ethylene relieved many of the stress symptoms. In fact, genes involved in photosynthesis, secondary metabolism and nitrate assimilation were up-regulated in transgenic plants compared with non-transformed plants in the presence of nickel, suggesting that ACC deaminase is effective at reducing the severe effects of this metal stress.

Global climate models predict that the current trend of warming in the Arctic will continue over the next century. The productivity of arctic plants is often limited by short growing seasons with relatively low temperatures such that a warmer climate could have large impacts on plants and plant communities. This study characterised alpine plant responses to changes in temperature at an alpine tundra site near Whitehorse, Yukon, Canada. I examined relationships between plant productivity and natural temperature variations and assessed responses of plants exposed to an experimental warming treatment. Non-destructive measurements of reproductive and growth characteristics of four target species (Dryas octopetala, Lupinus arcticus, Polygonum viviparum, and Salix arctica) were taken annually from 1999 to 2008. There was no significant effect of the warming treatment (OTCs) on average daily mean temperatures as midday warming of up to 1.4 °C was largely offset by night time cooling in the OTCs. Vegetative measurements of target species showed no significant responses to OTC treatments. However, peduncles of D. octopetala and sections of P. viviparum inflorescences that produced bulbils were an average of 34.6 % and 64.7 % longer in OTCs than in controls, respectively. These treatment responses were likely due to plants responding to a factor other than temperature that was modified by the chamber. One vegetative and five reproductive characteristics were significantly related to annual variation in temperature. The summer of 2004 was exceptionally hot, and some species that did not respond to smaller fluctuations in temperature showed large changes in growth or reproduction in this year, perhaps indicating a non-linear response to temperature. Among the larger responses to the warm summer of 2004 was a shift in P. viviparum allocation from predominantly asexual to sexual means of reproduction. Measurements of plant community composition assessed at five-year intervals showed no differences in community composition between experimental plots and controls, and changes in composition over the study period were not uni-directional. In general, both individual plants and community composition were highly resilient to observed variation in summer temperatures. Other factors, such as nutrient availability, may be more important in determining plant responses to environmental change at this site than the direct effects of summer temperature variation.

Genetically modified cotton (Gossypium hirsutum L.) acreage has increased dramatically over the last six years. Reports of variable results in fiber quality and yield have arisen in these cultivars. Some changes in production practices have occurred coincident with the introduction of transgenic technology, such as reduced use of broad-spectrum insecticides, including organophosphates (OP) that could potentially influence the growth and yield of cotton. One factor that might affect these parameters is the difference in the amount of foliarly-applied phosphorus (P) between an OP and non-phosphate (NP) insecticide regime. Therefore, a study was conducted to investigate selected growth characteristics, yield, and fiber quality of genetically modified and conventional cotton as influenced by OP and foliar phosphorus (FP) applications. A four replication strip-plot experimental design was utilized with cultivar serving as the whole plot and insecticide regime as the sub-plot. Three cultivars of the same recurrent parent (ST4892BR, ST4793R, and ST474) were planted in 2001 and 2002 under irrigated conditions in Burleson County, TX on a Weswood silty clay loam (fine-silty, mixed, thermic Fluventic Ustochrept). The insecticide regime consisted of NP, NP+FP, and OP treatments. The FP was applied at P2O5 weight equivalent to the P component in the concurrent OP application. ST4892BR had greater lint yield than ST4793R and ST474. The yield increase can be explained through plant mapping analysis which showed ST4892BR producing larger bolls and greater boll numbers. In addition, evaluation of fruiting distribution showed ST4892BR contained more lint on sympodial branches 6 through 10. The insecticide regime effect on lint yield resulted in higher yield (P=0.08) for the NP+FP regime. Examination of yield components revealed NP+FP increased second position bolls, predominantly at sympodial branches 6 through 10. Leaf tissue analysis revealed increased levels of P for the OP and NP+FP over that of the NP insecticide regime, which indicates a potential for plants to acquire P from OP insecticides. Furthermore, the considerable yield response to small amounts of FP is not clearly understood. While conclusive evidence exists regarding cultivar yield differences, this study does not provide sufficient evidence to conclude that OP insecticides influence growth, yield, or fiber quality characteristics of these cotton cultivars.

