Shoot tip cryotherapy for plant pathogen eradication

Diseases caused by plant pathogens such as viruses, viroids and phytoplasmas cause huge economic losses of agricultural production and limit the safe movement of plant materials across borders. The use of pathogen-free planting materials provides a strategy for efficient management of these diseases and facilitates the global exchange of genetic resources. Shoot tip cryotherapy is a novel biotechnology method that uses cryogenic procedures to eradicate plant pathogens from the diseased plants. Combining thermotherapy or chemotherapy with shoot tip cryotherapy has further enhanced pathogen eradication efficiency. This review provides updated and comprehensive information on shoot tip cryotherapy and the combination of thermotherapy or chemotherapy with shoot tip cryotherapy for pathogen eradication. Prospects are proposed for future studies.     

Periodicity in host availability does not account for evolutionary branching as observed in many plant pathogens: an application to Gaeumannomyces graminis var. tritici

Periodicity in host availability is common in agricultural systems. Although it is known to have profound effects on plant pathogen abundance, the evolutionary consequences of periodicity for the pathogen population have not previously been analyzed. An epidemiological model incorporating periodic absence of the host crop is combined with the theory of adaptive dynamics to determine whether or not seasonality in host presence plays a role in the occurrence of evolutionary branching, leading to coexisting yet genetically distinct pathogen phenotypes. The study is motivated and illustrated by the specific example of take-all disease of wheat, caused by the pathogen Gaeumannomyces graminis var. tritici, for which two coexisting but genetically distinct types and a trade-off related to seasonality in host presence have been identified. Numerical simulations are used to show that a trade-off between the pathogen transmission rate and the survival of the pathogen between cropping seasons cannot account for the evolutionary branching observed in many pathogens. Model elaborations show that this conclusion holds for a broad range of putative mechanisms. Although the analysis is motivated and illustrated by the specific example of take-all of wheat, the results apply to a broad range of pathogens.     

Exploiting generic platform technologies for the detection and identification of plant pathogens

The detection and identification of plant pathogens currently relies upon a very diverse range of techniques and skills, from traditional culturing and taxonomic skills to modern molecular-based methods. The wide range of methods employed reflects the great diversity of plant pathogens and the hosts they infect. The well-documented decline in taxonomic expertise, along with the need to develop ever more rapid and sensitive diagnostic methods has provided an impetus to develop technologies that are both generic and able to complement traditional skills and techniques. Real-time polymerase chain reaction (PCR) is emerging as one such generic platform technology and one that is well suited to high-throughput detection of a limited number of known target pathogens. Real-time PCR is now exploited as a front line diagnostic screening tool in human health, animal health, homeland security, biosecurity as well as plant health. Progress with developing generic techniques for plant pathogen identification, particularly of unknown samples, has been less rapid. Diagnostic microarrays and direct nucleic acid sequencing (de novo sequencing) both have potential as generic methods for the identification of unknown plant pathogens but are unlikely to be suitable as high-throughput detection techniques. This paper will review the application of generic technologies in the routine laboratory as well as highlighting some new techniques and the trend towards multi-disciplinary studies.     

A high-density random-oligonucleotide genome microarray for universal diagnostics

Microarrays offer virtually unlimited diagnostics capability, and have already been developed into diagnostic chips for many different plant pests. The full capacity of such chips, however, has lagged far behind their full potential. The main reason for this is that current chip design relies on a priori genetic information for target organisms and on a consensus on the genetic sequences to be used in particular organism groups. Such information is often unavailable and laborious to obtain. Thus, broad-application diagnostic microarrays have been limited to narrow organism groups focused on Genera of pests/pathogens or those affecting individual host crops, without applicability for simultaneous detection of diverse pests affecting many crops. This paper describes the development of a diagnostic microarray platform that has universal application based on genomic fingerprinting of any organism without a need for a priori sequence information. Taxon-specific hybridization patterns are obtained by unique hybridisation of genomic DNA to 100s–1000s of short random oligonucleotide probes. Taxon identification is then achieved by comparison of hybridisation patterns from an unknown sample against a reference-pattern database. Using bacteria as a model pathogen group, these methods deliver highly reproducible hybridisation patterns with high resolution power and enable discrimination at the species and subspecies level.     

