Circadian oscillations in Trichoderma atroviride and the role of core clock components in secondary metabolism, development, and mycoparasitism against the phytopathogen Botrytis cinerea.

Circadian clocks are important for an individual's fitness, and recent studies have underlined their role in the outcome of biological interactions. However, the relevance of circadian clocks in fungal-fungal interactions remains largely unexplored. We sought to characterize a functional clock in the biocontrol agent Trichoderma atroviride to assess its importance in the mycoparasitic interaction against the phytopathogen Botrytis cinerea. Thus, we confirmed the existence of circadian rhythms in T. atroviride, which are temperature-compensated and modulated by environmental cues such as light and temperature. Nevertheless, the presence of such molecular rhythms appears to be highly dependent on the nutritional composition of the media. Complementation of a clock null (Δfrq) Neurospora crassa strain with the T. atroviride-negative clock component (tafrq) restored core clock function, with the same period observed in the latter fungus, confirming the role of tafrq as a bona fide core clock component. Confrontation assays between wild-type and clock mutant strains of T. atroviride and B. cinerea, in constant light or darkness, revealed an inhibitory effect of light on T. atroviride's mycoparasitic capabilities. Interestingly, when confrontation assays were performed under light/dark cycles, T. atroviride's overgrowth capacity was enhanced when inoculations were at dawn compared to dusk. Deleting the core clock-negative element FRQ in B. cinerea, but not in T. atroviride, was vital for the daily differential phenotype, suggesting that the B. cinerea clock has a more significant influence on the result of this interaction. Additionally, we observed that T. atroviride clock components largely modulate development and secondary metabolism in this fungus, including the rhythmic production of distinct volatile organic compounds (VOCs). Thus, this study provides evidence on how clock components impact diverse aspects of T. atroviride lifestyle and how daily changes modulate fungal interactions and dynamics.

Fungal Evolution in Anthropogenic Environments: Botrytis cinerea Populations Infecting Small Fruit Hosts in the Pacific Northwest Rapidly Adapt to Human-Induced Selection Pressures.

Many fungal pathogens have short generation times, large population sizes, and mixed reproductive systems, providing high potential to adapt to heterogeneous environments of agroecosystems. Such adaptation complicates disease management and threatens food production. A better understanding of pathogen population biology in such environments is important to reveal key aspects of adaptive divergence processes to allow improved disease management. Here, we studied how evolutionary forces shape population structure of Botrytis cinerea, the causal agent of gray mold, in the Pacific Northwest agroecosystems. Populations of B. cinerea from adjacent fields of small fruit hosts were characterized by combining neutral markers (microsatellites) with markers that directly respond to human-induced selection pressures (fungicide resistance). Populations were diverse, without evidence for recombination and association of pathogen genotype with host. Populations were highly localized with limited migration even among adjacent fields within a farm. A fungicide resistance marker revealed strong selection on population structure due to fungicide use. We found no association of resistance allele with genetic background, suggesting de novo development of fungicide resistance and frequent extinction/recolonization events by different genotypes rather than the spread of resistance alleles among fields via migration of a dominant genotype. Overall our results showed that in agroecosystems, B. cinerea populations respond strongly to selection by fungicide use with greater effect on population structure compared to adaptation to host plant species. This knowledge will be used to improve disease management by developing strategies that limit pathogen local adaptation to fungicides and other human-induced selection pressures present in Pacific Northwest agroecosystems and elsewhere.IMPORTANCE Agroecosystems represent an efficient model for studying fungal adaptation and evolution in anthropogenic environments. In this work, we studied what evolutionary forces shape populations of one of the most important fungal plant pathogens, B. cinerea, in small fruit agroecosystems of the Pacific Northwest. We hypothesized that host, geographic, and anthropogenic factors of agroecosystems structure B. cinerea populations. By combining neutral markers with markers that directly respond to human-induced selection pressures, we show that pathogen populations are highly localized and that selection pressure caused by fungicide use can have a greater effect on population structure than adaptation to host. Our results give a better understanding of population biology and evolution of this important plant pathogen in heterogeneous environments but also provide a practical framework for the development of efficient management strategies by limiting pathogen adaptation to fungicides and other human-induced selection pressures present in agroecosystems of the Pacific Northwest and elsewhere.

