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During the infection of a host, plant pathogenic fungi secrete small proteins called effectors, which then modulate the defence response of the host. In the Fusarium oxysporum species complex (FOSC), the secreted in xylem (SIX) gene effectors are important for host-specific pathogenicity, and are also useful markers for identifying the various host-specific lineages. While the presence and diversity of the SIX genes has been explored in many of the pathogenic lineages of F. oxysporum, there is a limited understanding of these genes in non-pathogenic, endophytic isolates of F. oxysporum. In this study, universal primers for each of the known SIX genes are designed and used to screen a panel of endophytically-associated Fusarium species isolated from healthy, asymptomatic banana tissue. SIX gene orthologues are identified in the majority of the Fusarium isolates screened in this study. Furthermore, the SIX gene profiles of these endophytic isolates do not overlap with the SIX genes present in the pathogenic lineages of F. oxysporum that are assessed in this study. SIX gene orthologues have not been commonly identified in Fusarium species outside of the FOSC nor in non-pathogenic isolates of F. oxysporum. The results of this study indicate that the SIX gene effectors may be more broadly distributed throughout the Fusarium genus than previously thought. This has important implications for understanding the evolution of pathogenicity in the FOSC.

Banana is an important tropical crop and trading commodity in the world. However, Fusarium wilt is a severe risk for the world banana industry. Caused by the soil-borne fungal pathogen, Fusarium oxysporum f. sp. cubense (Foc), the disease is spreading insidiously and has been a source of enormous losses worldwide since the 1800s. Unfortunately, effective control methods are not available yet. The current study used an RNAi strategy, referred to as host-induced gene silencing (HIGS), to confer durable resistance against Fusarium by inhibiting in planta expression of Foc essential genes that are conserved in all Foc races and crucial for fungal virulence. In order to find efficient target genes, a wide search of literature has been conducted and four conserved target regions have been chosen after bioinformatics analysis. Preliminary findings from in vitro fungal inhibition assays showed significant reductions in fungal spore count, germination rate and mycelia growth after treating fungal cultures with double-stranded RNA (dsRNA) targeting four Foc genes, suggesting silencing of these genes was effective to reduce fungal growth. Intron hairpin constructs encoding dsRNA targeting selected Foc genes are in the process of being transformed into Arabidopsis thaliana for proof-of-concept of in planta fungal resistance.

It is hypothesized that the virulence of phytopathogenic fungi is mediated through the secretion of small effector proteins that interfere with the defence responses of the host plant. In Fusarium oxysporum, one family of effectors, the Secreted In Xylem (SIX) genes, has been identified. We sought to characterize the diversity and evolution of the SIX genes in the banana-infecting lineages of F. oxysporum f. sp. cubense (Foc). Whole-genome sequencing data were generated for the 23 genetic lineages of Foc, which were subsequently queried for the 14 known SIX genes (SIX1-SIX14). The sequences of the identified SIX genes were confirmed in a larger collection of Foc isolates. Genealogies were generated for each of the SIX genes identified in Foc to further investigate the evolution of the SIX genes in Foc. Within Foc, variation of the SIX gene profile, including the presence of specific SIX homologues, correlated with the pathogenic race structure of Foc. Furthermore, the topologies of the SIX gene trees were discordant with the topology of an infraspecies phylogeny inferred from EF-1α/RPB1/RPB2 (translation elongation factor-1α/RNA polymerase II subunit I/RNA polymerase II subunit II). A series of topological constraint models provided strong evidence for the horizontal transmission of SIX genes in Foc. The horizontal inheritance of pathogenicity genes in Foc counters previous assumptions that convergent evolution has driven the polyphyletic phylogeny of Foc. This work has significant implications for the management of Foc, including the improvement of diagnostics and breeding programmes.

Thirteen potentially new leaf rust resistance loci were identified in a Vavilov wheat diversity panel. We demonstrated the potential of allele stacking to strengthen resistance against this important pathogen. Leaf rust (LR) caused by Puccinia triticina is an important disease of wheat (Triticum aestivum L.), and the deployment of genetically resistant cultivars is the most viable strategy to minimise yield losses. In this study, we evaluated a diversity panel of 295 bread wheat accessions from the N. I. Vavilov Institute of Plant Genetic Resources (St Petersburg, Russia) for LR resistance and performed genome-wide association studies (GWAS) using 10,748 polymorphic DArT-seq markers. The diversity panel was evaluated at seedling and adult plant growth stages using three P. triticina pathotypes prevalent in Australia. GWAS was applied to 11 phenotypic data sets which identified a total of 52 significant marker–trait associations representing 31 quantitative trait loci (QTL). Among them, 29 QTL were associated with adult plant resistance (APR). Of the 31 QTL, 13 were considered potentially new loci, whereas 4 co-located with previously catalogued Lr genes and 14 aligned to regions reported in other GWAS and genomic prediction studies. One seedling LR resistance QTL located on chromosome 3A showed pronounced levels of linkage disequilibrium among markers (r 2 = 0.7), suggested a high allelic fixation. Subsequent haplotype analysis for this region found seven haplotype variants, of which two were strongly associated with LR resistance at seedling stage. Similarly, analysis of an APR QTL on chromosome 7B revealed 22 variants, of which 4 were associated with resistance at the adult plant stage. Furthermore, most of the tested lines in the diversity panel carried 10 or more combined resistance-associated marker alleles, highlighting the potential of allele stacking for long-lasting resistance.

