Tracking historic migrations of the Irish potato famine pathogen,Phytophthora infestans

The plant pathogen Phytophthora infestans causes late blight, a devastating disease on potato that led to the Irish potato famine during 1845-1847. The disease is considered a reemerging problem and still causes major epidemics on both potato and tomato crops worldwide. Theories on the origin of the disease based on an examination of the genetic diversity and structure of P. infestans populations and use of historic specimens to understand modern day epidemics are discussed.     

Genetic analysis of Phytophthora infestans populations in the Nordic European countries reveals high genetic variability

Late blight, caused by the oomycete Phytophthora infestans, is the most important disease of potato (Solanum tuberosum). The pathogen is highly adaptable and to get an overview of the genetic variation in the Nordic countries, Denmark, Finland, Norway and Sweden we have analyzed 200 isolates from different fields using nine simple-sequence repeat (SSR) markers. Forty-nine alleles were detected among the nine SSR loci and isolates from all four Nordic countries shared the most common alleles across the loci. In total 169 multilocus genotypes (based on seven loci) were identified among 191 isolates. The genotypic diversities, quantified by a normalized Shannon's diversity index (H(s)), were 0.95 for the four Nordic countries. The low F(ST) value of 0.04 indicates that the majority of variation is found within the four Nordic countries. The large number of genotypes and the frequency distribution of mating types (60% A1) support the hypothesis that sexual reproduction is contributing notably to the genetic variation of P. infestans in the Nordic countries.     

Oospore Germination and Formation by the Late Blight Pathogen Phytophthora infestans in vitro and under Field Conditions

The ability of the late blight pathogen Phytophthora infestans to form oospores in leaves of seven potato cultivars was examined at different incubation temperatures under controlled environmental conditions and under field conditions. At 10°C, the oospore formation in three intermediate-resistant cultivars all differed significantly from each other (P < 0.05), with the lowest amount formed in cv. Asterix. This latter cultivar did not form oospores at any other temperature. Under field conditions oospores were formed abundantly in a naturally infected field. A significant date by cultivar interaction showed that P. infestans increased the oospore formation in foliage by time in cvs Columbo, Hertha and Matilda, whereas no significant differences between dates were found for other cultivars. The genetic structure of P. infestans in the naturally infected field plot, where oospores formed abundantly, was studied by using amplified fragment length polymorphism and a high genetic diversity was revealed. Oospore germination from two Scandinavian (A1 and A2) P. infestans isolates was stimulated in visible light and in 1 : 2 and 1 : 10 soil extract. The effect of light and nutrients on oosporogenesis is discussed.     

Genotype and sex-based host variation in behaviour and susceptibility drives population disease dynamics

Host heterogeneity in pathogen transmission is widespread and presents a major hurdle to predicting and minimizing disease outbreaks. Using Drosophila melanogaster infected with Drosophila C virus as a model system, we integrated experimental measurements of social aggregation, virus shedding, and disease-induced mortality from different genetic lines and sexes into a disease modelling framework. The experimentally measured host heterogeneity produced substantial differences in simulated disease outbreaks, providing evidence for genetic and sex-specific effects on disease dynamics at a population level. While this was true for homogeneous populations of single sex/genetic line, the genetic background or sex of the index case did not alter outbreak dynamics in simulated, heterogeneous populations. Finally, to explore the relative effects of social aggregation, viral shedding and mortality, we compared simulations where we allowed these traits to vary, as measured experimentally, to simulations where we constrained variation in these traits to the population mean. In this context, variation in infectiousness, followed by social aggregation, was the most influential component of transmission. Overall, we show that host heterogeneity in three host traits dramatically affects population-level transmission, but the relative impact of this variation depends on both the susceptible population diversity and the distribution of population-level variation.     

