Ravenelia acaciae-arabicae and Ravenelia evansii are distinct species on Acacia nilotica subsp. indica in India

The two rust fungi, Ravenelia acaciae-arabicae and R. evansii, were both found on Acacia nilotica subsp. indica in southern (Tamil Nadu) and northern (Gujarat) India. R. acaciae-arabicae has been often incorrectly synonymised with R. evansii, although each has distinctive urediniospores, viz. echinulate in R. acaciae-arabicae and verruculose in R. evansii. Both species are re-described and illustrated from fresh specimens collected in India. Herbarium specimens of R. evansii from South Africa, including the holotype, were also examined. The difficulty in connecting different anamorphic spore stages to either of these teleomorphic rusts is highlighted by the presence of similar aecidia on plants of A. robusta infected with R. evansii in South Africa and on A. nilotica subsp. indica infected with R. acaciae-arabicae in India. It is not known whether these aecidial rusts represent the same species, nor is it known if they represent an aecidial stage of either R. acaciae-arabicae, R. evansii or other rusts.

Sequence variation in the putative effector gene SIX8 facilitates molecular differentiation of Fusarium oxysporum f. sp. cubense

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.

Sources and seasonal dynamics of Alternaria inoculum associated with leaf blotch and fruit spot of apples

Alternaria leaf blotch and fruit spot caused by Alternaria spp. cause annual losses to the Australian apple industry. Control options are limited, mainly due to a lack of understanding of the disease cycle. Therefore, this study aimed to determine potential sources of Alternaria spp. inoculum in the orchard and examine their relative contribution throughout the production season. Leaf residue from the orchard floor, canopy leaves, twigs and buds were collected monthly from three apple orchards for two years and examined for the number of spores on their surface. In addition, the effects of climatic factors on spore production dynamics in each plant part were examined. Although all four plant parts tested contributed to the Alternaria inoculum in the orchard, significant higher numbers of spores were obtained from leaf residue than the other plant parts supporting the hypothesis that overwintering of Alternaria spp. occurred mainly in leaf residue and minimally on twigs and buds. The most significant period of spore production on leaf residue occurred from dormancy until bloom and on canopy leaves and twigs during the fruit growth stage. Temperature was the single most significant factor influencing the amount of Alternaria inoculum and rainfall and relative humidity showed strong associations with temperature influencing the spore production dynamics in Australian orchards. The practical implications of this study include the eradication of leaf residue from the orchard floor and sanitation of the canopy after harvest to remove residual spores from the trees.

Molecular phylogenetic analysis reveals six new species of Diaporthe from Australia

Six new species of Diaporthe, D. beilharziae on Indigofera australis, D. fraxini-angustifoliae on Fraxinus angustifolia subsp. oxycarpa, D. litchicola on Litchi chinensis, D. nothofagi on Nothofagus cunninghamii, D. pascoei on Persea americana and D. salicicola on Salix purpurea from Australia are described and illustrated based on morphological characteristics and molecular analyses. Three of the new species no longer produced sporulating structures in culture and two of these were morphologically described from voucher specimens. Phylogenetic relationships of the new species with other Diaporthe species are revealed by DNA sequence analyses based on the internal transcribed spacer (ITS) region, and partial regions of the β-tubulin (BT) and translation elongation factor 1-alpha (TEF). © 2013 Mushroom Research Foundation.

Controlling Pythium in Ginger: Phase 2

The fungal disease Pythium Soft Rot is regarded by the ginger industry as their most serious disease threat. This project was developed to further investigate the factors contributing to the persistence and spread of Pythium Soft Rot on ginger farms and to identify measures for their control. The study demonstrated that the pathogen capable of causing Pythium Soft Rot in ginger was spread in contaminated ‘seed’, soil and water and that it can be managed through a combination of strategies that rely on early detection, reducing pathogen levels in soils, preventing water logging and restricting movement of contaminated ‘seed’, soil and water. However, in order to have an effective level of control, all strategies need to be integrated in an effective manner.

Infection of mungbean seed by Macrophomina phaseolina is more likely to result from localized pod infection than from systemic plant infection

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.

An inordinate fondness for Fusarium: Phylogenetic diversity of fusaria cultivated by ambrosia beetles in the genus Euwallacea on avocado and other plant hosts

