Analysis of proteins differentially accumulated during potato late blight resistance mediated by the RB resistance gene

The RB gene, cloned from the wild diploid potato species Solanum bulbocastanum, confers resistance against the late blight pathogen, Phytophthora infestans. We examined changes in the proteome of potato leaves in response to inoculation with P. infestans. A nearly isogenic system comprised of susceptible Solanum tuberosum cultivar ‘Katahdin’ and resistant transgenic potato (cv. ‘Katahdin’) carrying a single copy of RB was utilized for this study. Comparative proteomic analysis revealed the presence of only 12 protein spots with a significant difference (≥ twofold) in relative abundance in resistant and susceptible potato plants after inoculation with P. infestans. Five out of the 12 identified proteins have putative roles in photosynthesis and stress responses. Silencing of these genes in Nicotiana benthamiana had no impact on RB-mediated induction of the hypersensitive response (HR). However, we found that silencing of molecular chaperone HSP90 led to the inability of RB to induce the HR after recognition of the P. infestans effector IpiO in planta.     

Evidence for selection pressure from resistant potato genotypes but not from fungicide application within a clonal Phytophthora infestans population

Insight into pathogen population dynamics provides a key input for effective disease management of the potato late blight pathogen Phytophthora infestans. Phytophthora infestans populations vary from genetically complex to more simple with a few clonal lineages. The presence or absence of certain strains of P. infestans may impact the efficacy of fungicides or host resistance. Current evidence indicates that genetically, the Irish populations of P. infestans are relatively simple with a few clonal lineages. In this study, P. infestans populations were genetically characterized based on samples collected at the national centre for potato breeding during the period 2012–16. The dominance of clonal lineages within this P. infestans population was confirmed and the potential selection pressure of fungicide treatment (2013–15) and host resistance (2016) on this clonal P. infestans population was then investigated. It was found that fungicide products did not notably affect the genetic structure of sampled populations relative to samples from untreated control plants. In contrast, samples taken from several resistant potato genotypes were found to be more often of the EU_13_A2 lineage than those taken from control King Edward plants or potato genotypes with low resistance ratings. Resistant potato varieties Sarpo Mira and Bionica, containing characterized R genes, were found to strongly select for EU_13_A2 strains.     

The novel elicitor COS‐OGA enhances potato resistance to late blight

Phytophthora infestans is the causal agent of potato late blight. This pathogen is usually controlled by fungicides, but new European regulations have imposed changes in crop protection management that have led to a search for alternative control measures. The induction of plant defence responses by elicitors is a promising new strategy compatible with sustainable agriculture. This study investigated the effect of eliciting a defence response in potato against P. infestans using a formulation of the COS‐OGA elicitor that combines cationic chitosan oligomers (COS) and anionic pectin oligomers (OGA). Trials were conducted under greenhouse conditions to assess the ability of COS‐OGA to control P. infestans. The results showed that three foliar applications with this elicitor significantly increased potato protection against late blight in controlled conditions. The activation of potato defence genes was also evaluated by RT‐qPCR during these trials. Two pathogenesis‐related proteins, basic PR‐1 and acidic PR‐2, were rapidly and significantly up‐regulated by the elicitor treatment. Therefore, these results suggest that the COS‐OGA elicitor increases the protection of potato against P. infestans and that this protection could be explained by an increase in the expression of potato defence genes rather than by biocide activity.     

