Gibberella zeae (anamorph: Fusarium graminearum) is the causal agent of fusarium head blight (FHB) and maize ear rot, two major diseases of wheat and corn in Eastern Canada. / In Quebec, Microsphaeropsis sp., an antagonist of Venturia inaequalis, the causal agent of apple scab, was isolated from the apple leaf litter. This fungus, well adapted to Quebec climate, can reduce the initial inoculum of V. inaequalis. FHB and Gibberella ear rot are similar to apple scab because the major inoculum source comes from melanized structures produced on crop residues. Consequently, we evaluated the potential of Microsphaeropsis sp. (isolate P130A) to inhibit ascospore production of G. zeae when applied to crop residues as post harvest or pre-planting applications. Under in vitro conditions, the antagonist significantly reduced ascospore production on wheat and corn residues, when applied prior to (82% and 92% respectively) or at the same time as the pathogen (36% and 58% respectively). Under field conditions, the antagonist had no effect on the pattern of perithecia maturation but significantly reduced the number of ascospores produced on two sampling dates, May 1998 and July 1999. (Abstract shortened by UMI.)

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.

E’ stata approfondita l’espressione genica complessiva in spighe di mais, in seguito all’ infezione fungina. Nella prima parte del lavoro, sono stati valutati un genotipo di mais resistente ed uno suscettibile a F. verticillioides, campionando le cariossidi 48 ore dopo l’infezione. Sono state identificate circa 800 sequenze differenzialmente espresse e circa il 10% è stato assegnato alla categoria della difesa. Nel genotipo resistente, i geni coinvolti nella difesa hanno mostrato un tipo di risposta basale, mentre in quello suscettibile tali geni rispondevano specificamente all’infezione. Nella seconda parte del lavoro, l’analisi di espressione è stata estesa a fasi precoci e tardive dell’infezione utilizzando un ceppo normale ed uno mutante di F. verticillioides. Numerosi geni risultavano differenzialmente regolati 48 ore dopo l’infezione con entrambi i ceppi. Il ceppo normale era in grado di attivare i meccanismi di difesa prima del mutante. Nella terza parte del lavoro, 10 linee resistenti e suscettibili sono state infettate con 4 specie fungine. In tutti i genotipi l’espressione dei geni coinvolti nella difesa era indotta in seguito all’infezione, ma le linee resistenti presentavano una risposta basale di difesa. / We investigated global gene expression in maize ears at several time points after fungal infection. In the first part of the work, resistant and susceptible genotypes were tested in kernels sampled 48 h after infection with a wild type strain of F. verticillioides. About 800 differentially expressed sequences were identified and nearly 10% assigned to the category cell rescue, defense and virulence. In the resistant genotype, defense-related genes provided basic defense against the fungus, while in the susceptible genotype defense genes responded specifically to pathogen infection. In the second part of the work the expression analysis was extended to early and late phases of infection with a wild type and a mutant strains of F. verticillioides. Kernels were sampled in the area around the point of infection. Most of genes were differentially regulated 48 h after infection with both fungal strains. The wild type strain was able to activate host defense genes before the mutant strain. In the third part of the work, ten resistant and susceptible lines were infected by different fungal species. All genotypes were able to induce the expression of defense genes upon infection, but the resistant lines showed a basal defense response.