Historical records report Fusarium moniliforme sensu lato as the pathogen responsible for Fusarium diseases of sorghum; however, recent phylogenetic analysis has separated this complex into more than 25 species. During this study, surveys were undertaken in three major sorghum-producing regions in eastern Australia to assess the diversity and frequency of Fusarium species associated with stalk rot- and head blight-infected plants. A total of 523 isolates were collected from northern New South Wales, southern Queensland and central Queensland. Nine Fusarium species were isolated from diseased plants. Pathogenicity tests confirmed F. andiyazi and F. thapsinum were the dominant stalk rot pathogens, whilst F. thapsinum and species within the F. incarnatum–F. equiseti species complex were most frequently associated with head blight.

Fusarium pseudograminearum (Fp), the causative fungal pathogen of the diseases Fusarium crown rot, is an important constraint to cereals production in many countries including Australia. Fp produces a number of secondary metabolites throughout its life cycle. One of these metabolites, the cyclic lipopeptide fusaristatin A, is encoded by a specific gene cluster containing a polyketide synthase and a three-module non-ribosomal peptide synthetase. However, a recent survey of Fp populations across Australia suggests that this cluster may only be present in a subset of isolates from Western Australia (WA). In this study, we screened 319 Fp isolates from WA and 110 Fp isolates from the Australian eastern states of New South Wales, Victoria, Queensland and South Australia to examine the distribution of this gene cluster among Australian Fp populations. The fusaristatin A gene cluster was found to be present in ~50% of Fp isolates from WA but completely absent in Fp isolates from eastern states. To determine its potential function, mutants of the fusaristatin A gene cluster were generated by disrupting the non-ribosomal peptide synthetase and polyketide synthase genes simultaneously in two different parental backgrounds. The mutants showed increased growth rates and were significantly more aggressive than their respective parental strains on wheat in crown rot pathogenicity assays. This suggested that fusaristatin A has a negative effect on fungal development and aggressiveness. The possible reasons for the geographically restricted presence of the fusaristatin A gene cluster and its role in fungal biology are discussed.

Variation in the virulence of Fusarium oxysporum f. sp. fragariae (Fof) strains is important when evaluating the resistance of plants to this fungus. Twenty-five isolates of F. oxysporum harvested from strawberry (Fragaria ×ananassa) plants growing in Australia were characterized using pathogenicity tests, vegetative compatibility groups (VCGs), and genetic analysis of translation elongation factor 1 alpha (EF-1α). The level of disease varied depending on isolate used, indicating heterogeneous populations of Fof. Two distinct VCGs were identified and corresponded to two of the 10 lineages identified by partial EF-1α. Using a subset of Fof isolates, resistance in eight cultivars ranged from highly resistant to highly susceptible, with some cultivar × isolate interaction. ‘Strawberry Festival’, ‘QHI Sugarbaby’, and ‘DPI Rubygem’ had high levels of resistance across all isolates. Isolates from Western Australia (WA) were genetically distinct from those from Queensland (QLD) and were more virulent to ‘Camarosa’, a major cultivar grown in WA.

Intensive selection in wheat (Triticum aestivum L.) breeding programs over the past 100 years has led to a genetic bottleneck in modern bread wheat. Novel allelic variation is needed to break the yield plateau, particularly in the face of climate change and rapidly evolving pests and pathogens. Landraces preserved in seed banks likely harbour valuable sources of untapped genetic diversity because they were cultivated for thousands of years under diverse eco-geographical conditions prior to modern breeding. We performed the first genetic characterisation of bread wheat accessions sourced from the N. I. Vavilov Institute of Plant Genetic Resources (VIR) in St Petersburg, Russia. A panel comprising 295 accessions, including landraces, breeding lines and cultivars was subject to single seed descent (SSD) and genotyped using the genotyping-by-sequencing Diversity Arrays Technology platform (DArT-seq); returning a total of 34,311 polymorphic markers (14,228 mapped and 20,083 unmapped). Cluster analysis identified two distinct groups; one comprising mostly breeding lines and cultivars, and the other comprising landraces. Diversity was benchmarked in comparison to a set of standards, which revealed a high degree of genetic similarity among breeding material from Australia and the International Maize and Wheat Improvement Center (CIMMYT). Further, 11,025 markers (1888 mapped and 9137 unmapped) were polymorphic in the diversity panel only, thus representing allelic diversity potentially not present in Australian or CIMMYT germplasm. Open-access to DArT-seq markers and seed for SSD lines will empower researchers, pre-breeders and breeders to rediscover genetic diversity in the VIR collection and accelerate utilisation of novel alleles to improve wheat.

