The Nantucket pine tip moth, Rhyacionis frustrana (Comstock) Is a common pest of young pines. In the eastern United States all species of yellow pines are attacked to some degree except longleaf pine, Pinus palustrla Mill. (Yates 1960). Chemical control of the tip moth has been demonstrated to be effective, but It generally has been considered to be too expensive since the evidence was only Inconclusive as to any permanent additional height growth of pines resulting from complete control. There has been considerable speculation about silvicultural control and a number of methods have been suggested by various workers. Many of these methods, however, have not been backed by quantitative data. Some of the suggested silvicultural controls are mixing susceptible species with resistant species, starting. trees under an overstory, using close spacing, and starting trees In brush. To date, there have been no detailed investigations of silvicultural control for the tip moth. This study was designed to determine if stands of loblolly pine, Pinus taeda L., growing under various conditions show any differences in rate of attack by the tip moth that might, In the future, be a basis for silvicultural or integrated control. / Master of Science

Colonization of Pine Christmas trees by Verticicladiella procera Kendr. causes Procera root disease. Little is presently known regarding the pattern and effects of fungal development within colonized trees. The present studies were undertaken to elucidate the developmental pattern of the fungus in colonized trees, to gather information on possible mechanisms and physiological effects of disease development, and to explore the relationship between V. procera and other, well documented bluestain fungi. The presence of cellulose was demonstrated in the cell walls of X. procera, indicating the probable genetic relatedness of this fungus with Ophiostoma (Ceratocystis) bluestain fungi. Inoculation studies revealed that the fungus could penetrate wounded sapwood, and that colonized seedlings had lower water potentials than uncolonized seedlings. In addition, it was found that the fungus could persist in resinous stem lesions for 22 months without foliar symptoms, and resinous stem lesions with the fungus were significantly longer and deeper than wound lesions. An intensive isolation study revealed that the initial point of colonization in a tree is apparently at the root collar, progressing acropetally in both directions. Analysis of radial growth from increment cores showed that colonized trees had grown more slowly for the preceding three years than uncolonized trees. The sapwood moisture content of these cores was also significantly reduced in the colonized trees, indicating that the stem was drying out as symptoms developed. Histological examination of colonized sapwood showed that U fungal colonization of tissues progressed along rays and resin ducts, in a fashion similar to that of bluestain fungi. Permeability measurements demonstrated that symptomatic sapwood, either resin-soaked or black-stained, had significantly reduced water movement relative to asymptomatic sapwood. / Ph. D.

Summary: The future mass propagation of elite families of Pinus patula by cuttings is a realistic method of deployment if the short-term performance of cuttings and seedlings are confirmed at harvesting. This will impact significantly on the future outlook of forestry in South Africa as softwood yields are improved substantially through the introduction of material of high genetic value in commercial plantings. This, however, will require significant changes in future silviculture and other management practices as foresters and plantation staff learn to regenerate, maintain, and schedule the harvesting of cutting stands according to a different set of demands as a result of the change in plant type. Contrary to operational experience, cutting survival was similar to seedling survival in all field studies. This indicates that factors other than those that were studied and reported on, such as planting techniques, may be contributing to mortality. Also, due to the different root structure of cuttings they may be more fragile. The similar survival observed in these trials, therefore, may have been due to the close supervision given to the planting operations by the research staff. Although survival was similar, both plant types survived unacceptably poorly in the majority of studies with an average stocking of approximately 