Diagnostics,genetic diversity and pathogenic variation of ascochyta blight of cool season food and feed legumes

Molecular diagnostic techniques have been developed to differentiate the Ascochyta pathogens that infect cool season food and feed legumes, as well as to improve the sensitivity of detecting latent infection in plant tissues. A seed sampling technique was developed to detect a 1% level of infection by Ascochyta rabiei in commercial chickpea seed. The Ascochyta pathogens were shown to be genetically diverse in countries where the pathogen and host have coexisted for a long time. However, where the pathogen was recently introduced, such as A. rabiei to Australia, the level of diversity remained relatively low, even as the pathogen spread to all chickpea-growing areas. Pathogenic variability of A. rabiei and Ascochyta pinodes pathogens in chickpea and field pea respectively, appears to be quantitative, where measures of disease severity were based on aggressiveness (quantitative level of infection) rather than on true qualitative virulence. In contrast, qualitative differences in pathogenicity in lentil and faba bean genotypes indicated the existence of pathotypes of Ascochyta lentis and Ascochyta fabae. Therefore, reports of pathotype discrimination based on quantitative differences in pathogenicity in a set of specific genotypes is questionable for several of the ascochyta-legume pathosystems such as A. rabiei and A. pinodes. This is not surprising since host resistance to these pathogens has been reported to be mainly quantitative, making it difficult for the pathogen to overcome specific resistance genes and form pathotypes. For robust pathogenicity assessment, there needs to be consistency in selection of differential host genotypes, screening conditions and disease evaluation techniques for each of the Ascochyta sp. in legume-growing countries throughout the world. Nevertheless, knowledge of pathotype diversity and aggressiveness within populations is important in the selection of resistant genotypes.     

Target the core: durable plant resistance against filamentous plant pathogens through effector recognition

Plant pathogens colonize their host through the secretion of effector proteins that modulate plant metabolism and immune responses to their benefit. Plants evolve towards effector recognition, leading to host immunity. Typically, pathogen effectors are targets for recognition through plant receptors that are encoded by resistance genes. Resistance gene mediated crop immunity puts a tremendous pressure on pathogens to adapt and alter their effector repertoire to overcome recognition. We argue that the type of effector that is recognized by the host may have considerable implications on the durability of resistance against filamentous plant pathogens. Effector genes that are conserved among pathogens and reside in core genome regions are most likely to hold indispensable virulence functions. Consequently, the cost for the pathogen to overcome recognition by the host is higher than for diversified, host‐specific effectors with a quantitative impact on virulence. Consequently, resistance genes that directly target conserved effector proteins without the interception of other effector proteins are potentially excellent resistance resources. © 2019 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Aggressiveness and its role in the adaptation of plant pathogens

Aggressiveness, the quantitative component of pathogenicity, and its role in the adaptation of plant pathogens are still insufficiently investigated. Using mainly examples of biotrophic and necrotrophic fungal pathogens of cereals and Phytophthora infestans on potato, the empirical knowledge on the nature of aggressiveness components and their evolution in response to host and environment is reviewed. Means of measuring aggressiveness components are considered, as well as the sources of environmental variance in these traits. The adaptive potential of aggressiveness components is evaluated by reviewing evidence for their heritability, as well as for constraints on their evolution, including differential interactions between host and pathogen genotypes and trade-offs between components of pathogenicity. Adaptations of pathogen aggressiveness components to host and environment are analysed, showing that: (i) selection for aggressiveness in pathogen populations can be mediated by climatic parameters; (ii) global population changes or remarkable population structures may be explained by variation in aggressiveness; and (iii) selection for quantitative traits can influence pathogen evolution in agricultural pathosystems and can result in differential adaptation to host cultivars, sometimes leading to erosion of quantitative resistance. Possible links with concepts in evolutionary ecology are suggested.     