Ambrosia beetle fungiculture represents one of the most ecologically and evolutionarily successful symbioses, as evidenced by the 11 independent origins and 3500 species of ambrosia beetles. Here we document the evolution of a clade within Fusarium associated with ambrosia beetles in the genus Euwallacea (Coleoptera: Scolytinae). Ambrosia Fusarium Clade (AFC) symbionts are unusual in that some are plant pathogens that cause significant damage in naive natural and cultivated ecosystems, and currently threaten avocado production in the United States, Israel and Australia. Most AFC fusaria produce unusual clavate macroconidia that serve as a putative food source for their insect mutualists. AFC symbionts were abundant in the heads of four Euwallacea spp., which suggests that they are transported within and from the natal gallery in mandibular mycangia. In a four-locus phylogenetic analysis, the AFC was resolved in a strongly supported monophyletic group within the previously described Cade 3 of the Fusarium solani species complex (FSSC). Divergence-time estimates place the origin of the AFC in the early Miocene similar to 21.2 Mya, which coincides with the hypothesized adaptive radiation of the Xyleborini. Two strongly supported clades within the AFC (Clades A and B) were identified that include nine species lineages associated with ambrosia beetles, eight with Euwallacea spp. and one reportedly with Xyleborus ferrugineus, and two lineages with no known beetle association. More derived lineages within the AFC showed fixation of the clavate (club-shaped) macroconidial trait, while basal lineages showed a mix of clavate and more typical fusiform macroconidia. AFC lineages consisted mostly of genetically identical individuals associated with specific insect hosts in defined geographic locations, with at least three interspecific hybridization events inferred based on discordant placement in individual gene genealogies and detection of recombinant loci. Overall, these data are consistent with a strong evolutionary trend toward obligate symbiosis coupled with secondary contact and interspecific hybridization. (C) 2013 Elsevier Inc. All rights reserved.

The plasticity of NBS resistance genes in sorghum is driven by multiple evolutionary processes

Background Increased disease resistance is a key target of cereal breeding programs, with disease outbreaks continuing to threaten global food production, particularly in Africa. Of the disease resistance gene families, the nucleotide-binding site plus leucine-rich repeat (NBS-LRR) family is the most prevalent and ancient and is also one of the largest gene families known in plants. The sequence diversity in NBS-encoding genes was explored in sorghum, a critical food staple in Africa, with comparisons to rice and maize and with comparisons to fungal pathogen resistance QTL. Results In sorghum, NBS-encoding genes had significantly higher diversity in comparison to non NBS-encoding genes and were significantly enriched in regions of the genome under purifying and balancing selection, both through domestication and improvement. Ancestral genes, pre-dating species divergence, were more abundant in regions with signatures of selection than in regions not under selection. Sorghum NBS-encoding genes were also significantly enriched in the regions of the genome containing fungal pathogen disease resistance QTL; with the diversity of the NBS-encoding genes influenced by the type of co-locating biotic stress resistance QTL. Conclusions NBS-encoding genes are under strong selection pressure in sorghum, through the contrasting evolutionary processes of purifying and balancing selection. Such contrasting evolutionary processes have impacted ancestral genes more than species-specific genes. Fungal disease resistance hot-spots in the genome, with resistance against multiple pathogens, provides further insight into the mechanisms that cereals use in the “arms race” with rapidly evolving pathogens in addition to providing plant breeders with selection targets for fast-tracking the development of high performing varieties with more durable pathogen resistance.

Overcoming barriers to the successful implementation of a classical biological control strategy for the exotic invasive weed Mikania micrantha in the Asia-Pacific region

Mikania micrantha (Asteraceae) commonly known as mikania, is a major invasive alien plant (IAP) in the tropical humid agricultural and forest zones of the Asia-Pacific region. This fast-growing Neotropical vine is able to smother plants in agricultural ecosystems, agroforestry and natural habitats, reducing productivity and biodiversity. Fungal pathogens were first investigated for the classical biological control of this weed in 1996. This resulted in the selection and screening of the highly host-specific and damaging rust pathogen, Puccinia spegazzinii (Pucciniales). It was first released in India and China in 2005/6, although it is not believed to have established. Since then, it has been released successfully in Taiwan, Papua New Guinea (PNG), Fiji and most recently Vanuatu. The rust has established and is spreading rapidly after applying lessons learned from the first releases on the best rust pathotype and release strategy. In PNG, direct monitoring of vegetation change has demonstrated that the rust is having a significant impact on M. micrantha, with no unpredicted non-target impacts. Despite this, the authorities in many countries where mikania is a problem remain cautious about releasing the rust. In Western Samoa, introduction of the rust was not pursued because of a conflict of interest, and the perception that mikania suppresses even worse weeds. For some, ‘pathophobia’ is still a major obstacle. In Indonesia, where insects for weed CBC have been introduced, pathogens will currently not be considered. In other countries such as Bhutan and Myanmar, there are no baseline data on the presence and impact of IAPs and, with no history of CBC, no institutional framework for implementing this approach. Malaysia has a well-developed framework, but capacity needs to be built in the country. Overall, it remains critical to have champions at decision making levels. Hence, even with an effective ‘off-the-shelf’ agent available, implementation of mikania CBC still requires significant inputs tailored to the countries’ specific needs.

