Long‐distance dispersal and its influence on adaptation to host resistance in a heterogeneous landscape

Small propagules like pollen or fungal spores may be dispersed by the wind over distances of hundreds or thousands of kilometres, even though the median dispersal may be only a few metres. Such long‐distance dispersal is a stochastic event which may be exceptionally important in shaping a population. It has been found repeatedly in field studies that subpopulations of wind‐dispersed fungal pathogens virulent on cultivars with newly introduced, effective resistance genes are dominated by one or very few genotypes. The role of propagule dispersal distributions with distinct behaviour at long distances in generating this characteristic population structure was studied by computer simulation of dispersal of clonal organisms in a heterogeneous environment with fields of unselective and selective hosts. Power‐law distributions generated founder events in which new, virulent genotypes rapidly colonized fields of resistant crop varieties and subsequently dominated the pathogen population on both selective and unselective varieties, in agreement with data on rust and powdery mildew fungi. An exponential dispersal function, with extremely rare dispersal over long distances, resulted in slower colonization of resistant varieties by virulent pathogens or even no colonization if the distance between susceptible source and resistant target fields was sufficiently large. The founder events resulting from long‐distance dispersal were highly stochastic and exact quantitative prediction of genotype frequencies will therefore always be difficult.     

Virulence development and genetic polymorphism in Meloidogyne incognita (Kofoid & White) Chitwood after prolonged exposure to sublethal concentrations of nematicides and continuous growing of resistant tomato cultivars

BACKGROUND: The root‐knot nematode, Meloidogyne incognita (Kofoid & White) Chitwood, is an important plant pathogen damaging to tomato. Continuous use of resistant tomato cultivars and nematicides for its effective management might lead to resistance break‐up or nematicide failure. Genetic variability and virulence in M. incognita on susceptible Pusa Ruby tomato were analysed by bioassay, esterase and DNA polymorphism after a 5 year weekly exposure to carbofuran, carbosulfan, cadusafos and triazophos at 0.0125, 0.0250 and 0.0500 µg g^?1. Virulence in M. incognita after a 5 year multiplication on resistant tomatoes was assessed. RESULTS: The nematicidal treatments resulted in the development of virulent M. incognita populations. Their invasion potential increased significantly after continuous exposure to low concentrations of the nematicides. Also, growing resistant tomato cultivars for ten successive seasons resulted in a 6.6% increase in the invasion potential. These virulent populations exhibited 1–3 additional esterase and DNA bands compared with untreated populations. CONCLUSION: A 5 year exposure of M. incognita to sublethal concentrations of nematicides or resistant tomato cultivars exerted enough selection pressure to cause genomic alterations for virulence development. Isozyme markers can be used for rapid and precise diagnostics of field populations by advisory services, enabling judicious remedial management decisions. Copyright © 2009 Society of Chemical Industry     

Influence of spatial structure on pathogen colonization and extinction: a test using an experimental metapopulation

The Linum marginale–Melampsora lini plant–pathogen interaction has been studied extensively with regard to its epidemiology and population genetic structure (host resistance and pathogen virulence) in a natural metapopulation. In this study, this system was used in an experimental metapopulation approach to investigate explicitly how the distance (degree of isolation) between local population patches influences disease dynamics within a growing season, as well as the genetic structure of pathogen populations through stochastic colonization and extinction processes. The experimental design centred on four replicate sets of populations, within which patches were spaced at increasingly greater distances apart. Each patch consisted of an identical set of host and pathogen genotypes, with each pathogen genotype having the ability to attack only one of four host-resistance types. Over the 2 years of the experiment, the results showed clear 'boom-and-bust' epidemic patterns, with the strongest determinant of disease dynamics within a growing season being the identity of particular host–pathogen genotypic combinations. However, there were also significant effects of spatial structure, in that more isolated patches tended to exhibit lower levels of disease during epidemic peaks than patches that were close together. Extinction of pathogen genotypes from individual populations was positively related to the severity of disease during preceding epidemic peaks, but negatively related to the level of disease present at the final census prior to overwintering. The probability of recolonization of pathotypes into populations during the second growing season was most strongly related to the distance to the nearest neighbouring source population in which a given pathotype was present. Overall, these results highlight the importance of spatial scale in influencing the numerical and genetical dynamics of pathogen populations.     

