M.Sc. / Polygalacturonase-inhibiting proteins (PGIPs) are cell wall-associated plant proteins that inhibit endopolygalacturonases from phytopathogenic fungi. It has been proposed that pgip encoding genes could be utilised for engineering increased resistance in transgenic crops against important fungal pathogens such as Botrytis cinerea. During this study a pgip gene from Malus domestica cv Granny Smith apple fruit was cloned by the degenerate and inverse polymerase chain reaction (PCR) techniques. An alignment of the pear and bean PGIP sequences was used to design degenerate PCR primers in highly conserved regions. Degenerate PCR allowed the amplification of a 351bp internal fragment of the pgip gene, termed ipgip. The DNA sequence of ipgip was used to design inverse PCR primers. A Southern blot of apple genomic DNA probed with the ipgip fragment was used to identify restriction enzyme sites for inverse PCR. Inverse PCR enabled cloning of the remainder of the gene, from which a composite pgip gene sequence was constructed. The composite apple pgip gene comprised an open reading frame of 990bp that is predicted to encode a 330 amino acid polypeptide. The polypeptide contains a putative 24 amino acid N-terminal leader sequence that may function as a signal peptide for secretion. The deduced apple PGIP contains nine cysteine residues and seven potential N-linked glycosylation sites. Ten loosely conserved leucine-rich repeat motifs characteristic of PG1Ps were identified in the apple PGIP sequence. The apple PGIP showed 97% and 55% amino acid identity to the pear and bean PGIPs, respectively. The full-length apple pgip gene was re-isolated from genomic DNA by PCR using primers designed to the 5' and 3' ends of the composite pgip gene. The apple pgip gene was cloned into a plant transformation vector and transformed into tobacco by Agrobacterium-mediated transformation. Phenotypically normal transgenic tobacco plants were produced. Stable transgene insertion into the transgenic tobacco genomes was verified by PCR and Southern blot analyses. Sequence analysis of the pgip construct used for transformation revealed two potential mutations in the deduced amino acid sequence. The substitutions of Asp residues with Asn and Tyr at positions 43 and 196, respectively, could interfere with the secondary structure of the expressed transgene protein. To test whether the apple PGIP was effective against Botrytis cinerea, protein extracts were prepared from apple fruit and transgenic tobacco and tested for inhibitory activity against B. cinerea polygalacturonases. Biochemical assays showed that a heat-denaturable PGIP extract prepared from apple fruit inhibited the polygalacturonases produced by a virulent isolate of Botrytis cinerea grown on pectin and apple cell walls. Protein extracts prepared from transgenic tobacco did not show any inhibitory activity towards Botrytis polygalacturonases. This suggests the absence of active PGIP in the extracts possibly due to inefficient transcription of the transgene or due to the introduced mutations.

Plant fungal pathogens are a major threat to food security worldwide. The most important method of protecting plants against fungal attack is the use of fungicides, but the development of resistance towards synthetic fungicides is of great concern. Moreover, the health risks associated with the use of chemical fungicides increase the need to search for safe, efficacious and environmentally friendly fungicides. Plants produce antifungal agents by secondary metabolism to protect themselves from fungal attack, and therefore many plant species have substantial antifungal activity. The use of plant extracts could enable the development of inexpensive and environmentally acceptable fungicides based on locally available natural products. This study was undertaken to investigate weedy and invasive plant species for antifungal activity against plant pathogens in order to develop a useful product using a widely available resource. Acetone leaf extracts of seven invasive species (Chromoleana odorata, Ipomoea alba, Tecoma stans, Passiflora suberosa, Passiflora subpeltata, Aristolochia sp, Solanum seaforthianum) were screened against eight plant fungal pathogens viz Rhizoctonia solani, Fusarium oxysporium, Penicillum janthinellum, Penicillum expansum, Aspergillus parasiticus, Aspergillus niger, Pythium ultimum and Phytophthora nicotiana, using microdilution assay and bioautography. The acetone extract of Tecoma stans had reasonable antifungal activity with