Fusarium oxysporum f. sp. cubense (Foc) is a fungal pathogen of banana and is a significant threat to global banana production. A novel strain of Foc (named tropical race 4) first emerged in Southeast Asia and has recently been detected in Africa and Jordan. Tropical race 4 of Foc is of significant concern to the banana industry as it is extremely pathogenic towards all commercial banana cultivars. Currently, very few genes involved in pathogenicity have been identified in Foc. However, many pathogenicity-associated genes have been studied in the F oxysporum species complex. This study sought to identify the orthologues of genes previously demonstrated to be involved in the pathogenicity of F oxysporum in a collection of Foc isolates. Of those genes assessed, we detected all pathogenicity-associated genes in all Foc isolates, with the exception of SIX8, which was only detected in race 4 isolates. Our results indicate that many genes are required for general pathogenicity towards plants and are widely distributed across the F oxysporum species complex. However, the host-specific pathogenicity of Foc may be attributed to fungal effectors, such as SIX8. Previous work in the tomato-infecting lineage of E oxysporum has demonstrated that the SIX gene effector family is localized on a small, dispensable chromosome which has also been associated with the host-specific pathogenicity of the tomato infecting lineages. The current work sheds light on the pathogenomics of Fusarium. The results of this study are significant for the diagnosis and management of Foc. The identification and characterization of the genes controlling pathogenicity in Foc should be an ongoing field of research. It will also be of substantial value to better understand the genomic organization of each VCG of Foc for the development a pangenome of Foc and the E oxysporum species complex. An improved understanding of the pathogenomics of Foc is critical for the ongoing improvement of disease management strategies for Fusarium wilt of banana.

Leaf rust (LR) caused by Puccinia triticina, is among the most important diseases of wheat (Triticum aestivum L.) crops globally. Deployment of cultivars incorporating genetic resistance, such as adult plant resistance (APR) or all-stage resistance, is considered the most sustainable control method. APR is preferred for durability because it places lower selection pressure on the pathogen and is often polygenic. In the search for new sources of APR, here we explored a diversity panel sourced from the N. I. Vavilov Institute of Plant Genetic Resources. Based on DNA marker screening, 83 of the 300 lines were deemed to carry known APR genes; namely, Lr34, Lr46, and Lr67. Interestingly, lines carrying Lr67 were mostly landraces from India and Pakistan, reconfirming the likely origin of the gene. Rapid phenotypic screening using a method that integrates assessment at both seedling and adult growth stages under accelerated growth conditions (i.e., constant light and controlled temperature) identified 50 lines carrying APR. Levels of APR corresponded well with phenotypes obtained in a field nursery inoculated using the same pathotype (R2 = 0.82). The second year of field testing, using a mixture of pathotypes with additional virulence for race-specific APR genes (Lr13 and Lr37), identified a subset of 13 lines that consistently displayed high levels of APR across years and pathotypes. These lines provide useful sources of resistance for future research. A strategy combining rapid generation advance coupled with phenotyping under controlled conditions could accelerate introgression of these potentially novel alleles into adapted genetic backgrounds.

This study examines the CO2-mediated influence of plant resistance on crown rot dynamics under continuous cropping of partially resistant wheat line 249 and the susceptible cultivar Tamaroi. Disease incidence, severity, deoxynivalenol and Fusarium biomass were assessed after each cycle in microcosms established at ambient and 700 mg kg-1 CO2 using soil and stubble of these wheat lines from a field experiment with free to air CO2 enrichment. Monoconidial isolates from wheat stubble were collected initially, and after five cropping cycles, to compare the frequency and aggressiveness of Fusarium species in the two populations. Aggressiveness was measured using a high-throughput seedling bioassay. At elevated CO2, the higher initial incidence in Tamaroi increased with cropping cycles, but incidence in 249 remained unchanged. Incidence at ambient CO2 did not change for either line. Elevated CO2 induced partial resistance in Tamaroi, but not in 249. Increased Fusarium biomass in wheat tissue at elevated CO2 matched raised deoxynivalenol of the stem base in both lines. After five cycles of continuous wheat cropping, aggressiveness increased in pathogenic F. culmorum and F. pseudograminearum by 110%, but decreased in weakly pathogenic F. equiseti and F. oxysporum by 50%. CO2 and host resistance interactively influenced species frequency, and the highly aggressive F. pseudograminearum became dominant on Tamaroi irrespective of CO2 concentration, while its frequency declined on 249. This study shows that induced resistance at elevated CO2 will not reduce crown rot severity, or impede the selection and enrichment of Fusarium populations with increased aggressiveness. © 2013 British Society for Plant Pathology.

Fusarium oxysporum f. sp. cubense (Foc), causal agent of fusarium wilt of banana, is among the most destructive pathogens of banana and plantain. The development of a molecular diagnostic capable of reliably distinguishing between the various races of the pathogen is of key importance to disease management. However, attempts to distinguish isolates using the standard molecular loci typically used for fungal phylogenetics have been complicated by a poor correlation between phylogeny and pathogenicity. Among the available alternative loci are several putative effector genes, known as SIX genes, which have been successfully used to differentiate the three races of F. oxysporum f. sp. lycopersici. In this study, an international collection of Foc isolates was screened for the presence of the putative effector SIX8. Using a PCR and sequencing approach, variation in Foc-SIX8 was identified which allowed race 4 to be differentiated from race 1 and 2 isolates, and tropical and subtropical race 4 isolates to be distinguished from one another.