Population genetic analysis of Phytophthora infestans in northwestern China

Late blight caused by Phytophthora infestans is the most devastating disease of potato worldwide. To understand the P. infestans population structure and dynamics in northwestern China, 959 single‐lesion isolates were purified in three consecutive years (2009–2011) and were characterized for mating type, pathotype, mtDNA haplotype and molecular variation at eight SSR loci. The results showed that the distribution of mating types changed significantly over years, with self‐fertile isolates dominant in 2010 and 2011. SSR genotyping distinguished 959 isolates into 151 genotypes, and association analysis indicated that P. infestans populations in 2010 and 2011 were strictly asexual while in 2009 they showed signs of sexual reproduction. Population analysis showed that the majority of genetic variation was within P. infestans populations. Isolates sharing identical SSR genotypes were detected in distant regions, indicating that migration of P. infestans could have occurred between regions. Pathogenicity assays on a set of potato differential lines containing R1 to R11 resistance genes detected four pathotypes from 74 selected isolates, with the pathotype virulent against all 11 R genes being dominant. Three mtDNA haplotypes (Ia, IIa, IIb) were detected with Ia being dominant among 507 isolates examined. Phylogenetic analysis indicated that P. infestans populations in northwestern China are distant from European lineages including 13‐A2 (blue‐13) at the time of this survey. The results have implications for the trade of healthy seed tubers as a means of managing late blight.     

Reduced efficacy of fluazinam against <Emphasis FontCategory="SansSerif" Type="Italic">Phytophthora infestans</Emphasis> in the Netherlands

Phytophthora infestans is the causal organism of potato late blight, the most important disease in potato, the second most important arable crop in Europe. The P. infestans population in Europe is well known for its sudden changes in composition. Currently it is composed of a wide variety of genotypes, some of which are dominant clonal lines while others are rare or even unique to a year or location. Fungicides play a crucial role in the integrated control of late blight. Since its introduction in the Netherlands in 1992, fluazinam has been used in late blight control strategies in ware and starch potatoes. It has a broad spectrum of activity and is effective against a range of diseases including potato late blight. Fluazinam interrupts the pathogen cell’s energy production process by an uncoupling effect on oxidative phosphorylation. It is considered to have a low resistance risk. Until recently, reduced efficacy against fluazinam was not detected in P. infestans surveys in Europe. In this paper we present the finding of a new clonal lineage (EU_33_A2) of P. infestans in the Netherlands and the reduced efficacy of fluazinam to control one of the EU_33_A2 isolates in field experiments carried out in 2011 and 2015 under high disease pressure. The potential effects of this finding on practical late blight control strategies are discussed.     

SGT1 and HSP90 are essential for age-related non-host resistance of Nicotiana benthamiana against the oomycete pathogen Phytophthora infestans

The oomycete pathogen, Phytophthora infestans, is the causal agent of potato late blight, which is one of the most destructive and economically important plant diseases. We investigated the interaction between P. infestans and Solanaceous model plant Nicotiana benthamiana. Mature N. benthamiana plants were resistant to 8 isolates of P. infestans, whereas relatively young plants were susceptible to all isolates. Analysis with virus-induced gene silencing (VIGS) indicated that NbSGT1 and NbHSP90, genes essential for the function of R proteins, are required for the resistance of N. benthamiana to P. infestans. NbSGT1 was also required for the production of reactive oxygen species (ROS), hypersensitive cell death and expression of NbEAS, a gene for phytoalexin biosynthesis, induced by INF1, a secretory protein derived from P. infestans. These results suggested that N. benthamiana possibly possesses a broad-spectrum R protein against P. infestans, which requires an SGT1/HSP90-dependent mechanism, for the recognition of a conserved molecular pattern of P. infestans.     

Inheritance of resistance in <Emphasis Type="Italic">Solanum nigrum</Emphasis> to <Emphasis Type="Italic">Phytophthora infestans</Emphasis>