The plant pathogen Phytophthora andina emerged via hybridization of an unknown Phytophthora species and the Irish potato famine pathogen, P. infestans

Emerging plant pathogens have largely been a consequence of the movement of pathogens to new geographic regions. Another documented mechanism for the emergence of plant pathogens is hybridization between individuals of different species or subspecies, which may allow rapid evolution and adaptation to new hosts or environments. Hybrid plant pathogens have traditionally been difficult to detect or confirm, but the increasing ease of cloning and sequencing PCR products now makes the identification of species that consistently have genes or alleles with phylogenetically divergent origins relatively straightforward. We investigated the genetic origin of Phytophthora andina, an increasingly common pathogen of Andean crops Solanum betaceum, S. muricatum, S. quitoense, and several wild Solanum spp. It has been hypothesized that P. andina is a hybrid between the potato late blight pathogen P. infestans and another Phytophthora species. We tested this hypothesis by cloning four nuclear loci to obtain haplotypes and using these loci to infer the phylogenetic relationships of P. andina to P. infestans and other related species. Sequencing of cloned PCR products in every case revealed two distinct haplotypes for each locus in P. andina, such that each isolate had one allele derived from a P. infestans parent and a second divergent allele derived from an unknown species that is closely related but distinct from P. infestans, P. mirabilis, and P. ipomoeae. To the best of our knowledge, the unknown parent has not yet been collected. We also observed sequence polymorphism among P. andina isolates at three of the four loci, many of which segregate between previously described P. andina clonal lineages. These results provide strong support that P. andina emerged via hybridization between P. infestans and another unknown Phytophthora species also belonging to Phytophthora clade 1c.     

Genetic diversity of Triatoma infestans (Hemiptera: Reduviidae) in Chuquisaca, Bolivia based on the mitochondrial cytochrome b gene

Partial cytochrome b DNA sequences for 62 Triatoma infestans were analyzed to determine the degree of genetic variation present in populations of this insect in the northwest region of Chuquisaca, Bolivia. A total of seven haplotypes were detected in the localities sampled. The phylogenetic relationship and population genetic structure of the haplotypes found in this region, indicate that there is greater variation in this relatively small region of Bolivia than what has been previously reported by studies using the same gene fragment, for more distant geographic areas of this country. In addition, a comparison of rural and peri-urban localities, indicate that there is no difference in the genetic variation of T. infestans between these two environments.     

Genetic Variation of Sclerotinia sclerotiorum from Multiple Crops in the North Central United States

Sclerotinia sclerotiorum is an important pathogen of numerous crops in the North Central region of the United States. The objective of this study was to examine the genetic diversity of 145 isolates of the pathogen from multiple hosts in the region. Mycelial compatibility groups (MCG) and microsatellite haplotypes were determined and analyzed for standard estimates of population genetic diversity and the importance of host and distance for genetic variation was examined. MCG tests indicated there were 49 different MCGs in the population and 52 unique microsatellite haplotypes were identified. There was an association between MCG and haplotype such that isolates belonging to the same MCG either shared identical haplotypes or differed at no more than 2 of the 12 polymorphic loci. For the majority of isolates, there was a one-to-one correspondence between MCG and haplotype. Eleven MCGs shared haplotypes. A single haplotype was found to be prevalent throughout the region. The majority of genetic variation in the isolate collection was found within rather than among host crops, suggesting little genetic divergence of S. sclerotiorum among hosts. There was only weak evidence of isolation by distance. Pairwise population comparisons among isolates from canola, dry bean, soybean and sunflower suggested that gene flow between host-populations is more common for some crops than others. Analysis of linkage disequilibrium in the isolates from the four major crops indicated primarily clonal reproduction, but also evidence of genetic recombination for isolates from canola and sunflower. Accordingly, genetic diversity was highest for populations from canola and sunflower. Distribution of microsatellite haplotypes across the study region strongly suggest that specific haplotypes of S. sclerotiorum are often found on multiple crops, movement of individual haplotypes among crops is common and host identity is not a barrier to gene flow for S. sclerotiorum in the north central United States.     