Sorghum [Sorghum bicolor (L.) Moench] is a major food security crop for millions of people in sub-Saharan Africa and the fourth most important crop in Africa. The potential sorghum yields are limited due to a number of abiotic, biotic and socio-economic constraints. Among the biotic stresses is the parasitic weed, Striga hermonthica, which inflicts yield losses ranging from 30-100%. Various control options have been recommended to reduce levels of Striga damage. However, these techniques need to be integrated for effective control and to boost sorghum productivity. A series of experiments was conducted to integrate host resistance improvement and the use of a biological control agent, Fusarium oxysporum f.sp. strigae to control Striga hermonthica. These studies were also focused on improving breeders‟ awareness of the traits that farmers‟ desire, on the assumption that farmers‟ variety preference traits are the missing link in technology development and adoption process for S. hermonthica management. The objectives of the study were to: 1) determine farmers‟ views on sorghum production opportunities; threats; indigenous knowledge and perceptions; breeding priorities; Striga infestation; and the coping mechanisms of farmers in the north eastern and north western Ethiopia, 2) evaluate sorghum genotypes for compatibility to F. oxysporum inoculation where grown in Striga infested soil in controlled environments, 3) determine field responses of sorghum genotypes and F. oxysporum compatibility for integrated Striga management (ISM), 4) determine the variability present among selected sorghum genotypes exhibiting S. hermonthica resistance, and compatibility with the biological control agent using phenotypic and simple sequence repeat (SSR) markers, 5) identify F. oxysporum compatible sorghum parents and hybrids with high combining ability for grain yield, yield components, and Striga resistance for ISM, and 6) undertake farmers‟ participatory assessment, and identify their preferred traits for sorghum genotypes under ISM, simultaneously with the breeders‟ evaluation. A participatory rural appraisal (PRA) research was conducted involving 315 farmers in nine districts of three administrative zones within two provinces in Ethiopia. Sorghum landraces were preferred by >85% of participants rather than previously improved released varieties. The participating farmers listed and prioritized their sorghum production constraints. In the North Shewa and North Wello zones drought was the most important constraint, followed by Striga. In the Metekel zone Striga was the number one constraint followed by a lack of genotypes with high grain quality. Controlled environment experiments were conducted involving greenhouse and laboratory tests in order to evaluate 50 sorghum genotypes for their compatibility with F. oxysporum and for possible deployment of the bio-control agent to control Striga. Striga population was reduced by 92% through the application of F. oxysporum, resulting in yield increment of 144%. Twelve sorghum genotypes were identified as promising parents for breeding and to control Striga through integration of host resistance and F. oxysporum seed treatment. During field and sick plot plot evaluations differential responses to F. oxysporum application among the sorghum genotypes were observed for various attributes including Striga plant height. Most traits showed highly significant (p<0.001) genotype X site interactions. Similarly, the main effects of F.oxysporum application were highly significant (p<0.001) across sites for most of the traits. The genotype and genotype X environment biplot identified 13 genotypes that consistently performed well following Fusarium application. The variability present among 14 selected sorghum genotypes exhibiting S. hermonthica resistance, and compatibility with a biological control agent, Fusarium oxysporum, were determined using phenotypic and 20 polymorphic simple sequence repeat (SSR) markers. Highly significant (p<0.001) differences were detected among genotypes for phenotypic traits. Principal component analysis showed three components that accounted for 73.99% of the total variability exhibited among genotypes. Cluster analysis allocated the genotypes into two major groups, one with a further two subgroups based on morphological traits, showing clear demarcations between the genotypes. The SSR markers revealed high levels of polymorphisms among genotypes, with the mean number of alleles per locus being 6.95 and the mean polymorphic information content being 0.80. The observed genetic diversity was relatively wide, with the allele sizes ranging from 203.6-334 bp. The SSR markers allocated genotypes into two distinct clusters close to the phenotypic markers. Forty sorghum hybrids were developed through a line by tester mating design involving 10 lines selected for their compatibility with F. oxysporum and high agronomic performances and four Striga resistant tester parents. The F1s and their parents were field evaluated with complementary in-vitro tests. Field evaluations were conducted at two locations: Kobo and Shewa Robit in Ethiopia, which are well known for their severe Striga infestation. Significant (p<0.05) general combining ability (GCA) effects were observed among testers and lines at both sites for days to 50% flowering and maturity, plant height, biomass, number of Striga plants and Striga plant height. Furthermore, significant (p<0.05) specific combining ability (SCA) effects were detected for days to 50% flowering, biomass, grain yield and number of Striga plants. From the complementary in-vitro experiment, highly significant variation (p<0.01) was exhibited due to line x tester interaction for maximum Striga germination distance. The study identified paternal parents with high GCA effects including SRN-39 and Birhan and maternals 235761, 2384443, IC9830, 235466, 237289,235763, and 235929 to be useful for breeding for ISM in sorghum. At Kobo, cross 235763 x N-13 and Shewa Robit IC9830 x SRN-39 had significantly negative SCA effects for the numbers of Striga plants. Progenies of these crosses will be selected in the Striga resistance breeding program. In the participatory sorghum genotypes assessment, farmers were invited to assess and select the genotypes based on their preferences at maturity and harvesting. The standard agronomic traits and Striga parameters relevant for breeding were collected by the breeders. Earliness, Striga resistance, high yield and high grain quality and threshability were the most important farmers‟-preferred traits for sorghum genotypes. Comparative analyses between farmers‟ and breeders‟ evaluations revealed highly significant correlations (p<0.01) except between Striga resistance and Striga damage and pest resistance and insect damage. Repeatability of scoring genotypes among farmers was consistent (>0.80) for all traits except Striga and pest resistance. The prioritized traits through farmers‟ participation are important for further breeding program. Overall, the study established farmers‟ preferred traits, the effectiveness of ISM to boost sorghum productivity, and identified useful parents and crosses for effective sorghum breeding to control Striga in Ethiopia. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2013.