Solanum nigrum, black nightshade, is a wild non-tuber bearing hexaploid species with a high level of resistance to Phytophthora infestans (Colon et al. 1993), the causal agent of potato late blight, the most devastating disease in potato production. However, the genetic mode of resistance in S. nigrum is still poorly understood. In the present study, two S. nigrum accessions, 984750019 (N19) and #13, resistant (R) and susceptible (S), respectively, to three different isolates of P. infestans, were sexually crossed. The various kinds of progeny including F1, F2, F3, and backcross populations (BC₁; F1 × S), as well as two populations produced by self-pollinating the R parent and S parent, were each screened for susceptibility to P. infestans isolate MP 324 using detached leaf assays. Fifty seedling plant individuals of the F1 progeny were each resistant to this specific isolate, similarly to the seedling plants resulting from self-pollination of the resistant R parent. Thirty seedling plants obtained from self-pollination of the S parent were susceptible. Among a total of 180 F2 plants, the segregation ratio between resistant and susceptible plants was approximately 3: 1. Among the 66 seedling plants of the BC₁ progeny originating from crossing an F1 plant with the susceptible S parent, there were 26 susceptible and 40 resistant plants to P. infestans. The segregation patterns obtained indicated monogenic dominant inheritance of resistance to P. infestans isolate MP 324 in S. nigrum acc. 984750019. This gene, conferring resistance to P. infestans, may be useful for the transformation of potato cultivars susceptible to late blight.     

Population structure of Phytophthora infestans in Turkey reveals expansion and spread of dominant clonal lineages and virulence

Late blight, caused by the oomycete Phytophthora infestans, is considered the most important and destructive disease of potato in Turkey. In this study, characterization of 367 isolates of P. infestans obtained from the potato-growing areas of the country was carried out to evaluate the pathogen population structure over the 2017–2019 production seasons. The isolates were characterized by numerous features including mating type, in vitro mefenoxam sensitivity, simple-sequence repeat (SSR) markers, and virulence against a set of potato differential lines. Most isolates were A2 mating type (353 isolates). Also, 68% of isolates were resistant to mefenoxam; the remainder were intermediate in their sensitivity and there were no sensitive isolates. SSR-based genotypic analysis of P. infestans populations showed a low genetic diversity. The 13_A2 clonal lineage predominated with a frequency of 92.1%, followed by 34_A1 (3.3%) and 37_A2 (2.7%). Genotypes 34_A1 and 37_A2 were detected only in 2019. This is the first report of 34_A1 and 37_A2 clonal lineages causing late blight disease of potato in Turkey. The most abundant virulence type was one overcoming resistance genes R1, R2, R3, R4, R6, R7, R10, and R11. These results emphasized that the migration of individuals and the asexual generation of subclonal differences were the main factors driving the population structure of P. infestans in Turkey.     

Diversity of Avr‐vnt1 and AvrSmira1 effector genes in Polish and Norwegian populations of Phytophthora infestans

The oomycete Phytophthora infestans, the cause of late blight, is one of the most important potato pathogens. During infection, it secretes effector proteins that manipulate host cell function, thus contributing to pathogenicity. This study examines sequence differentiation of two P. infestans effectors from 91 isolates collected in Poland and Norway and five reference isolates. A gene encoding the Avr‐vnt1 effector, recognized by the potato Rpi‐phu1 resistance gene product, is conserved. In contrast, the second effector, AvrSmira1 recognized by Rpi‐Smira1, is highly diverse. Both effectors contain positively selected amino acids. A majority of the polymorphisms and all selected sites are located in the effector C‐terminal region, which is responsible for their function inside host cells. Hence it is concluded that they are associated with a response to diversified target protein or recognition avoidance. Diversification of the AvrSmira1 effector sequences, which existed prior to the large‐scale cultivation of plants containing the Rpi‐Smira1 gene, may reduce the predicted durability of resistance provided by this gene. Although no isolates virulent to plants with the Rpi‐phu1 gene were found, the corresponding Avr‐vnt1 effector has undergone selection, providing evidence for an ongoing ‘arms race’ between the host and pathogen. Both genes remain valuable components for resistance gene pyramiding.     