The ubiquitous fungal pathogen Macrophomina phaseolina is best known as causing charcoal rot and premature death when host plants are subject to post-flowering stress. Overseas reports of M.phaseolina causing a rapid rot during the sprouting of Australian mungbean seed resulted in an investigation of the possible modes of infection of seed. Isolations from serial portions of 10 mungbean plants naturally infected with the pathogen revealed that on most plants there were discrete portions of infected tissue separated by apparently healthy tissue. The results from these studies, together with molecular analysis of isolates collected from infected tissue on two of the plants, suggested that aerial infection of aboveground parts by different isolates is common. Inoculations of roots and aboveground parts of mungbean plants at nine temperaturexsoil moisture incubation combinations and of detached green pods strongly supported the concept that seed infection results from infection of pods by microsclerotia, rather than from hyphae growing systemically through the plant after root or stem infection. This proposal is reinforced by anecdotal evidence that high levels of seed infection are common when rainfall occurs during pod fill, and by the isolation of M.phaseolina from soil peds collected on pods of mungbean plants in the field. However, other experiments showed that when inoculum was placed within 130mm of a green developing pod and a herbicide containing paraquat and diquat was sprayed on the inoculated plants, M.phaseolina was capable of some systemic growth from vegetative tissue into the pods and seeds.

The initial infection stages of Phyllosticta maculata on banana were studied using scanning electron microscopy. Conidial germination on the banana leaf surface commenced within 3h postinoculation to produce a long and slender germ tube. The hyphae developed secondary branches and mostly grew randomly across the leaf surface. Appressoria were formed at the apex of the germ tubes within 18h postinoculation and were variable in shape. A layer of an extracellular matrix surrounded the appressoria at the pathogenhost interface. On the fruit surface, conidia germinated to produce predominantly swollen germ tubes which functioned as lateral appressoria together with some slender ones. These germ tubes were formed within 3h postinoculation. There was no stomatal penetration apparent on the leaf; instead, direct penetration through the cuticle with and without the formation of appressoria was observed. Cuticular degradation on the leaf surface was evident with a circular, darkened area around the point of penetration by hyphae or appressoria. The significant role of pycnidia and conidia in the epidemiology of the disease was further demonstrated in naturally infected leaf samples.

Since the 1980s a new disease has been affecting Australian lychee. Pepper spot appears as small, black superficial lesions on fruit, leaves, petioles and pedicels and is caused by Colletotrichum gloeosporioides, the same fungus that causes postharvest anthracnose of lychee fruit. The aim of this study was to determine if a new genotype of C.gloeosporioides is responsible for the pepper spot symptom. Morphological assessments, arbitrarily-primed PCR (ap-PCR) and DNA sequencing studies did not differentiate isolates of C.gloeosporioides from anthracnose and pepper spot lesions. The ap-PCR identified 21 different genotypes of C.gloeosporioides, three of which were predominant. A specific genotype identified using ap-PCR was associated with the production of the teleomorph in culture. Analysis of sequence data of ITS and -tubulin regions of representative isolates did not group the lychee isolates into a monophyletic clade; however, given the majority of the isolates were from one of three genotypes found using ap-PCR, the possibility of a lychee specific group of C.gloeosporioides is discussed.

Stripe or yellow rust (YR) is a significant problem in wheat crops worldwide. The deployment of adult-plant resistance (APR) genes in wheat cultivars is considered a sustainable management strategy, as these genes confer partial resistance that is usually non-race specific. Screening for APR typically involves assessment of adult plants in the field, where expression may be influenced by environmental factors. We report a high-throughput screening method for YR APR that can be used to assess fixed lines or segregating populations grown under controlled environmental conditions (CEC). Inoculation of 3-week-old wheat plants from lines with known APR responses to YR, when grown under constant light and temperature, provided disease responses typical of adult plants. Two F-2 populations ('H45' x 'ST93' and 'Wyalkatchem' x 'ST93') segregating for APR were assessed under both CEC and field conditions. These populations showed similar variation in disease response and lines assessed in both environments attained similar rankings. Phenotypic screening using CEC and continuous light provides an opportunity to accelerate the development of new wheat cultivars with durable resistance.

The effect of acibenzolar-S-methyl ( ASM) applied in the form of Bion (R) as a seed soak or a foliar spray, on transcript accumulation of six defence related genes, and activities of four defence related enzymes, was assessed after cotton was inoculated with Fusarium oxysporum f. sp. vasinfectum ( Fov). Significant upregulation of chitinase class I, peroxidase, and beta-1, 3-glucanase transcripts occurred in the Bion (R) seed soak treatment with Fov inoculation compared to all other treatments. Bion (R) alone did not significantly increase transcripts compared with water control treatments in the majority of cases. Phenylalanine ammonia lyase transcript in Bion (R) treated plants following fungal inoculation was greater than in the respective Bion (R) alone treatments, and osmotin-like protein transcript was significantly greater in inoculated plants sprayed with Bion (R) than in all other treatments. Transcripts of 4-coumarate-Co Aligase-like protein were generally lower than for all other genes, and significant upregulation compared with water control occurred in extracts from Fov inoculated plants receiving no Bion (R) treatment, and Fov inoculated plants cultivated from Bion (R) treated seed. The greatest increases in chitinase, beta-1, 3-glucanase and peroxidase activity occurred in root and shoot tissue from Bion (R) seed treated material inoculated with Fov. Bion (R) treatment alone did not consistently enhance enzyme activities above those measured in water treated controls. Polyphenol oxidase activities were enhanced by various Bion (R) or Fov treatments at 3 days after inoculation only. (C) 2009 Elsevier Ltd. All rights reserved.