Commercial-scale Alternaria brown spot resistance screening as the first step in breeding new mandarins for Australia

Rapid screening tests and an appreciation of the simple genetic control of Alternaria brown spot (ABS) susceptibility have existed for many years, and yet the application of this knowledge to commercial-scale breeding programs has been limited. Detached leaf assays were first demonstrated more than 40 years ago and reliable data suggesting a single gene determining susceptibility has been emerging for at least 20 years. However it is only recently that the requirement for genetic resistance in new hybrids has become a priority, following increased disease prevalence in Australian mandarin production areas previously considered too dry for the pathogen. Almost all of the high-fruit-quality parents developed so far by the Queensland-based breeding program are susceptible to ABS necessitating the screening of their progeny to avoid commercialisation of susceptible hybrids. This is done effectively and efficiently by spraying 3-6 month old hybrid seedlings with a spore suspension derived from a toxin-producing field isolate of Alternaria alternate, then incubating these seedlings in a cool room at 25°C and high humidity for 5 days. Susceptible seedlings show clear disease symptoms and are discarded. Analysis of observed and expected segregation ratios loosely support the hypothesis for a single dominant gene for susceptibility, but do not rule out the possibility of alternative genetic models. After implementing the routine screening for ABS resistance for three seasons we now have more than 20,000 hybrids growing in field progeny blocks that have been screened for resistance to the ABS disease.

Novel species of Cercospora and Pseudocercospora (Capnodiales, Mycosphaerellaceae) from Australia

Novel species of Cercospora and Pseudocercospora are described from Australian native plant species. These taxa are Cercospora ischaemi sp. nov. on Ischaemum australe (Poaceae); Pseudocercospora airliensis sp. nov. on Polyalthia nitidissima (Annonaceae); Pseudocercospora proiphydis sp. nov. on Proiphys amboinensis (Amaryllidaceae); and Pseudocercospora jagerae sp. nov. on Jagera pseudorhus var. pseudorhus (Sapindaceae). These species were characterised by morphology and an analysis of partial nucleotide sequence data for the three gene loci, ITS, LSU and EF-1α. Recent divergence of closely related Australian species of Pseudocercospora on native plants is proposed.

Polyphasic characterization of four new plant pathogenic Phyllosticta species from China, Japan, and the United States

The black rot disease of Vitis species and other host genera of Vitacease is caused by Phyllosticta ampelicida and allied taxa which is considered to be a species complex. In this paper, we introduce four new species of Phyllosticta, including two from the P. ampelicida complex, based on a polyphasic characterization including disease symptoms and host association, morphology, and molecular phylogeny. The phylogenetic analysis was conducted based on the ribosomal internal transcribed spacer (ITS) region and a combined multi-locus alignment of the ITS, actin (ACT), partial translation elongation factor 1-alpha (TEF-1), and glyceraldehydes 3-phosphate dehydrogenase (GPDH) gene regions. Our study confirms the phylogenetic distinctions of the four new species, as well as their phenotypic differences with known species in the genus.

Benzyl isothiocyanate: maximising production in papaya tissue extracts

Abstract: Although mainly grown for its sweet flavoured fruit, papaya (Carica papaya) has also been used for pharmacological purposes for many years. The reasons for use are varied with one of the best known being its anti-fungal action. Benzyl isothiocyanate (BITC) is the constituent most often implicated in this activity. Isothiocyanates are formed when the enzyme myrosinase catalyses the hydrolysis of the non-bioactive glucosinolates. This occurs when cellular contents come into contact through chewing, cutting or during extraction processes in the laboratory. While this is common in Brassica vegetables, the glucosinolate-myrosinase system is rare in fruit, papaya being a notable exception. It contains benzyl glucosinolate (BG), the glucosinolate precursor of BITC, in significant quantities. Parameters that determine the amount of BITC formed are duration of hydrolysis, presence/absence of nitrile-specifier proteins and BG content of different cultivars and tissues. We experimented with differing BITC extraction solvents, with the intention of developing a low cost, natural anti-fungal extract based on under-utilised papaya tissues. The findings suggest that papaya seeds, particularly from quarter-ripe fruit, have the potential to produce the highest levels of BITC necessary. Furthermore, they compare well with the nitrile-specifier protein-containing garden cress seeds (Lepidium sativum). To utilise the papaya seeds as a BITC source, an organic solvent such as ethanol is required to extract the largely water-insoluble BITC from the hydrolysed papaya seed mixture.

A co-evolutionary relationship exists between Endoraecium (Pucciniales) and its Acacia hosts in Australia

Endoraecium is a genus of rust fungi that infects several species of Acacia in Australia, South-East Asia and Hawaii. This study investigated the systematics of Endoraecium from 55 specimens in Australia based on a combined morphological and molecular approach. Phylogenetic analyses were conducted on partitioned datasets of loci from ribosomal and mitochondrial DNA. The recovered molecular phylogeny supported a recently published taxonomy based on morphology and host range that divided Endoraecium digitatum into five species. Spore morphology is synapomorphic and there is evidence Endoraecium co-evolved with its Acacia hosts. The broad host ranges of E. digitatum, E. parvum, E. phyllodiorum and E. violae-faustiae are revised in light of this study, and nine new species of Endoraecium are described from Australia based on host taxonomy, morphology and phylogenetic concordance.