Effects of host and pathogen genotypes on inducibility of resistance in tomato (Solanum lycopersicum) to Phytophthora infestans

Thirteen tomato (Solanum lycopersicum) accessions were tested for inducibility of resistance against two isolates of Phytophthora infestans using BABA (dl ‐3‐amino butyric acid) as the inducing agent. In a more detailed trial, six of the accessions were assessed for inducibility of resistance to six P. infestans isolates on three leaves of different age per plant. Plants were inoculated 1 week after treatment with BABA. Area under the disease progress curve (AUDPC), sporulation capacity (SC) and infection efficiency (IE) were all affected by treatment with BABA. On leaves of all ages AUDPC was most affected by induction (43–100% reduction on the youngest leaves) followed by SC (14–100%) and IE (0–100% reduction). Tomato genotypes varied significantly in inducibility of resistance against P. infestans and the degree of induction generally decreased with increasing leaf age, whilst the absolute susceptibility with respect to AUDPC and SC rarely changed. The level of induction was not always related to the resistance level of the tomato accession and it was significantly influenced by the pathogen isolate used for challenge inoculation. The results show that inducibility of resistance is a selectable trait that is, however, isolate‐specific.     

甘肃省大麦条纹病菌遗传多样性及致病力差异

为了解甘肃省大麦条纹病病原菌Pyrenophora graminea的遗传多样性及致病力差异,运用RAPD分子标记技术对大麦条纹病菌不同菌株进行遗传多样性分析,并采用三明治法进行菌株致病力差异研究。结果表明:17个RAPD标记从45个菌株中扩增出126条带,平均每个标记7.41条带,遗传相似系数范围为0.468 3～0.984 1,平均值为0.830 8,当遗传相似系数为0.723 6时,可将供试菌株划分为4个类群,分别包含41、2、1和1个菌株;致病力测定结果显示菌株QWC较菌株QQ致病力强,两菌株除在品种‘甘啤2号’和‘GP-3’上无致病力外,在其他供试品种上致病力均存在差异。表明大麦条纹病菌不同菌株间存在遗传差异,且菌株QWC和菌株QQ存在致病力差异。     

Genetic diversity of the pea root pathogen Aphanomyces euteiches in Europe

The oomycete pathogen Aphanomyces euteiches causes root rot in various legume species. In this study we focused on A. euteiches causing root rot in pea (Pisum sativum), thereby being responsible for severe yield losses in pea production. We aimed to understand the genetic diversity of A. euteiches in Europe, covering a north-to-south gradient spanning from Sweden, Norway and Finland to the UK, France and Italy. A collection of 85 European A. euteiches strains was obtained, all isolated from infected pea roots from commercial vining pea cultivation fields. The strains were genotyped using 22 simple-sequence repeat markers. Multilocus genotypes were compiled and the genetic diversity between individual strains and population structure between countries was analysed. The population comprising strains from Italy was genetically different and did not share ancestry with any other population. Also, strains originating from Finland and the eastern parts of Sweden were found to be significantly different from the other populations, while strains from the rest of Europe were more closely related. A subset of 10 A. euteiches strains from four countries was further phenotyped on two susceptible pea genotypes, as well as on one genotype with partial resistance towards A. euteiches. All strains were pathogenic on all pea genotypes, but with varying levels of disease severity. No correlation between the genetic relatedness of strains and virulence levels was found. In summary, our study identified three genetically distinct groups of A. euteiches in Europe along a north-to-south gradient, indicating local pathogen differentiation.     

Phenotype and spectrum of action of the Pvr4 resistance in pepper against potyviruses, and selection for virulent variants

The dominant Pvr4 gene identified in Capsicum annuum cv. Criollo de Morelos 334 (CM334) is frequently used in pepper cultivars because it possesses one of the largest spectra of action among plant virus resistance genes. This gene was previously shown to confer efficient resistance to all known Potato virus Y isolates, to Pepper mottle virus , to Pepper yellow mosaic virus and to Ecuadorian rocoto virus. This study showed that the W4 line, derived from CM334 and carrying Pvr4 , was also resistant to Peru tomato mosaic virus and Pepper severe mosaic virus , but not to Pepper veinal mottle virus , Chilli veinal mottle virus or Tobacco etch virus . It was noticed that the phenotype of the resistance was atypical since, in the W4 line, hypersensitive reaction or extreme resistance could be observed, depending on virus isolates and inoculated organs. Despite the large deployment of Pvr4 in hybrid cultivars, the numerous tests performed in controlled conditions and the use of W4 serial back-inoculations with potyvirus isolates controlled by this line, no virulent variant isolates were obtained. However, it was shown that the use of graft inoculation experiments allow PVY virulent variants to be selected.     