Innovative approach to sunlight activated biofungicides for strawberry crop protection: ZnO nanoparticles.

Strawberries are one of the most common and important fruits in the world, widely investigated for their nutritional and nutraceutical properties. However, after the emergence of several outbreaks of foodborne diseases some concerns regarding the microbiological safety of fresh strawberries have increased in recent years. In this paper new insights, based on application of ZnO nanoparticles (NPs) as alternative to chemical fungicides in the fields for preharvest preservation of strawberries are presented. Antifungal activity of ZnO NPs was tested on main strawberry plant pathogen Botrytis cinerea. Obtained data indicated that used ZnO NPs (5 × 10-3 M) in the dark just insignificantly (12%) inhibited the radial growth of B. cinerea. But photoactivated ZnO NPs (5 × 10-3 M, 405 nm, 34 J/cm2) inhibited the growth of B. cinera by 80%. Real-time field experiments revelead, that spraying of ZnO NPs in the strawberry field in sunny day reduced Botrytis incidences by 43%, enhanced the crop production by 28.5% and stoped the spoilage of harvested fruits during storage by 8 days, if compare with control. No harm to crowns and leaves of strawberry plant have been found, however this treatment increased the growth of inflorescence (37.5%) and reduced the growth of runners (32.8%). For comparison, spraying of conventionqal chemical fungicide fenhexamid (FEN) reduced Botrytis incidences in the same level as ZnO NPs, increased the harvest by 21.9% and delayed the spoilage of fruits by 8 days. The presented results look highly promising, since ZnO NPs in the presence of sunlight, activated by UV and visible light can protect strawberry fruits from Botrytis infection more effectively than conventional fungicide fenhexamid. This treatment significantly increased crop production and reduced spoilage of strawberries. It looks like ZnO NPs have great potential in the future to replace chemical fungicides.

Botrytis cinerea response to pulsed light: Cultivability, physiological state, ultrastructure and growth ability on strawberry fruit.

Botrytis cinerea causes postharvest spoilage in important crops such as strawberry and other berries. Pulsed light (PL) treatment could be an environmentally friendly postharvest alternative to synthetic fungicides in berries. Cultivability, physiological state, ultrastructure of Botrytis cinerea suspended in peptone water and irradiated with PL (fluence = 1.2 to 47.8 J/cm2) were investigated by using conventional plate count technique, flow cytometry analysis (FCM) and transmission electron microscopy. In addition, PL effect on B. cinerea development in artificially contaminated strawberries throughout storage at (5 ±â€¯1) °C was evaluated. PL reduced fungus' ability to form colonies on agarized culture media. Survival curve fitted with the Weibullian model evidenced a wide distribution of conidia sensitivity to PL. FCM showed that most of irradiated conidia entered in a viable non-culturable state, although a subpopulation without esterase activity and compromised membranes and a subpopulation with active esterase and intact membranes were also detected. PL attacked multiple targets in B. cinerea. Ultrastructural changes varied with the dose and within the conidia population, supporting FCM results. Damage included plasmalemma detachment from cell wall, cytoplasm collapse, and vacuolization of cytoplasm, disruption of cell wall and plasmalemma with massive loss of cytoplasm and/or disruption of organelles. In strawberries artificially contaminated with B. cinerea, a 2-day delay on the onset of the infection and a lower incidence in PL-treated strawberries (11.9 and 23.9 J/cm2) compared to control (16-20%) up to 10 days of cold storage was observed. Results indicated that PL significantly reduces B. cinerea growth in peptone water and in inoculated strawberries. However, other preservation factor(s) in combination would be needed to increase PL action for a better control of this fungus.

Effects of cold plasma, UV-C or aqueous ozone treatment on Botrytis cinerea and their potential application in preserving blueberry.