A study was undertaken to investigate the potential sources of Pythium inoculum in greenhouse soils. About 7% of fallow soils were found to harbour Pythium before being introduced into greenhouses. When replacing the top layer (30–60 cm) of cultivated soil in greenhouses with fallow soil, Pythium inoculum was still recovered from the bottom layer of soil left in the greenhouse. Other potential sources of Pythium were found to be potting mixtures and contaminated soil adhering to cultivation equipment, growers' shoes and reused irrigation pipes. Pythium isolates from different sources were from two species: Pythium aphanidermatum (88%) and P. spinosum (12%). This appears to be the first report of transmission of Pythium via contaminated soil adhering to reused irrigation pipes. It also represents the first report in Oman of transmission of Pythium into greenhouses via potting mixtures and fallow soils.

Sorghum ergot, caused predominantly by Claviceps africana Frederickson, Mantle, de Milliano, is a significant threat to the sorghum industry worldwide. The objectives of this study were firstly, to identify molecular markers linked to ergot resistance and to two pollen traits, pollen quantity (PQ) and pollen viability (PV), and secondly, to assess the relationship between the two pollen traits and ergot resistance in sorghum. A genetic linkage map of sorghum RIL population R931945-2-2 × IS 8525 (resistance source) was constructed using 303 markers including 36 SSR, 117 AFLP™, 148 DArT™ and two morphological trait loci. Composite interval mapping identified nine, five, and four QTL linked to molecular markers for percentage ergot infection (PCERGOT), PQ and PV, respectively, at a LOD >2.0. Co-location/linkage of QTL were identified on four chromosomes while other QTL for the three traits mapped independently, indicating that both pollen and non pollen-based mechanisms of ergot resistance were operating in this sorghum population. Of the nine QTL identified for PCERGOT, five were identified using the overall data set while four were specific to the group data sets defined by temperature and humidity. QTL identified on SBI-02 and SBI-06 were further validated in additional populations. This is the first report of QTL associated with ergot resistance in sorghum. The markers reported herein could be used for marker-assisted selection for this important disease of sorghum.

A total of 24 isolates of Pythium spinosum from cucumber obtained from five regions in Oman were characterized for genetic diversity using amplified fragment length polymorphism (AFLP) fingerprinting and three isolates from the Netherlands, South Africa and Japan were included for comparison. Isolates from Oman were also characterized for aggressiveness on cucumber seedlings and sensitivity to metalaxyl. Identity of all isolates was confirmed using sequences of the internal transcribed spacer (ITS) region of the ribosomal DNA (rDNA), which showed more than 99% nucleotide similarity among all isolates. Using six primer-pair combinations, AFLP fingerprinting resolved 295 AFLP markers of which 193 were polymorphic among isolates from other countries and only six were polymorphic among isolates of P. spinosum from Oman. Seven different AFLP phenotypes of P. spinosum were recovered in Oman; two of them were found to contain over 79% of isolates and one was recovered from all regions in Oman. Phenotypes from Oman showed very high (≥99%) levels of genetic similarity to each other compared to moderate (mean =53%) levels of genetic similarity with phenotypes from other countries. In addition, all isolates from Oman were found to be highly sensitive to metalaxyl and all were aggressive on cucumber seedlings at 25°C. The high genetic similarity among phenotypes of P. spinosum in Oman as well as recovering two major clones across regions may suggest that P. spinosum has been recently introduced in Oman via a common source.

Sclerotinia species are sexually reproducing ascomycetes. In the past S. minor and S. sclerotiorum, have been assumed to be homothallic because of the self-fertility of colonies derived from single ascospores. S. trifoliorum has previously been shown to be bipolar heterothallic due to the presence of four self-fertile and four self-sterile ascospores within a single ascus [Uhm, J.Y., Fujii, H., 1983a. Ascospore dimorphism in Sclerotinia trifoliorum and cultural characters of strains from different-sized spores. Phytopathology 73: 565-569]. However, isolates of S. minor and S. sclerotiorum were proven to be homothallic ascomycetes, by self-fertility of all eight ascospores within an ascus. Apothecia were raised from all eight ascospores of a single tetrad from four isolates of S. minor and from an isolate of S. sclerotiorum, indicating that inbreeding may be the predominant breeding mechanism of S. minor. Ascospores from asci of S. minor and S. sclerotiorum were predominantly monomorphic, but rare examples of ascospore dimorphism similar to S. trifoliorum were found. (c) 2006 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Fusarium wilt of banana is a potentially devastating disease throughout the world. Options for control of the causal organism, Fusarium oxysporum f.sp. cubense (Foc) are limited. Suppressive soil sites have previously been identified where, despite the presence of Foc, Fusarium wilt does not develop. In order to understand some aspects of this disease suppression, endophytic Fusarium oxysporum isolates were obtained from banana roots. These isolates were genetically characterized and compared with an isolate of Fusarium oxysporum previously identified as being capable of suppressing Fusarium wilt of banana in glasshouse trials. Three additional isolates were selected for glasshouse trials to assess suppression of Fusarium wilt in two different cultivars of banana, Cavendish and Lady Finger. One isolate (BRIP 29089) was identified as a potential biocontrol organism, reducing the disease severity of Fusarium wilt in Lady Finger and Cavendish cultivars. Interestingly, one isolate (BRIP 45952) increased Fusarium wilt disease severity on Cavendish. The implications of an isolate of Fusarium oxysporum, non-pathogenic on banana, increasing disease severity and the potential role of non-pathogenic isolates of Fusarium oxysporum in disease complexes are discussed.