La fusariosi della spiga è tra le malattie del grano più distruttive e diffuse al mondo. Alcune specie responsabili di questa malattia sono micotossigene. Ci sono state segnalazioni della presenza di nivalenolo (NIV) e tossine T-2 e HT-2 nel nord Italia, micotossine prodotte da Fusarium poae, F. sporotrichioides e F. langsethiae. I limiti massimi di T-2 e HT-2 ammessi nei cereali non trasformati e nei prodotti a base di cereali non sono ancora stati individuati (Regolamento CE 856/2005). Il programma di dottorato del candidato è inserito in questa linea di ricerca. Sono stati condotti studi in vitro sulla biologia ed ecologia dei funghi produttori di NIV, T-2 e HT-2, considerando quattro isolati di F. poae, due di F. sporotrichioides e due di F. langsethiae. Le prove hanno riguardato la crescita delle colonie, la produzione di spore, la loro germinazione e la produzione di micotossine. La gravità dell'infezione, l'invasione (quantità di DNA nelle spighe), la resa in granella e la produzione di tossine sono state misurate a diversi tempi, temperature (in planta) e stadi fenologici (in campo) dopo l'inoculazione artificiale. Al fine di verificare l'attendibilità dei dati, tutti gli esperimenti in planta e in campo sono stati ripetuti per due anni. / The Fusarium head blight is one of the most destructive diseases of wheat in different parts of the world. Some of the species responsible for Fusarium head blight are potentially mycotoxigenic. There are reports of nivalenol (NIV), T-2 toxin and HT-2 in northern Italy. These are metabolites produced by F. poae, F. sporotrichioides, and F. langsethiae. The maximum limits of T-2 and HT-2 permitted in unprocessed cereals and cereal-based products, included in EC Regulation 856/2005 has not been identified. The PhD program of the candidate is placed in this line of the research. In vitro studies conducted on the biological and ecological of pathogenic fungi producer NIV, T-2 and HT-2 including four isolates of F. poae, two of F. sporotrichioides and two isolate of F. langsethiae. Tests were carried out to investigate the conidia germination, colony growth, spore production and mycotoxins production. The infection severity, invasion (amount of DNA in spikes), grain yield and toxin production were measured at different temperatures, different time (in planta) and different growth stages (in field) after artificial inoculation.

Banana is the most important fruit crop in the world but ironically one of the crops least studied. This fruit constitutes a major staple food for millions of people in developing countries and also it is considered the highest selling fruit in the world market making this crop a very important export commodity for the producing countries. At the present time, one of the most significant constraints of banana production that causes significant economical losses are fungal diseases. Among these, Panama disease, also known as Fusarium wilt has been the most catastrophic. Panama disease is caused by the soil-borne fungus Fusarium oxysporum formae specialis (f.sp) cubense (FOC), which infects susceptible bananas through the roots causing a lethal vascular wilt. To date, the race 4 of this pathogen represents the most serious threat to banana production worldwide since most of the commercial cultivars are highly susceptible to this pathogen. Introduction of FOC resistance into commercial cultivars by conventional breeding has been difficult because edible bananas are sterile polyploids without seeds. Genetic transformation of banana, which has already been established in various laboratories around the world has the potential to solve this problem by transferring a FOC race 4 resistance gene into susceptible banana cultivars (eg. Cavendish cultivars). However, a FOC resistant (R) gene has not been isolated. Genes that confer resistance to Fusarium oxysporum have been isolated from tomato and melon using a map-based positional cloning approach. The tomato I2 and melon Fom-2 genes belong to the non-Toll/interleukin like receptors (TIR) subclass of nucleotide-binding site and leucine-rich repeat (NBS-LRR) R genes. These genes confer resistance only to certain races of F. oxysporum in their corresponding plant families limiting their use in other plant families. The fact that these two Fusarium resistance genes share the same basic non-TIR-NBS-LRR structure suggests