The integrity of the actin cytoskeleton is essential for plant immune signalling. Consequently, it is generally assumed that actin disruption reduces plant resistance to pathogen attack. Here, we demonstrate that actin depolymerization induced a dramatic increase in salicylic acid (SA) levels in Arabidopsis thaliana . Transcriptomic analysis showed that the SA pathway was activated due to the action of isochorismate synthase (ICS). The effect was also confirmed in Brassica napus . This raises the question of whether actin depolymerization could, under particular conditions, lead to increased resistance to pathogens. Thus, we explored the effect of pretreatment with actin-depolymerizing drugs on the resistance of Arabidopsis thaliana to the bacterial pathogen Pseudomonas syringae , and on the resistance of an important crop Brassica napus to its natural fungal pathogen Leptosphaeria maculans . In both pathosystems, actin depolymerization activated the SA pathway, leading to increased plant resistance. To our best knowledge, we herein provide the first direct evidence that disruption of the actin cytoskeleton can actually lead to increased plant resistance to pathogens, and that SA is crucial to this process.

Being natural plant antimicrobials, saponins have potential for use as biopesticides. Nevertheless, their activity in plant–pathogen interaction is poorly understood. We performed a comparative study of saponins' antifungal activities on important crop pathogens based on their effective dose (EC50) values. Among those saponins tested, aescin showed itself to be the strongest antifungal agent. The antifungal effect of aescin could be reversed by ergosterol, thus suggesting that aescin interferes with fungal sterols. We tested the effect of aescin on plant–pathogen interaction in two different pathosystems: Brassica napus versus (fungus) Leptosphaeria maculans and Arabidopsis thaliana versus (bacterium) Pseudomonas syringae pv tomato DC3000 (Pst DC3000). We analyzed resistance assays, defense gene transcription, phytohormonal production, and reactive oxygen species production. Aescin activated B. napus defense through induction of the salicylic acid pathway and oxidative burst. This defense response led finally to highly efficient plant protection against L. maculans that was comparable to the effect of fungicides. Aescin also inhibited colonization of A. thaliana by Pst DC3000, the effect being based on active elicitation of salicylic acid (SA)-dependent immune mechanisms and without any direct antibacterial effect detected. Therefore, this study brings the first report on the ability of saponins to trigger plant immune responses. Taken together, aescin in addition to its antifungal properties activates plant immunity in two different plant species and provides SA-dependent resistance against both fungal and bacterial pathogens.

The phytohormone salicylic acid (SA) has a crucial role in plant physiology. Its role is best described in the context of plant response to pathogen attack. During infection, SA is rapidly accumulated throughout the green tissues and is important for both local and systemic defences. However, some genetic/metabolic variations can also result in SA overaccumulation in plants, even in basal conditions. To date, more than forty Arabidopsis thaliana mutants have been described as having enhanced endogenous SA levels or constitutively activated SA signalling pathways. In this study, we established a collection of mutants containing different SA levels due to diverse genetic modifications and distinct gene functions. We chose prototypic SA-overaccumulators (SA-OAs), such as bon1-1, but also “non-typical” ones such as exo70b1-1; the selection of OA is accompanied by their crosses with SA-deficient lines. Here, we extensively studied the plant development and SA level/signalling under various growth conditions in soil and in vitro, and showed a strong negative correlation between rosette size, SA content and PR1/ICS1 transcript signature. SA-OAs (namely cpr5, acd6, bon1-1, fah1/fah2 and pi4kβ1β2) had bigger rosettes under high light conditions, whereas WT plants did not. Our data provide new insights clarifying a link between SA and plant behaviour under environmental stresses. The presented SA mutant collection is thus a suitable tool to shed light on the mechanisms underlying trade-offs between growth and defence in plants.