AIMS: This study aimed to compare the effects of cold plasma (CP), ultraviolet (UV-C) and aqueous ozone (AO) on Botrytis cinerea and explore their application in preserving blueberry. METHODS AND RESULTS: The effects of CP, UV-C or AO on B. cinerea were investigated. Results showed that three treatments effectively inhibited the growth of B. cinerea, increasing cell membrane penetrability and causing the leakage of cytoplasm and nucleic acid. Scanning electron microscopy showed that CP caused the mycelium fold and collapse depression; UV-C caused the mycelium shrivelled; mycelium treated with AO appeared to fold and break. In the in vivo test in blueberry, all treatments effectively inhibited the growth of micro-organisms, maintained qualities and enhanced antioxidant activities in blueberry during postharvest storage. CONCLUSION: Cold plasma, UV-C and AO effectively inhibited the growth of B. cinerea and significantly extended the shelf life of blueberry. Based on in vitro and in vivo tests, CP showed better effects to preserve blueberry compared with other treatments. SIGNIFICANCE AND IMPACT OF THE STUDY: Our findings revealed the effective mechanisms of CP, UV-C and AO to inhibit B. cinerea in vitro. CP exhibited better application potential to preserve fresh fruit than traditional sterilization methods such as UV-C and AO.

Characterization of a Vivid Homolog in Botrytis cinerea.

Blue light-signaling pathways regulated by members of the light-oxygen-voltage (LOV) domain family integrate stress responses, circadian rhythms and pathogenesis in fungi. The canonical signaling mechanism involves two LOV-containing proteins that maintain homology to Neurospora crassa Vivid (NcVVD) and White Collar 1 (NcWC1). These proteins engage in homo- and heterodimerization events that modulate gene transcription in response to light. Here, we clone and characterize the VVD homolog in Botrytis cinerea (BcVVD). BcVVD retains divergent photocycle kinetics and is incapable of LOV mediated homodimerization, indicating modification of the classical hetero/homodimerization mechanism of photoadaptation in fungi.

The Two Cryptochrome/Photolyase Family Proteins Fulfill Distinct Roles in DNA Photorepair and Regulation of Conidiation in the Gray Mold Fungus Botrytis cinerea.

The plant-pathogenic leotiomycete Botrytis cinerea is known for the strict regulation of its asexual differentiation programs by environmental light conditions. Sclerotia are formed in constant darkness; black/near-UV (NUV) light induces conidiation; and blue light represses both differentiation programs. Sensing of black/NUV light is attributed to proteins of the cryptochrome/photolyase family (CPF). To elucidate the molecular basis of the photoinduction of conidiation, we functionally characterized the two CPF proteins encoded in the genome of B. cinerea as putative positive-acting components. B. cinerea CRY1 (BcCRY1), a cyclobutane pyrimidine dimer (CPD) photolyase, acts as the major enzyme of light-driven DNA repair (photoreactivation) and has no obvious role in signaling. In contrast, BcCRY2, belonging to the cry-DASH proteins, is dispensable for photorepair but performs regulatory functions by repressing conidiation in white and especially black/NUV light. The transcription of bccry1 and bccry2 is induced by light in a White Collar complex (WCC)-dependent manner, but neither light nor the WCC is essential for the repression of conidiation through BcCRY2 when bccry2 is constitutively expressed. Further, BcCRY2 affects the transcript levels of both WCC-induced and WCC-repressed genes, suggesting a signaling function downstream of the WCC. Since both CPF proteins are dispensable for photoinduction by black/NUV light, the origin of this effect remains elusive and may be connected to a yet unknown UV-light-responsive system.IMPORTANCEBotrytis cinerea is an economically important plant pathogen that causes gray mold diseases in a wide variety of plant species, including high-value crops and ornamental flowers. The spread of disease in the field relies on the formation of conidia, a process that is regulated by different light qualities. While this feature has been known for a long time, we are just starting to understand the underlying molecular mechanisms. Conidiation in B. cinerea is induced by black/near-UV light, whose sensing is attributed to the action of cryptochrome/photolyase family (CPF) proteins. Here we report on the distinct functions of two CPF proteins in the photoresponse of B. cinerea While BcCRY1 acts as the major photolyase in photoprotection, BcCRY2 acts as a cryptochrome with a signaling function in regulating photomorphogenesis (repression of conidiation).