Plant growth-promoting bacteria (PGPB) have been studied in many agriculturally interesting plants, but never in pitcher plants. Sarracenia oreophila (the green pitcher plant) is an endangered species in Georgia, Alabama, and North Carolina (Rice 2010). With the help of Dr. Jim Spain's lab, a previous student in Dr. Gerald Pullman's lab discovered evidence that nitrogen-fixing bacteria (Burkholderia spp.) live within these pitcher plants. This study aims to determine whether these nitrogen-fixing bacteria confer a benefit to their host plants by providing fixed nitrogen. To do this, pitcher plants were inoculated with the Burkholderia and grown on a control medium, a medium without sugar (as the sugar causes the bacteria to grow until they hinder the plants), various media that are missing nitrogen-containing compounds usually provided in growth media, and a medium completely lacking nitrogen. These plants were compared to control plants on the same media that had not been inoculated with Burkholderia. The plants' biomass and root growth were measured. The data suggest that Burkholderia may stimulate plant biomass growth when sufficient nitrogen is present and there may be a nitrogen-threshold that needs to be met in order to sustain the Burkholderia-Sarracenia symbiosis. Also, the Burkholderia has a negative effect on roots grown in high-nitrogen media, possibly due to competition for nutrients.

Cucumber root rot caused by Pythium aphanidermatum can be suppressed by introduced plant growth-promoting rhizobacteria (PGPR). Preliminary experiments clarified that this root disease could be suppressed by strains of Pseudomonas aureofaciens, P. corrugata, and P. fluorescens. To determine whether the mechanism was a systemic resistance induced by PGPR, a split root technique was employed on greenhouse cucumbers grown in soilless substrates. On the split roots, bacteria which were introduced into one side of the root were completely separated from pathogen challenged-inoculated roots-on the other side of the roots. Results from the series of experiments conducted with this design demonstrated that (i) the resistance against root rot induced by PGPR was systemic, (ii) germination of P. aphanidermatum zoospores was reduced in extracts from bacterized roots compared to non-treated control, and (iii) spread of Pythium mycelia was delayed and zoospore germination was inhibited on the distant induced root, compared to the non-bacterized control. Furthermore, enzyme analysis indicated that phenylalanine ammonia lyase, peroxidase and polyphenoloxidase increased on cucumber roots two days after they were bacterized with Pseudomonas strains 13 or 63--28. When the bacterized roots were challenged with P. aphanidermatum, these plant defense enzymes increased as the symptoms appeared, but this accumulation of enzymes was not any higher on roots induced with each of the Pseudomonas strains compared to the Pythium inoculated control. This enzyme stimulation was also systemically induced by PGPR or P. aphanidermatum on cucumber roots. The patterns of iso-peroxidase induced with the PGPR and P. aphanidermatum treatments were different. High levels of salicylic acid (SA) accumulated in bacteria-induced roots, as well as in pathogen-infected roots, which suggests that SA may be associated with cucumber resistance response. But exogenous application of SA did not induce any systemi

