Molecular aspects of the potato — Phytophthora infestans interaction

The fungal pathogenPhytophthora infestans is the causal organism of late blight, one of the most devastating diseases of potato. In the past, various aspects of the potato-P. infestans interaction have been studied extensively. In this paper we briefly review the current knowledge of the molecular events associated with the interaction and in addition we discuss a new approach for analyzing the molecular basis of pathogenicity ofP. infestans.     

Can weed hosts increase aggressiveness of Phytophthora infestans on potato?

Potato late blight, caused by Phytophthora infestans, is a major disease in potato production throughout the world. In southern Sweden, hairy nightshade (Solanum physalifolium), an alternative non-crop host to the pathogen, is an increasing weed problem. Single-lesion leaves infected by P. infestans were collected from potato and hairy nightshade to determine phenotypic and genotypic population differentiation of P. infestans between the two hosts. Genotypic variation was estimated using microsatellites as markers. The results showed no genotypic differentiation in the samples between the two hosts. Aggressiveness tests were performed using the sampled isolates to cross-inoculate potato and hairy nightshade. The proportion of infected leaves, latency period, lesion growth rate, and sporulation capacity were measured. For isolates from hairy nightshade, the odds of infection were higher on both hosts combined. When tested on potato leaves, isolates from hairy nightshade showed a significantly shorter latency period and higher sporulation capacity compared with isolates from potato. This indicates that an alternative host can filter populations of P. infestans toward a higher aggressiveness, which could lead to increasing problems in controlling potato late blight.     

Quantification of late blight resistance of potato using transgenic Phytophthora infestans expressing β-glucuronidase

A transgenic Phytophthora infestans strain that constitutively produces and secretes -glucuronidase (GUS) was used in detached leaf assays to quantify the levels of resistance to late blight in potato cultivars Surprise, Irene, Pimpernel, Alpha and Bintje. Four days after inoculation levels of GUS activity were determined in infected leaves. Significant differences between the various cultivars were observed. Discrimination between resistant and susceptible cultivars was possible based solely on levels of GUS activity. Regression analysis revealed a positive correlation between in planta GUS levels and field resistance expressed as Area under Disease Progress Curve (ADPC).     

Phenotypic and genotypic structure of Phytophthora infestans populations on tomato and potato in the North of Thailand in 2000–2002

A total of 80 single–lesion isolates of Phytophthora infestans were collected from tomatoes and potatoes in several locations in Chiang Mai and Tak provinces in 2000–2002. These isolates were analyzed for mating type, metalaxyl sensitivity, mitochondrial DNA (mtDNA) haplotype RFLP pattern as determined by probe RG57, and for microsatellite markers. All isolates were A1 mating type. Isolates from tomato were usually sensitive to metalaxyl, but isolates from potato were usually resistant to metalaxyl. With one exception, all tomato isolates were related to the US-1 clonal lineage. With two exceptions, all potato isolates were related to two European lineages. In these two provinces, the populations of P. infestans on tomatoes are clearly different from those on potatoes.     

What is the evidence for sexual reproduction of Phytophthora infestans in Europe?

The biology of late blight of potato and tomato, caused by Phytophthora infestans, changed when sexual reproduction by the pathogen became possible in many parts of the world, including Europe. In northern Europe, especially Scandinavia, there is increasing evidence that the pathogen is reproducing sexually on a regular basis, although in other regions further south or to the west it appears to reproduce primarily in a clonal manner. The presence of both mating types, the production of viable oospores, and observations of fields with soilborne sources of inoculum are consistent with sexual reproduction. Studies with different marker systems have revealed a population structure without any dominating clonal lineages in Scandinavia, and that is most easily explained by sexual reproduction. Phytophthora infestans recovered from the soil can also be linked to parental genotypes using likelihood‐based methods when codominant markers are used. A synthesis of all the available data points to a second centre of sexual reproduction in northern Europe.     

The dynamics of Phytophthora infestans populations in the major potato-growing regions of Asia – A review

Asia is now the largest potato-producing region of the world and late blight, caused by Phytophthora infestans, is the most important pathogen limiting production. This review documents, in both the historical and the current context, the population structure of P. infestans in the major areas of potato production in Asia. Information from diverse sources regarding the stated or inferred clonal pathogen lineages present, population changes, and possible migration routes of the pathogen into the countries of this region have been reviewed to aid researchers and those involved in managing late blight in Asia. The single most important factor for population change and resultant epidemics in this region has been found to be migration of pathogen genotypes from Europe and the Americas. Reducing the impact of such migration in the future will necessitate putting in place improved phytosanitary measures. To achieve this, data sharing using global networks such as AsiaBlight and EuroBlight is imperative.     