50% at one year. It is therefore anticipated that commercial stands will require several blanking operations in order to achieve an acceptable stocking in excess of 85% by the following planting season. The reduction in expected profitability as a result of blanking costs, delayed establishment, and the loss of improved genetic plant material, indicates that this is an area that still requires further research irrespective of what plant type is being planted. The pathogen, Fusarium circinatum, was commonly isolated from the planting stock before and after planting in two studies. Due to its virulent nature, it was assumed that mortality on the trees on which F. circinatum was isolated was principally due to this pathogen. At planting all plants were observed to be healthy and free of disease indicating that this pathogen maybe carried from the nursery to the field in a cryptic form, either inside or outside the plant tissue , which results in the death of the newly planted tree. In two field studies, where F. circinatum was commonly isolated, the application of Benomyl fungicide and to some extent the biological control agent Trichoderma harzianum at planting appeared to improve survival although this improvement was not significant. Laboratory studies, designed to determine alternatives to Benomyl fungicide, indicated that three fungicides (Octave, Folicur and Tilt), three sterilants (Sporekill®, Prasin®and Citex®) , as well as a biological control agent (T.harzianum), were all highly successful in controlling F. circinatum colony growth in vitro. It is recommended that these products undergo nursery testing , where the plant material is inoculated with F. circinatum spores, in order to test their efficacy and possible phytotoxicity in vivo before commercial application. Post-planting survival was also affected by site climate . Greater temperature extremes, as well as lower humidity and less rainfall resulted in poor survival. Plant dimension at planting was found to interact with site quality where it was a significant factor on a poor quality site. Optimal cutting dimensions at planting was a root collar diameter of 2.8 - 3.2 mm, and a stem height greater than 7 cm at planting for cuttings produced in cavities 90 ml in volume. Optimal seedling dimensions at planting were a root collar diameter of 1.8 - 2 mm, and a stem height of 10 - 15 cm for seedlings produced in cavities 80 ml in volume. In a separate study, plant morphological criteria influenced medium-term growth, where greater root mass and thicker cutting root collar diameters at planting improved field growth performance for seven years after planting. A greater root mass at planting was achieved by raising cuttings in containers that could support greater medium volume. From the study it was concluded that cuttings should be raised for an approximate period of 9 months in container cavities no smaller than 80 ml in volume and possess an oven-dry root mass of 0.3 - 0.5 g at planting. In addition to similar survival, the cuttings in this study grew either similarly to, or in some cases out-performed, the seedlings that were used as a control. Several other published studies indicate that hedge maturation poses the greatest threat to the success of softwood cutting deployment. This is especially true in clonal forestry and methods to maintain juvenility, such as cold storage of shoots and cryopreservation, require further research before clonal plantations of P. patula can be realised. In the studies carried out on family hedges in this report, the effect of donor hedge maturation was found to influence nursery management practice and the characteristics of rooted cuttings. The nursery data indicates that rooting efficiency, root system quality, and stem size and form, all decline with increasing hedge age particularly from two years after the date of sowing. A decline in root system quality was particularly apparent and was observed prior to a decline in rooting efficiency. If field trials indicate poorer performance from older hedges , it may be necessary to determine whether the causes are purely ontogenetic, morphological, or both before drawing final conclusions about hedge longevity. Until such results are known, it is recommended that P. patula cuttings should be propagated from seedling donors maintained as hedges , approximately 15 cm high, for a period not more than three years from the date of sowing. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.