Genetic Diversity of Rice False Smut Fungus, Ustilaginoidea virens and its Pronounced Differentiation of Populations in North China

Rice false smut caused by Ustilaginoidea virens is an important constraint affecting rice yield and quality in Asia. In China, rice false smut is especially severe in the japonica rice-growing areas in the North China. Nothing is known neither about the diversity of the pathogen in this region nor about the characteristic of its local population. In this study, 110 U. virens isolates sampled from Liaoning and Beijing of North China were analysed using amplified fragment length polymorphism (AFLP) markers to primarily understand the genetic diversity of this pathogen and its population characteristics. At the genetic distance of 0.32, all isolates were divided into two groups. Group A consists of two subgroups differentiated at the genetic distance of 0.55, subgroup 1 included all isolates from Liaoning with an average genetic similarity over 0.82 and subgroup 2 included 27 isolates from Beijing with similarity of 0.74. Group B consists of 28 isolates from Beijing having a diversity of 0.054. The isolates from the Liaoning province, where rice false smut has generated different take-all epidemic for 20 years, showed a genetic diversity of 0.305, which was approximately equally distributed within and among populations. Whereas genetic diversity was 0.458 among isolates from Beijing, an extremely high level of genetic differentiation among 55 isolates was observed in this disease hotspot. Our results suggested that the populations among different locations where sexual stage of the pathogen was rare to be found within ecological region were similar, and the variation of this pathogen has mainly arisen via asexual mechanisms. The migration through human activities in breeding perhaps provides a means of transporting the pathogen from one region to another.     

Effects of the foliar-applied protein "Harpin(Ea)" (messenger) on tomatoes infected with Phytophthora infestans

The active ingredient in Messenger, is Harpin(Ea), a naturally occurring protein derived from Erwinia amylovora, a causal agent of fire blight. When Messenger is applied to a plant, the protein Harpin(Ea) binds foliar receptors to it. The receptors recognize the presence of Harpin(Ea), sending a signal that a pathogen is present, actually "tricking" the plant into thinking that it is under attack. This binding process triggers a cascade of responses affecting a global change of gene expressions, stimulating several distinct biochemical pathways within the plant responsible for growth and disease and insect resistance. The objective of this work is to characterize the development of an induced resistance against Phytophthora infestans. No effective treatment is currently available against this pathogenic agent, which causes the loss of complete harvests of different crops. Tomato plants with and without Messenger applications were inoculated with Phytophthora infestans in the same way. In addition, some plants with and without Messenger applications were not inoculated. Inoculated plants were symptomatologically checked for local and systemic symptoms. Evaluations of the number of tomatoes produced, with or without damage, and their growth, were also carried out. Based on the data obtained from the assays, significant changes were observed in the parameters measured due to Messenger treatment. The severe damage of this disease was reduced in the plants which received Messenger applications. These results open up new pathways in the control of diseases like Phytophthora infestans, in which effective means to combat them still do not exist, or these means are harmful to the environment.     

Inhibitory effect of a defensin gene from the Andean crop maca (Lepidium meyenii) against Phytophthora infestans

In this study, we report the isolation of a defensin gene, lm-def, isolated from the Andean crop 'maca' (Lepidium meyenii) with activity against the pathogen Phytophthora infestans responsible of late blight disease of the potato and tomato crops. The lm-def gene has been isolated by polymerase chain reaction (PCR) using degenerate primers corresponding to conserved regions of 13 plant defensin genes of the Brassicaceae family assuming that defensin genes are highly conserved among cruciferous species. The lm-def gene belongs to a small multigene family of at least 10 members possibly including pseudogenes as assessed by genomic hybridization and nucleotide sequence analyses. The deduced mature Lm-Def peptide is 51 amino acids in length and has 74-94% sequence identity with other plant defensins of the Brassicaceae family. The Lm-Def peptide was produced as a fusion protein using the pET-44a expression vector and purified using an immobilized metal ion affinity chromatography. The recombinant protein (NusA:Lm-Def) exhibited in vitro activity against P. infestans. The NusA:Lm-Def protein caused growth inhibition and hyphal damage at concentration not greater than 0.4 microM. In contrast, the NusA protein alone expressed and purified similarly did not show any activity against P. infestans. Therefore, these results indicate that the lm-def gene isolated from maca belong to the plant defensin family with activity against P. infestans. Its expression in potato, as a transgene, might help to control the late blight disease caused by P. infestans with the advantage of being of plant origin.     