Potato <Emphasis Type="Italic">R1</Emphasis> resistance gene confers resistance against <Emphasis Type="Italic">Phytophthora infestans</Emphasis> in transgenic tomato plants

Tomato is challenged by several pathogens which cause loss of production. One such pathogen is the oomycete Phytophthora infestans which is able to attack all the aerial parts of the plant. Although a wide range of resistance sources are available, genetic control of this disease is not yet successful. Pyramiding R-genes through genetic transformation could be a straightforward way to produce tomato and potato lines carrying durable resistance to P. infestans. In this work the R1 potato gene was transferred into tomato lines. The tomato transgenic lines were analyzed by using q-RT-PCR and progeny segregation to determine the gene copy number. To test the hypothesis that R1 represents a specifically regulated R-gene, transgenic tomato plants were inoculated with P. infestans isolate 88133 and IPO. All the plants containing the R1 gene were resistant to the late blight isolate IPO-0 and susceptible to isolate 88133. These results provide evidence for specific activation of the R1 gene during pathogen challenge. Furthermore, evidence for enhancement of PR-1 gene expression during P. infestans resistance response was obtained.     

Resistance to <Emphasis Type="Italic">Phytophthora infestans</Emphasis>: exploring genes required for disease resistance in Solanaceae plants

The oomycete Phytophthora infestans is the causal agent of potato late blight, one of the most destructive and historically significant pathogens in agricultural production. A virus-induced gene silencing-based screening of the solanaceous model plant N. benthamiana resulted in revealing a wide range of resistance mechanisms of solanaceous plants against this pathogen. In this article, we present an overview of the various pathways involved in the N. benthamiana–P. infestans pathosystem, including some of the follow-up work that was triggered by these findings. The purpose of this review is to assemble these findings and integrate them into our current understanding of plant pathogen defense mechanisms and discuss their potential application for the development of potato resistance to P. infestans.     

Characterization of Egyptian <Emphasis Type="Italic">Phytophthora infestans</Emphasis> population using simple sequence repeat markers

The plant pathogenic oomycete, Phytophthora infestans, is the causal agent of late blight disease in tomato and potato. For characterizing Egyptian P. infestans isolates by DNA marker analysis, 40 isolates of P. infestans were collected from different locations in Egypt during two growing seasons (2012/2013 and 2013/2014). The 40 isolates were grouped into seven genotypes, in which 24 alleles were detected. The identified genotypes were not completely associated with geographic location and sample collection years. These results provide genetic and geographical information for developing a program to manage late blight disease.     

29个马铃薯品种(品系)抗晚疫病基因组成与抗病相关性分析

由致病疫霉引起的马铃薯晚疫病是马铃薯生产中最具危害性的病害,明确现有马铃薯品种(品系)中抗病基因组成情况对于指导抗病品种合理布局及抗病育种具有重要意义。本研究利用农杆菌介导的无毒基因瞬时表达技术分析了致病疫霉8个无毒基因在29个马铃薯品种(品系)中的分布情况。结果表明,品种间抗病基因组成各异,平均每个品种含有4个抗病基因。同时,不同抗病基因在品种中的分布频率也不同,抗病基因R1(76%)、Rpi-blb1(66%)和Rpi-blb2(66%)在品种中分布较普遍;而含抗病基因R3b(45%)、R4(38%)和Rpi-vnt1(34%)的品种较少。此外,品种中所含抗性基因的数量和品种室内抗性程度呈显著正相关(r=0.915 6,P=0.029 1)。     

Identification and characterization of RB-orthologous genes from the late blight resistant wild potato species Solanum verrucosum

Late blight, caused by the oomycete pathogen, Phytophthora infestans, is a devastating disease of potatoes and tomatoes. A key long-term management strategy for combating this disease is to develop potato cultivars with high levels of durable resistance through identification and integration of major resistance genes. The RBgene, cloned from the Mexican diploid potato species Solanum bulbocastanum, confers broad-spectrum resistance to potato late blight. Here, we have determined the late blight resistance phenotypes of eight accessions of Solanum verrucosum, another wild diploid potato species, using greenhouse inoculations and discovered variability among the accessions. While most accessions were resistant, one accession was notably more susceptible. Transcribed orthologs of the RB gene from the eight S. verrucosum accessions were cloned using a homology-based PCR approach. Sequence analysis revealed that the RB^ver orthologs share up to 83.5% nucleotide identity with RB^blb. Stable introduction of the RB ortholog from late blight resistant S. verrucosum PI275260 into susceptible S. tuberosum confers resistance to P. infestans. Interestingly, this functional RB^ver ortholog contains an insertion of a complete leucine rich repeat when compared to RB^blb, and differs from the RB^ver ortholog from a susceptible accession at only four amino acid residues.     