Weather damage reduces the value of commercial mungbean. but hard-seededness can reduce the level of damage. However. attempts lo breed large- and hard-seeded mungbean varieties have been unsuccessful. To understand the relationship between seed weight and hard-seededness. these trails were investigated using a quantitative trail loci (QTL) mapping approach with a recombinant inbred population derived from a cross between a completely soft-seeded variety and a completely hard-seeded genotype. The two parental genotypes also had a sixfold difference in seed weight. QTL analyses revealed four loci for hard-seededness and 11 loci for seed weight. Two of the hard-seeded ness loci co-localized with seed weight QTL. When seed weight was used as a covariate in the analysis of hard-seededness from the field data, two of the four hard-seeded QTL remained significant with the effect al one of these remaining unchanged. These results explain why retaining hard-seededness in large seeded mungbean lines has been unsuccessful. The existence of a persistent locus, however. indicated that breeding large and persistently hard-seeded varieties of mungbean may be possible.

Single-copy restriction fragment length polymorphism (RFLP) markers were used to determine the genetic structure of #Mycosphaerella fijiensis#, the cause of black leaf streak (black Sigatoka) disease of banana and plantain, in the Torres Strait, Papua New Guinea (PNG), and the Pacific Islands. A moderate level of genetic variation was observed in all populations with genotypic diversity values of 60-78% of the theoretical maximum, and gene diversity (H) values between 0.269 and 0.336. All populations were at gametic equilibrium, and with the high level of genotypic diversity observed this indicated that sexual reproduction has a major role in the genetic structure of the #M. fijiensis# populations examined. Population differentiation was tested on several hierarchical scales. No evidence of population differentiation was observed between sites on Mer Island. A moderate level of population differentiation was observed within the Torres Strait, between Badu and Met Islands (Fst = 0.097). On a regional scale, the greatest differentiation was found between the populations of the Torres Strait and the Pacific. Populations from these regions were more closely related to the PNG population than to each other, suggesting they were founded in separate events from the same population.

Single-copy restriction fragment length polymorphism (RFLP) markers were used to determine the genetic structure of the global population of #Mycosphaerella musicola#, the cause of Sigatoka (yellow Sigatoka) disease of banana. The isolates of #M. musicola# examined were grouped into four geographic populations representing Africa, Latin America and the Caribbean, Australia and Indonesia. Moderate levels of genetic diversity were observed for most of the populations (H = 0.22-0.44). The greatest genetic diversity was found in the Indonesian population (H = 0.44). Genotypic diversity was close to 50% in all populations. Population differentiation tests showed that the geographic populations of Africa, Latin America and the Caribbean, Australia and Indonesia were genetically different populations. Using tests, very high levels of genetic differentiation were detected between all the population pairs (Fst > 0.40), with the exception of the Africa and Latin America -Caribbean population pair. These two populations differed by only 3% (Fst = 0.03), and were significantly different (P < 0.05) from all other population pairs. The high level of genetic diversity detected in Indonesia in comparison to the other populations provides some support for the theory that #M. musicola# originated in South-east Asia and that #M. musicola# populations in other regions were founded by isolates from the South-east Asian region. The results also suggest the migration of M. musicola between Africa and the Latin America-Caribbean region.

A major locus conferring resistance to the causal organism of powdery mildew, Erysiphe polygoni DC, in mungbean (Vigna radiata L. Wilczek) was identified using QTL analysis with a population of 147 recombinant inbred individuals. The population was derived from a cross between 'Berken', a highly susceptible variety, and ATF 3640, a highly resistant line. To test for response to powdery mildew, F7and F8lines were inoculated by dispersing decaying mungbean leaves with residual conidia of E. polygoni amongst the young plants to create an artificial epidemic and assayed in a glasshouse facility. To generate a linkage map, 322 RFLP clones were tested against the two parents and 51 of these were selected to screen the mapping population. The 51 probes generated 52 mapped loci, which were used to construct a linkage map spanning 350 cM of the mungbean genome over 10 linkage groups. Using these markers, a single locus was identified that explained up to a maximum of 86% of the total variation in the resistance response to the pathogen.Key words: mungbean, powdery mildew, Erysiphe polygoni, QTL, molecular markers.