an average minimal inhibitory concentration (MIC) value against all the fungi of 550 ìg/ml and clear zones on bioautograms indicating inhibition of fungal growth of a compounds with an Rf of 0.082 in BEA against several of the fungal pathogens. Due to the clear compound on bioautography and availability of Tecoma stans, this species was selected for further work. Bioassay-guided fractionation of the leaves of the Tecoma stans dichloromethane (DCM) extract obtained from solvent-solvent fractionation resulted in one major compound, oleanolic acid. The isolated compound had antifungal activity with an average MIC value of 130 ìg/ml against the 10 plant pathogenic fungi and clear bands with an Rf value of 0.082 on bioautograms, indicating fungal growth inhibition. It was surprising that the MIC value of the crude DCM extract was as high as that of the only compound with antifungal activity based on bioautography. These results clearly indicated the possibility of synergisms especially since the average total activity of the extract was nearly 6.5 times higher than that of oleanolic acid with total activity values of 60154 ml for the extract and 9262 ml for oleanolic acid. Cellular cytotoxicity of DCM extract and oleanolic acid was investigated using tetrazoliumbased colorimetric assay (MTT) on Vero monkey kidney cells. The toxicity of the extract and oleanolic acid was determined by LC50 values. The DCM extract and oleanolic acid were toxic with and LC50 of 0.413 mg/ml and 0.129 mg/ml respectively, lower than that of berberine the toxic compound used as control. However therapeutic index which can be defined here as the LC50 in (ìg/ml)/MIC in (ìg/ml), indicated that though the extract and oleanolic acid were toxic, they could be used under controlled conditions against infections of certain of the fungal pathogens. The crude extract had a high therapeutic index value of 21 against microorganisms T. harzianum, R. solani, F. oxysporium and P. expansum; and oleanolic acid had high therapeutic index values of 16 and 64 of against T. harzianum and R.solani respectively. This high therapeutic index value of crude extract and oleanolic acid means that, crude extract and oleanolic acid may be used for treatment of infections by these tested fungi with very little toxicity under controlled conditions. Oleanolic acid had very low antibacterial activity (MIC >250 ìg/ml). against two Grampositive (Staphylococcus aureus, ATCC 29213 and Enterococcus faecalis, ATCC 29212) and two Gram-negative bacteria (Escherichia coli, ATCC 27853 and Pseudomonas aeruginosa, ATCC 25922). Animal pathogenic fungi were more resistant than the plant fungal pathogens. Based on the good activity of the DCM crude extract, the surprising selectivity in activity against different fungi coupled with reasonably good therapeutic indexes and the wide availability of T stans leaves opens up the possibility that a commercial product to protect plants against certain pathogens may be developed from T. stans leaves. Copyright / Dissertation (MSc (Veterinary Science))--University of Pretoria, 2008. / Paraclinical Sciences / unrestricted

The ability to sense and respond to environmental stimuli is essential for the survival of all living things. As a result, nature has evolved an uncountable number of ways to detect environmental signals. At the cellular level, G protein-coupled receptors (GPCRs) are used by eukaryotes, including fungi and humans, to convert extracellular molecular binding events into intracellular responses. Recently, synthetic biologists have shown that biological sensing systems can be repurposed to suit human needs, developing tools such as diagnostic devices and drug screening platforms. In this thesis, I present work exploring the potential of fungal GPCRs to be used as sensing elements in yeast-based biosensors. Chapter 1 gives background information related to synthetic biology, biosensors, and yeast signaling pathways. Chapter 2 describes the development of the baker's yeast Saccharomyces cerevisiae into a diagnostic device for detection of fungal pathogens, using fungal GPCRs. In Chapter 3 I demonstrate that the substrate specificity of fungal GPCRs can be altered using directed evolution. Chapter 4 describes experiments further probing the native binding abilities of fungal GPCRs, specifically examining protein ligands. Finally, in Chapter 5 we move beyond fungal GPCRs and engineer yeast to detect other stimuli, in the context of an engineered living material.

No description available.