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.     

The occurrence of the A2 mating type of Phytophthora infestans in the Netherlands; significance and consequences

Phytophthora infestans (Mont.) de Bary, the causal agent of potato late blight, was first discovered in Europe in 1845. Until 1980, only A1 mating type isolates were known to occur in Europe. The absence of A2 mating type isolates restrained the fungus from sexual reproduction. In the early 1980s, A2 mating type isolates were discovered in Europe. Presumably, a new introduction ofP. infestans isolates originating from Mexico had taken place. In this paper, the significance of the presence of A1 and A2 mating type isolates in the Netherlands is reviewed. Now that both mating types are present, sexual reproduction can occur and its consequences for the control of potato late blight are discussed.     

Chemical late blight control in Sweden1

The climatological conditions for attack of late blight caused by Phytophthora infestans vary considerably in different parts of Sweden. This is due to the fact that the country is long from south to north covering 14 degrees of latitude from 55 to 69°. One important factor for blight development in Sweden is that potato production is dominated by cultivars susceptible to P. infestans. Approximately 80% of table potatoes are of cvs Bintje and King Edward VII. Cultivars with race-specific resistance such as Bellona, Ukama and Provita are attacked in the whole country. During recent years, many experiments have been carried out to investigate possible reductions in the usage of chemical agents for late blight control. These experiments include choice of fungicide, number of applications, dose rates and treatments according to blight warning systems.     

Involvement of actin filament association in hypersensitive reactions in potato cells

In potato tissues after either penetration by an incompatible race ofPhytophthora infestans, the potato late blight fungus, or treatment with hyphal wall components (HWC) prepared fromP. infestans, cytoplasmic aggregation is observed as one of the earliest resistance reactions. In potato protoplasts, the number of aggregated protoplasts increased after treatment with HWC. The increase was repressed by treatment with cytochalasin D, staurosporine, verapamil, ophiobolin and quinacrine prior to HWC treatment, suggesting that cytoplasmic aggregation in potato protoplasts was connected with actin filament association, protein phosphorylation, Ca²⁺, calmodulin and phospholipase reactions. The ratio of monomer-actin in potato tissues was decreased by treatment with HWC, while the decrease was repressed by treatment with cytochalasin D, staurosporine, ophiobolin and quinacrine prior to HWC treatment. These inhibitors inhibited the accumulation of rishitin, a potato phytoalexin, which occurs in potato tissues treated with HWC. The results imply that the factors involved in signal transduction might be connected with the cytoskeleton, especially actin filament association, during the initiation of defence responses in potato cells.     

Restriction of potato and tomato late blight development by sub‐phytotoxic concentrations of boron

Boron is a microelement required for normal growth and development of plants but its positive effect is restricted to a narrow range of concentrations. The gradual increase in use of recycled water, which contains high concentrations of boron for irrigation, has already raised the level of boron in soils and plants in southern Israel. This research was conducted to examine the direct effects of sub‐phytotoxic boron concentrations on potato late blight epidemics and to explore the mode of action of boron against Phytophthora infestans. When boron was applied alone to field grown potato plants it did not affect the epidemic. However, together with a reduced rate of the fungicide Melody Duo (propineb + iprovalicarb), boron improved late blight suppression compared to plants treated with the fungicide alone. The ED₅₀ of boron against P. infestans (256·4 mg L^?1) was about 6400 times higher than the ED₅₀ value of the fungicide chlorothalonil (0·04 mg L^?1), indicating that boron does not have a direct fungicidal activity that would explain the level of protection seen in the field. In greenhouse experiments conducted with potted tomato plants, boron decreased late blight severity in both treated leaves and distant leaves not treated with boron. The results suggest that boron is active locally but also may induce systemic acquired resistance against P. infestans.