Positive regulatory role of sound vibration treatment in Arabidopsis thaliana against Botrytis cinerea infection.

Sound vibration (SV), a mechanical stimulus, can trigger various molecular and physiological changes in plants like gene expression, hormonal modulation, induced antioxidant activity and calcium spiking. It also alters the seed germination and growth of plants. In this study, we investigated the effects of SV on the resistance of Arabidopsis thaliana against Botrytis cinerea infection. The microarray analysis was performed on infected Arabidopsis plants pre-exposed to SV of 1000 Hertz with 100 decibels. Broadly, the transcriptomic analysis revealed up-regulation of several defense and SA-responsive and/or signaling genes. Quantitative real-time PCR (qRT-PCR) analysis of selected genes also validated the induction of SA-mediated response in the infected Arabidopsis plants pre-exposed to SV. Corroboratively, hormonal analysis identified the increased concentration of salicylic acid (SA) in the SV-treated plants after pathogen inoculation. In contrast, jasmonic acid (JA) level in the SV-treated plants remained stable but lower than control plants during the infection. Based on these findings, we propose that SV treatment invigorates the plant defense system by regulating the SA-mediated priming effect, consequently promoting the SV-induced resistance in Arabidopsis against B. cinerea.

How light affects the life of Botrytis.

Fungi, like other organisms, actively sense the environmental light conditions in order to drive adaptive responses, including protective mechanisms against the light-associated stresses, and to regulate development. Ecological niches are characterized by different light regimes, for instance light is absent underground, and light spectra from the sunlight are changed underwater or under the canopy of foliage due to the absorption of distinct wavelengths by bacterial, algal and plant pigments. Considering the fact that fungi have evolved to adapt to their habitats, the complexities of their 'visual' systems may vary significantly. Fungi that are pathogenic on plants experience a special light regime because the host always seeks the optimum light conditions for photosynthesis - and the pathogen has to cope with this environment. When the pathogen lives under the canopy and is indirectly exposed to sunlight, it is confronted with an altered light spectrum enriched for green and far-red light. Botrytis cinerea, the gray mold fungus, is an aggressive plant pathogen mainly infecting the above-ground parts of the plant. As outlined in this review, the Leotiomycete maintains a highly sophisticated light signaling machinery, integrating (near)-UV, blue, green, red and far-red light signals by use of at least eleven potential photoreceptors to trigger a variety of responses, i.e. protection (pigmentation, enzymatic systems), morphogenesis (conidiation, apothecial development), entrainment of a circadian clock, and positive and negative tropism of multicellular (conidiophores, apothecia) and unicellular structures (conidial germ tubes). In that sense, 'looking through the eyes' of this plant pathogen will expand our knowledge of fungal photobiology.

Regulation of conidiation in Botrytis cinerea involves the light-responsive transcriptional regulators BcLTF3 and BcREG1.

Botrytis cinerea is a plant pathogenic fungus with a broad host range. Due to its rapid growth and reproduction by asexual spores (conidia), which increases the inoculum pressure, the fungus is a serious problem in different fields of agriculture. The formation of the conidia is promoted by light, whereas the formation of sclerotia as survival structures occurs in its absence. Based on this observation, putative transcription factors (TFs) whose expression is induced upon light exposure have been considered as candidates for activating conidiation and/or repressing sclerotial development. Previous studies reported on the identification of six light-responsive TFs (LTFs), and two of them have been confirmed as crucial developmental regulators: BcLTF2 is the positive regulator of conidiation, whose expression is negatively regulated by BcLTF1. Here, the functional characterization of the four remaining LTFs is reported. BcLTF3 has a dual function, as it represses conidiophore development by repressing bcltf2 in light and darkness, and is moreover essential for conidiogenesis. In bcltf3 deletion mutants conidium initials grow out to hyphae, which develop secondary conidiophores. In contrast, no obvious functions could be assigned to BcLTF4, BcLTF5 and BcLTF6 in these experiments. BcREG1, previously reported to be required for virulence and conidiogenesis, has been re-identified as light-responsive transcriptional regulator. Studies with bcreg1 overexpression strains indicated that BcREG1 differentially affects conidiation by acting as a repressor of BcLTF2-induced conidiation in the light and as an activator of a BcLTF2-independent conidiation program in the dark.