The LATERAL ORGAN BOUNDARIES (LOB) DOMAIN (LBD) gene family encodes plant-specific transcriptional regulators functioning in organ development. In a screen of Arabidopsis (Arabidopsis thaliana) sequence-indexed transferred DNA insertion mutants, we found disruption of the LOB DOMAIN-CONTAINING PROTEIN20 (LBD20) gene led to increased resistance to the root-infecting vascular wilt pathogen Fusarium oxysporum. In wild-type plants, LBD20 transcripts were barely detectable in leaves but abundant in roots, where they were further induced after F. oxysporum inoculation or methyl jasmonate treatment. Induction of LBD20 expression in roots was abolished in coronatine insensitive1 (coi1) and myc2 (allelic to jasmonate insensitive1) mutants, suggesting LBD20 may function in jasmonate (JA) signaling. Consistent with this, expression of the JA-regulated THIONIN2.1 (Thi2.1) and VEGETATIVE STORAGE PROTEIN2 (VSP2) genes were up-regulated in shoots of lbd20 following treatment of roots with F. oxysporum or methyl jasmonate. However, PLANT DEFENSIN1.2 expression was unaltered, indicating a repressor role for LBD20 in a branch of the JA-signaling pathway. Plants overexpressing LBD20 (LBD20-OX) had reduced Thi2.1 and VSP2 expression. There was a significant correlation between increased LBD20 expression in the LBD20-OX lines with both Thi2.1 and VSP2 repression, and reduced survival following F. oxysporum infection. Chlorosis resulting from application of F. oxysporum culture filtrate was also reduced in lbd20 leaves relative to the wild type. Taken together, LBD20 is a F. oxysporum susceptibility gene that appears to regulate components of JA signaling downstream of COI1 and MYC2 that are required for full elicitation of F. oxysporum- and JA-dependent responses. To our knowledge, this is the first demonstration of a role for a LBD gene family member in either biotic stress or JA signaling.

A transposon-tagging strategy has been employed in order to allow the isolation of a race-specific rust resistance gene from Antirrhinum. Six mutant (susceptible) plants have been produced. Evidence that the R gene has been tagged is: i) repeats of the tagging protocol in which transposons are not active have yielded no susceptible mutants, ii) on selling, the progeny of all six plants include some revertants to resistance, and iii) for the two mutants so far studied, the mutations map at, or very close to, the R gene. Molecular analyses, using cloned Antirrhinum sequences, are now under way. This work is being published in a fuller form in The Plant Journal vol. 2(5).

Jasmonate signaling plays an important role in both plant defense and development. Here, we have identified a subunit of the Mediator complex as a regulator of the jasmonate signaling pathway in Arabidopsis thaliana. The Mediator complex is a conserved multiprotein complex that acts as a universal adaptor between transcription factors and the RNA polymerase II transcriptional machinery. We report that the PHYTOCHROME AND FLOWERING TIME1 (PFT1) gene, which encodes the MEDIATOR25 subunit of Mediator, is required for jasmonate-dependent defense gene expression and resistance to leaf-infecting necrotrophic fungal pathogens. Conversely, PFT1 appears to confer susceptibility to Fusarium oxysporum, a root-infecting hemibiotrophic fungal pathogen known to hijack jasmonate responses for disease development. Consistent with this, jasmonate gene expression was suppressed in the pft1 mutant during infection with F. oxysporum. In addition, a wheat (Triticum aestivum) homolog of PFT1 complemented the defense and the developmental phenotypes of the pft1 mutant, suggesting that the jasmonate signaling functions of PFT1 may be conserved in higher plants. Overall, our results identify an important control point in the regulation of the jasmonate signaling pathway within the transcriptional machinery.

A study was undertaken in 2004 and 2005 to characterize pathogens associated with damping-off of greenhouse-grown cucumber seedlings in 13 districts in Oman. Identification of Pythium to the species level was based on sequences of the internal transcribed spacer (ITS) of the ribosomal DNA. Of the 98 Pythium isolates collected during the survey, Pythium aphanidermatum, P. spinosum, P. splendens and P. oligandrum accounted for 76%, 22%, 1% and 1%, respectively. Pythium aphanidermatum was isolated from all of the districts, while P. spinosum was isolated from seven districts. Pathogenicity tests showed inter- and intraspecific variation in aggressiveness between Pythium species. Pythium aphanidermatum, P. spinosum and P. splendens were found to be highly aggressive at 25°C. However, the aggressiveness of P. spinosum decreased when the temperature was raised to 30°C, which was found to correspond to the lower frequency of isolation of P. spinosum in the warmer seasons, compared to the cooler time of the year. Pythium aphanidermatum exhibited limited intraspecific variation in the sequences of the ITS region of the rDNA and showed 100% similarity to the corresponding P. aphanidermatum sequences from GenBank. The ITS sequence data, as well as morphological characteristics of P. spinosum isolates, showed a high level of similarity within and between P. spinosum and P. kunmingense, and suggested that the two species were synonymous. This study represents the first report of P. spinosum, P. splendens and P. oligandrum in Oman.

A genetic linkage map of mungbean ( Vigna radiata, 2n = 2 x = 22) consisting of 255 RFLP loci was developed using a recombinant inbred population of 80 individuals. The population was derived from an inter-subspecific cross between the cultivated mungbean variety 'Berken' and a wild mungbean genotype 'ACC 41' ( V. radiata subsp. sublobata). The total length of the map, which comprised 13 linkage groups, spanned 737.9 cM with an average distance between markers of 3.0 cM and a maximum distance between linked markers of 15.4 cM. The mungbean map was compared to a previously published map of lablab ( Lablab purpureus, 2n = 2 x = 24) using a common set of 65 RFLP probes. In contrast to some other comparative mapping studies among members of the Fabaceae, where a high level of chromosomal rearrangement has been observed, marker order between mungbean and lablab was found to be highly conserved. However, the two genomes have apparently accumulated a large number of duplications/deletions after they diverged.