Effect of light and dark on the growth and development of downy mildew pathogen Hyaloperonospora arabidopsidis

Disease development in plants requires a susceptible host, a virulent pathogen, and a favourable environment. Oomycete pathogens cause many important diseases and have evolved sophisticated molecular mechanisms to manipulate their hosts. Day length has been shown to impact plant–oomycete interactions but a need exists for a tractable reference system to understand the mechanistic interplay between light regulation, oomycete pathogen virulence, and plant host immunity. Here we present data demonstrating that light is a critical factor in the interaction between Arabidopsis thaliana and its naturally occurring downy mildew pathogen Hyaloperonospora arabidopsidis (Hpa). We investigated the role of light on spore germination, mycelium development, sporulation, and oospore formation of Hpa, along with defence responses in the host. We observed abundant Hpa sporulation on compatible Arabidopsis under day lengths ranging from 10 to 14 hr. In contrast, exposure to constant light or constant dark suppressed sporulation. Exposure to constant dark suppressed spore germination, mycelial development, and oospore formation, whereas exposure to constant light stimulated these three stages of development. A biomarker of plant immune system activation was induced under both constant light and constant dark. Altogether, these findings demonstrate that Hpa has the molecular mechanisms to perceive and respond to light and that both the host and pathogen responses are influenced by the light regime. Therefore, this pathosystem can be used for investigations to understand the molecular mechanisms through which oomycete pathogens like Hpa perceive and integrate light signals, and how light influences pathogen virulence and host immunity during their interactions.     

What proteomic analysis of the apoplast tells us about plant–pathogen interactions

Plant pathogens have developed different strategies during their evolution to infect and colonize their hosts. In the same way, plants have evolved different mechanisms acting against potential pathogens trying to infect and colonize their tissues. Regulation of a wide variety of proteins is required in order to perceive the pathogen and to activate the plant defence mechanisms. The apoplast is the first compartment where these recognition phenomena occur in most plant–pathogen interactions, allowing the exchange of different molecules and facilitating inter‐ and intracellular communication in plant cells. Proteomic analysis of the apoplast in recent years has found the initial biochemical responses involved in pathogen recognition and early defence responses. However, this proteomic approach requires some specific experimental conditions to obtain an extract free of cytoplasmic proteins and nonprotein contaminants that affect the subsequent stages of separation and quantification. Obtaining the highest proportion of proteins from the apoplastic space in infected tissues requires different steps such as extraction of apoplastic washing fluids and preparation of total secreted proteins (protein precipitation, solubilization, separation and digestion). Protein identification using mass spectrometry techniques and bioinformatics tools identifying peptides for the extracellular exportation is required to confirm the apoplastic location. This review compiles the most commonly used techniques for proteomic studies, focusing on the early biochemical changes occurring in the apoplast of plants infected by a wide range of pathogens. The scope of this approach to discover the molecular mechanisms involved in the plant–pathogen interaction is discussed.     

Direct and host‐mediated interactions between Fusarium pathogens and herbivorous arthropods in cereals

Fusarium head blight and fusarium ear rot diseases of cereal crops are significant global problems, causing yield and grain quality losses and accumulation of harmful mycotoxins. Safety limits have been set by the European Commission for several Fusarium‐produced mycotoxins; mitigating the risk of breaching these limits is of great importance to crop producers as part of an integrated approach to disease management. This review examines current knowledge regarding the role of arthropods in disease epidemiology. In the field, diseased host plants are likely to interact with arthropods that may substantially impact the disease by influencing spread or condition of the shared host. For example, disease progress by Fusarium graminearum can be doubled if wheat plants are aphid‐infested. Arthropods have been implicated in disease epidemiology in several cases and the evidence ranges from observed correlations between arthropod infestation and increased disease severity and mycotoxin accumulation, to experimental evidence for arthropod infestation causing heightened pathogen prevalence in hosts. Fusarium pathogens differ in spore production and impact on host volatile chemistry, which influences their suitability for arthropod dispersal. Herbivores may allow secondary fungal infection after wounding a plant or they may alter host susceptibility by inducing changes in plant defence pathways. Post‐harvest, during storage, arthropods may also interact with Fusarium pathogens, with instances of fungivory and altered behaviour by arthropods towards volatile chemicals from infected grain. Host‐mediated indirect pathogen–arthropod interactions are discussed alongside a comprehensive review of evidence for direct interactions where arthropods act as vectors for inoculum.     