Phenotypic and genotypic changes in French populations of Phytophthora infestans: are invasive clones the most aggressive?

The main objective of this investigation was to test the ‘always more aggressive’ hypothesis, often advocated to explain lineage replacements in clonal populations of the potato late blight oomycete Phytophthora infestans. To this end, genotypic and pathogenicity data on 1274 French isolates of P. infestans, collected over the period 2001–2008, were analysed. Overall, the populations sampled showed limited genetic diversity, with four multilocus lineages (1_A1, 2_A1, 8_A1 and 13_A2) accounting for over 80% of the isolates collected. As in other West European countries, drastic changes in these dominant clonal lineages were observed over the course of the 8 years, particularly in the appearance and propagation of the clone 13_A2. However, invasiveness of clones was not associated with higher aggressiveness; on the contrary, dominant clones had generally low or moderate aggressiveness relative to others present at the same time within the same populations. This finding challenges the link between invasive behaviour and increased aggressiveness often assumed for this biotrophic pathogen, and could reflect the existence of a trade‐off between intra‐season and inter‐season transmission. This would be consistent with the concept of inclusive fitness, which involves the abilities to both reproduce and survive.     

Host–pathogen interaction between Phytophthora infestans and Solanum nigrum, S. villosum, and S. scabrum

Potato and tomato are the two major hosts for Phytophthora infestans causing late blight. The susceptibility of leaves and whole plants of Solanum nigrum, S. villosum, and S. scabrum to infection by P. infestans was tested under laboratory conditions. Out of 39 plants representing 38 different S. nigrum accessions, 16 were highly resistant (seven accessions did not show any symptoms of infection, nine were highly resistant showing necrotic lesions in the place of infection), and 23 plants of S. nigrum were colonized by, at least, 1 of the 2 isolates of P. infestans (17 accessions were infected with two P. infestans isolates, and 6 accessions showed different reactions depending on the isolate used for inoculation). Three accessions of S. villosum, and one accession of S. scabrum were tested and did not show any symptoms of infection. The majority of S. nigrum accessions infected by P. infestans in a detached leaf assay were also infected in the whole plant assay. The reaction of field- and greenhouse-grown plants to inoculation with P. infestans in detached leaf assays was similar, but in some cases leaves from field-grown plants reacted as resistant in comparison with the leaves from greenhouse-grown plants, which were susceptible.     

Invasion of Phytophthora infestans at the landscape level: how do spatial scale and weather modulate the consequences of spatial heterogeneity in host resistance?

Strategic spatial patterning of crop species and cultivars could make agricultural landscapes less vulnerable to plant disease epidemics, but experimentation to explore effective disease-suppressive landscape designs is problematic. Here, we present a realistic, multiscale, spatiotemporal, integrodifference equation model of potato late blight epidemics to determine the relationship between spatial heterogeneity and disease spread, and determine the effectiveness of mixing resistant and susceptible cultivars at different spatial scales under the influence of weather. The model framework comprised a landscape generator, a potato late blight model that includes host and pathogen life cycles and fungicide management at the field scale, and an atmospheric dispersion model that calculates spore dispersal at the landscape scale. Landscapes consisted of one or two distinct potato-growing regions (6.4-by-6.4-km) embedded within a nonhost matrix. The characteristics of fields and growing regions and the separation distance between two growing regions were investigated for their effects on disease incidence, measured as the proportion of fields with ≥1% severity, after inoculation of a single potato grid cell with a low initial level of disease. The most effective spatial strategies for suppressing disease spread in a region were those that reduced the acreage of potato or increased the proportion of a resistant potato cultivar. Clustering potato cultivation in some parts of a region, either by planting in large fields or clustering small fields, enhanced the spread within such a cluster while it delayed spread from one cluster to another; however, the net effect of clustering was an increase in disease at the landscape scale. The planting of mixtures of a resistant and susceptible cultivar was a consistently effective option for creating potato-growing regions that suppressed disease spread. It was more effective to mix susceptible and resistant cultivars within fields than plant some fields entirely with a susceptible cultivar and other fields with a resistant cultivar, at the same ratio of susceptible to resistant potato plants at the landscape level. Separation distances of at least 16 km were needed to completely prevent epidemic spread from one potato-growing region to another. Effects of spatial placement of resistant and susceptible potato cultivars depended strongly on meteorological conditions, indicating that landscape connectivity for the spread of plant disease depends on the particular coincidence between direction of spread, location of fields, distance between the fields, and survival of the spores depending on the weather. Therefore, in the simulation of (airborne) pathogen invasions, it is important to consider the large variability of atmospheric dispersion conditions.     

