Endophytic fungal isolates from apple tissue: Latent pathogens lurking within?

Fungal endophytes inhabit a similar ecological niche to that occupied by many phytopathogens, with several pathogens isolated from healthy tissues in their latent phase. This study aimed to evaluate the pathogenicity, the colonisation ability, and the enzyme activity of 37 endophytic fungal isolates recovered from apparently healthy apple shoot and leaf tissues. The pathogenicity of the isolates was assessed on 'Royal Gala' and 'Braeburn' fruit and detached 'Royal Gala' shoots. For the non-pathogenic isolates, their ability to endophytically colonise detached 'Royal Gala' shoots was evaluated. Enzyme activity assays were undertaken to determine whether the pathogenicity of the endophytes was related to the production of the extracellular enzymes, amylase, cellulase, pectinase, protease, and xylanase. Of the 37 isolates studied, eight isolates, representing the genera Colletotrichum, Diaporthe, Fusarium, and Penicillium, were shown to be pathogenic on both apple shoots and fruit. Two isolates identified as Trichoderma atroviride, were pathogenic only on shoots, and three isolates, representing the genus Diaporthe, were pathogenic only on fruit. Of the remaining 24 isolates, 22 (Biscogniauxia (n = 8), Chaetomium (n = 4), Trichoderma (n = 3), Epicoccum (n = 2), Neosetophoma (n = 2), Xylaria (n = 1), Daldinia (n = 1), and Paraphaeosphaeria (n = 1)) were recovered from the inoculated apple shoots but two failed to colonise the shoot tissues. Of the isolates tested, 20 produced amylase, 15 cellulase, 25 pectinase, 26 protease, and 13 xylanase. There was no correlation between the range and type of enzymes produced by the isolates and their pathogenicity or ability to endophytically colonise the shoot tissue. The study showed that approximately one-third (13/37) of the isolates recovered from the apparently healthy apple shoot tissues were observed as latent pathogens. The isolates that did not cause disease symptoms may have the ability to reduce colonisation of apple tissues by pathogens including Neonectria ditissima associated with European canker of apple.

Californian thistle (Cirsium arvense): endophytes and Puccinia punctiformis.

Californian thistle (Cirisum arvense) is a troublesome weed in pastures and cropping systems. The fungal biocontrol agent Puccinia punctiformis, commonly referred to as thistle rust, performs inconsistently on C. arvense. Problems with P. punctiformis establishment and control of C. arvense may be attributable to differing plant endophytic populations in various environments. This article provides an overview of the relationships between endophytes and their host, but also between endophytes and pathogens with a focus on rust pathogens. This review provides insights into reasons why P. punctiformis performs inconsistently and identifies gaps in our knowledge. Filling these gaps may help to improve performance of this classical fungal biocontrol agent. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Broad host-range pathogens as bioherbicides: managing nontarget plant disease risk.

Plant pathogens with a broad host range are commercially more attractive as microbial bioherbicides than strictly host-specific pathogens as a result of the wider market potential of a product capable of controlling multiple species. However, the perceived spatiotemporal disease risk to nontarget plants is a barrier to their adoption for weed control. We consider two approaches to managing this risk. First, we consider safety zones and withholding periods for bioherbicide treatment sites. These must ensure inoculum spreading from, or surviving at the site, exposes nontarget plants to no more inoculum than from natural sources. They can be determined using simple dispersal models. We show that a ratio of added:natural inoculum of 1.0 is biologically reasonable as an 'acceptable risk' and a sound basis for safety zones and withholding periods. These would be analogous to the 'conditions of use' for synthetic chemical herbicides aimed at minimizing collateral damage to susceptible plants from spray drift and persistent soil residues. Second, weed-specific isolates of broad host-range pathogens may avoid the need for safety zones and withholding periods. Such isolates have been found in many broad host-range pathogen species. Their utilization as bioherbicides may more easily meet the requirements of regulators. Mixtures of different weed-specific isolates of a pathogen could provide bioherbicides with commercially attractive spectrums of weed control activity. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

In vitro inhibition of Sclerotinia sclerotiorum mycelial growth and reduction of sclerotial viability by the volatile bioactive compounds of Brassicaceae crops.

AIMS: Sclerotinia sclerotiorum is an important pathogen of a wide range of crops, with current control mostly relying on the use of fungicides. This study assessed the effect of biofumigation on in vitro inhibition of mycelial growth and reduction of sclerotial viability of S. sclerotiorum as an attempt to seek an alternative management strategy. METHODS AND RESULTS: The effect of different biofumigant crop types to inhibit mycelial growth of ten S. sclerotiorum isolates was investigated, with Brassica juncea 'Caliente 199' being the most effective biofumigant crop. The efficacy of 'Caliente 199' to inhibit mycelial growth and reduce sclerotial viability was influenced by different crop factors. Plant tissue of 'Caliente 199' harvested at 50% or 100% flowering and adjusted to 80% (w/w) moisture resulted in greater mycelial inhibition and a reduction in the sclerotial viability compared with the vegetative tissue with the same plant moisture. Mycelial inhibition and reduction of sclerotial viability were affected by tissue quantity. Whole plant tissue and shoots only resulted in a similar inhibition of mycelial growth, but whole plant tissue resulted in a greater reduction of sclerotial viability. The S. sclerotiorum isolates differed in sensitivity to the volatile bioactive compounds released by the biofumigant plant tissue. CONCLUSIONS: The volatile bioactive compounds released by 'Caliente 199' resulted in effective mycelial inhibition but did not kill sclerotia completely.

