Activation of defence responses to Phytophthora infestans in potato by BABA

Late blight caused by Phytophthora infestans is one of the most devastating diseases of the potato crop. Resistance breeding and current fungicides are unable to control the rapidly evolving P. infestans and new control strategies are urgently needed. This study examined mechanisms of dl ‐β‐aminobutyric acid (BABA)‐induced resistance (IR) in the potato–P. infestans system. Leaves from two cultivars that differ in their degree of resistance, Bintje and Ovatio, were analysed after foliar treatment with BABA. Rapid activation of various defence responses and a significant reduction in P. infestans growth were observed in leaves treated with BABA. In the more resistant cultivar, Ovatio, the activation was both faster and stronger than in Bintje. Microscopic analysis of leaves treated with BABA revealed induction of small hypersensitive response (HR)‐like lesions surrounded by callose, as well as production of hydrogen peroxide (H₂O₂). Molecular and chemical analyses revealed soluble phenols such as arbutin and chlorogenic acid and activation of PR‐1. These results show a direct activation of defence responses in potato, rather than priming as reported for other plant species. They also show that the efficiency of BABA‐IR differs between cultivars, which highlights the importance of taking all aspects into consideration when establishing new methods for disease management.     

BABA effects on the behaviour of potato cultivars infected by <Emphasis Type="Italic">Phytophthora infestans</Emphasis> and <Emphasis Type="Italic">Fusarium solani</Emphasis>

Since most plants possess resistance mechanisms which can be induced upon pre-treatment with a variety of chemical compounds, the use of β-aminobutyric acid (BABA) as a defence inducer without reported toxic effect on the environment was studied. The aim of this work was to analyse the effectiveness of BABA to induce resistance against Phytophthora infestans and Fusarium solani in potato cultivars differing in their level of resistance to late blight. The behaviour of some components of biochemical mechanisms by which BABA increases resistance against P. infestans, as well as the effect of BABA on the activity of a potential pathogenic factor of F. solani, were studied. Plants with four applications of BABA throughout the crop cycle produced tubers more resistant to P. infestans and F. solani than non-treated plants. In addition, tuber slices from treated plants, inoculated with P. infestans, showed an increase in phenol and phytoalexin content. The aspartyl protease StAP1 accumulation was also higher in tubers obtained from treated plants and inoculated with P. infestans. This result was observed only in the more resistant potato cv. Pampeana, early after infection. In the potato–F. solani interaction, infected tubers coming from BABA-treated plants showed minor fungal proteolytic activity than infected, non-treated ones. For potato cvs Pampeana and Bintje, the BABA treatment improved the yield of harvested tubers. The number of tubers per plant and total weight of harvested tubers was greater for those obtained from treated plants with two early or four applications of BABA. The results show that the BABA treatment increases the resistance of potatoes but the degree of increase depends on the original level of resistance present in each cultivar.     

Synergistic interaction between BABA and mancozeb in controlling<Emphasis Type="Italic">Phytophthora infestans</Emphasis> in potato and tomato and<Emphasis Type="Italic">Pseudoperonospora cubensis</Emphasis> in cucumber

Spray mixtures consisting of the plant activator BABA (DL-3-aminobutyric acid) and the protectant fungicide mancozeb were significantly more effective than BABA or mancozeb alone in controlling late blight (Phytophthora infestans) in potato and tomato and downy mildew (Pseudoperonospora cubensis) in cucumber. A mixture composed of 5 parts BABA and 1 part mancozeb (w/w, a.i.) exhibited a higher synergy factor than the 1+1 or the 1+5 (BABA + mancozeb) mixtures. No synergistic interaction was detected between BABA plus mancozeb in controlling sporangial or cystospore germination, nor mycelial growth ofP. infestans in vitro. The results showed enhanced effect of mancozeb in BABA-induced plants, suggesting, therefore, that lower dosages of this fungicide may be sufficient to control late blight or downy mildew under field conditions. http://www.phytoparasitica.org posting July 15, 2003.     

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.     

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.     