Regulation of semiochemicals — global aspects

A general awareness of environmental concerns, an increasing chemophobic populace and the reregistration of pesticides, both in the United States and in the European Community, with the concomitant loss of many pest control chemicals through cancellation of their registrations has provided an impetus for the research, development and commercialization of alternative, environmentally benign and safer plant protection products. Amongst such alternatives are pheromones, a class of semiochemicals whose commercial development is being championed by a neophyte industry made up, for the most part, of undercapitalized entrepreneurial companies for whom the major hurdle to the introduction of products into the marketplace is the time and cost required to complete the regulatory process. That pheromones and other semiochemicals are different from chemical insecticides has been recognized by several national regulatory agencies and the Food and Agricultural Organization of the United Nations [FAO]. Although these regulatory authorities have made significant contributions towards reducing the data requirements and hence the cost of pheromone product registration, it is widely felt that more can, and should be done to further expedite and harmonize pheromone regulation. The adoption of a structure/activity approach to the evaluation of health and environmental risks for types of lepidopteran pheromones together with a 90-day evaluation time-frame. This approach would satisfy the regulatory authorities' risk assessment for food and environmental safety while substantially reducing the cost to the registrants and expediting the registration. It is proposed that this approach be tested first in an actual product application to the US Environmental Protection Agency [EPA], with subsequent applications to other regulatory authorities. A database which could be modified already exists within the EPA and as other databases are incorporated, and shared with other regulatory authorities, harmonization of the regulation of pheromones may be possible. The application of this structure/activity approach should be expanded to other types of semiochemicals as the database is enlarged.     

Enkele waarnemingen over versmelten van zoösporen bij Phytophthora infestans (Mont.) de Bary

Summary Zoospores ofPhytophthora infestans were seen to fuse in pairs. Generally within 30 min. after zoospore discharge, fusion started by the formation of a connection between touching zoospores. Gradually the connection became shorter and thicker (fig. 1,2 and 3), and within 30 min. the two zoospores united to form a spherical body (fig. 4). The flagella were thrown off just before the compound zoospore assumed the spherical shape. The compound zoospores germinated by a germ tube within 30 min. after their formation. The analogy withCritopoulos' observations on zoospore fusion inP. capsici is discussed.     

Genotypic and phenotypic characterization of Phytophthora infestans in South Korea during 2009–2016 reveals clonal reproduction and absence of EU_13_A2 genotype

In order to better understand the Phytophthora infestans population structure in South Korea, 172 isolates were collected between 2009 and 2016 from four major potato cultivation areas. Fungicide (metalaxyl and dimethomorph) response, mating type, and microsatellite (SSR) genetic fingerprints were analysed to characterize these isolates. Ten isolates collected in Gyeongnam Province, which specializes in protected winter cultivation in polytunnels, were A2 mating type. All other isolates were A1 mating type. Overall, 42% of the isolates were resistant to metalaxyl, and 43% were sensitive. All isolates were sensitive to dimethomorph. From the SSR fingerprints, 45 distinct genotypes were identified, which could be clustered into four clonal lineages: KR_1_A1, KR_2_A2, SIB-1, and US-11. KR_1_A1 was the predominant P. infestans genotype in South Korea. KR_2_A2 was only found in Gyeongnam Province; all isolates were A2 mating type and resistant to metalaxyl. SIB-1 was dominant until 2013 but its frequency has gradually decreased in more recent years. US-11 was first found in 2014, after which its frequency has increased to become codominant with KR_1_A1. The calculated standardized index of association (I_A) suggests that the South Korean P. infestans population is undergoing clonal reproduction. When compared with populations of P. infestans from the Netherlands, it has less genetic diversity and the dominant Netherlands P. infestans genotype, EU_13_A2 (Blue_13), was not found in South Korea. Such monitoring of the pathogen population contributes to a more efficient integrated pest management-based control strategy for potato late blight control in South Korea.     