Understanding pathogen population structure and virulence variation for efficient resistance breeding to control cucurbit powdery mildews

Cucurbit powdery mildew (CPM) is caused most frequently by well-differentiated obligate erysiphaceous ectoparasites Golovinomyces orontii and Podosphaera xanthii, which vary in their ecology and virulence. All economically important cucurbit crops host both of these CPM species. Breeding of cucurbits for CPM resistance is highly important worldwide, but adequate knowledge of CPM species determination, as well as virulence structure, population dynamics, and spatiotemporal variation of these pathogens, has not yet been achieved. New tools have been developed to enhance research on CPM virulence variation for more efficient breeding and seed and crop production. A set of differential genotypes of Cucumis melo, with high differentiation capacity, may contribute substantially to understanding of variation in CPM virulence at both individual and population levels. Long-term observations (2001–2012) of CPM pathogens in the Czech Republic were used to analyse virulence variation within and among annual CPM populations and demonstrate the utility of recently developed tools for studying species variability and virulence variation of CPM pathogens worldwide. Detailed analyses of diversity and spatiotemporal fluctuations in the composition of CPM populations provide crucial information for shaping breeding programmes and predicting the most effective sources of race-specific resistance. The primary aim of this work was to create a uniform framework for determination of CPM species structure and diversity, virulence phenotypes, virulence and phenotype frequencies, phenotype complexity, dynamics, and variation within and among CPM populations. In addition, practical advice is presented on how to select the most relevant data and interpret them for use in cucurbit resistance breeding.     

Bacterial wilt disease: Host resistance and pathogen virulence mechanisms

Bacterial wilt (BW), caused by Ralstonia solanacearum, is one of the most destructive bacterial diseases of Solanaceous species worldwide. The species infects plants in more than 200 species and 50 families and was ranked second in a list of the top 10 most scientifically and economically important bacterial plant pathogens [1]. The molecular mechanisms underlying resistance and the functions of R. solanacearum effectors are beginning to be uncovered, and much remains to be discovered. In this mini-review, we provide a summary of host resistance and R. solanacearum virulence mechanisms, with a focus on tomato.     

Characterisation of the European pathogen population of Magnaporthe grisea by DNA fingerprinting and pathotype analysis

The genetic variability among 41 isolates of the blast pathogen (Magnaporthe grisea) from five European rice growing countries was studied. The genealogy of the isolates was investigated by DNA fingerprinting and the results compared to the degree of similarity for (a)virulence factors. Fingerprinting grouped the isolates into five discrete lineages, that typically showed less than 65% band similarity. Within each lineage, two or more haplotypes were detected with a band similarity of 80% or higher. Each lineage showed a characteristic virulence pattern. All isolates of lineage E5 belonged to the same pathotype. The other lineages were composed of clusters of closely related pathotypes that showed variation for virulence to cultivars with certain known resistance genes, while remaining invariably (a)virulent to others. In most cases, lineage classification of an isolate could be easily inferred by its pathotype. Certain resistance genes and certain lineage-excluding resistance gene combinations appear to provide protection against all of the virulence factors sampled.     

Population genetic diversity and structure of the pecan scab pathogen,Venturia effusa,on cv. Desirable and native seedlings,and the impact of marker number

Scab (Venturia effusa) is the major cause of economic loss in pecan in the south-eastern USA. We explored population genetic diversity and structure among orchards of cv. Desirable and native seedlings, and within-orchard variability among trees of all cultivars sampled. We compared the ability of 30, 15 and 7 previously developed microsatellites to characterize the population genetic diversity and structure of V. effusa. Analyses of molecular variance (AMOVA) provided little evidence of structure dependent on cultivar, but there was some evidence of structure between orchards of a cultivar based on distance. Individual orchard AMOVA showed that three of 11 orchards had between-tree population structure. Among six populations from cv. Desirable, a Mantel test showed that geographic distance was related to the pairwise genetic divergence (R² = 0.84). Among 11 orchards of various cultivars there was little difference in diversity using 30, 15 or 7 markers, or population structure based on AMOVA. Some minor differences in population structure were seen based on discriminant analysis of principal components, or dendrograms. Thus, depending on the objectives, future studies may use as few as 15 or 7 markers without losing ability to discern population genetic diversity or structure. More populations exhibited linkage disequilibrium when using 15 or 30 markers compared to when using seven markers. Knowledge of population genetics of V. effusa in relation to host genotype is needed to understand pathogen population interactions and gene flow, knowledge that will help underpin future breeding efforts to develop durable resistance in this long-lived orchard tree.     