Long‐lasting β‐aminobutyric acid‐induced resistance protects tomato fruit against Botrytis cinerea

Minimizing losses to pests and diseases is essential for producing sufficient food to feed the world's rapidly growing population. The necrotrophic fungus Botrytis cinerea triggers devastating pre‐ and post‐harvest yield losses in tomato (Solanum lycopersicum). Current control methods are based on the pre‐harvest use of fungicides, which are limited by strict legislation. This investigation tested whether induction of resistance by β‐aminobutyric acid (BABA) at different developmental stages provides an alternative strategy to protect post‐harvest tomato fruit against B. cinerea. Soil‐drenching plants with BABA once fruit had already formed had no impact on tomato susceptibility to B. cinerea. However, BABA application to seedlings significantly reduced post‐harvest infection of fruit. This resistance response was not associated with a yield reduction; however, there was a delay in fruit ripening. Untargeted metabolomics revealed differences between fruit from water‐ and BABA‐treated plants, demonstrating that BABA triggered a defence‐associated metabolomics profile that was long lasting. Targeted analysis of defence hormones suggested a role of abscisic acid (ABA) in the resistance phenotype. Post‐harvest application of ABA to the fruit of water‐treated plants induced susceptibility to B. cinerea. This phenotype was absent from the ABA‐exposed fruit of BABA‐treated plants, suggesting a complex role of ABA in BABA‐induced resistance. A final targeted metabolomic analysis detected trace residues of BABA accumulated in the red fruit. Overall, it was demonstrated that BABA induces post‐harvest resistance in tomato fruit against B. cinerea with no penalties in yield.     

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.     

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.     

The role of aggressiveness and competition in the selection of Phytophthora infestans populations

Selection within populations of Phytophthora infestans was investigated by comparing the aggressiveness of single‐lesion isolates on detached leaflets of four potato cultivars with differing levels of race‐nonspecific resistance to P. infestans. The isolates included 23 representative of Northern Ireland genotypes from the early 2000s, used to inoculate previously reported field trials on competitive selection (2003–2005), plus 12 isolates recovered from the 2003 trial. The cultivars were those planted in the previous trials: Atlantic (blight‐susceptible) and Santé, Milagro and Stirling (partially resistant). Very highly significant variation for latent period, infection frequency and lesion area was found between genotypes and cultivars; differences between genotypes were more marked on the more resistant cultivars, but no one genotype was the most aggressive across all. Detached leaflets were also inoculated with mixtures of isolates from each genotype group at three sporangial concentrations: differences in aggressiveness between genotypes were more apparent at lower concentrations and on the more resistant cultivars. Genotype groups that were the most aggressive on the more resistant cultivars tended to be those selected by the same cultivars in the field. A mixture of all isolates of all genotypes was used to inoculate detached leaflets of the same cultivars. With one exception, single spore isolates recovered from any one leaflet belonged to a single genotype, but different genotypes were recovered from different cultivars. Phytophthora infestans isolates from Northern Ireland showed significant variation for foliar aggressiveness, and pathogen genotypes exhibited differential aggressiveness to partially resistant cultivars and interacted competitively in genotype selection.     

Activation of tomato plant defence responses against bacterial wilt caused by Ralstonia solanacearum using DL-3-aminobutyric acid (BABA)

In this study, we investigated the ability of DL-3-aminobutyric acid (BABA) to protect tomato against bacterial wilt caused by Ralstonia solanacearum. This was combined with studies of accumulation of total phenolic compounds, free and total salicylic acid (SA), and activity of enzymes related to plant defence, i.e., polyphenol oxidase (PPO) and catalase (CAT). Under greenhouse conditions, tomato plants pre-treated by soil drenching with BABA profoundly reduced disease severity of bacterial wilt compared to plants receiving a soil drench with water. Thus, BABA reduced leaf wilting index by 75.3 % and vascular browning index by 69.9 %, without any in vitro inhibitory activity on the pathogen. BABA treatment significantly reduced the population of R. solanacearum in stems of tomato plants and additionally also significantly increased both fresh and dry weight of roots and shoots of tomato plants compared with the inoculated control. Application of BABA resulted in a high increase in PPO activity both in plants with and without inoculation. Compared to water-treated plants, treatment with BABA also induced a significant increase of total phenolic compounds as well as of free and total SA in leaves of both inoculated and non-inoculated tomato plants at all sampling times. CAT activity decreased in tomato plants treated with BABA in comparison with the water-treated control plants and the decrease in activity correlated with an increasing total SA accumulation. These findings suggest that BABA treatment resulted in induction of resistance to bacterial wilt in tomato.     