Genomic advances will herald new insights into the Brassica: Leptosphaeria maculans pathosystem

The study of the relationship between plants and phytopathogenic fungi is one of the most rapidly moving fields in the plant sciences, the findings of which have contributed to the development of new strategies and technologies to protect crops. Plants employ sophisticated mechanisms to perceive and appropriately defend themselves against pathogens. A good example of plant and pathogen evolution is the gene-for-gene interaction between the fungal pathogen Leptosphaeria maculans, the causal agent of blackleg disease, and Brassica crops. This interaction has been studied at the genetic and physiological level due to its agro-economic importance. The newly available genome sequence for Brassica spp. and L.?maculans will provide the resources to study the co-evolution of this plant and pathogen. Particularly, an understanding of the co-evolution of genes responsible for virulence and resistance will lead to improved plant protection strategies for Brassica canola and provide a model to understand plant-pathogen interactions in other major crops. This review summarises the research-to-date in the study of the Brassica-L.?maculans gene-for-gene interaction, with a focus on the genetics of resistance in Brassica and the wealth of information to be gained from genome sequencing efforts.     

Linkage Between Virulence Genes, Compatibility Types and Sexual Recombination in the Swedish Population of Bremia lactucae

The host pathogen interaction between Lactuca sativa and Bremia lactucae fits a gene-for-gene model well. Twelve resistance genes of the host are matched by twelve genes for virulence in the pathogen. The evolution of the parasite involves drastic changes in virulence frequencies, and a great diversity in virulence even on a sub-poipulation level. Bremia is a heterothallic, obligate parasite, in which presence of two mating types is needed for sexual reproduction. Sexual recombination probably occurs frequently, indicated by simultaneous occurrence of mating types in commercial lettuce crops, zygote formation, and sufficiently high oospore germination. The pattern of variation agrees well with that of a diploid, out- crossing organism with frequent sexual recombination. Unexpected high frequencies of some of the unnecessary v-genes are probably due to genetic linkage with another "necessary" v-gene.     

Mitochondrial 16S DNA variation in populations of Triatoma infestans from Argentina

Abstract.  Variation in the mtDNA 16S ribosomal RNA gene in populations of Triatoma infestans (Klug) was surveyed. DNA sequence comparisons yielded 18 haplotypes among 130 individuals from 16 localities that represent a large proportion of the range of T. infestans in Argentina. The most common genotype in all populations was found in 76.9% of individuals and two other haplotypes were shared among different populations. The remaining 15 haplotypes were present exclusively in one of the populations, suggesting currently low levels of genetic exchange. Analysis of mtDNA 16S sequences uncovered substantial genetic variation among T. infestans populations. Haplotype and nucleotide diversities varied among populations, from 0% to 0.84% and 0% to 0.29%, respectively. It appears that this locus has a low mutation rate. Uncorrected pairwise differences of T. infestans haplotypes ranged from 0% to 1.2%. The molecular phylogeny supported the monophyly of T. infestans haplotypes and clustered two different pairs of haplotypes with a moderate degree of bootstrap support (∼ 60%). Mitochondrial DNA phylogeographic differentiation was not evident, suggesting a recent rapid spread of the species. Analysis of molecular variance showed hierarchical structure in the data. Considerably less variation was found among T. infestans populations from the northwest and northeast regions than among those belonging to the central area. Such a lack of variation may be indicative of one or more past population bottlenecks.     