Seventy three isolates of Pythium aphanidermatum obtained from cucumber from four different regions of Oman and 16 isolates of muskmelon from the Batinah region in Oman were characterized for aggressiveness, sensitivity to metalaxyl and genetic diversity using AFLP fingerprinting. Twenty isolates of P. aphanidermatum from diverse hosts from different countries were also included in the study. Most isolates from Oman were found to be aggressive on cucumber seedlings and all were highly sensitive to metalaxyl (EC50 0.05), which implies a lack of host specialization in P. aphanidermatum on these two hosts in Oman. AFLP analysis of all isolates using four primer-pair combinations resolved 152 bands, of which 61 (~40%) were polymorphic. Isolates of P. aphanidermatum from Oman and other countries exhibited high genetic similarity (mean = 94.1%) and produced 59 different AFLP profiles. Analysis of molecular variance indicated that most AFLP variation among populations of P. aphanidermatum in Oman was associated with geographical regions (FST = 0.118; P < 0.0001), not hosts (FST = -0.004; P = 0.4323). These data were supported by the high rate of recovery (24%) of identical phenotypes between cucumber and muskmelon fields in the same region as compared to the low recovery (10%) across regions in Oman, which suggests more frequent movement of Pythium inoculum among muskmelon and cucumber fields in the same region compared to movement across geographically separated regions. However, recovering clones among regions and different countries may imply circulation of Pythium inoculum via common sources in Oman and also intercontinental spread of isolates.

Genetic variation among Australian isolates of the fungus Fusarium oxysporum f. sp. cubense (Foc), which causes Fusarium wilt in banana, was examined using DNA amplification fingerprinting (DAF). Ninety-four isolates which represented Races 1, 2, 3, and 4, and vegetative compatibility groups (VCGs) 0120, 0124, 0125, 0128, 0129, 01211, 01213/16, and 01220 were analysed. The genetic relatedness among isolates within each VCG, and between the 8 different VCGs of Foc present in Australia was determined. The DNA fingerprint patterns were VCG-specific, with each VCG representing a unique genotype. The genetic similarity among isolates within each VCG ranged from 97% to 100%. Among the different VCGs of Foc, 3 major clusters were distinguished which corresponded with race. All Race 1 and 2 isolates (VCGs 0124, 0125, 0128, and 01220) were closely related and clustered together, the Race 3 isolates from Heliconia clustered separately, and all Race 4 isolates (VCGs 0120, 0129, 01211, and 01213/16) clustered together. Fifteen isolates from Alstonville, NSW, were characterised because although they were classified as Race 2 based on their recovery from cooking banana cultivars, they belonged in VCG 0124, which had previously contained only Race 1 isolates. The occurrence of more than one race within a VCG means that vegetative compatibility grouping cannot be used to assign pathotype to pathogenic race as previously thought. It was possible to distinguish the Race 1 and Race 2 isolates within VCG 0124 using DNA fingerprinting, as each race produced a unique DNA fingerprint pattern. Among the Australian isolates, DNA fingerprinting analysis identified 9 different VCGs and genotypes of Foc.

Fifty-five isolates of P. antirrhini urediospores were collected from around the world. These isolates were used to inoculate the leaves of 10 cultivars of Antirrhinum majus. Two races of P. antirrhini were identified (α and β). None of the cultivars challenged exhibited resistance to isolates of Race β. Some cultivars were resistant to Race a. Resistance was inherited in a manner consistent with the existence of a dominant, nuclear allele at a single locus.

Background Leaf rust (LR), caused by Puccinia triticina and is an important disease of wheat (Triticum aestivum L.). The most sustainable method for controlling rust diseases is deployment of cultivars incorporating adult plant resistance (APR). However, phenotyping breeding populations or germplasm collections for resistance in the field is dependent on weather conditions and limited to once a year. In this study, we explored the ability to phenotype APR to LR under accelerated growth conditions (AGC; i.e. constant light and controlled temperature) using a method that integrates assessment at both seedling and adult growth stages. A panel of 21 spring wheat genotypes, including disease standards carrying known APR genes (i.e. Lr34 and Lr46) were characterised under AGC and in the field. Results Disease response displayed by adult wheat plants grown under AGC (i.e. flag-2 leaf) was highly correlated with field-based measures (R2 = 0.77). The integrated method is more efficient—requiring less time, space, and labour compared to traditional approaches that perform seedling and adult plant assays separately. Further, this method enables up to seven consecutive adult plant LR assays compared to one in the field. Conclusion The integrated seedling and adult plant phenotyping method reported in this study provides a great tool for identifying APR to LR. Assessing plants at early growth stages can enable selection for desirable gene combinations and crossing of the selected plants in the same plant generation. The method has the potential to be scaled-up for screening large numbers of fixed lines and segregating populations. This strategy would reduce the time required for moving APR genes into adapted germplasm or combining traits in top crosses in breeding programs. This method could accelerate selection for resistance factors effective across diverse climates by conducting successive cycles of screening performed at different temperature regimes. Electronic supplementary material The online version of this article (doi:10.1186/s13007-016-0117-7) contains supplementary material, which is available to authorized users.