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.     

The enhanced disease susceptibility phenotype of ethylene-insensitive tobacco cannot be counteracted by inducing resistance or application of bacterial antagonists

In an attempt to overcome the enhanced disease susceptibility phenotype that is typical for transgenic ethylene-insensitive tobacco (Tetr), Tetr plants were treated with chemical agents that induce resistance or with antagonistic rhizobacteria. Treatments with β-aminobutyric acid (BABA), benzo-(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester (BTH), methyl jasmonate (MeJA), or salicylic acid (SA) induced PR-genes generally to a lesser extent than in non-transformed plants and did not reduce wilting symptoms upon infection with Pythium sp., except for a marginal effect of SA. In Tetr lines overexpressing PR-1g, PR-5c, or both, no significant reduction in disease development was apparent. Also treatment of Tetr plants with the antagonistic rhizobacteria Bacillus cereus UW85, Pseudomonas aeruginosa 7NSK2, Pseudomonas fluorescens WCS417r or Q8r-196, Pseudomonas putida WCS358r, or antibiotic-producing derivatives of WCS358r, did not reduce symptoms caused by Pythium.     

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.     

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.     

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.     

Induced resistance againstPhytophthora capsici in pepper plants in response to DL-β-amino-n-butyric acid

Treatment of pepper plants with the nonprotein amino acid, DL-ß-amino-n-butyric acid (BABA) induced resistance to subsequent infection byPhytophthora capsici. In contrast, theα-, andγ-isomers of aminobutyric acid were ineffective as inducers of resistance. A relatively high concentration of BABA at 1,000μg ml^?1, which had no antifungal activityin vitro againstP. capsici, was required to induce resistance against Phytophthora blight with a foliar and stem spray, thus leading to complete control of the disease. About 1 day interval between BABA-treatment and challenge inoculation was sufficient to induce resistance in pepper plants. High inoculum levels ofP. capsici caused Phytophthora development slowly in pepper stems treated with BABA, especially at early plant growth stage, which suggests that the induced resistance in pepper plants may be more quantitative rather than qualitative. BABA applied to the root system also protected pepper stems fromP. capsici infection.     

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.     

Effect of foliar applications of phosphite on post-harvest potato tubers

The utilization of phosphites (Phi) could be considered as another strategy to be included in integrated disease management programmes to reduce the intensive use of fungicides and production costs. The aim of the present work was to analyze whether the beneficial effects of phosphite treatment previously observed in potato plants grown under greenhouse conditions, were reflected after harvest of field grown potatoes, both in disease protection and in yield. In addition, biochemical compounds possibly involved in induced defence responses by Phi, like phytoalexins, pathogenesis related proteins and oxidative stress enzymes were measured. Foliar applications of KPhi to field grown crops resulted in post-harvest tubers with a reduced susceptibility to Phytophthora infestans, Fusarium solani and Erwinia carotovora infections, suggesting that this compound induced a systemic defence response. An increase in phytoalexin content in P. infestans inoculated tubers obtained from Phi-treated plants suggests their participation in the defence response. Chitinase content increased 72 h after wounding or inoculation with P. infestans in tubers from KPhi-treated plants compared to wounded or infected tubers from non-treated plants. Contrary to this, the isoforms of β-1,3-glucanases analyzed did not increase in the tubers of Phi-treated plants. The increment in peroxidase and polyphenol oxidase activities indicated that these enzymes could be part of the Phi defence mechanism. No negative effects were observed in potato yield at harvest, measured as total tuber weight and dry matter, after foliar KPhi treatment. This suggests that the energetic cost involved in the defence response activation would not be detrimental to plant growth.