Preharvest UV-C radiation influences physiological, biochemical, and transcriptional changes in strawberry cv. Camarosa.

Ultraviolet C (UV-C) radiation is known for preventing fungal decay and enhancing phytochemical content in fruit when applied postharvest. However, limited knowledge is available regarding fruit responses to preharvest application of UV-C radiation. Thus, the effects of UV-C radiation on photosynthetic efficiency, dry matter accumulation and partitioning, fruit yield and decay, phytochemical content, and relative transcript accumulation of genes associated with these metabolic pathways were monitored in strawberry (Fragaria x ananassa Duch.) cv. Camarosa. A reduction in photosynthetic efficiency was followed by a decrease in light harvesting complex LhcIIb-1 mRNA accumulation as well as a decrease in yield per plant. Phenylalanine ammonia lyase activity, phenolic, anthocyanin, and L-ascorbic acid contents were higher in UV-C treated fruit. In addition, preharvest UV-C treatment reduced microorganism incidence in the greenhouse and on the fruit surface, increased the accumulation of ß-1,3-Gluc and PR-1 mRNA, and prevented fruit decay.

Light governs asexual differentiation in the grey mould fungus Botrytis cinerea via the putative transcription factor BcLTF2.

Botrytis cinerea is a plant pathogenic fungus known for its utilization of light as environmental cue to regulate asexual differentiation: conidia are formed in the light, while sclerotia are formed in the dark. As no orthologues of known regulators of conidiation (e.g., Aspergillus nidulans BrlA, Neurospora crassa FL) exist in the Leotiomycetes, we initiated a de novo approach to identify the functional counterpart in B. cinerea. The search revealed the light-responsive C2H2 transcription factor BcLTF2 whose expression - usually restricted to light conditions - is necessary and sufficient to induce conidiation and simultaneously to suppress sclerotial development. Light-induced expression of bcltf2 is mediated via a so far unknown pathway, and is attenuated in a (blue) light-dependent fashion by the White Collar complex, BcLTF1 and the VELVET complex. Mutation of either component leads to increased bcltf2 expression and causes light-independent conidiation (always conidia phenotype). Hence, the tight regulation of bcltf2 governs the balance between vegetative growth that allows for the colonization of the substrate and subsequent reproduction via conidia in the light. The orthologue ssltf2 in the closely related species Sclerotinia sclerotiorum is not significantly expressed suggesting that its deregulation may cause the lack of the conidiation program in this fungus.

Dark Period Following UV-C Treatment Enhances Killing of Botrytis cinerea Conidia and Controls Gray Mold of Strawberries.

Strawberries are available throughout the year either from production in the field or from high and low tunnel culture. Diversity of production conditions results in new challenges in controlling diseases before and after harvest. Fungicides have traditionally been used to control these diseases; however, their limitations necessitate a search for new approaches. We found that UV-C irradiation of Botrytis cinerea, a major pathogen of strawberry, can effectively kill this fungus if a dark period follows the treatment. The inclusion of a 4-h dark period resulted in almost complete kill of B. cinerea conidia on agar media at a dose of 12.36 J/m2. The UV-C dose did not cause a reduction in photosynthesis in strawberry leaves or discoloration of sepals, even after exposing plants repeatedly (twice a week) for 7 weeks. Although irradiation of dry conidia of B. cinerea with this dose resulted in some survival, the conidia were not infective and not able to cause decay even when inoculated onto a highly susceptible mature apple fruit. Irradiation of strawberry pollen at 12.36 J/m2 did not affect pollen germination, tube growth and length in vitro, or germination and tube growth in the style of hand-pollinated emasculated strawberry flowers. No negative effect of the UV-C treatment was observed on fruit yield and quality in high tunnel culture. In the fruit and flower petal inoculation tests, the UV-C treatment was highly effective in reducing fruit decay and petal infection. This UV-C treatment with an exposure time of 60 s may be useful in controlling gray mold in tunnel production of strawberries and may also have the potential for use in intensive field and indoor production of other fruits and vegetables providing that a 4-h dark period follows the irradiation.