Fusarium oxysporum (Fo) is a soil-borne fungal pathogen that causes vascular wilt disease on a broad range of plants, including agricultural crops and the model plant Arabidopsis thaliana. There are non-pathogenic members of the Fo species complex that confer defense benefits against other pathogens to the host plant, however alteration to the host’s physiology through interaction with one of these strains, Fo47, have not been described. In this study, we aimed to establish the Fo47-A. thaliana interaction and determine if Fo47 reduces disease severity of a pathogenic Fo isolate, Fo5176. Additionally, we sought to use bioinformatics to mine transcriptomic data of the infection between Fo47 and A. thaliana for putative effectors from the non-pathogenic isolate using a pipeline that is validated by identifying known effectors in the interaction between Fo5176 and A. thaliana. Phenotypic characterization of A. thaliana plants inoculated with Fo47 or Fo5176 has revealed a significant increase in rosette biomass of Fo47 inoculated plants when compared to mock (sterile water) inoculated plants. As is observed in other systems, treatment of plants with Fo47 prior to challenging with pathogenic Fo significantly reduces the disease severity over time. The results of this study suggest that Fo47 is a possible biocontrol agent against Fo5176, and that inoculation with non-pathogenic Fo alters the physiology of A. thaliana such that it has a higher rosette biomass without alterations to the water status of the plant. Our pipeline for extracting putative effectors using transcriptomic data as a critical filter generated 13 candidate genes for further experimentation to determine their role in the Fo47-A. thaliana interaction. This research reports the first known observation that Fo47 increases the shoot biomass of the host plant it is interacting with, and that the model plant A. thaliana can be used as a host to examine the spectrum of interactions capable within the Fusarium oxysporum species complex.

A antracnose é uma das principais doenças que afetam a goiaba no Estado de São Paulo. Tanto Colletotrichum gloeosporioides quanto Colletotrichum acutatum, são relatados como sendo os agentes causais da doença. Os objetivos do trabalho foram caracterizar e identificar 54 isolados de Colletotrichum oriundos de lesões de goiaba, baseados nos aspectos culturais, morfológicos, moleculares e enzimáticos, além da caracterização patogênica de isolados representativos de cada espécie de Colletotrichum identificadas. A caracterização cultural foi avaliada mediante a mensuração do crescimento micelial dos isolados a 25 ºC, além dos aspectos culturais, como a coloração e a topografia das colônias. Na caracterização morfológica foram mensurados comprimento e largura de conídios, bem como avaliados os seus formatos. Oligonucleotídeos específicos foram utilizados na caracterização molecular, visando identificar as espécies dos isolados de Colletotrichum. A caracterização enzimática envolveu a mensuração, in vitro, do halo de degradação dos substratos da amilase, proteinase, celulase, pectinase e lipase. Por fim, alguns isolados representativos e identificados foram utilizados na caracterização patogênica, sendo avaliados os períodos de latência e incubação, a área lesionadas dos frutos e a esporulação. Baseados na coloração das colônias, os isolados foram reunidos em 9 grupos diferentes. Os mesmos puderam se reunidos em dois grupos distintos de acordo com a taxa do crescimento micelial. Os conídios apresentaram os formatos: (i) reto, fusiforme, com ápices afilados, (ii) reto, oblongo, com ápices arredondados, (iii) reto, clavado, afilado em uma extremidade e (iv) reto, com constrição. As dimensões variaram de 11,4 a 16,8 µm de comprimento por 2,6 a 4,9 µm de largura. O uso de oligonucleotídeos específicos permitiu identificar C. acutatum e C. gloeosporioides entre os isolados avaliados. Grande parte dos isolados, 94%, foram identificados como pertencendo à espécie C. gloeosporioides, enquanto que apenas 4% foram identificados como C. acutatum. Em relação à caracterização enzimática, apenas a atividade celulolítica proporcionou diferenças significativas entre C. gloeosporioides e C. acutatum. A patogenicidade dos isolados avaliados mostrou alta variabilidade na severidade da doença nos frutos, contudo não foi possível evidenciar diferenças significativas que distinguissem C. acutatum de C. gloeosporioides. Os períodos de incubação e latência foram menores para os isolados de C. acutatum em relação aos isolados de C. gloeosporioides. C. acutatum produziu quantidade superior de esporos nos frutos inoculados quando comparados a C. gloeosporioides. Observou-se, ainda, correlação positiva entre a área do halo de degradação de pectina, lipídio e amido e a área lesionada dos frutos afetados pelos isolados avaliados. / Anthracnose is one of the major diseases affecting guava in the State of São Paulo. Both Colletotrichum gloeosporioides and Colletotrichum acutatum are reported as the causal agents of the disease. The objectives of this work were to characterize and to identify 54 Colletotrichum