Fusarium circinatum is a fungal pathogen that has had a serious impact on pine production throughout the world. It attacks most Pinus species including Pinus elliottii, Pinus patula and Pinus radiata. Infections in South Africa (SA) are largely on seedlings, and result in fatal seedling wilt. Accurate and quick detection systems suitable for field use are needed to monitor the spread of the disease and optimize fungicide applications. Detection of F. circinatum is currently based on visual observations of typical symptoms. However, symptoms are not unique to the pathogen and can be caused by other biotic and abiotic stress factors. Nucleic acid-based identification techniques using PCR are available for different fungal species. These are sensitive and accurate, but they are expensive and require skilled biotechnologists to conduct the assays. In this study an enzyme-linked immunosorbent assay (ELISA) was developed to identify F. circinatum in infected seedlings. This optimized ELISA is able to discriminate between F. circinatum and two other fungi that frequently affect pine. This method has advantages over other assays because of its ease of operation and sample preparation, sensitivity and the ability to run multiple tests simultaneously. Mycelium-soluble antigens from Diplodia pinea (=Sphaeropsis sapinea), F. circinatum and F. oxysporum were prepared in nutrient broth. Analysis of these antigens on SDS-PAGE indicated the presence of common antigens between the different fungal pathogens. Some antigens were expressed more by some isolates than by others. Separate groups of chickens were immunised with mycelium-soluble antigens from D. pinea, F. circinatum and F. oxysporum and exo-antigen from F. circinatum prepared in nutrient broth. A 34 kDa protein purified from SDS-PAGE specific for D. pinea was also used for immunisation. Five sets of antibodies were obtained including anti-D. pinea, anti-F. circinatum, anti-F. oxysporum, anti-F. circinatumexo and anti-D. pinea 34 kDa antibodies, respectively. Reactivity of these antibodies was evaluated against antigens prepared in nutrient broth using western blotting and ELISA. Western blot analysis indicated that immuno-dominant antigens for F. circinatum were larger than 34 kDa and their reactivity was not the same between different isolates. Each of the antibodies prepared using mycelium-soluble antigens showed increased reactivity when detecting its own specific pathogen, but cross-reactivity was observed. Anti-D.pineaantibodies showed minimal cross-reactivity with antigens from F. circinatum and F. oxysporum. Anti-F. circinatum antibodies cross-reacted with antigens from F. oxysporum but showed little cross-reactivity with D. pinea antigens. Anti-F. oxysporum antibodies showed more cross-reactivity towards antigens from F. circinatum than those from D. pinea. No reactivity was observed when anti-F. circinatum-exo antigen and anti-D. pinea 34 kDa antibodies were used in immuno-blotting analysis. Evaluation of antibody reactivity using indirect ELISA showed patterns similar to those observed on western blotting, where anti-D. pinea, anti-F. circinatum and anti-F. oxysporum antibodies showed the same cross-reactivity relationships. Anti-F. circinatum and anti-F. oxysporumantibodies showed a significant difference when reacting with antigens isolated from other pathogens including D. pinea, F. circinatum, F. oxysporum, F. solani, F. graminearum and F. culmorum (P = 0.001). No significant difference was observed when the antigens were detected with anti-D. pinea antibodies. Reactivity of anti-F. circinatum-exo and anti-D. pinea34 kDa antibodies was mostly similar to that of non-immune antibodies and showed no significant difference between detection of different antigens. Pine seedlings were artificially infected with the three fungal pathogens using a spore concentration of 1 – 1 x 106conidiaml-1.Infection was monitored using scanning electron microscopy. Results showed increased levels of mycelium growth on the stem and roots of the F. circinatum and F. oxysporum infected seedlings and on the leaves and stem in the case of D. pinea infected seedlings. These plant parts were used in ELISA tests for the detection of antigens. Isolation of antigens from the plant materials involved crushing plant parts in buffer and centrifugation of the suspension. The supernatant obtained was directly used in the assay. ELISA tests prepared in this study were sensitive enough to detect infection caused by 1 conidium ml-1at two weeks post inoculation. A positive reaction for detection of F. circinatum and F. oxysporum was indicated by an ELISA reading above an optical density at 405 nm. The plant material used in ELISA tests were further analysed using PCR. Results indicated that there was no cross-infection between seedlings and served as a confirmation of the disease-causing pathogen. This indicated that cross-reactivity observed was due to other factors such as common epitopes on the major antigens. Use of an ELISA dip-stick or ELISA using these antibodies should provide an easy, fast field test to identify infections of pine, discriminating between F. circinatum, F. oxysporum and D. pinea. / M.Sc.Agric. University of KwaZulu-Natal, Pietermaritzburg 2013.