Antifungal effect of 405-nm light on Botrytis cinerea.

UNLABELLED: There is very little information on the fungistatic or fungicidal effect of visible light. This study investigated the effect of 405-nm light, generated by a light-emitting diode array, on the economically important fungus Botrytis cinerea. The mycelial growth of B. cinerea was inhibited to the greatest extent by light at 405 and 415 nm and was negligibly inactivated at 450 nm, suggesting the presence of a photosensitizing compound that absorbs light mainly at wavelengths of 405-415 nm. Delta-aminolevulinic acid, a precursor of endogenous photosensitizer porphyrins, was used to determine the role of these porphyrins in 405-nm light-mediated photoinactivation of the fungus. Concentration-dependent inhibition of spore germination by delta-aminolevulinic acid and accumulation of singlet oxygen in the spores was observed when the spores were exposed to 405-nm light. These results suggest that the excitation of endogenous porphyrins and subsequent accumulation of singlet oxygen could partially explain the 405-nm light-mediated photoinactivation of B. cinerea. The development of symptoms in detached tomato leaves inoculated with B. cinerea spores was significantly reduced by irradiation with 405-nm light, indicating that 405-nm light has a potential use for controlling plant diseases caused by B. cinerea. SIGNIFICANCE AND IMPACT OF THE STUDY: Grey mould (Botrytis cinerea) is a very successful necrotroph, causing serious losses in more than 200 crop hosts. This study investigated the antifungal effect of 405-nm light on this pathogen. Our results suggest that the excitation of endogenous porphyrins and subsequent accumulation of singlet oxygen contribute to the 405-nm light-mediated photoinactivation of grey mould. The development of symptoms in detached tomato leaves inoculated with B. cinerea spores was significantly inhibited by irradiation with 405-nm light, indicating that this wavelength of light has a potential use in controlling plant diseases caused by B. cinerea.

Heat-induced oxidative injury contributes to inhibition of Botrytis cinerea spore germination and growth.

The inhibitory effect of heat treatment (HT) on Botrytis cinerea, a major postharvest fungal pathogen, and the possible mode of action were investigated. Spore germination and germ tube elongation of B. cinerea were both increasingly and significantly inhibited by HT (43 °C) for 10, 20 or 30 min. HT-induced gene expression of NADPH oxidase A, resulted in the intracellular accumulation of reactive oxygen species. HT-treated B. cinerea spores exhibited higher levels of oxidative damage to proteins and lipids, compared to the non-HT control. These findings indicate that HT resulted in oxidative damage which then played an important role in the inhibitory effect on B. cinerea. In the current study, HT was effective in controlling gray mold, caused by B. cinerea, in pear fruits. Understanding the mode of action by which HT inhibits fungal pathogens will help in the application of HT for management of postharvest fungal diseases of fruits and vegetables.

Assessing the effects of light on differentiation and virulence of the plant pathogen Botrytis cinerea: characterization of the White Collar Complex.