A novel Phytophthora infestans haustorium-specific membrane protein is required for infection of potato

Phytophthora infestans causes late-blight, a devastating and re-emerging disease of potato crops. During the early stages of infection, P. infestans differentiates infection-specific structures such as appressoria for host epidermal cell penetration, followed by infection vesicles, and haustoria to establish a biotrophic phase of interaction. Here we report the cloning, from a suppression subtractive hybridization library, of a P. infestans gene called Pihmp1 encoding a putative glycosylated protein with four closely spaced trans-membrane helices. Pihmp1 expression is upregulated in germinating cysts and in germinating cysts with appressoria, and significantly upregulated throughout infection of potato. Transient gene silencing of Pihmp1 led to loss of pathogenicity and indicated involvement of this gene in the penetration and early infection processes of P. infestans. P. infestans transformants expressing a Pihmp1::monomeric red fluorescent protein (mRFP) fusion demonstrated that Pihmp1 was translated in germinating sporangia, germinating cysts and appressoria, accumulated in the appressorium, and was located at the haustorial membrane during infection. Furthermore, we discovered that haustorial structures are formed over a 3 h period, maturing for up to 12 h, and that their formation is initiated only at sites on the surface of intercellular hyphae where Pihmp1::mRFP is localized. We propose that Pihmp1 is an integral membrane protein that provides physical stability to the plasma membrane of P. infestans infection structures. We have provided the first evidence that the surface of oomycete haustoria possess proteins specific to these biotrophic structures, and that formation of biotrophic structures (infection vesicles and haustoria) is essential to successful host colonization by P. infestans.     

Macrogeographic and microgeographic genetic structure of the Chagas’ disease vector <Emphasis Type="Italic">Triatoma infestans</Emphasis> (Hemiptera: Reduviidae) from Catamarca,Argentina

The genetic structure in populations of the Chagas' disease vector Triatoma infestans from six localities belonging to areas under the same insecticide treatment conditions of Catamarca province (Argentina) was examined at macrogeographical and microgeographical scales. A total of 238 insects were typed for 10 polymorphic microsatellite loci. The average observed and expected heterozygosities ranged from 0.319 to 0.549 and from 0.389 to 0.689, respectively. The present results confirm that populations of T. infestans are highly structured. Spatial genetic structure was detectable at macrogeographical and microgeographical levels. Comparisons of the levels of genetic variability between two temporal samples were carried out to assess the impact of the insecticide treatment. The genetic diversity of the population was not significantly affected after insecticide use since different genetic parameters (allele number, observed and expected heterozygosities) remained stable. However, loss of low frequency alleles and not previously found alleles were detected. The effective population size (N(e)) estimated was substantially lower in the second temporal sample than in the first; nevertheless, it is possible that the size of the remnant population after insecticide treatment was still large enough to retain the genetic diversity. Very few individuals did not belong to the local T. infestans populations as determined by assignment analyses, suggesting a low level of immigration in the population. The results of the assignment and first-generation migrant tests suggest male-biased dispersal at microgeographical level.     

七筋菇自然居群的遗传结构分析

采用ISSR分子标记,对七筋菇(Clintonia udensis)17个居群的遗传多样性与遗传结构进行了研究。结果表明:七筋菇不同居群的多态位点百分率PPB为11.90%~59.52%,总的多态位点百分率PPB为98.8%,具有高的遗传多样性。Shannon多样性指数(0.6903)和基因分化系数(GST=0.6944)均揭示出七筋菇居群间存在明显的遗传差异,AMOVA分析结果也显示遗传变异主要发生在居群之间(81.47%),而居群内部的遗传变异仅为18.53%。七筋菇居群间的遗传距离从0.1871~0.6632,平均为0.3838,大于同一物种居群间的平均遗传距离值(0.05),同样表明七筋菇居群间的遗传多样性存在较大差异。七筋菇居群间的基因流Nm=0.2200,远远低于一般广布种植物的基因流(Nm=1.881)。Mantel检测显示居群间的遗传距离与地理距离之间没有显著相关性(r=0.029,P=0.3196)。七筋菇分布范围广以及其进化历史是其具有高遗传多样性的原因;居群间存在较高遗传变异可能是由于七筋菇本身的生物学特性、有限的基因流以及遗传漂变等原因造成的。