小麦条锈菌有性生殖与毒性变异的研究进展

小麦条锈病是由条形柄锈菌小麦专化型Puccinia striiformis f.sp.tritici Erikss.et Henn.引起的、最具毁灭性的小麦真菌病害之一,在全世界小麦种植的国家或地区均有发生,其流行常常导致小麦严重的产量损失。利用抗病品种是防治小麦条锈病最为经济、有效的措施。然而,由于病原菌毒性变异不断产生新小种,频繁导致品种抗病性"丧失",继而引发病害流行成灾。小麦条锈菌的有性阶段过去一直未被发现,有性生殖与条锈菌变异及其在病害发生中的作用研究一直处于空白状态。近年来,随着小麦条锈菌转主寄主的发现,国内外学者开展了相关的研究工作,取得了一系列重要的研究进展。本文综述了小麦条锈病的发生症状、病菌的生物学特性、转主寄主、有性生殖与条锈菌毒性变异、转主寄主在条锈病发生中的作用、影响有性生殖发生的因素等方面的最新进展,并展望了深入开展小麦条锈菌有性生殖的研究思路。     

Evolution of a pathogen population in host mixtures: simple race–complex race competition

The evolution of the genetic structure of a pathogen population was studied in a varietal mixture with an epidemic simulator based on the model EPIMUL. The pathogen population was composed of simple races able to develop on only one component of the mixture and a complex race which developed on all mixture components. The effects on the simple race–complex race competition of a cost of virulence, of density dependence and of differential adaptation were studied. The selection for simple or complex races in the pathogen population did not depend on initial race frequencies. For a given multiplication rate, complex race frequency increased faster when the spore dispersal gradient was shallow, when distribution of initial disease was generalized, when amount of initial disease was reduced and when the number of mixture components was increased. This was attributed to a better efficacy of the mixture in controlling simple races, resulting in a higher relative fitness of the complex race. For measured values of density dependence or differential adaptation effects, the complex race was at a higher frequency after a mean number of pathogen cycles between 2.5 and 5. The effect of the cost of virulence was stronger and, in certain situations, could result in selection for simple races. In the conditions of our simulations and with the effects tested, stabilization of the pathogen population in host mixtures was unlikely to occur. However, more information is needed concerning the rate at which complex races could evolve and how quickly mixture resistance could be eroded.     

Production of hydrogen peroxide and expression of ROS‐generating genes in peach flower petals in response to host and non‐host fungal pathogens

Reactive oxygen species (ROS) play dual roles in plant–microbe interactions in that they can either stimulate host resistance or enhance pathogen virulence. Innate resistance in peach (Prunus persica) to the brown rot fungal pathogen Monilinia fructicola is very limited, and knowledge of the mechanism of virulence is rudimentary. In this study, production of hydrogen peroxide, a major component of ROS, was determined in peach flower petals in response to M. fructicola (a host pathogen) and Penicillium digitatum (a non‐host pathogen). Monilinia fructicola was able to infect flower petals while P. digitatum was not. During the host‐specific interaction, M. fructicola induced hydrogen peroxide accumulation in flower petals. Application of exogenous antioxidants significantly reduced hydrogen peroxide accumulation as well as the incidence of brown rot disease. Application of M. fructicola spores to the surface of intact flower petals induced gene expression and increased enzyme activity of NADPH oxidase and cell wall peroxidase in host tissues, resulting in the production of hydrogen peroxide. Petals inoculated with M. fructicola exhibited high levels of protein carbonylation and lipid peroxidation. No significant response in gene expression, enzyme activity or hydrogen peroxide levels was observed in peach flower petals treated with P. digitatum. These results suggest that M. fructicola, as with other necrotrophic fungi, uses the strong oxidative response as part of a virulence mechanism.     