a similar Fusarium resistance mechanism is shared between the families Solanaceae and Cucurbitaceae. This observation opens the possibility to find similar Fusarium resistance genes in other plant families including the Musaceae. A remarkable discovery of a population of the wild banana Musa acuminata subspecies (ssp.) malaccensis segregating for FOC race 4 resistance was made by Dr. Ivan Buddenhagen (University of California, Davis) in Southeast Asia. Research carried out at Queensland Department of Primary Industries (Australia) using this plant material has demonstrated that a single dominant gene is involved in FOC race 4 resistance (Dr. Mike Smith, unpublished results). Tissue-culture plantlets of this FOC race 4 segregating population were kindly provided to the Plant Biotechnology Program (Queensland University of Technology) by Dr. Mike Smith to be used in our research. This population holds the potential to assist in the isolation of a FOC race 4 resistance gene and other potential Fusarium resistance genes. The overall aims of this research were to isolate and characterise resistance gene candidates of the NBS-type from M. acuminata ssp. malaccensis and to identify and characterise potential Fusarium resistance genes using a combination of bioinformatics and gene expression analysis. Chapter 4 describes the isolation by degenerate PCR of five different classes of NBS sequences from banana (Musa acuminata ssp malaccensis) designated as resistance gene candidates (RGCs). Deduced amino acid sequences of the RGCs revealed the typical motifs present in the majority of known plant NBS-LRR resistance genes. Structural and phylogenetic analyses showed that the banana RGCs are related to non-TIR subclass of NBS sequences. The copy number of each class was estimated by Southern hybridisation and each RGC was found to be in low copy number. The expression of the RGCs was assessed by RT-PCR in leaf and root tissues of plants resistant or susceptible to Fusarium oxysporum f. sp. cubense (FOC) race 4. Four classes showed a constitutive expression profile whereas no expression was detected for one class in either tissue. Interestingly, a transcriptional polymorphism was found for RGC2 whose expression correlated with resistance to FOC race 4 suggesting a possible role of this gene in resistance to this devastating FOC race. Moreover, RGC2 along with RGC5 showed significant sequence similarity to the Fusarium resistance gene I2 from tomato and were chosen for further characterisation. The NBS sequences isolated in this study represent a valuable source of information that could be used to assist the cloning of functional R genes in banana. Chapter 5 describes the isolation and characterisation of the full open reading frame (ORF) of RGC2 and RGC5 cDNAs. The ORFs of these two banana RGCs were predicted to encode proteins that showed the typical structure of non-TIR-NBS-LRR resistance proteins. Homology searches using the entire ORF of RGC2 and RGC5 revealed significant sequence similarity to the Fusarium resistance gene I2 from tomato. Interestingly, the phylogenetic analysis showed that RGC2 and RGC5 were grouped within the same phylogenetic clade, along with the Fusarium resistance genes l2 and Fom-2. These findings suggest that the banana RGC2 and RGC5 are potential resistance gene candidates that could be associated with Fusarium resistance. The case of RGC2 is more remarkable because its expression was correlated to FOC race 4 resistance (Chapter 4). As a first step to test whether RGC2 has a role in FOC race 4 resistance, different expression constructs were made with the ORF of this sequence. One of the constructs contains a RGC2 putative promoter region that was successfully cloned in this work. These constructs will be used to transform susceptible banana plants that can then be challenged with FOC race 4 to assess whether resistance has been acquired by genetic complementation. The results of this thesis provide interesting insights about the structure, expression and phylogeny of two potential Fusarium resistance genes in banana, and provide a rational starting point for their functional characterisation. The information generated in this thesis may lead to the identification of a Fusarium resistance gene in banana in further studies and may also assist the cloning of Fusarium resistance genes in other plant species.