Development of a One-Step Real-Time Polymerase Chain Reaction Assay for Diagnosis of Phytophthora ramorum

ABSTRACT Phytophthora ramorum is a recently described pathogen causing bleeding cankers, dieback, and leaf blight on trees and shrubs in parts of Europe and North America, where the disease is commonly known as sudden oak death. This article describes the development of a single-round real-time polymerase chain reaction (PCR) assay based on TaqMan chemistry, designed within the internal transcribed spacer 1 region of the nuclear ribosomal (nr)RNA gene for detection of P. ramorum in plant material. Unlike previously described methods for the molecular detection of P. ramorum, this assay involves no post amplification steps or multiple rounds of PCR. The assay was found to have a limit of detection of 10 pg of P. ramorum DNA, and could detect P. ramorum in plant material containing 1% infected material by weight within 36 cycles of PCR. The assay also was used to test DNA from 28 other Phytophthora spp. to establish its specificity for P. ramorum. A quick and simple method was used to extract DNA directly from host plant material, and detection of P. ramorum was carried out in multiplex with an assay for a gene from the host plant in order to demonstrate whether amplifiable DNA had been extracted. Amplifiable DNA was extracted from 84.4% of samples, as demonstrated by amplification of host plant DNA. The real-time protocol was used to test 320 plant samples (from 19 different plant species) from which DNA extraction had been successful, and was shown to give results comparable with a traditional isolation technique for diagnosis of P. ramorum in plant material from common U.K. hosts.     

Developing smarter host mixtures to control plant disease

Adaptation of plant pathogens to disease control measures (both chemical and genetic) is facilitated by the genetic uniformity underlying modern agroecosystems. One path to sustainable disease control lies in increasing genetic diversity at the field scale by using genetically diverse host mixtures. In this study, a robust population dynamics approach was used to model how host mixtures could improve disease control. It was found that when pathogens exhibit host specialization, the overall disease severity decreases with the number of components in the mixture; this finding makes it possible to determine an optimal number of components to use. In a simple case, where two host varieties are exposed to two host‐specialized pathogen species or strains, quantitative criteria for optimal mixing ratios are determined. Using these model outcomes, ways to optimize the use of host mixtures to decrease disease in agroecosystems are proposed.     

Climate change impacts on plant canopy architecture: implications for pest and pathogen management

Climate change influences on pests and pathogens are mainly plant-mediated. Rising carbon dioxide and temperature and altered precipitation modifies plant growth and development with concomitant changes in canopy architecture, size, density, microclimate and the quantity of susceptible tissue. The modified host physiology and canopy microclimate at elevated carbon dioxide influences production, dispersal and survival of pathogen inoculum and feeding behaviour of insect pests. Elevated temperature accelerates plant growth and developmental rates to modify canopy architecture and pest and pathogen development. Altered precipitation affects canopy architecture through either drought or flooding stress with corresponding effects on pests and pathogens. But canopy-level interactions are largely ignored in epidemiology models used to project climate change impacts. Nevertheless, models based on rules of plant morphogenesis have been used to explore pest and pathogen dynamics and their trophic interactions under elevated carbon dioxide. The prospect of modifying canopy architecture for pest and disease management has also been raised. We offer a conceptual framework incorporating canopy characteristics in the traditional disease triangle concept to advance understanding of host-pathogen-environment interactions and explore how climate change may influence these interactions. From a review of recent literature we summarize interrelationships between canopy architecture of cultivated crops, pest and pathogen biology and climate change under four areas of research: (a) relationships between canopy architecture, microclimate and host-pathogen interaction; (b) effect of climate change related variables on canopy architecture; (c) development of pests and pathogens in modified canopy under climate change; and (d) pests and pathogen management under climate change.     