Various Trichoderma and Bacillus spp. have been documented as being antagonistic to a wide range of soilborne plant pathogens, as well as being plant growth stimulants. Successes in biological control and plant growth promotion research has led to the development of various Trichoderma and Bacillus products, which are available commercially. This study was conducted to evaluate the effect of six Trichoderma spp. and three Bacillus spp. and their respective combinations, for the biological control of Rhizoctonia solani damping-off of cucumber and plant growth promotion of dry bean (Phaseolus vulgaris L.). In vivo biological control and growth promotion studies were carried out under greenhouse and shadehouse conditions with the use of seed treatment as the method of application. In vitro and in vivo screening was undertaken to select the best Trichoderma isolates from 20 Trichoderma isolated from composted soil. For in vitro screening, dual culture bioassays were undertaken and assessed for antagonisms/antibiosis using the Bell test ratings and a proposed Invasive Ability rating based on a scale of 1-4 for possible mycoparasitic/hyperparasitic activity. The isolates were further screened in vivo under greenhouse conditions for antagonistic activity against R. solani damping-off of cucumber (Cucumis sativus L.) cv. Ashley seedlings. The data generated from the in vivo greenhouse screening with cucumber plants were analysed and grouped according to performance of isolates using Ward‟s Cluster Analysis based on a four cluster solution to select the best isolates in vivo. Isolates exhibiting marked mycoparasitism of R. solani (during ultrastructural studies) viz, T. atroviride SY3A and T. harzianum SYN, were found to be the best biological control agents in vivo with 62.50 and 60.06% control of R. solani damping-off of cucumber respectively. The in vitro mode of action of the commercial Trichoderma product, Eco-T®, and Bacillus B69 and B81 suggested the production of antimicrobial substances active against R. solani. In vitro interaction studies on V8 tomato juice medium showed that the Trichoderma and Bacillus isolates did not antagonise each other, indicating the possibility of using the two organisms together for biological control and plant growth promotion studies. Greenhouse studies indicated that combined inoculation of T. atroviride SYN6 and Bacillus B69 gave the greatest plant growth promotion (43.0% over the uninoculated control) of bean seedlings in terms of seedling dry biomass. This was confirmed during in vivo rhizotron studies. However, results obtained from two successive bean yield trials in the greenhouse did not correlate with the seedling trials. Moreover, no increase in protein or fat content of bean seed for selected treatments was observed. In the biological control trials with cucumber seedlings, none of the Trichoderma and Bacillus combinations was better than single inoculations of Eco-T®, T. atroviride SY3A and T. harzianum SYN. Under nutrient limiting conditions, dry bean plants treated with single and dual inoculations of Trichoderma and Bacillus isolates exhibited a greater photosynthetic efficiency that the unfertilized control plants. Bacillus B77, under nutrient limiting conditions, caused 126.0% increase in dry biomass of bean seedlings after a 35-day period. Nitrogen concentrations significantly increased in leaves of plants treated with Trichoderma-Bacillus isolates. However, no significant differences in potassium and calcium concentrations were found. Integrated control (i.e. combining chemical and biological treatments) of R. solani damping-off of cucumber seedlings proved successful. In vitro bioassays with three Rizolex® concentrations, viz., 0.01g.l-1, 0.1g.l-1 and 0.25g.l-1 indicated that the selected Trichoderma isolates were partly sensitive to these concentrations whereas the Bacillus isolates were not at all affected. In a greenhouse trial, up to 86% control was achieved by integrating 0.1g.l-1 Rizolex® with T. harzianum SYN, which was comparable to the full strength Rizolex® (1g.l-1) application. Irrespective of either a single or dual inoculations of Trichoderma and/or Bacillus isolates used, improved percentage seedling survival as achieved with the integrated system, indicating a synergistic effect. The results presented in this thesis further reinforce the concept of biological control by Trichoderma and Bacillus spp. as an alternative disease control strategy. Furthermore, this thesis forms a basis for Trichoderma-Bacillus interaction studies and proposes that the two organisms could be used together to enhance biological control and plant growth promotion. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.