The Eurasian wood wasp, Sirex noctilio, is considered a secondary pest in its native range; however, it has caused significant economic damage when introduced to pine plantations in the Southern Hemisphere. Sirex noctilio was recently introduced to the northeastern U.S., which has raised concerns about its potential impact on Southeastern pine plantations. This research was conducted to understand how silvicultural management affects populations of a native wood wasp, Sirex nigricornis, a wood wasp with similar ecosystem functions as S. noctilio. Sirex nigricornis abundance was higher in un-managed pine plantations than in managed plantations, mixed, and old growth forests. Additionally, geospatial models were built displaying S. noctilio hazard for the Southeastern U.S. based on oviposition host preference assays and historical outbreak information. Sirex noctilio hazard models will inform land managers about areas of greatest concern under various scenarios and should be used to decrease susceptibility of pine forests to this pest.

Studies were undertaken to determine the dispersal mechanisms of Verticicladiella procera Kendrick, the causal agent of Procera Root Disease (PRD). Propagule germinability in artificially infested soil decrease rapidly under natural and controlled conditions. Colonization of seedlings in artificially infested soil was rare and symptoms were not displayed by colonized seedlings. Natural populations of V. procera were closely associated with colonized root tissue. Colonization of field planted seedlings was related to proximity to root collars of diseased trees and insect activity on the seedlings. Insects (Coleoptera) contaminated with V. procera were found in plantations both with and without PRD. The percent of weevils and bark beetles contaminated with V. procera was 64 and 0.76 respectively. Verticicladiella procera was transmitted to white pine bolts in the field and under controlled conditions following visitation by contaminated insects. Verticicladiella procera was associated with larval galleries and frass in trap bolts and was observed fruiting in insect galleries in root systems of diseased trees. This evidence suggests that transmission by insects, especially weevils, is the more important mechanism for dispersal and that soil-borne propagules have a minor role in pathogen spread. / M.S.

Sirex noctilio is causing considerable mortality in commercial pine forests in KwaZulu- Natal, South Africa. The ability to remotely detect S. noctilio infestations remains crucial for monitoring the spread of the wasp and for the effective deployment of suppression activities. This thesis advocates the development of techniques based on remote sensing technology to accurately detect and map S. noctilio infestations. To date, no research has examined the potential of remote sensing technologies for the detection and mapping of Pinus patula trees infested by S. noctilio. In the first part of this thesis, the focus was on whether high spatial resolution imagery could characterize S. noctilio induced stress in P. patula forests. Results showed that, the normalized difference vegetation index derived from high spatial resolution imagery has the potential to accurately detect and map the later stages of S. noctilio infestations. Additionally, operational guidelines for the optimal spatial resolutions that are suitable for detecting and mapping varying levels of sustained S. noctilio mortality were defined. Results showed that a pixel size of 2.3 m is recommended to detect high (11-15%) infestation levels, and a pixel size of 1.75 m is recommended for detecting low to medium infestation levels (1-10%). In the second part of this thesis, the focus was on the ability of high spectral resolution (hyperspectral) data to discriminate between healthy trees and the early stages of S. noctilio infestation. Results showed that specific wavelengths located in the visible and near infrared region have the greatest potential for discriminating between healthy trees and the early stages of S. noctilio infestation. The researcher also evaluated the robustness and accuracy of various machine learning algorithms in identifying spectral parameters that allowed for the successful detection of S. noctilio infestations. Results showed that the random forest algorithm simplified the process by identifying the minimum number of spectral parameters that provided the best overall accuracies. In the final part of this thesis spatial modelling techniques were used to proactively identify pine forests that are highly susceptible to S. noctilio infestations. For the first time the random forest algorithm was used in conjunction with geographic information systems for mapping pine forests that are susceptible to S. noctilio infestations. Overall, there is a high probability of S. noctilio infestation for the majority (63%) of pine forest plantations located in Mpumalanga, South Africa. Compared to previous studies, the random forest model identified highly susceptible pine forests at a more regional scale and provided an understanding of localized variations of environmental conditions in relation to the distribution of the wasps. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2008.