In previous studies silicon has been associated with reduced disease severity and incidence, the enhanced accumulation of phenolic compounds and lignin, and with changes in the defence-related enzyme activity and transcript abundance of defence and stress related genes. All of these aspects of plant defence were considered in this study on cotton infected with Fusarium oxysporum f. sp. vasinfectum (Fov), and the results obtained have greatly enhanced our understanding of the effects of silicon on this interaction. In all experiments conducted, defence responses were only significantly enhanced by silicon treatment following inoculation with Fov, strongly suggesting that silicon can prime defence responses in cotton infected with Fov. Sicot F-1 was the cultivar most resistant to Fov infection at the commencement of this research, whilst Sicot 189 was considered to have moderate resistance to the pathogen. Vascular discolouration was significantly reduced in the more resistant cultivar, Sicot F-1 following treatment with potassium silicate, compared to mock inoculated plants and inoculated plants treated with potassium sulphate or calcium sulphate. No significant differences between treatments were observed in the moderately resistant cultivar, Sicot 189, though further trials may need to be conducted to confirm this result. In both cultivars, silicon content was significantly greater in plants which had been treated regularly with liquid potassium silicate, rather than with calcium silicate powder. Histological investigation of cotton infected with Fov, with and without silicon treatment, was conducted to ascertain the effects of this element on the accumulation of fungitoxic phenolic compounds, cell ultrastructural changes and fungal infection structures. Fov proliferated through the cortex and stele of plants from both the resistant (Sicot F-1), and moderately resistant (Sicot 189) cultivars, regardless of silicon treatment. However, defences were more rapidly and intensely induced in endodermal and vascular regions of inoculated, potassium silicate treated Sicot F-1 plants. Significantly more phenolic compounds were present at seven days post infection (dpi) in root extracts of inoculated, potassium silicate treated Sicot F-1 plants. Phenolic compounds were not significantly increased in inoculated, potassium silicate treated root extracts of Sicot 189 plants at three or seven dpi. Lignin assays demonstrated that the dry weight percentage of lignin in root material from inoculated, potassium silicate treated Sicot F-1 plants was significantly higher than that of extracts from inoculated plants not receiving silicon treatment at three dpi. This trend was also observed at seven dpi; however lignin content was not significantly different in this case. Percentage lignin content in the roots of Sicot 189 plants was not significantly different between inoculated potassium silicate treated plants and those not treated with silicon. Histological alterations were not observed in mock inoculated water or potassium silicate treated plants, nor were any significant increases in phenolic compounds or lignin accumulation detected in control treatments not inoculated with the pathogen. The expression of several defence related genes was assessed with quantitative reverse transcriptase real-time polymerase chain reaction. The results obtained verify that potassium silicate can enhance defence responses in Sicot 189 and Sicot F-1 plants inoculated with Fov, with silicon having a more pronounced effect on the more resistant cultivar, Sicot F-1. Genes upregulated at three and four dpi in potassium silicate treated, Fov inoculated Sicot F-1 plants included peroxidase, cadinene synthase and polygalacturonase inhibiting protein (PGIP), with peroxidase associated with phenol oxidation and lignification and cadinene synthase with phytoalexin biosynthesis. Osmotin-like protein and chitinase class I were consistently upregulated in potassium silicate treated, inoculated Sicot 189 plants; both genes coding for pathogenesis related (PR) proteins, with chitinase also classified as an antifungal protein. In both cultivars, silicon treatment without Fov inoculation did not result in the significant up-regulation of any of the defence genes assessed, providing further evidence for the role of silicon in priming in this interaction. The activities of three defence related enzymes, peroxidase, chitinase and β-1, 3- glucanase was assessed in root and shoot material by colourimetric assays. Regular application of potassium silicate significantly increased the activity of peroxidase in root extracts from the highly resistant cultivar Sicot F-1, at three, four and seven dpi with Fov, and in root extracts from the moderately resistant Sicot 189 at three and four dpi. Significant increases in chitinase activity in inoculated, silicon treated Sicot 189 plants were observed in root extracts at three dpi, and in shoot extracts at four dpi. Soluble potassium silicate treatment resulted in significant increases in β-1, 3- glucanase activity in Sicot 189 root extracts at four dpi. Few significant differences between treatments in terms of chitinase and β-1, 3- glucanase activity were detected in Sicot F-1 plants, though higher levels of each of these enzymes were present in root and shoot extracts from this cultivar. In this study the effects of acibenzolar-S-methyl, applied in the form of Bion®, on defence gene expression and enzyme activity in cotton infected with Fov were more pronounced in plants cultivated from treated seed, rather than in plants treated via foliar spray; a finding which is particularly relevant to the industry presently. Significant up-regulation of chitinase class I, peroxidase, and β-1, 3-glucanase transcripts and enzyme activities occurred in the Bion® seed soak treatment with Fov inoculation compared to all other treatments. It was possible to compare the actions of silicon with those of Bion® in this study. Bion® primed defence responses in cotton infected with Fov, in a manner similar to that observed in silicon treated cotton. The use of silicon and Bion® treatments, both alone and in combination as part of integrated disease management programmes, may potentially contribute to increased protection against this pathogen in Australian cotton fields in the future.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv., 2007. / Härtill 5 uppsatser.

No description available.