isolates from guava, based on cultural, morphological, molecular, enzymatic, and pathogenic aspects. Cultural characterization was achieved by measuring the mycelial growth at 25 ° C, as well as reporting cultural aspects, such as color and topography of the colonies. In the morphological characterization it was measured length and width of conidia, and rated their shapes. CaInt2 and CgInt specific primers were used in the molecular identification of the Colletotrichum isolates. The enzymatic characterization was performed by measuring, in vitro degradation of starch, protein, cellulose, pectin and lipid. Finally some representative and identified isolates were used in the pathogenic characterization, evaluated by latency and incubation periods, diseased area and sporulation. Based on the color of the colonies, the isolates were grouped in 9 different groups. These same isolates showed two distinct growth paterns according to the mycelial growth rates. Conidia showed shapes: (i) straight, fusiform, with acute ends, (ii) straight, oblong, with round ends, (iii) straight, clavate, tapered at one end and (iv) straight, with a constriction in the middle. Conidia size ranged from 11.4 to 16.8 µm in length by 2.6 to 4.9 µm in width. The use of specific primers identified C. acutatum and C. gloeosporioides among the isolates. Most of the isolates (94%) were identified as C. gloeosporioides, while only (6%) were identified as C. acutatum. In the enzymatic characterization, only cellulolytic activity revealed significant differences between C. gloeosporioides and C. acutatum. Pathogenicity of the isolates was highly variable, but could not help to distinguish between C. acutatum and C. gloeosporioides. The incubation and latency periods were shorter for C. acutatum in relation to C. gloeosporioides. C. acutatum produced higher amounts of spores on inoculated fruits compared to C. gloeosporioides. There was also a positive correlation between in vitro degradation of pectin, lipid and starch, and the diseased area for tested isolates.

As condições climáticas presentes nas regiões produtoras de trigo, do Brasil e dos principais países do qual o produto é importado, favorecem o aparecimento de doenças importantes desta cultura, dentre elas a fusariose, causada principalmente pelo fungo Fusarium graminearum Schwabe. Além dos danos diretos causados pela doença, os grãos infectados podem ser tóxicos para o homem e animais devido à presença de micotoxinas especialmente o desoxinivalenol (DON). Um total de 100 amostras de trigo, sendo 50 de trigo nacional (provenientes do Estado de São Paulo, Paraná e Rio grande do Sul) e 50 de trigo importado (Argentina e Paraguai) foram coletadas de empresas que normalmente comercializam ou processam trigo durante o período de maio a dezembro de 2005. Foram avaliados o percentual de freqüência de fungos, especialmente Fusarium graminearum, a contaminação com DON, percentual de grãos giberelados e realizadas correlações entre os parâmetros avaliados. Os resultados indicaram que freqüência média de Fusarium graminearum e F. spp. foram baixas (&#8804;2,6 e &#8804;3,6%, respectivamente), porém foi maior no trigo nacional do que no trigo importado. Do total de amostras avaliadas 94% do trigo nacional e 88% do trigo importado apresentaram-se contaminadas com DON em níveis médios de 332 µg.kg-1 (nacional) e 90 µg.kg-1 (importado). Existiu correlação positiva e significativa entre contaminação com DON e percentual de grãos giberelados (r = 0,83; p<0,0001), freqüência de Fusarium graminearum (r = 0,92; p<0,0001) e freqüência de Fusarium spp. (r = 0,86; p<0,0001). / The climatic conditions present in wheat producing areas from Brazil and main countries from which the product is imported favor the occurrence of important diseases in this crop, among them the Fusarium Head Blight or scab. It is mainly caused by the fungus Fusarium graminearum Besides, direct damages caused by this disease, the infected kernels may be toxic for humans and animals due to presence of mycotoxins (e.g deoxynivalenol). A total of 100 wheat samples, being 50 from national production (São Paulo, Paraná and Rio Grande Do Sul states) and 50 from imported one (Argentina and Paraguay), were collected during the period of May to December 2005 from companies that normally commercialize or process wheat. Frequency (%) of fungi occurrence, specially Fusarium graminearum and Fusarium spp., DON contamination and Fusarium damaged kernels (%) were evaluated. Correlations between the evaluated parameters were carried out. Frequency of Fusarium graminearum and Fusarium spp. were low (&#8804;2.6 and &#8804;3.6%, respectively), however it was higher in Brazilian wheat when compared with imported wheat. Ninety-four percent of national wheat samples and 88% of the imported samples were DON contaminated (mean levels, 332 µg.kg-1 and 90 µg.kg-1, respectively). The occurrence of DON was highly correlated with percentage of Fusarium damaged kernels, (r = 0,83; p<0.0001), percentual frequency of Fusarium graminearum (r = 0,92; p<0,0001) and percentual frequency of Fusarium spp. (r = 0,86; p<0,0001).