Analytical models of weed biocontrol with sterilizing fungi: the consequences of differences in weed and pathogen life-histories

A model of the population dynamics of healthy weed plants, weed seeds in the soil, pathogen-infected weed plants and pathogen spores in the soil, was devised to investigate interactions that are important for the success of biocontrol with pathogens that prevent seed set. Three particular features of the host-pathogen interaction were examined in detail: the form of the density dependent relationship which determined seed and spore production, the host life stage at which infection could occur, and the relative competitive abilities of healthy and infected host plants. It was found that, when both weed and pathogen coexist, the equilibrium abundance of the weed in the presence of the pathogen was independent of the form of the relationship between plant density and seed or spore production. However, the form of this relationship did affect estimated equilibrium densities in the absence of biocontrol, and also affected the parametrizations under which both host and pathogen could coexist. Parameters derived from experiments with isolated host plants may therefore be sufficient to assess the biocontrol potential of new pathogens along with knowledge of densities achievable when the weed is uncontrolled. The form of the relationship used to control seed and spore production also had a marked influence on the range of parameter values over which the pathogen could persist. Other control measures were represented by changes to appropriate parameter values, e.g. weeding was represented by a change in weed death rate. With one exception, the use of additional control measures was not antagonistic to biocontrol. Often, however, the combined effect was less than additive, and the existence of synergy (where the combined effects are more than additive) was critically dependent on the form of the relationship of the rate of seed production per plant with density and the efficiency of the other control measures.     

Opportunities and problems of control of foliar pathogens with micro-organisms

Three different strategies in biological control of diseases of above-ground plant pans are discussed. (1) Microbial suppression of infection. Undisturbed naturally occurring yeast populations will create unfavourable nutritional conditions for leaf infection by necrotrophic pathogens. Application of biological control agents, operating through nutrient competition, to healthy leaves, however, is generally not very effective, because the pathogen rapidly penetrates the leaf and escapes competition. In contrast, microbial protection of man-made wounds on fruits looks promising. Field applications of bacterial preparations or aqueous compost extract to leaves reduce disease by necrotrophs as well as biotrophs. The mechanisms are not clear, but involvement of induced resistance may make the establishment of the biological control agents in the phyllosphere less important. (2) Microbial suppression of pathogen sporulation. Suppression of the dissemination of the pathogen is, in principle, effective against diseases with many infection cycles per season. This approach allows a long interaction period between the antagonist and the pathogen. Research on the use of mycoparasites for the control of mildews and rusts is well known in this respect. Promising, and relatively new. is this approach in relation to necrotrophic pathogens sporulating on dead plant material. (3) Microbial suppression of pathogen survival. Mycoparasites may also interfere with the formation and vitality of sclerotia in infected above-ground plant tissue and crop remains. Depending on the importance of initial inoculum, originating from sclerotia, treatment of infected plant material may reduce the severity of the disease in the following season.     

植物真菌和卵菌病暴发的起源和传播

传染病暴发在植物、动物和人群中很常见。除了少数已发展为流行病和大流行病外,在很大程度上大多数传染病暴发的原因仍未知,植物真菌和卵菌病暴发尤其如此。所有流行病和大流行病都是从局部暴发开始,然后蔓延到更广泛的地理区域,因此了解其初始暴发的原因对于有效预防和控制植物病害流行病和大流行病至关重要。该文首先描述疾病暴发的定义和检测,随后简要描述导致植物传染病暴发的主要原因,包括寄主植物、病原体及其相关的环境因素,以一种真菌和一种卵菌病原体为例简要概述宿主病原体系统,并强调分子工具在帮助揭示病原体的起源和传播及其暴发及大流行方面的作用。由于人为活动及气候的加速变化,植物病害暴发的可能性越来越大,最后提出应该如何应对其暴发。     

Evolution of gene-for-gene systems in metapopulations: the effect of spatial scale of host and pathogen dispersal

The concept of gene-for-gene coevolution is a major model for research on disease resistance in crop plants. However, few theoretical or empirical studies have examined such systems in natural situations, and as a consequence, there is little knowledge of how spatial effects are likely to influence the evolution of host resistance and pathogen virulence in gene-for-gene interactions. In this work, a simulation approach was used to investigate the epidemiological and genetic consequences of varying host and pathogen dispersal in metapopulation situations. The results demonstrate clear impacts of dispersal distance on the total number of host and pathogen genotypes that are maintained, as well as on genetic variation at individual host resistance and pathogen virulence loci. Several other important results also emerged from this study. In contrast to the predictions of many earlier nonspatial models, so-called 'super-races' of pathogens do not always evolve and dominate, indicating that it is not necessary to assume costs of resistance or virulence to maintain high levels of polymorphism in biologically realistic situations. The rate of evolution of both resistance and virulence depend on the scale of dispersal, with greater mixing (as a function of dispersal scale) resulting in a faster approach to a dynamic endpoint. The model in this paper also predicts that, despite the greater total genotypic diversity of pathogens across the metapopulation, variation in host resistance will generally be greater than variation in pathogen virulence within local populations.     