Quantification of within‐season focal spread of wheat take‐all in relation to pathogen genotype and host spatial distribution

Within season gradient of wheat take‐all was measured in field experiments according to line or random sowing host spatial distribution, and two Gaeumannomyces graminis var. tritici (Ggt) isolates (G1i and G2i), representative of the G1 and G2 genotype groups in terms of aggressiveness. Root disease incidence and severity were assessed at six dates from early March to late June on plants located at regular distances from the inoculum sources. Simple models relating disease intensity at different levels of hierarchy fitted observed data well, and indicated a strong disease aggregation both within and among plants. Disease severity on source plants placed nearby the inoculum source increased over time, ranging from 5 to 46% at the first assessment, and from 55 to 98% at the last assessment, being in general larger for G2i than G1i. In line‐sown plots, disease progressed steadily along the line but did not extend beyond 20 cm, seldom reaching the neighbour line. Disease rarely reached 25 cm in the direct‐seeded crop stands. These results indicate that Ggt intensifies but does not spread to a large extent during a cropping season. Distance from the source, pathogen genotype and assessment date had a significant effect on disease severity according to mixed model analyses, disease spread being larger for G2i than G1i. However, no significant effect of host spatial distribution could be detected. Yield loss within 20 cm of the source plant ranged between 20 and 40%, and was not significantly affected by pathogen genotype or host spatial distribution.     

甜菜夜蛾核型多角体病毒对宿主种群的控制作用

室内试验表明,甜菜夜蛾核型多角体病毒对宿主种群有明显的控制作用.当饲毒浓度在1.14×108PIBs/mL以上时,能引起97%的甜菜夜蛾幼虫死亡;饲毒后1～8天内,致死中浓度LC50值在8.4718×105～2.6098×10 12PIBs/mL之间;在饲毒浓度范围内,致死中时间LT50在1.98～6.99天之间;感病幼虫每日病死时间分布可用时间-剂量-条件死亡率模型较好地拟合,累计病死时间分布可用Gompertz模型拟合.室内试验还表明,病毒对甜菜夜蛾种群有明显的亚致死作用.免于病死蛹重量减轻,羽化所得雌成虫产卵量下降,产卵期延长;从感病高龄幼虫发育的成虫能将病毒传播至子代,引起20%～48%的子代幼虫死亡,并使子代成虫产卵量下降.田间试验表明,病毒除具有明显的控制作用外,还有一定的后效作用.     

Plant—pathogen Interactions: Genetic and Comparative Analyses

The interactions between plants and pathogens can be shifted to favor either plant of pathogen by small changes in the environment, primarily temperature and plant nutrition, and it leaves a quandary as to whether the plant on pathogen is most affected by the change in the environment. The stage of development of a plant can affect the resistance or susceptibility to a pathogen. A plant may be susceptible to a given pathogen a one stage of development but resistant at another stage of development. The view of the gene-for-gene hypotheses as a one-for-one relationship is not supported by experiments that ask whether avirulence genes and resistance genes function alone. The term genomics has been intemreted several different ways, but its most useful impact on studies of host-pathogen interactions will, most likely, be to find all the pieces to the puzzle of how plants and pathogens communicate.     

Spatial genetic structure and dispersal of the cacao pathogen Moniliophthora perniciosa in the Brazilian Amazon

Moniliophthora perniciosa is the causal agent of witches’ broom in Theobroma cacao (cacao). Three biotypes of M. perniciosa are recognized, differing in host specificity, with two causing symptoms on cacao or Solanaceae species (C‐ and S‐biotypes), and the third found growing endophytically on lianas (L‐biotype). The objectives of this study were to clarify the genetic relationship between the three biotypes, and to identify those regions in the Brazilian Amazon with the greatest genetic diversity for the C‐biotype. Phylogenetic reconstruction based on the rRNA ITS regions showed that the C‐ and S‐biotypes formed a well‐supported clade separated from the L‐biotype. Analysis of 131 isolates genotyped at 11 microsatellite loci found that S‐ and especially L‐biotypes showed a higher genetic diversity. A significant spatial genetic structure was detected for the C‐biotype populations in Amazonia for up to 137 km, suggesting ‘isolation by distance’ mode of dispersal. However, in regions containing extensive cacao plantings, C‐biotype populations were essentially ‘clonal’, as evidenced by high frequency of repeated multilocus genotypes. Among the Amazonian C‐biotype populations, Acre and West Amazon displayed the largest genotypic diversity and might be part of the centre of diversity of the fungus. The pathogen dispersal may have followed the direction of river flow downstream from Acre, Rondônia and West Amazon eastward to the rest of the Amazon valley, where cacao is not endemic. The Bahia population exhibited the lowest genotypic diversity, but high allele richness, suggesting multiple invasions, with origin assigned to Rondônia and West Amazon, possibly through isolates from the Lower Amazon population.