Olufemi A. Akinsanmi

Identity and pathogenicity of Fusarium spp. isolated from wheat fields in Queensland and northern New South Wales

To establish the identity of Fusarium species associated with head blight (FHB) and crown rot (CR) of wheat, samples were collected from wheat paddocks with different cropping history in southern Queensland and northern New South Wales during 2001. CR was more widespread but FHB was only evident in northern NSW and often occurred with CR in the same paddock. Twenty different Fusarium spp. were identified from monoconidial isolates originating from different plant parts by using morphology and species-specific PCR assays. Fusarium pseudograminearum constituted 48% of all isolates and was more frequently obtained from the crown, whereas Fusarium graminearum made up 28% of all isolates and came mostly from the head. All 17 Fusarium species tested caused FHB and all 10 tested caused CR in plant infection assays, with significant (P < 0.001) difference in aggressiveness among species and among isolates within species for both diseases. Overall, isolates from stubble and crown were more aggressive for CR, whereas isolates from the flag leaf node were more aggressive for FHB. Isolates that were highly aggressive in causing CR were those originating from paddocks with wheat following wheat, whereas those from fields with wheat following maize or sorghum were highly aggressive for FHB. Although 20% of isolates caused severe to highly severe FHB and CR, there was no significant (P < 0.32) correlation between aggressiveness for FHB and CR. Given the ability of F. graminearum to colonise crowns in the field and to cause severe CR in bioassays, it is unclear why this pathogen is not more widely distributed in Australia.

Pathogen population structure and epidemiology are keys to wheat crown rot and Fusarium head blight management

This paper summarises the key findings from recent research on the population genetics and epidemiology of Fusarium pathogens causing head blight and crown rot of wheat in Australia and how this information has enabled the screening and selection of wheat germplasm with improved resistance to Fusarium. By relating new findings to the current state of knowledge, the paper serves as a timely and critical review of the international literature. In Australia, both Fusarium pseudograminearum and F. graminearum can cause both crown rot and Fusarium head blight under artificial inoculation. However, the former species is more widespread and is predominantly associated with crown rot whereas F. graminearum is mainly associated with Fusarium head blight, with limited geographical distribution in and around the Liverpool Plains in northern New South Wales. Studies of population structure and genetics have revealed that both species are genotypically diverse with similar levels of genetic recombination despite Gibberella zeae, the teleomorph of F. graminearum, being homothallic and G. coronicola, the teleomorph of F. pseudograminearum, being heterothallic. A high-throughput and reliable crown rot bioassay has been developed and used to screen over 1500 wheat germplasms to select 17 lines with putative crown rot resistance. Key differences in pathogen biology and epidemiology between Australia and the USA have emerged from other recent collaborative studies, which show that macroconidia constitute the bulk of aerial Fusarium head blight inoculum in Australia, whereas ascospores are the dominant primary inoculum for Fusarium head blight worldwide. The limited spread of splash-dispersed macroconidia of F. graminearum probably explains the restricted geographical distribution of this species in Australia. Other research collaboration has compared the aggressiveness, mycotoxin production and genotypic polymorphisms of the pathogen population from Australia and the USA. These and other differences in pathogen adaptation emphasise that research outcomes from elsewhere must be tested for relevance before applying them to Australian farming systems.

Timing of Fungicide Applications for Control of Husk Spot Caused by Pseudocercospora macadamiae in Macadamia

Pseudocercospora macadamiae is an important pathogen of macadamia in Australia, causing a disease known as husk spot. Growers strive to control the disease with a number of carbendazim and copper treatments. The aim of this study was to consider the macadamia fruit developmental stage at which fungicide application is most effective against husk spot, and whether application of copper-only applications at full-size fruit developmental stage toward the end of the season contributed to effective disease control. Fungicides were applied to macadamia trees at four developmental stages in three orchards in two subsequent production seasons. The effects of the treatments on disease incidence and severity were quantified using area under disease progress curve (AUDPC) and logistic regression models. Although disease incidence varied between cultivars, incidence and severity on cv. A16 showed consistent differences between the treatments. Most significant reduction in husk spot incidence occurred when spraying commenced at match-head sized-fruit developmental stage. All treatments significantly reduced husk spot incidence and severity compared with the untreated controls, and a significant positive linear relationship (R2 = 73%) between AUDPC and severity showed that timing of the first fungicide application is important for effective disease control. Application of fungicide at full-size fruit stage reduced disease incidence but had no impact on premature fruit drop.