Local maladaptation of the anther-smut fungus parasitizing Dianthus carthusianorum

Pathogens are generally expected to evolve faster than their hosts and are therefore likely to be locally adapted. However, some pathogens might lag behind in the co-evolutionary arms race because they do not have some of the advantages shared by most other pathogens (e.g., high mutation or recombination rates, short generation time, high dispersal ability). This is the case of Microbotryum fungi that cause the anther smut disease in plants of the family Caryophyllaceae. We investigated the patterns of local adaptation and maladaptation in Microbotryum carthusianorum and its host plant Dianthus carthusianorum. We performed a full cross-inoculation experiment using half-sib plant families and fungal samples originating from three naturally infected populations in the Czech Republic. We specifically asked, which components of pathogen fitness (i.e., infectivity and host manipulation) are affected by local (mal)adaptation. The pathogen was on average 1.6 times more successful in infecting plants from foreign populations compared to plants from its home population. Once the infection was successful, the pathogen accelerated the plant’s flowering and thus increased the opportunity for transmission to new hosts. However, the level of manipulation of host flowering did not differ between home and foreign populations. This study showed that the pathogen’s infectivity followed a clear pattern of local maladaptation, whereas the host manipulation did not. Our study taken together with previous studies of a related anther smut species reveals the pervasiveness of local maladaptation in this group of pathogens that arises as the result of their restricted gene flow and reduced recombination rates.

     

Pathogenicity and virulence factors of Pseudomonas syringae

In 1994, Oku reported that plant pathogens, mainly fungal pathogens, require three essential abilities to infect plants: to enter plants, to overcome host resistance, and to evoke disease. Because the infectious process of phytopathogenic bacteria differs from that of fungal pathogens, we have attempted to characterize pathogenicity, the ability of a pathogen to cause disease, using the phytopathogenic bacterium Pseudomonas syringae as a representative pathogen. To establish infection and incite disease development, bacteria first have to enter a plant. This process requires flagella- and type IV pili-mediated motility, and active taxis is probably necessary for effective infection. After bacteria enter a plant’s apoplastic spaces, they need to overcome host plant resistance. To do this, they secrete a wide variety of hypersensitive response and pathogenicity (Hrp) effector proteins into the plant cytoplasm to interfere with pathogen/microbe-associated molecular pattern- and effector-triggered immunity, produce phytohormones and/or phytotoxins to suppress plant defense responses and extracellular polysaccharides to prevent access by antibiotics and to chelate Ca²⁺, and activate the multidrug resistance efflux pump to extrude antimicrobial compounds for successful colonization. Furthermore, to evoke disease, bacteria produce toxins and Hrp effectors that compromise a plant’s homeostasis and injure plant cells. The expression of these virulence factors depends on the infection processes and environmental conditions. Thus, the expression and function of virulence factors interact with each other, creating complex networks in the regulation of bacterial virulence-related genes.     

Recent developments in pathogen detection arrays: implications for fungal plant pathogens and use in practice

ABSTRACT The failure to adequately identify plant pathogens from culture-based morphological techniques has led to the development of culture-independent molecular approaches. Increasingly, diagnostic laboratories are pursuing fast routine methods that provide reliable identification, sensitive detection, and accurate quantification of plant pathogens. In addition, since plants or parts thereof can be infected by multiple pathogens, multiplex assays that can detect and quantify different pathogens simultaneously are highly desirable. Technologies that can meet these requirements, especially those involving polymerase chain reaction, are being developed and implemented in horticultural and agricultural practice. Currently, DNA array technology is the most suitable technique for multiplex detection of plant pathogens. Recently, a quantitative aspect was added to this technology, making DNA arrays highly attractive for various research and practical applications. Here, we review the most important recent advances in molecular plant pathogen diagnostics, with special attention to fungal molecular diagnostics. In addition to their applicability in practice, the different criteria that have to be fulfilled for developing robust detection procedures that can routinely be used by diagnostic laboratories are discussed.