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No. of bitstreams: 1 losnak_do_me_bot.pdf: 3093790 bytes, checksum: f1b838682d27c1345ac29cab3b2bfc9a (MD5) Previous issue date: 2017-02-21 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A emergência e reemergência de doenças infecciosas é impulsionada por vários fatores e a busca de patógenos em amostras animais podem oferecer oportunidades para estudos eco-epidemiológicos e também dados sobre a evolução dos patógenos. O objetivo deste estudo foi avaliar a ocorrência de fungos patogênicos importantes em saúde pública, em exemplares de animais silvestres mortos por atropelamento no estado de Santa Catarina e identificar e mapear áreas de risco para a infecção humana. Grande parte destes fungos apresenta em comum dimorfismo, distribuição geográfica restrita e produção de conídios infectantes que são aspirados pelo hospedeiro por meio das vias respiratórias. Cães e tatus são apontados como transmissores de Paracoccidioides brasiliensis, os morcegos ao Histoplasma spp., assim como as fezes de pombos ao Cryptococcus spp.. No presente trabalho foram analisadas 1063 amostras de pulmão, fígado, baço, pele e coração de 297 animais silvestres, para detecção de Paracoccidioides brasiliensis, Histoplasma capsulatum e Cryptococcus spp. pela técnica de Reação em Cadeia de Polimerase (PCR). Utilizou-se primers universais para detecção de fungos em geral e obteve-se positividade em 102 amostras de 59 animais. Para a análise de P. brasiliensis, utilizou-se os primers específicos, obtendo oito amostras positivas em cinco animais (quatro Oxymycterus spp. e um Euryoryzomys russatus). Não houve a detecção molecular para Histoplasma spp.. Foi possível a identificação de três amostras para Cryptococcus spp.. O sequenciamento foi realizado, porém em 89 amostras de 49 animais foi possível somente a identificação em Fungal sp. (GenBank KT923226.1), duas amostras para Cryptococcus neoformans (GenBank KY107218.1) obtidas de Oxymycterus spp. e Akodon spp. e três amostras de Aspergillus penicillioides (GenBank KP131612.1 e KP997215.1) obtidas de Gracilinanus spp., Oxymycterus spp. e Philander spp. Importante salientar que houve coinfecção de P. brasiliensis e Cryptococcus neoformans em amostra de um Oxymycterus spp. Esta pesquisa mostra a importância dos animais silvestres na transmissão de doenças e auxilia no mapeamento dos locais de ocorrência de determinados patógenos e doenças em uma região ainda não avaliada. / The emergence and reemergence of infectious diseases is propelled by many diverse factors and the search for pathogens in animal samples may offer opportunity for eco-epidemiologic studies as well as data on the evolution of pathogens. The objective of this study was to evaluate in samples of road-killed wild animals the occurrence of pathogenic fungi of importance for public health. A great part of these fungi presented, in common, dimorphism, restricted geographic distribution and production of conidia infecting, which are aspirated by the host by means of their respiratory tract. Dogs and armadillos are normally related to the transmission of Paracoccidioides brasiliensis, bats to Histoplasma spp., as well as pigeons feces to Cryptococcus spp.. In this study we analyzed 1063 samples of organs of 297 wild animals for the detection of Paracoccidioides brasiliensis, Histoplasma capsulatum and Cryptococcus spp. by the technique of Polymerase Chain Reaction (PCR). Universal primers were employed for the detection of fungi in general and positivity was obtained in 102 samples from 59 animals. For the P. brasiliensis analysis was used specific primers, resulting in eight positive samples from five animals (four Oxymycterus spp. and one Euryoryzomys russatus). There was no molecular detection to Histoplasma spp.. Was possible the identification of three samples to Cryptococcus spp.. The sequencing was performed, however in 89 samples from 49 animals was possible to identify Fungal sp. (GenBank KT923226.1), two samples for Cryptococcus neoformans (GenBank KY107218.1) obtained from Oxymycterus spp. and Akodon spp. and three samples from Gracilinanus spp., Oxymycterus spp. and Philander spp. were positive for Aspergillus penicillioides (GenBank KP131612.1 e KP997215.1). Is important emphasize the coinfection with P. brasiliensis and Cryptococcus neoformans in a sample from Oxymycterus spp.. This research shows the importance of the wild animals in transmissions of diseases and assists in the mapping of pathogen and disease sites in a region that has not yet been evaluated. / FAPESP: 2015/17519-4