Infection, colonisation and sporulation by Pseudocercospora macadamiae on macadamia fruit

The structures and manner with which Pseudocercospora macadamiae penetrates, colonises and proliferates from the pericarp of macadamia fruit was studied using scanning electron microscopy and fluorescence light microscopy. Germ tubes arising from conidia penetrated open stomata within 20 h of inoculation, without observation of specialised infection structures such as appressoria. Colonisation of the pericarp was intercellular, without observation of specialised intracellular infection structures such as haustoria, and was complete from the epidermis to the mesocarp. The fungus proliferated at the epidermis by the formation of conidiophores and conidia on substomatal and protuberant subepidermal stromata. These structures were not observed on the mesocarp surface. The onset of visual husk spot symptoms coincided with an increase in pathogen biomass on the pericarp surface. The progression of symptoms from tan-coloured spots to larger red-brown lesions coincided with the production of conidiophores from substomatal and protuberant subepidermal stromata. The darker the colour of the husk spot lesion, the more frequently protuberant subepidermal stromata were observed. These findings are discussed in the context of observation of other cercosporoid fungi.

Effect of Fusarium oxysporum f. sp. glycines and Sclerotium rolfsii on the pathogenicity of Meloidogyne incognita race 2 to soybean

Screenhouse studies were conducted to investigate the effects of Fusarium oxysporum f. sp. glycines and Sclerotium rolfsii on the pathogenicity of Meloidogyne incognita race 2 on soybean and the influence of the nematode on wilt incidence and growth of soybean. The interaction of each fungus with the nematode resulted in reduced shoot and root growth. Final nematode population was also reduced with concomitant inoculation of nematode and fungus or inoculation of fungus before nematode. While M. incognita suppressed wilt incidence in two nematode-susceptible cultivars of soybean (TGX 1485-2D and TGX 1440-IE), it had limited effect on wilt incidence in the nematode resistant cultivar of soybean (TGX 1448-2E). When F. oxysporumwas inoculated with the nematode, the mean number of nematodes that penetrated soybean roots decreased by 75% in TGX 1448-2E, 68% in TGX 1485-1D and 65% in TGX 1440-1E. Similarly when the soil was treated with S. rolfsii, the number decreased by 78% in TGX 1448-2E, 77% in TGX 1485-1D and 68% in TGX 1440-1E. The nematode did not develop beyond second-stage juvenile in TGX-1448-2E.

Alternative fungicides for controlling husk spot caused by Pseudocercospora macadamiae in macadamia

Husk spot, caused by Pseudocercospora macadamiae is amajor fungal disease of macadamia in Australia. Chemicals to control the disease are limited and frequent failure to control the disease is a major concern to growers. The overall goal of this research was to improve the chemical control strategy of P. macadamiae through the provision of fungicides with different modes of action to carbendazim, which is the current industry standard. Husk spot incidence, premature fruit abscission, kernel quality and yield were evaluated following application of different fungicide products in replicated field experiments at three different sites. Results showed significant differences in disease incidence and premature fruit abscission between fungicide treatments, field sites and years. Generally, disease incidence and premature fruit abscission on trees treated with fungicide were significantly (P < 0.05) lower than the untreated control. Pyraclostrobin conferred significantly better protection than trifloxystrobin, reducing disease severity by 70% compared with a 50% reduction by trifloxystrobin. The pyraclostrobin treatment had a similar efficacy to the current industry standard (70% reduction cf. 73% reduction by tank-mixed carbendazim and copper). Higher amounts of immature kernels occurred in the untreated control, followed by difenoconazole and trifloxystrobin. Diseased fruit accounted for 78% of premature fruit abscission, which indicates that husk spot enhances fruit abscission in macadamia. Our results suggest that pyraclostrobin provided similar efficacy to the industry standard and could, therefore, play a key role in the management of husk spot.