Influence of light on the Monilinia laxa–stone fruit interaction

Light represents a signal for the regulation of virulence in many microbial pathogens. Two stone fruits, nectarines and cherries, were used to investigate the influence of light on brown rot caused by Monilinia laxa. Three single-spore isolates were inoculated on nectarines and incubated under different white lights, red light, blue light, green light, and black light with two photoperiods. In addition, to understand the effect of daylight irradiance on brown rot, M. laxa was inoculated on different cherry cultivars and incubated under two simulated solar irradiations. Significantly higher disease severity and sporulation were reported on inoculated nectarines incubated under 58 W white light for 12 hr light/12 hr darkness than on nectarines incubated in continuous darkness. Only red light caused a significant increase in the incidence and severity of the disease in nectarines inoculated with the three M. laxa isolates, compared to fruit incubated under white light. High light intensity (185.45 W/m²), caused greater brown rot severity in all cherry cultivars, both early and late varieties, than low irradiance (145.85 W/m²). Significant up-regulation of the pathogenicity-related MlPNL2 gene was observed as an early response after cherry inoculation under high-intensity light, especially in late cherry cultivars, while MlPG1 expression did not show any changes under different light irradiances. M. laxa was shown to be a light-responsive fungal pathogen, and light seemed to play an active role in brown rot development.     

Characterisation of <Emphasis Type="Italic">Phytophthora infestans</Emphasis> isolates collected from potato in Estonia during 2002–2003

A collection of 101 isolates of Phytophthora infestans, obtained from seven sampling sites representing central, east and south-east Estonia during 2002 and 2003 were assessed for several phenotypic and genotypic markers. All 101 isolates were assessed for virulence and resistance to metalaxyl. Virulence to each of the 11 classic resistance genes was found among the tested isolates. The mean number of virulences per isolate was 6.3, with a very low frequency of virulence against resistance genes R5 (5%) and R9 (14%). The most common pathotypes were 1.3.4.7.8.10.11 and 1.3.4.7.10.11, representing altogether 12% of the studied strains. In terms of metalaxyl resistance, 30 resistant, 52 intermediate and 19 sensitive isolates were found. A subgroup of 50 isolates was assessed for mating type, allozymes [glucose-6-phosphate isomerase (Gpi) and peptidase (Pep)], DNA fingerprints with probe RG57 and mtDNA haplotype. Of this subset, 30 were A1 and 20 were A2. Collections from three of the seven fields contained both mating types. Allozyme analysis did not reveal any polymorphism. However, 19 diverse RG57 fingerprints were detected, and two mitochondrial DNA haplotypes, Ia and IIa, were detected. By combining the mating type, mtDNA haplotype and RG57 fingerprint data, 26 multilocus genotypes were identified, of which 18 were detected only once. Genotypic diversity measured by the normalised Shannon diversity index was high (0.76). The large number of multilocus genotypes and the presence of both mating types in some fields indicate that sexual reproduction may take place in Estonian populations of P. infestans.     

Induced resistance in potato to <Emphasis Type="Italic">Phytphthora infestans</Emphasis>—effects of BABA in greenhouse and field tests with different potato varieties

We have investigated to what degree induced resistance with β-aminobutyric acid (BABA) can protect potato from late blight infection under Swedish field conditions and if synergistic interactions occur if BABA is applied in combination with a commonly used fungicide, Shirlan. In greenhouse experiments we also investigated the durability of BABA induced resistance, the dose-response relationships in susceptible (Bintje) and partially resistant (Ovatio, Suberb) cultivars and effects of combined applications of BABA and fungicides. We found a clear effect of BABA on P. infestans infection of greenhouse grown potato plants. The lesion sizes were reduced by on average 40–50% compared to untreated control. However, this effect lasted for only 4–5 days after BABA treatment and then the efficacy was lower. When BABA was given in combination with the fungicides it appeared to have an additive effect both in greenhouse and field experiments. Higher concentrations of BABA gave a stronger protective effect. The partially resistant cultivars Ovatio and Superb reacted to lower concentrations of BABA where no effect was found in susceptible Bintje. According to our field data, 20–25% reduction of the fungicide dose in combination with BABA gave on average the same result on late blight development as full dose Shirlan alone; while reduced dose of Shirlan alone sometimes resulted in less effective protection. Our results indicate that induced resistance could be used in practice in combinations with fungicides in order to reduce the amount of toxic compounds under north European conditions.