Organisms are exposed to a tough environment, where acute daily challenges, like light, can strongly affect several aspects of an individual's physiology, including pathogenesis. While several fungal models have been widely employed to understand the physiological and molecular events associated with light perception, various other agricultural-relevant fungi still remain, in terms of their responsiveness to light, in the dark. The fungus Botrytis cinerea is an aggressive pathogen able to cause disease on a wide range of plant species. Natural B. cinerea isolates exhibit a high degree of diversity in their predominant mode of reproduction. Thus, the majority of naturally occurring strains are known to reproduce asexually via conidia and sclerotia, and sexually via apothecia. Studies from the 1970's reported on specific developmental responses to treatments with near-UV, blue, red and far-red light. To unravel the signaling machinery triggering development--and possibly also connected with virulence--we initiated the functional characterization of the transcription factor/photoreceptor BcWCL1 and its partner BcWCL2, that form the White Collar Complex (WCC) in B. cinerea. Using mutants either abolished in or exhibiting enhanced WCC signaling (overexpression of both bcwcl1 and bcwcl2), we demonstrate that the WCC is an integral part of the mentioned machinery by mediating transcriptional responses to white light and the inhibition of conidiation in response to this stimulus. Furthermore, the WCC is required for coping with excessive light, oxidative stress and also to achieve full virulence. Although several transcriptional responses are abolished in the absence of bcwcl1, the expression of some genes is still light induced and a distinct conidiation pattern in response to daily light oscillations is enhanced, revealing a complex underlying photobiology. Though overlaps with well-studied fungal systems exist, the light-associated machinery of B. cinerea appears more complex than those of Neurospora crassa and Aspergillus nidulans.

UVR8 mediates UV-B-induced Arabidopsis defense responses against Botrytis cinerea by controlling sinapate accumulation.

Light is emerging as a central regulator of plant immune responses against herbivores and pathogens. Solar UV-B radiation plays an important role as a positive modulator of plant defense. However, since UV-B photons can interact with a wide spectrum of molecular targets in plant tissues, the mechanisms that mediate their effects on plant defense have remained elusive. Here, we show that ecologically meaningful doses of UV-B radiation increase Arabidopsis resistance to the necrotrophic fungus Botrytis cinerea and that this effect is mediated by the photoreceptor UVR8. The UV-B effect on plant resistance was conserved in mutants impaired in jasmonate (JA) signaling (jar1-1 and P35S:JAZ10.4) or metabolism of tryptophan-derived defense compounds (pen2-1, pad3-1, pen2 pad3), suggesting that neither regulation of the JA pathway nor changes in levels of indolic glucosinolates (iGS) or camalexin are involved in this response. UV-B radiation, acting through UVR8, increased the levels of flavonoids and sinapates in leaf tissue. The UV-B effect on pathogen resistance was still detectable in tt4-1, a mutant deficient in chalcone synthase and therefore impaired in the synthesis of flavonoids, but was absent in fah1-7, a mutant deficient in ferulic acid 5-hydroxylase, which is essential for sinapate biosynthesis. Collectively, these results indicate that UVR8 plays an important role in mediating the effects of UV-B radiation on pathogen resistance by controlling the expression of the sinapate biosynthetic pathway.

Low red/far-red ratios reduce Arabidopsis resistance to Botrytis cinerea and jasmonate responses via a COI1-JAZ10-dependent, salicylic acid-independent mechanism.

Light is an important modulator of plant immune responses. Here, we show that inactivation of the photoreceptor phytochrome B (phyB) by a low red/far-red ratio (R:FR), which is a signal of competition in plant canopies, down-regulates the expression of defense markers induced by the necrotrophic fungus Botrytis cinerea, including the genes that encode the transcription factor ETHYLENE RESPONSE FACTOR1 (ERF1) and the plant defensin PLANT DEFENSIN1.2 (PDF1.2). This effect of low R:FR correlated with a reduced sensitivity to jasmonate (JA), thus resembling the antagonistic effects of salicylic acid (SA) on JA responses. Low R:FR failed to depress PDF1.2 mRNA levels in a transgenic line in which PDF1.2 transcription was up-regulated by constitutive expression of ERF1 in a coronatine insensitive1 (coi1) mutant background (35S::ERF1/coi1). These results suggest that the low R:FR effect, in contrast to the SA effect, requires a functional SCFCOI1-JASMONATE ZIM-DOMAIN (JAZ) JA receptor module. Furthermore, the effect of low R:FR depressing the JA response was conserved in mutants impaired in SA signaling (sid2-1 and npr1-1). Plant exposure to low R:FR ratios and the phyB mutation markedly increased plant susceptibility to B. cinerea; the effect of low R:FR was (1) independent of the activation of the shade-avoidance syndrome, (2) conserved in the sid2-1 and npr1-1 mutants, and (3) absent in two RNA interference lines disrupted for the expression of the JAZ10 gene. Collectively, our results suggest that low R:FR ratios depress Arabidopsis (Arabidopsis thaliana) immune responses against necrotrophic microorganisms via a SA-independent mechanism that requires the JAZ10 transcriptional repressor and that this effect may increase plant susceptibility to fungal infection in dense canopies.