For millennia, scions have been grafted onto dwarfing apple rootstocks to reduce final tree size. However, it is unclear how scion architecture is first modified by the dwarfing apple rootstock, the time from grafting when this occurs and the endogenous hormonal signalling mechanisms that may cause the initial modifications in growth that then define the future architecture of the scion. In this study, the dwarfing (M.9) rootstock significantly decreased the mean total shoot length and node number of ‘Royal Gala’ apple scions by the end of the first year of growth from grafting when compared with rootstock(s) of greater vigour (MM.106, M.793 and a ‘Royal Gala’ rootstock control). Similarly, the auxin transport inhibitor 1-N-naphthylphthalamic acid (NPA) applied to the stem of vigorous rootstocks significantly decreased mean total shoot length and node number of the scion, and the architectural changes imposed were generally similar to those imposed by M.9. For example, both treatments decreased the mean length and node number of the primary shoot, reduced the formation of secondary axes on the primary shoot and caused a greater proportion of primary and secondary shoots (if present) to terminate growth early. Decreased formation of secondary axes imposed by both treatments was reversed by applying the cytokinin benzylaminopurine (BAP) repeatedly to the scion, whilst applications of gibberellins (GA4+7) reduced the proportion of primary and secondary shoots that terminated growth early, therefore increasing the final mean length and node number of these shoot types. Both M.9 and NPA also significantly decreased the final mean dry mass and length of the root system. Given these general similarities, it is proposed that the basipetal IAA signal is of central importance in rootstock-induced scion dwarfing, and that a shoot/root/shoot signalling mechanism may exist whereby the stem tissue of the M.9 rootstock decreases the basipetal transport of IAA to the root during summer, thereby decreasing root growth and the amount of rootproduced cytokinin and gibberellin transported to scion. Reduced amounts of cytokinin transported to the scion may decrease branching, whilst reduced amounts of gibberellins may decrease the duration for which a large proportion of primary and secondary shoots grow. Analysis of endogenous hormones for newly grafted composite ‘Royal Gala’ apple trees on rootstocks of different vigour provided some additional support for these ideas. It is recommended that future studies elucidate what unique properties of the M.9 bark act to restrict IAA transport, whilst it is concluded that gene(s) regulating rootstock-induced scion dwarfing are likely to control processes within the rootstock that modify the metabolism of IAA, its basipetal transport and the subsequent synthesis of root-produced vigour-inducing hormones including cytokinins and gibberellins.

Willow and poplar are the main trees used for soil erosion control in New Zealand (NZ) with successful establishment critical to greater use of this technology. Five experiments were conducted at the Pasture and Crop Research Unit, Massey University Palmerston North, NZ to examine the ability of willow and poplar to establish as a willow/poplar pasture system in NZ over a period of 3 years from December 2002 to April 2005. With an objective to select the best size for low cost planting, stem diameters (10 mm, 25 mm and 35 mm) were planted vertically and stem lengths (50 mm, 200 mm and 600 mm) were planted horizontally to determine their growth, establishment, biomass production and regrowth after browsing. From December 2002 to March 2003, three management treatments, mowing, herbicide and control (no weed control) treatments, were designed and applied to determine their effects on growth and shoot biomass production. From September 2004 to April 2005, mowing was replaced by sheep browsing and the effect of browsing, herbicide and control treatments on soil and tree water status (soil water content, soil water loss and deficit, and stem water potential) were examined. Longer (600 mm) and thicker stem diameters (35 mm) produced the greatest shoot biomass (edible biomass, total biomass and root biomass) compared to the thin stem cuttings (50 mm length and 10 mm diameter). Higher tree survival was also found for longer (600 mm) and thicker stems diameter (35 mm). The mown treatment produced significantly more edible and total shoot dry matter (DM) than the herbicide and control treatments, with willow clone 'Kinuyanagi' producing higher total shoot DM than 'Tangoio'. Pasture management had no significant effect on soil water content during spring 2004 and late summer/autumn 2005, however, it was significant during early and mid summer. Strong relationships between (i) soil water content and stem water potential (SWP), and (ii) SWP and soil water deficit were found and could help growers to predict the amount of water required during the growing season. The results clearly demonstrated that sheep grazing had negligible damaging effect on willow and poplar (main stem, branch breakage and tree leaning) and tree mortality, and that young trees can be browsed during pasture shortages in summer drought. Cutting size and understorey control for establishing willow and poplar into pasture have been better defined by this research. It is recommended that farmers establishing willow and poplar for fodder plant thick (vertical planting) and long stems (horizontal planting) for higher growth and biomass production.