Thesis (M.Sc. (Plant Protection )) --University of Limpopo, 2011 / Crude extracts of wild watermelon (Cucumis africanus) and wild cucumber (C. myriocarpus) fruits are widely used for both medicinal and ritual purposes in South Africa. Fruits are collected fresh from the wild, but have high incidence of post-harvest decay. A study was conducted to isolate and identify the pathogen responsible for post-harvest fruit decay, followed by the pathogenicity tests. Decayed fruits were individually surface-sterilised using 0.5% NaOCl, incubated at 25ºC to allow for decay, small rotten pieces were severed and placed on solidified plates of potato dextrose agar and incubated. At harvest, seven days after incubation, isolated fungus was repeatedly cultured for 21 days for verification of diagnostic characteristics. Based on the morphological characteristics, the pathogen associated with fruit rot of both Cucumis species was identified through the assistance of an expert as Penicillium simplicissimum (Oudem) Thom. Pathogenicity results suggested that P. simplicissimum was responsible for the observed fruit decay in both species, with the higher incidence being in C. africanus, probably due to its low pH. Due to the antibiotics that P. simplicissimum releases and its reduction of medium pH, the culture retained its purity, without any contamination. In conclusion, the pathogen that induces post-harvest fruit decay in C. africanus and C. myriocarpus is P. simplicissimum, which has the ability to reduce the pH of the growing medium and also produce antibiotics. / National Research Foundation

Phoma medicaginis is a necrotrophic fungal pathogen, commonly found infecting Medicago truncatula and M. sativa in temperate regions of Australia. To identify, characterize and differentiate eight P. medicaginis isolates from Western Australia, morphological phenotypes and five gene regions (actin, â- tubulin, calmodulin, internal transcribed spacer, translation elongation factor 1-á) were examined. Sequence comparisons showed that specimens isolated from M. truncatula in Western Australia formed a group that was consistently different from, but closely allied to, a P. medicaginis var. medicaginis type specimen. Characterization of three P. medicaginis genotypes showed that all exhibited a narrow host range, causing disease only in M. sativa and M. truncatula among eight commonly cultivated legume species sampled. Infection of 85 M. truncatula accessions showed a continuous distribution in disease phenotypes, with the majority of accessions susceptible. Differences in disease phenotypes suggest that M. truncatula harbours specific and diverse sources of resistance to individual P. medicaginis genotypes. To characterize the genetic basis of resistance to P. medicaginis two F2 populations derived from crosses between the resistant accession SA27063 and the susceptible accessions SA3054 and A17 were phenotyped for disease symptoms. Highly significant recessive QTLs for resistance to P. medicaginis OMT5 were identified in each mapping population. In SA27063 x A17 a QTL named resistance to the necrotroph Phoma medicaginis one (rnpm1) was identified on the short arm of LG4. In SA27063 x SA3054 a QTL (rnpm2) was identified on the long arm of LG8. Further fine mapping of the areas surrounding the QTLs is underway to identify the genes underlying rnpm1 and rnpm2. Examination of the recombination frequencies between genetic markers on the long arms of chromosomes 4 and 8 in the SA27063 x A17 cross revealed an apparent genetic linkage between these chromosomes. Subsequent analysis of other crosses showed this unexpected linkage relationship is characteristic for genetic maps derived from A17. Furthermore F1 individuals derived from crosses involving A17 showed 50% pollen viability or less. This semisterility and the unexpected linkage relationships provide good evidence for a reciprocal translocation in A17 between chromosomes four and eight. The implications of the distinctive chromosomal rearrangement in A17 on genetic mapping, genome sequencing and comparative mapping are discussed. The Mt16kOLI1plus microarray was used to identify transcriptional changes in M. truncatula expressed in defence against P. medicaginis. Three-hundred-and-thirty-four differentially expressed transcripts showed a change of two-fold or more in either the resistant or susceptible interaction, and most of the Phoma-regulated genes could be assigned to functional categories which have been reported to be involved in plant defence responses. RT-qPCR and HPLCUV confirmed involvement of the octadecanoid and phenylpropanoid pathways in response to P. medicaginis infection. Faster induction of lipoxygenase genes and constitutively higher levels of certain phenolic metabolites were observed in resistant plants.