Plasmopara halstedii was isolated from diseased sunflowers collected from eight locations in the Czech Republic from 2007 to 2014. Races of the pathogen were determined based on 84 isolates collected during the study. In total, eight races of P. halstedii were detected using a set of nine sunflower differential lines. Races 700, 704, 705, 710, 714 and 715 were proven by soil drench inoculation, and two additional races (730 and 770) proposed by the previously applied leaf disc inoculation method. Race 700 was the most dominant in the Czech P. halstedii populations, with race 710 being the second most frequent. Races 704 and 714 were found over three seasons, while other races were recorded only in one growing season (race 730 in 2010, and the new races 705 and 715 in 2014). A comprehensive study was further conducted for isolates collected in 2013-14 using an extended differential set consisting of 15 sunflower lines. According to the latter methodology which marks races with five-digit virulence codes, races 70060, 70471, 70571, 71060, 71461 and 71571 were recorded. The growing complexity of P. halstedii pathogenicity exhibited by the ability to infect higher numbers of differential genotypes and resulting in determination of the new pathogen races (virulence profiles) 70571, 71461 and 71571 is alarming. Although the limited number of isolates studied cannot characterize the entire pathogen diversity in the Czech Republic, the trend towards more diverse virulence in P. halstedii populations is clearly demonstrated by the new records of races 704, 705, 714 and 715, all capable of overcoming the resistance gene Pl 6 .

28Actin cytoskeleton is indispensable for plant cell integrity. Besides, increasing evidences illustrate 29 its crucial role in plant responses to environment, including defence against pathogens. Recently, 30 we have demonstrated that pre-treatment with actin disrupting drugs latrunculin B (latB) and 31 cytochalasin E can enhance plant resistance against bacterial and fungal pathogens via activation 32 of salicylic acid (SA) pathway. Here, we show that actin depolymerization in Arabidopsis thaliana 33 seedlings not only triggers SA biosynthesis by ICS1, but also induces callose deposition via callose 34 synthase PMR4. This effect is SA-independent since still present in mutants that do not accumulate 35 SA. LatB also triggers the expression of several defence related genes. We could distinguish genes, 36 induced in a SA-dependent manner (PR1, WRKY38, ICS1) and those that are SA-independent 37 (PR2, PAD4, BAP1). As actin cytoskeleton is tightly connected with membrane trafficking, we 38 assayed the effect of latB on mutant plants invalidated in phosphatidylinositol 4-kinase beta1 and 39 beta2 (PI4Kβ1β2). Deficiency in PI4Kβ1β2 enhanced latB-triggered actin filaments 40 depolymerisation. Yet, it did not lead to a stronger callose deposition or SA biosynthesis in 41 response to latB. Surprisingly, introduction of NahG construct or pad4 mutation in pi4kß1ß2 42 background had much lower effect on SA induction and SA-dependent gene expression changes

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Phosphatidylinositol 4-kinases (PI4Ks) are the first enzymes that commit phosphatidylinositol into the phosphoinositide pathway. Here, we show that Arabidopsis thaliana seedlings deficient in PI4Kβ1 and β2 have several developmental defects including shorter roots and unfinished cytokinesis. The pi4kβ1β2 double mutant was insensitive to exogenous auxin concerning inhibition of root length and cell elongation; it also responded more slowly to gravistimulation. The pi4kß1ß2 root transcriptome displayed some similarities to a wild type plant response to auxin. Yet, not all the genes displayed such a constitutive auxin-like response. Besides, most assessed genes did not respond to exogenous auxin. This is consistent with data with the transcriptional reporter DR5-GUS. The content of bioactive auxin in the pi4kß1ß2 roots was similar to that in wild-type ones. Yet, an enhanced auxin-conjugating activity was detected and the auxin level reporter DII-VENUS did not respond to exogenous auxin in pi4kß1ß2 mutant. The mutant exhibited altered subcellular trafficking behavior including the trapping of PIN-FORMED 2 protein in rapidly moving vesicles. Bigger and less fragmented vacuoles were observed in pi4kß1ß2 roots when compared to the wild type. Furthermore, the actin filament web of the pi4kß1ß2 double mutant was less dense than in wild-type seedling roots, and less prone to rebuilding after treatment with latrunculin B. A mechanistic model is proposed in which an altered PI4K activity leads to actin filament disorganization, changes in vesicle trafficking, and altered auxin homeostasis and response resulting in a pleiotropic root phenotypes.

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