Phenotypic characterization of the Hordeum bulbosum derived leaf rust resistance genes Rph22 and Rph26 in barley.

AIMS: Two introgression lines (ILs), 182Q20 and 200A12, which had chromosomal segments introgressed from Hordeum bulbosum in H. vulgare backgrounds, were identified to show seedling resistance against Puccinia hordei, possibly attributed to two resistance genes, Rph22 and Rph26, respectively. This study characterized the phenotypic responses of the two genes against P. hordei over different plant development stages. METHODS AND RESULTS: Using visual and fungal biomass assessments, responses of ILs 182Q20, 200A12 and four other barley cultivars against P. hordei were determined at seedling, tillering, stem elongation and booting stages. Plants carrying either Rph22 or Rph26 were found to confer gradually increasing resistance over the course of different development stages, with partial resistant phenotypes (i.e. prolonged rust latency periods, reduced uredinia numbers but with susceptible infection types) observed at seedling stage and adult plant resistance (APR) at booting stage. A definitive switch between the two types of resistance occurred at tillering stage. CONCLUSIONS: Rph22 and Rph26 derived from H. bulbosum were well characterized and had typical APR phenotypes against P. hordei. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides important insights on the effectiveness and expression of Rph22 and Rph26 against P. hordei during plant development and underpins future barley breeding programmes using non-host as a genetic resource for leaf rust management.

Short Communication: Latent Detection of Downy Mildew (Peronospora pisi) in Bioassays against Pisum sativum.

Downy mildew of peas is caused by the obligate parasite Peronospora pisi, which occurs sporadically throughout temperate pea-growing regions across the world. To screen pea lines against this biotrophic pathogen, a suitable and reproducible in vitro method using living plant material is required. Field screening can be influenced by environmental factors, thus giving variable results. The aim of this study was to develop a method that could reliably be used to screen pea cultivars against P. pisi in a laboratory setting. A range of bioassays were used to test various methods of inoculation, utilizing sporangia and naturally infested soil. Latent infection was achieved by planting seeds in soil collected from a site with a known history of P. pisi infection and directly inoculating young pea plants with sporangia. Out of the 108 plants which survived the experimental period, only two plants expressed visible signs of disease; however, through a two-step nested PCR process we detected latent infection in 24 plants. This research highlights the importance of considering the presence of latent infection when screening pea lines against downy mildew.

Genetic mapping of a barley leaf rust resistance gene Rph26 introgressed from Hordeum bulbosum.

KEY MESSAGE: The quantitative barley leaf rust resistance gene, Rph26, was fine mapped within a H. bulbosum introgression on barley chromosome 1HL. This provides the tools for pyramiding with other resistance genes. A novel quantitative resistance gene, Rph26, effective against barley leaf rust (Puccinia hordei) was introgressed from Hordeum bulbosum into the barley (Hordeum vulgare) cultivar 'Emir'. The effect of Rph26 was to reduce the observed symptoms of leaf rust infection (uredinium number and infection type). In addition, this resistance also increased the fungal latency period and reduced the fungal biomass within infected leaves. The resulting introgression line 200A12, containing Rph26, was backcrossed to its barley parental cultivar 'Emir' to create an F2 population focused on detecting interspecific recombination within the introgressed segment. A total of 1368 individuals from this F2 population were genotyped with flanking markers at either end of the 1HL introgression, resulting in the identification of 19 genotypes, which had undergone interspecific recombination within the original introgression. F3 seeds that were homozygous for the introgressions of reduced size were selected from each F2 recombinant and were used for subsequent genotyping and phenotyping. Rph26 was genetically mapped to the proximal end of the introgressed segment located at the distal end of chromosome 1HL. Molecular markers closely linked to Rph26 were identified and will enable this disease resistance gene to be combined with other sources of quantitative resistance to maximize the effectiveness and durability of leaf rust resistance in barley breeding. Heterozygous genotypes containing a single copy of Rph26 had an intermediate phenotype when compared with the homozygous resistant and susceptible genotypes, indicating an incompletely dominant inheritance.

Process of infection of armored scale insects (Diaspididae) by an entomopathogenic Cosmospora sp.

Several species in the fungal genus Cosmospora (synonym Nectria) (anamorph Fusarium) are specialist entomopathogens of armored scale insects (Diaspididae), known to cause periodic epizootics in host populations. Inconsistent mortality rates recorded under laboratory conditions prompted a study into the process of infection of armored scale insects by this fungus. Scale insect mortality following exposure to a Cosmospora sp. (Culture Collection Number: CC89) from New Zealand was related to insect age, with reproductively mature insects having a significantly higher infection rate than immature insects. Examination using scanning electron microscopy found no evidence that the fungus penetrated directly through the wax test (cap) of the scale insect or through the un-lifted interface between the test and the substrate on which the insect resided. However, fungal hyphae were observed growing beneath the test when the test of the reproductively mature insect lifted away from the substrate for the purpose of releasing crawlers, the mobile pre-settled juveniles. Once the hyphae of CC89 advanced under the test, germ-tubes readily penetrated the insect body through a number of natural openings (e.g. spiracles, vulva, stylet), with mycosis observed within seven days after inoculation. Direct penetration through the cuticle of the scale insect was not observed.