Spatial pattern and the effects of climatic factors on husk spot disease in macadamia

The relative importance of Pseudocercospora macadamiae conidia produced from the subepidermal stromata of diseased husk and the effect of climatic factors on husk spot epidemics were assessed on managed (fungicide-protected) and unmanaged macadamia trees. The spatial pattern of diseased fruit in the tree canopy was quantified using the binary form of Taylor’s power law on 54 trees for 4 years. The a and b parameters of the power law showed year-to-year variations in the unmanaged epidemic. This was attributed to differences in climatic conditions and the amount of early infection in the tree canopy. Overall, diseased fruit aggregated, heterogeneity was in excess of the levels expected for a random pattern (a >1; b >1), and the degree of aggregation increased with the incidence mean. The rate of increase in disease incidence was significantly lower in the managed rather than the unmanaged epidemic. However, the rates of increase of disease severity were comparable, suggesting that other factors contribute to abscission of diseased fruit. The effect of each climatic factor examined on disease severity was <50%. The number of rainy days (R2 = 74%) was the single-most important factor influencing disease incidence. The results indicate that P. macadamiae conidia are mostly rain-splash dispersed, thus proximity of new fruit to the source of inoculum (diseased husk) significantly influences disease incidence. Practical implications for effective husk spot control are discussed.

Timing of infection of macadamia fruit by Pseudocercospora macadamiae and climatic effects on growth and spore germination.

Pseudocercospora macadamiae causes husk spot of macadamia. Husk spot control would be improved by verifying the stages in fruit development susceptible to infection, and determine some of the climatic conditions likely to lead to high disease pressure periods in the field. Our results showed that the percent conidia germination and growth of germ tubes and mycelia of P. macadamiae were greatest at 26°C, with better conidia germination associated with high relative humidity and free water. The exposure of match-head-sized and pea-sized fruit stages to natural P. macadamiae inoculum in the field led to 2–5-fold increases in husk spot incidence, and up to 8.5-fold increases in premature abscission, compared with unexposed fruit. Exposure of fruit stages later than match-head-sized and pea-sized fruit generally caused no further increases in disease incidence or premature abscission. Climatic conditions were found to have a strong influence on the behaviour of P. macadamiae, the host, oil accumulation, and the subsequent impact of husk spot on premature abscission. Our findings suggest that fungicide application should target fruit at the match-head-sized stage of development in order to best reduce yield losses, particularly in seasons where oil accumulation in fruit is prolonged and climatic conditions are optimal for P. macadamiae.

Timing of Infection and Development of Alternaria Diseases in the Canopy of Apple Trees

Alternaria leaf blotch and fruit spot of apple caused by Alternaria spp. cause annual losses to the Australian apple industry. Erratic control using protectant fungicides is often experienced and may be due to the lack of understanding of the timing of infection and epidemiology of the diseases. We found that Alternaria leaf blotch infection began about 20 days after bloom (DAB) and the highest disease incidence occurred from 70 to 110 DAB. Alternaria fruit spot infection occurred about 100 DAB in the orchard. Fruit inoculations in planta showed that there was no specific susceptible stage of fruit. Leaves and fruit in the lower canopy of trees showed higher levels of leaf blotch and fruit spot incidence than those in the upper canopy and the incidence of leaf blotch in shoot leaves was higher than in spur leaves. Temperature, relative humidity, and rainfall affected leaf blotch and fruit spot incidence. The gained knowledge on the timing of infection and development of disease may aid in the development of more effective disease management strategies.

Pathogenic variation of Alternaria species associated with leaf blotch and fruit spot of apple in Australia

Four Alternaria species groups (A. longipes, A. arborescens, A. alternata/A. tenuissima and A. tenuissima/A. mali) are associated with leaf blotch and fruit spot of apple in Australia. There is no information on the variability of pathogenicity among the species and isolates within each species causing leaf blotch or fruit spot. We used a detached leaf assay and an in planta fruit inoculation assay to determine the pathogenicity and virulence of the four Alternaria species. Our results showed that isolates within the same species were not specific to either leaf or fruit tissue and showed great variability in pathogenicity and virulence, indicating cross-pathogenicity, which may be isolate dependent rather than species dependent. Generally, virulence of A. tenuissima and A. alternata isolates on leaf and fruit was higher than other species. Isolates of all species groups were pathogenic on leaves of different cultivars, but pathogenicity on fruit of different cultivars varied among isolates and species. Implications of our findings on prevalence of the diseases in different apple-producing regions in Australia and the development of targeted disease management of the diseases are discussed.