Postharvest physio-pathological disorders in table grapes as affected by UV-C light.

To gain knowledge on the influence of postharvest treatments with ultraviolet-C light upon the keeping quality of table grapes, a trail was performed employing commercially mature 'Corina', 'Dawn Seedless', 'Centennial Seedless' and 'Gran Perlon' grape cultivars (cvs). After grading, bunches were subjected to 0.0, 0.5, 1.5 or 3 kJm(-2) and then stored at 5 degrees C and 90% relative humidity (RH) for 6 weeks followed by a 2 day shelf-life at 25 degrees C and 70% RH. A weekly inspection was performed and a visual evaluation of the appearance, treatment damage, stems browning and berry shrivelling was performed. Weight loss, decay and shatter were quantified at the end of storage and shelf-life. Regardless the cv and UV-C dose, fruit appearance was acceptable until the end of storage and shelf-life. Among the cvs, the highest score was held by 'G. Perlon'. After the fourth week of storage, the berries of 'Centennial S.' turned light brown and darkened over time when treated with 3.0 kJm(-2). Stem browning was not induced by the light treatment, but resulted cv depended and was pronounced for 'Centennial S.' and 'Dawn S.'. Berry shrivelling was insignificant, while shatter was very high in 'Corina' and did not depend upon UV-C treatment. Regarding weight loss, differences could not be attributed to the light treatment and after storage it ranged from 3 up to 5%. Decay was significantly reduced by light treatment and the efficacy increased by raising the dose. Botrytis cinerea was the main cause of decay with 'Corina' being the most jeopardized, followed by 'Dawn S.' and 'Centennial S.', whereas 'G. Perlon' resulted the less affected. In conclusion, hormetic effects of postharvest light treatment on table grapes were observed in almost all cvs with 'G. Perlon' having the best performance.

Conservation and clade-specific diversification of pathogen-inducible tryptophan and indole glucosinolate metabolism in Arabidopsis thaliana relatives.

• A hallmark of the innate immune system of plants is the biosynthesis of low-molecular-weight compounds referred to as secondary metabolites. Tryptophan-derived branch pathways contribute to the capacity for chemical defense against microbes in Arabidopsis thaliana. • Here, we investigated phylogenetic patterns of this metabolic pathway in relatives of A. thaliana following inoculation with filamentous fungal pathogens that employ contrasting infection strategies. • The study revealed unexpected phylogenetic conservation of the pathogen-induced indole glucosinolate (IG) metabolic pathway, including a metabolic shift of IG biosynthesis to 4-methoxyindol-3-ylmethylglucosinolate and IG metabolization. By contrast, indole-3-carboxylic acid and camalexin biosyntheses are clade-specific innovations within this metabolic framework. A Capsella rubella accession was found to be devoid of any IG metabolites and to lack orthologs of two A. thaliana genes needed for 4-methoxyindol-3-ylmethylglucosinolate biosynthesis or hydrolysis. However, C. rubella was found to retain the capacity to deposit callose after treatment with the bacterial flagellin-derived epitope flg22 and pre-invasive resistance against a nonadapted powdery mildew fungus. • We conclude that pathogen-inducible IG metabolism in the Brassicaceae is evolutionarily ancient, while other tryptophan-derived branch pathways represent relatively recent manifestations of a plant-pathogen arms race. Moreover, at least one Brassicaceae lineage appears to have evolved IG-independent defense signaling and/or output pathway(s).